Gymnosperm

The Gymnosperms
THE GYMNOSPERMS
Chhaya Biswas D B.M. Johri
Springer-Verlag Berlin Heidelberg GmbH

Cover photograph: Taxus baccata. Courtesy Prof. B.D. Sharma,
JNV University, Jodhpur, India
Dr. Cbhaya Biswas

Formerly Principal, Gargi College
University of Delhi South Campus
New Delhi 110 049, India
Prof. (Retired) B.M. Johri
Central Reference Library
University of Delhi,
Delhi 110 007, India
Copyright © I 997 Springer-Verlag Berlin Heidelberg

Originally published by Springer-Verlag Berlin Heidelberg New York in 1997
All rights reserved. No part of this publication may be reproduced,

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Unauthorised export is a violation of Copyright Law and is subject to legal action.
ISBN 978-3-662-13166-4 ISBN 978-3-662-13164-0 (eBook)

DOI 10.1007/978-3-662-13164-0
To
Our Teacher
Professor Panchanan Maheshwari

(1904-1966)
Preface
The Gymnosperms is a well-illustrated comprehensive account of living

and fossil plants of this group. Chapters 1 and 2 give a general account, and
describe similarities and dissimilarities with pteridophytes and angiosperms.
Chapter 3 deals with classification. The next 18 chapters (4-21) deal
sequentially with fossil and living taxa. Phylogenetic relationships are
considered for each order. Chapter 22 discusses the in vitro experimental
studies on the growth, development and differentiation of vegetative and
reproductive organs and tissues. Chapter 23 summarizes the economic
importance of gymnosperms. Chapter 24 gives the conciuding remarks.
Thus, there is a complete coverage of significant findings concerning
morphology, anatomy, reproduction, development of embryo and seed,
cytology, and -evolutionary trends and phylogeny. Ultrastructural and
histochemical details are given wherever considered necessary.
There is a comprehensive list of literature citations, and a plant index.
This book is essentially meant for the postgraduate students in India and
abroad. Undergraduate students can also use it profitably. The entire course
should be taught in 25-30 lectures/hours and about 75 hours of field and
laboratory work.
CHHAYA BISWAS
BRu MoHAN JoHRI
Acknowledgements
For the preparation of this book we are grateful to:

Dr. B.S. Venkatachala, former Director, Birbal Sahni Institute of
Palaeobotany, Lucknow, for allowing Dr. Chhaya Biswas to work in the
Library of the Institute, and to Dr. H.K. Maheshwari, Dr. Sheila Chandra,
Dr. A.K. Srivastava, Dr. Usha Bajpai, Dr. Sukh Devi, Dr. S.C. Srivastava
and Dr. Jayashree Bannerjee for helping her to update the information,
fruitful discussions and suggestions concerning fossil members of the
gymnosperms, to the Librarian of the Institute for providing the literature.
(The Late) Professor P.N. Mehra, Department of Botany, Punjab University,
Chandigarh, for scrutinizing the outlines of this book and giving valuable
advice, for providing his monograph on the Indian Conifers, Gnetophytes
and Phylogeny of Gymnosperms; Professor B.D. Sharma, Department of
Botany, JNV University, Jodhpur, for a critical review of several chapters
of the manuscript; to Professor S.P. Bhatnagar, and Dr. S.S. Bhojwani,
Department of Botany, University of Delhi, for providing relevant literature
and fruitful discussions; Dr. P.S. Srivastava, Department of Botany, Hamdard
(deemed) University, New Delhi, for important literature and valuable
suggestions; eur colleagues in Gargi College, University of Delhi, Dr. Bharati
Bhattacharyya for useful suggestions, Dr. Asha Juneja and Dr. Lalita Sehgal
for making available several publications from their personal collections,
and Dr. Gita Mathur for scrutinizing some of the chapters; our students
Arindam Bhattacharyya and Somdutta Sinha Roy for technical assistance
in the preparation of the manuscript.
Professor Upendra Baxi (former Vice-Chancellor), Professor A.L. Nagar
(former Pro-Vice-Chancellor), Professor A.P. Srivastava (former Librarian)
and Mr. Sher Singh (Deputy Librarian), University of Delhi, for providing
an office in the Library where this work was completed.
Mr. Kishen Lal and Mr. S.K. Das for preparing the photographs,
Mr. R.K. Gupta for typing the manuscript, and Professor N.N. Bhandari
(then Head of the Department of Botany, University of Delhi) for allowing
us to use the departmental facilities during the preparation of this book.
Our family members, especially Mr. Alok Biswas and Mrs. Raj Johri,
for their encouragement and interest in this work.
We wish to express our gratitude to all the persons and institutions
mentioned above.
CHHAYA BISWAS
B.M. JOHRI
Contents
Preface vii
Acknowledgements ix
1. Introduction 1-11
Antiquity and Fossil History 1
Geographical Distribution 4
Cycadales and Ginkgoales 4
Pinaceae 6
Taxodiaceae 6
Cupressaceae 7
Podocarpaceae 7
Araucariaceae 7
Cephalotaxaceae 7
Taxales 8
Ephedrales 8
Welwitschiales 8
Gnetales 8
Characteristic Features 8
Gymnosperms and Pteridophytes 9
Gymnosperms and Angiosperms 10
2. Seed Development 12-19

Microsporangium 12
Microsporogenesis 12
Male Gametophyte 13
Ovule 14
Megasporogenesis 14
Megaspore Wall 15
Female Gametophyte 16
Archegonia 16
Pollination and Fertilization 17
Pollination 17
Fertilization 18
Embryo 19
Seed 19
Seed-Coat 19
Temporal Considerations 19
3. Classification 20-23
xii Contents
4. Progymnospermopsida 24-26
Phylogenetic Considerations of Archaeopteris 25
5. Gymnospermopsida 27
Gymnosperm opsida-Cycad ophytes

(Chapters 6-8)
6. Pteridospermales 28-35
Lyginopteridaceae 29
Lyginopteris oldhamia (Calymmatotheca hoeninghausi) 29
Habit 29
Root 30
Stem 30
Leaf 30
Sporangia 33
Seed 33
Phylogenetic Considerations 34
7. Cycadeoidales 36-43
Cycadeoidaceae 38
Cycadeoidea 38
Stem 38
Leaves 38
Reproduction 39
8. Cycadales 44-81
Fossil Cycads 44
Nilssonia 46
Baenia 47
Androstrobus manis 47
Living Cycads 47
Root 47
Stem 48
Leaf 49
Reproduction 50
Spermatozoid 51
Ovule 52
Pollen Tube 52
Embryo 54
Chromosome Number 55
Cycadaceae 56
Cycas 56
Contents xiii
Morphology 56
Anatomy 59
Root 59
Stem 59
Leaf 61
Reproduction 63
Male Cone 63
Microsporangium 66
Male Gametophyte 67
Megasporophyll 68
Ovule 68
Megasporogenesis 71
Archegonium 71
Pollination 73
Fertilization 77
Embryogeny 77
Differentiation of Embryo 79
Seed 79
Temporal Consideration 80
Gymnospermopsida-Gymnosperms of Uncertain
Relationship (Chapters 9-12)
9. Caytoniales 82-85
Caytoniaceae-Caytonia 82
Leaf 82
Pollen-bearing Organs 82
Ovule-bearing Organs 84
10. Glossopteridales 86-93

Glossopteridaceae-Glossopteris 86
Root 86
Leaf 88
Male Fructifications 88
Ovulate Structures 89
11. Pentoxylales 94-97

Pentoxylon 94
Habit 94
xiv Contents
Stem 94
Leaf 95
Microsporangiate Organs 96
Ovulate Cone 96
12. Ginkgoales 98-126

Fossil Taxa 98
Ginkgoaceae 100
Ginkgo 100
Morphology 100
Anatomy 104
Root 104
Stem 104
Leaf 105
Reproduction 105
Male Cone 105
Microsporangium 105
Male Gametophyte 107
Ovule 107
Megasporogenesis 108
Pollination 117
Post-Pollination Development of Male Gametophyte 117
Fertilization 121
Embryogeny 122
Seed 124
Germination 125
Gymnospermopsida-Coniferophytes
(Chapters 13-16)
13. Cordaitales 127-136
Cordaitaceae-Cordaites 127
Habit 127
Root 127
Stem 128
Leaf 131
Fructifications 131
Cordaianthus concinus 133
Cordaianthus pseudofluitans 133
Contents xv
14. Voltziales 137-143

Morphology 137
Anatomy 138
Reproductive Organs 139
Lebachiaceae 139
Voltziaceae 139
Ullmania 142
Glyptolepis 142
Voltziopsis 142
15. Coniferales 144-146

15.1 Pinaceae 146-188
Pinus 146
Morphology 146
An~tomy 150
Root 150
Stem: Shoot Apex 150
Compression Wood 156
Dwarf Shoot 156
Leaf 156
Reproduction 157
Male Cone 157
Microsporangium 158
Pre-meiotic Changes 158
Meiotic Changes 160
Post-meiotic Changes 160
Female Cone 166
Ovule 166
Female Gametophyte (First period of growth) 168
Pollination 168
Post-pollination Development of Male Gametophyte 170
Female Gametophyte (Second period of growth) 171
Fertilization 175
Embryogeny 178
Proembryo 178
Seed 181
Germination 184
15.2 Taxodiaceae 188-202

Cryptomeria japonica 189
xvi Contents
Morphology 189
Anatomy 189
Stem 189
Leaf 191
Reproduction 191
Male Cone 191
Microsporangium 192
Female Cone 194
Ovule 194
Fertilization 199
Embryogeny 199
Diferentiation of Embryo 201
Seed 201
Germination 201
15.3 Cupressaceae 202-221

Biota orienta/is 203
Morphology 203
Reproduction 205
Male Cone 205
Microsporangium 205
Female Cone 207
Ovule 207
Archegonial Complex 209
Pollination 212
Fertilization 213
Embryogeny 215
Seed 218
Germination 218
15.4 Podocarpaceae 221-251

Podocarpus 222
Morphology 222
Contents xvii
Anatomy 222
Root 222
Stem. Shoot Apex 222
Leaf 224
Reproduction 227
Male Cone 227
Microsporangium 227
Female Cone 234
Ovule 234
Pollination 237
Post-Pollination Development of Male Gametophyte 239
Fertilization 242
Embryogeny 242
Seed 248
Germination 250
15.5 Araucariaceae 251-265

Araucaria 252
Morphology 252
Anatomy 254
Stem 254
Leaf 254
Reproduction 257
Male Cone 257
Female Cone 259
Pollination 262
Embryogeny 262
15.6 Cephalotaxaceae 265-290

Cephalotaxus 265
Morphology 267
Anatomy 268
Leaf 268
Reproduction 270
Male Cone 270
01iii Contents
Microsporangium 270
Female Cone 273
Ovule 274
Pollination 282
Fertilization 283
Embryogeny 284
Seed 287
Germination 289
15.7 Phylogenetic Considerations of Coniferales 290-291
16. Taxales 292-310

Taxaceae 292
Taxus 292
Morphology 292
Anatomy 293
Stem 293
Leaf 294
Reproduction 295
Male Cone 295
Microsporangium 295
Ovule 296
Pollination 302
Fertilization 303
Embryogeny 304
Seed 308
Germination 308
17. Gnetopsida 311
18. Ephedrales 312-342

Ephedraceae 312
Ephedra 312
Contents xix
Morphology 312
Anatomy 313
Root 313
Stem 314
Reproduction 321
Male Strobilus 321
Microsporangium 321
Female Strobilus 327
Ovule 327
Pollination 335
Fertilization 336
Embryogeny 337
Seed 338
19. Welwitschiales 343-365

Welwitschiaceae 343
Welwitschia 343
Morphology 343
Anatomy 346
Root 346
Stem 347
Leaf 349
Reproduction 352
Cone Bracts 352
Male Strobilus 354
Microsporangium 355
Pollination 359
Fertilization 360
Embryogeny 360
Seed 364
Germination 364
20. Gnetales 366-404

Gnetaceae 366
Gnetum 366
xx Contents
Morphology 366
Anatomy 366
Root 366
Stem 367
Leaf 375
Reproduction 378
Male Strobilus 378
Microsporangium 378
Ovule 384
Pollination 391
Fertilization 393
Embryogeny 396
Seed 400
Germination 402
Temporal Consideration 403
21. Phylogenetic Considerations: Ephedra,

Welwitschia and Gnetum 405-407
22. In Vitro Experimental Studies 408-439

Vegetative Tissues and Organs 408
Microspore and Male Gametophyte 411
Embryo 423
Conifer Biotechnolgy 433
Somatic Embryogenesis 433
Genetic Transformation 436
Protoplast Culture 436
Micropropagation 437
23. Economic Importance 440-456

General Aspects 440
Woods 441
AJaucariaceae 442
Cupressaceae 442
Pinaceae 443
Podocarpaceae 444
Taxodiaceae 445
Taxaceae 445
Ginkgoaceae 445
Contents xxi
Paper and Board 445

Resins 446
Hard Resins 447
Copal 447
Kauri Copal 447
Manila Copal 447
Amber 448
Sandarc 448
Oleoresins 449
Turpentine 449
Canada Balsam 450
Venice Turpentine 450
Tannins 451
Essential Oils 451
Fatty Oils 451
Food Supplements 452
Pharmaceuticals 454
24. Concluding Remarks 457-459

Seasonal Changes in Secondary Growth 457
Population and Wood Requirement 457
Pollution 458
Symbiosis 458
Reproductive Biology 458
Pollen Biology 459
Life History 459
References 460-489
Plant Index 491-494
The Gymnosperms
1. Introduction
Indubitable seeds first appeared in the Devonian (395-359 my B.P. =million

years Before Present). These seeds are not enclosed in ovaries, as in flowering
plants, and are designated naked. This character is the basis for grouping
a large number of varied plants as a natural greup, the Gymnospermae
(gymnos = naked, sperm = seed). Palaeobotanical evidence suggests that
the seeds evolved independently in more than one group of Palaeozoic
plants and diversified rapidly during the Lower Carboniferous (345 my
B.P.). In the course of their evolution, the seed plants have tended to
evolve a variety of structures to protect the ovules. Some of these evolutionary
"experiments" were successful, and provide insight into the origin of the
carpel, while others ended in extinction (see Stewart 1983).
Presently, with the advanced knowledge of fossil plants, the gymnosperms
are regarded as a heterogeneous group of seed plants. These plants constituted
most of the world's dominant vegetation throughout the late Paleozoic and
Mesozoic, and steadily declined thereafter. A modern approach is to interpret
the phylogeny of gymnosperms (as already in flowering plants) on the
basis of phytogeographical and ecological conditions in which the taxa
survived.
Antiquity and Fossil History

The gymnosperms are an ancient group which dates back to the Devonian
(Figs. 1.1, 1.2). They began ca. 395 my B.P., lasted for ca. 50 my, and gave
rise to aneurophytes, progymnosperms, archaeopterids and pteridosperms.
During the Carboniferous (ca. 345-280 my B.P.), a large variety of cordaitels
and seed ferns existed. In the Permian and Triassic (ca. 280 and 225 my
B.P.) the Carboniferous pteridosperms became extinct. The early conifers
(Voltziales) diversified and the cycads and cycadeoids became evident for
the first time. Glossopteridales formed a conspicuous flora of the Southern
Hemisphere during the Permian. The ginkgophytes appeared some time
during the Permian and became more widely spread in the Triassic. In the
Jurassic (ca. 195-141 my B.P.), the cycads, cycadeoids, conifers and
ginkgophytes reached their peak of diversification, and the glossopterids
became extinct. During the Upper Cretaceous (ca. 141-65 my B.P.), the
!!! Z)>-ZO<mo cnc;:O:llm-n-ZO!Il:ll)>O z )>- 3: :ll m..., ln-cncn)>-:ll-iln-cncn)>:llC<-1 en c CO> m n )> -i m :ll n
r- N
c
:ll
i> ~
z ~
:om;:EOr
Gl !!l m m Gl
-i
0 b c
'C
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;:::
4' ~ c: 'C c
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c: :!. 3 <' !!!
0 5' i .. .
o;· o;· ~ ~ ~ i !!! ~
o;· ~
..!!! ." ..3
iii' -· ;;;·
ii'
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"~· iii'
" I" " " " ~
"
t
........................
"' ... _
Fig. 1.1 The distribution in geological time of major groups of Cycadophytes and their presumed relatives; their origin and relationship is also
suggested. Ane Aneurophytales. Arch Archaeopteridales. Calam Calamopityaceae. Call Callistophytaceae. Czek Czekanowskiales, Pen Pentoxylales.
Trim Trimerophytopsida. (After Stewart 1983)
CI>CO:Dm"Tl-ZO!D:D)>O
i
f
~· i
...;;--
C:l
~
Q.
Fig. 1.2 The distribution in geological time of Ginkgoales, Gnetopsida, and major groups of Coniferophytes and their suggested origin and §"
relationships. Ane Aneurophytales, Arch Archaeopteridales, Calam Calamopityaceae, Trim Trimerophytopsida. (After Stewart 1983) (M
4 The Gymnosperms
angiosperms appeared and diversified rapidly. They began to replace the

already declining cycads, cycadeoids, conifers and ginkgophytes. Mesozoic
pteridosperms and other smaller groups became extinct. However, the majority
of the conifer families have continued up to the present. In the Tertiary (65
my B.P.) the angiosperms evolved steadily while the conifers declined in
diversity. Thereafter, the angiosperms became dominant and the gymnosperms
occupied a secondary position, although they still dominated landscapes
that angiosperms have yet to conquer. The long evolutionary history of
gymnosperms provides many examples of taxa which flourished, and finally
became completely or nearly extinct. Presently, there are ca. 13 000 genera
and 240 000 spp. of angiosperms (Takhtajan 1980), while the gymnosperms
comprise only 69 genera and 750-760 spp. These are grouped in seven
orders: Cycadales, Ginkgoales, Coniferales, Taxales, Ephedrales,
Welwitschiales, and Gnetales.
Geographical Distribution
The Cycadales and Ginkgoales represent the surviving members of extremely
ancient groups. The cycads were once a large and dominant group with
widest distribution in the Mesozoic (Fig. 1.1 ), which is sometimes referred
to as the age of cycads (Scott 1923). Now there are only 11 living taxa
(including Chigua, a new genus; see Stevenson 1990a, b) confined to limited
areas of the tropics and subtropics. They form neither extensive nor
conspicuous features of the vegetation. Six taxa occur in the Eastern
Hemisphere, and five in the Western Hemisphere (Fig. 1.3A); no single
genus is represented in both hemispheres. Among the eastern cycads,
Macrozamia, Lepidozamia and Bowenia are confined to Australia, and
Encephalartos and Stangeria exclusively to South Africa. The genus Cycas
occurs from Australia to Japan, touching India and China. Of the western
genera, Dioon and Ceratozamia are confined entirely to Mexico, Microcycas
to western Cuba, Zamia to both areas, and Chigua in Colombia (the only
endemic cycad in south America). Stangeria resembles the ferns so much
that it was placed, for a long time, in the family Polypodiaceae. According
to Arnold (1953), these isolated genera are "leftovers" from the widely
distributed cycads of the Mesozoic.
The order Ginkgoales included many genera and species, reported from
the Permian; from the Triassic onwards and during the Mesozoic they had
worldwide distribution (Fig. 1.2), but, before the end of the Jurassic, ,they
began to disappear, and all the members of this order, except Ginkgo, became
extinct by the Cretaceous. The sole surviving member, G. biloba, is restricted
in geographical distribution. At present its natural occurrence is confined
to a small, inaccessible region in southeastern China. There is, however,
some uncertainty whether or not such specimens are escapes from gardens
(see Andrews 1961). It is doubtful whether it actually exists in the wild/
natural state today. One of the reasons for its survival from extinction is
Introduction 5
AMERICAN
~
eERATOZAMIA
DIOOH
AFRICAN
AUSTiROA~~£NIA
MICIIOCYCAS ~~ ENCEPHAUIP.TOS (ere~
[ZAMIA) 0 STANG£RIA MACROlAMIA
Fig.1.3 A, B. World distribution. A Living cycad genera. B Gnetum. (A After Schuster

1932, B after P. Maheshwari and V. Vasil196la)
that for centuries priests in China and Japan cultivated and worshipped it.
The modern Ginkgo is remarkably resistant to attacks of insects and fungi.
The reason probably is its immense vigour, which enabled it to survive for
millions of years. Ginkgo is to be regarded as one of the wonders of the
world because it has persisted with very ~.ttle change through a long succession
of ages inhabited by plants and animals quite different from those present
in the modern age.
The cycads and Ginkgo are designated living fossils because they still
6 The Gymnosperms
exhibit many of their ancestral features without much change. One such
character is the presence of motile or swimming sperms, displayed only in
these plants among the existing seed plants.
The Coniferales form 75% of the modem gymnosperms, and are an
important constituent of the flora today. There are ca. 52 genera and 560
spp. (Mehra 1988), usually grouped into six families. They have a markedly
disjunct geographical distribution and several taxa are endemic. Each family
is widely distributed, although individual taxa often show extremely limited
distribution. Such a distribution pattern indicates antiquity. This is evident
in the evolutionary history of each of the six families, which extend back
to the Mesozoic (Fig. 1.2). Most modem taxa of the Coniferales were
already present by the onset of the Tertiary (Stewart 1983).
The conifers are plants of the more temperate regions of the world, and
only a few taxa are strictly tropical. Western North America, Eastern and
Central China, and parts of Australia and New Zealand have abundant
conifers, which show exceptional diversity.
Pinaceae. The Pinaceae have ten genera, which form pi·ominent coniferous
forests of the Northern Hemisphere (Fig. 1.4). This family is totally
unrepresented in the Southern Hemisphere (Coulter and Chamberlain 1917).
Fig. 1.4. World distribution of pines. (After P. Maheshwari and Konar 1971)
Taxodiaceae. There are ten genera, seven monotypic. In the past, these
taxa were an important constituent of the forests of the Northern Hemisphere.
At present, they show a relict distribution. Of the ten taxa, only three occur
in the USA; Sequoia sempervirens (Californian redwood) restricted to a
narrow coastal belt in California, Sequoiadendron giganteum (big tree) in
central California, and Taxodium spp. which grow in lowland swamps of
Introduction 7
the Southeastern USA and Mexico. Of the remaining seven taxa, Cryptomeria
japonica (Japanese cedar) and Cunninghamia spp. are distributed in Japan
and parts of China, Sciadopitys verticillata (Japanese um!Tella pine) in Japan,
Glyptostrobus (Chinese deciduous cypress) in China, and Taiwania in
Formosa. Metasequoia glyptostroboides (Dawn redwood) was known only
as a fossil from Pliocene deposits until 1948, when living specimens were
discovered from Szechuan Province, China (Hu and Cheng 1948). Athrotaxis
spp. (Tasmanian cedars) is the only southern taxon confined to Tasmania.
Cupressaceae. The largest family of the conifers includes ca. 19 genera,

8 monotypic. Of these 19 genera 9 are distributed in the Northern and 10
in the Southern Hemisphere. Of the northern taxa, Cupressus (cypress),
Chamaecyparis (false cypress) and Thuja (arbor vitae) have a disjunct
distribution. Juniperus (Juniper) is distributed in a broad belt round the
Northern Hemisphere. This may be due to its female cones which are
berry-like and distributed by birds. Some of the southern taxa are Callitris
(Cypress pines) confined to Australia, Tasmania, and New Caledonia,
Libocedrus to New Zealand and New Caledonia, and Papuacedrus to New
Guinea, extending somewhat across the equator into the Northern Hemisphere.
Podocarpaceae. Podocarpaceae is the most important family of the Southern

Hemisphere, where it originated. It comprises seven taxa, two monotypic.
Some of the taxa are represented in the Northern Hemisphere also, and are
assumed to be recent arrivals. The largest genus, Podocarpus (yellow woods),
occurs in the mountain forests of warm temperate and subtropical regions
of the Southern Hemisphere. Some species occur in Japan, China, India,
Malaya and The Philippines. Dacrydium and Phyllocladus are distributed
chiefly in New Zealand and Tasmania, Dacrydium also occurs in Malaysia,
Burma, Southern China, Patagonia and Chile, while Phyllocladus is found
in New Guinea, North Borneo and the Philippines. They have representatives
north of the Equator. Saxegothaea (Prince Alberts' yew) and Microstrobus
(= Pherosphera) are monotypic. The former is confined to Chile and the
latter to Tasmania. Acmopyle is confined to New Caledonia and Fiji.
Araucariaceae. An extremely old family with fossil record extending

back to the Triassic of both North and South Hemispheres. Both the present-
day taxa are restricted to the Southern Hemisphere. Araucaria occurs in South
America, Australia, New Guinea and New Caledonia, and Agathis (kauri
pines) is exclusively eastern, extending from The Philippines to New Zealand
and Malaya to Fiji.
Cephalotaxacae. A single taxon, Cephalotaxus, is restricted to eastern

Asia. It is distributed from the Eastern IJimalayas to Japan in subtropical
forests.
8 The Gymnosperms
Taxales. The order Taxales includes a single family, Taxaceae, with five
genera. Taxus (Yew) occurs in North America, Europe and Asia, and extends
up to Malaya. This wide geographical distribution is partly due to birds.
Pseudotaxus (=Nothotaxus) grows in a small part of east China. Amentotaxus
presently occurs in east Asia, but fossil remains have been reported from
Europe and western America. Austrotaxus is restricted to New Caledonia.
Torreya occurs only in California, Florida and eastern Asia.
Ephedrales. The Ephedrales comprise a single mono generic family,

Ephedraceae, and Ephedra with ca. 42 spp. has a worldwide distribution.
None of the species is common to the Old and the New Worlds or Eastern
and Western Hemispheres. Most of the species inhabit arid or desert regions
including saline tracts where they act as a sand-binder. The plants grow up
to 5000 m above sea level.
Welwitschiales. Welwitschia is a monotypic genus of limited distribution.

It is restricted to a strip ca. 1200 km long along the west coast of southern
Africa from Angola (Nicolau River) to South-West Africa, Namibia (Kuiseb
River). In the Welwitschia Flats (ca. 45 km east of Swakopmund) 5000-
6000 specimens grow (von Willart 1985). Welwitschia never reaches the
coast. It occupies only the northern and central parts of the Namib desert
in its south-north extension and stretches over subtropical grassland, entering
the Mopane Savanna in its west-east extension. Thus, it occupies an area
with a wide -ecological range. It grows mainly where the annual rainfall is
between 0.0 and 10n rnm and precipitation (except dew and fog) is only in
the surrtr,.;r months, January to ¥arch. In the Mopane Savanna, annual
rainfall can exceed 200 mm, and fog is absent.
~netales. The single family, Gnetaceae, includes only one genus, Gnetum.
-~ inhabits moist tropical forests in parts of Asia, Africa, northern South
/ merica, and certain islands between Asia and Australia (Fig. 1.3 B). Most
~: :ies, ca 30, are endemic within the areas of their distribution. There is
no species common to both the hemisphere, and none of the Asian species
occurs in Africa or America. The center of the present diversification appears
to be Eastern Malayasia (seeP. Maheshwari and V. Vasil 196la).
Characteristic Features
The seed plants are usually classified into two major groups: gymonsperms
and angiosperms, based on the protection to the ovule (at the time of
pollination). The gymnosperms bear naked ovules, i.e. the ovules are borne
directly on the sporophyll or an equivalent structure, and are exposed. In
some of the gymnosperms, overlapping scales and sporophylls may protect
the ovules but they are freely exposed at pollination. In the angiosperms,
the ovules develop within an ovary. This distinction between naked and
enclosed ovules and later seeds is of considerable importance.
Introduction 9
Some of the other features .typical of gymnosperms are: There are no

herbaceous gymnosperms, and the plants whether trees, shrubs or lianas are
woody and evergreen. They have a tap root which usually persists for a
long time. The xylem consists of tracheids, parenchyma and rays. Vessels
are present in Ephedra, Welwitschia and Gnetum. They have evolved from
pitted tracheids, as shown by intermediate stages between pits and perforations.
In phloem only sieve cells differentiate; sieve areas commonly occur on the
radial walls as well, and are numerous where the end of one sieve element
overlaps that of another. The companion cells are absent (see Chaps. 8, 12,
15, 18, 19, 20 for details.). Secondary growth takes place in all gymnosperms;
mature metaxylem shows bordered pits of various types; Stangeria and Zamia
show scalariform thickenings (Pant 1973). The anther has an exothecium.
There are numerous light pollen grains which land directly on the surface
of the nucellus during pollination. Prothallial cells are formed in the male
gametophyte. The ovule is unitegmic and orthotropous. There is a prolonged
free-nuclear phase in the development of the female gametophyte, a long
interval between pollination and fertilization, a free-nuclear phase in the
development of the proembryo, and haploid nutritive and storage tissue
(the female gametophyte).
Gymnosperms and Pteridophytes

The gymnosperms and pteridophytes share some common features: (a) Both
the groups have independent sporophyte, mostly differentiated into root,
stem and leaf. (b) A well-developed vascular tissue is present, the xylem
lacks true vessels, and phloem is without companion cells. (c) The leaves
of cycads are compound with circinate vemation, while in Ginkgo the venation
is dichotomous (cf. ferns). (d) Motile male gametes are present in cycads
and in Ginkgo, and (e) Archegonia are present in the female gametophyte
of gymnosperms except Welwitschia and Gnetum.
The gymnosperms show a definite advance over pteridophytes: (a) There
is a continuously growing, long-lasting tap root system in gymnosperms
which provides better anchorage to a perennial plant. In pteridophytes the
roots are mostly adventitious. (b) The majority of gymnosperms are
monostelic. The pteridophytes have a wide range of primary vascular system-
protostele, actinostele, siphonostele, solenostele, dictyostele, polycyclic stele
and polystele. (c) All gymnosperms show secondary growth, but it is absent
in pteridophytes. In Isoetes and Botrychium, secondary growth occurs, but
is not as extensive as in gymnosperms. The mature metaxylem, in most
gymnosperms, shows bordered pits. In most pteridophytes, the xylem is
typically scalariform. The phloem of gymnosperms differs from the
pteridophytes in the type of conducting cells, their cellular composition,
and the degree and nature of functional interrelationships between the
conducting and parenchyma cells (see Paliwal 1992). A typical pteridophytic
sieve element is identical to the elongate parenchyma cells but is longer;
10 The Gymnosperms
the gymnospermous sieve cells are shorter. The sieve elements have a
smaller diameter, their endwalls are horizontal or slightly oblique. The
sieve areas are small and their outline varies. The sieve pores occur in the
sieve areas as well as scattered on the vertical walls. Callose deposition has
been observed in Psilotum, lycopods and the ferns (see Paliwal 1992).
(d) All gymnosperms are heterosporous. At pollination the microspores are
usually carried to the megasporangia by wind; entomophily is reported in
Ephedra aphylla Forsk. (Bino et al. 1984), Gnetum sp. (Kato et al. 1995),
and Welwitschia (Kubitzki 1990). The need for water to bring about
fertilization has been eliminated in gymnosperms. A major difference between
the motile spermatozoids of cryptogams and those of cycads and Ginkgo is
that the former swim freely in water, whereas the latter swim in a fluid
medium enclosed within the developing ovule (see Chaps. 8, 12). The
majority of the pteridophytes are homosporous with monoecious
gametophytes. With this combination there is a greater possibility of self-
fertilization; this is disadvantageous since the sporophytes produced are
mostly homozygous for all characters and are, therefore, relatively slow in
evolutionary progress (see Sporne 1965). A few pteridophytes, e.g. Selaginella,
Isoetes, Stylites, Marsilea, Pilularia, Regnellidium, Salvinia and Azolla are
heterosporous. (e) The female gametophyte in gymnosperms is retained
inside the megaspore; and is dependent on the sporophyte for nutrition.
There is progressive reduction in the archegonia which is without neck
canal cells and, occasionally, ventral canal cell. Gymnosperms are seed-
bearing plants. Seed formation is absent in pteridophytes.
Gymnosperms and Angiosperms

Both gymno- and angiosperms are seeded plants, and there are many
differences in vegetative and reproductive structures: (a) The gymnosperms
are slow-growing perennial plants with limited vegetative reproduction. In
angiosperms, the plants are annual, biennial or perennial, and have varied
means of vegetative multiplication. (b) In angiosperms, both tracheids and
vessels are present. The sieve-tube elements with the sieve plates are confined
to the end walls, and companion cells are present.
The wood in gymnosperms is: (i) Manoxylic-relatively meager xylem
with wide parenchymatous rays, and of no economic use. (ii) Pycnoxylic-
xylem relatively dense with small and narrow wood rays, making up most
of the trunk and branches. The wood is commercially very useful. The
angiosperms do not show such a distinction. (iii) Pollination and dispersal
of seeds in gymnosperms is mostly anemophilous; in angiosperms
entomophilous, hydrophilous and zoophilous. (iv) The ovules are naked
and. unitegmic in gymnosperms. During pollination, the pollen grains enter
the ovules directly through the micropylar canal and rest on the nucellus.
However, in Gnetum (P. Maheshwari and V. Vasil 1961a), Tsuga,
Pseudotsuga, Abies (Doyle 1945), Torreya (Favre-Duchartre 1967) and the
Introduction 11
extinct Caytoniales (Thomas 1925, Harris 1951), the pollen grains germinate
(a short distance) away from the nucellus (an angiospermic tendency) or
inside the micropyle. In angiosperms the carpel is closed and its upper part
is differentiated into style and stigma. Pollen grains land on the stigma and
the pollen tubes grow all the way down through the style before they can
reach the ovules, which are uni- or bitegmic. Johri (1936), in a few carpels
of Butomopsis lanceolata, reported, germinated pollen grains and bits of
pollen tubes in the stylar canal and ovary. In one case, a pollen grain had
germinated on an ovule. Sahni (see Sahni and Johri 1936) considered these
features to be very significant, since a typically gymnospermous character
has been discovered in a confirmed angiosperm. Sahni also pointed out that
the pollen grains must have been sucked in (into the style) as in the micropyles
of ovules in gymnosperms. The pollen grains were three-celled and positively
of Butomopsis. Sahni recalls the inter-micropylar pollen· grains in Caytonia
(see Harris 1951; Chap. 9).
It may be added that (entire) intercarpellary pollen grains (but no pollen
tubes comparable to the condition in Butomopsis) have been reported in
several angiosperms (see Johri and Ambegaokar 1984). (v) The development
of female gametophyte in gymnosperms is monosporic (tetrasporic in Gnetum
and Welwitschia ). It undergoes a variable number of free-nuclear divisions
followed by wall formation, and remains haploid. It bears archegonia at the
micropylar end. In angiosperms the development of the female gametophyte
may be mono-, bi- or tetrasporic, and a 4-16-nucleate embryo sac is formed.
(vi) The haploid female gametophyte in gymnosperms functions as endosperm,
after fertilization. In angiosperms the endosperm is triploid, formed by the
fusion of the two polar nuclei and the second male gamete (double
fertilization). (vii) In gymnosperms, the zygote undergoes free nuclear
divisions (except in Sequoia, Gnetum and Welwitschia ) followed by wall
formation. Later, only the apical cells give rise to the embryo. Cleavage
polyembryony is known in several taxa. In angiosperms, the division of
zygote is invariably followed by wall formation (except in Paeonia ). Cleavage
polyembryony is absent.
The angiosperms are much more advanced than gymnosperms, and have
diverse habit and vegetative form. They grow successfully in all habitats
and have an efficient means of vegetative propagation which ensures rapid
multiplication. There is a greater reduction in the gametophytes-prothallial
cells are absent in the male gametophyte, and the female gametophyte
lacks archegonia. The ovules are enclosed in an ovary which protects the
ovules. This protection gives biological advantages to the angiosperms over
the gymnosperms. The seeds develop within the ovary; when mature, these
are effectively and efficiently dispersed by insects, birds, wind and water.
The antiquity and the fascination of a group of plants which stand between
cryptogams and flowering plants (in their reproductive development) make
the gymnosperms a very interesting and fruitful area for investigations.
2. Seed Development
While the plant form is quite diverse, most gymnosperms show a more or
less similar mode of reproduction. The developmental aspects and important
variations are dealt with under each order. Gnetum and Welwitschia show
remarkable deviations from the rest of the gymnosperms.
Microsporangium
There are two main patterns of initiation of microsporangium (Fagerlind
1961, 1971): (a) In most taxa it is initiated from a meristem which is
differentiated into the epidermis and the hypodermal archesporium, e.g.
Cephalotaxus (Fig. 15.6.5 A, B). (b) It is initiated from a meristem with
exposed (surface) initial cells which divide to form an outer epidermis and
an inner archesporium, e.g. Cedrus, Larix, Picea, Pinus and Pseudotsuga
(see H. Singh 1978). The archesporia! cells show a higher protein content,
RNA and histone.
Microsporogenesis
The archesporia! cells divide periclinally to form the primary parietal and
primary sporogenous layers. The primary parietal layer undergoes periclinal
divisions and gives rise to a multilayered microsporangial wall, the innermost
layer functions as tapetum. The cells of the epidermal layer develop thick
walls (except along the line of dehiscence) and is designated exothecium.
The middle layers become crushed during enlargement and maturation of
the microsporangium. The tapetum encloses the sporogenous tissue, and
comprises a single (occasionally double) layer of large, richly cytoplasmic
multinucleate cells. The tapetum is mostly of the secretory type. The walls
of tapetal and sporogenous cells are mostly pectinaceous, those of the
epidermis and wall layers predominantly cellulosic. The tapetal cells show
maximal activity during meiosis in microspore mother cells, and degenerate
after the spores are released from the tetrads. The main function of the
tapetum is to nourish the sporogenous cells and young microspores. In
addition the tapetum is concerned in the production and release of callase,
transmits PAS-positive material into the locule, forms an acetolysis-resistant
membrane (see Pacini et al. 1985), and finally the exine of the microspores.
Seed Development 13
In most gymnosperms, cytokinesis following meiosis is simultaneous

and centripetal callose walls are laid down between the four microspore
nuclei. The Golgi bodies become active and produce a variety of vesicles
during anaphase II and telophase II. These vescicles become arranged in
the region of the future wall. A two-layered membrane of ER origin separates
the four microspores. Eventually, the callose spreads between the two layers.
Plasmodesmata! connections do not exist either between the dividing mother
cells or between the microspores in a tetrad. Callose with a low permeability
isolates the mother cells and their products from the influence of adjoining
tissues. This helps the micro.spore mother cells to follow an independent
course of development during meiosis.
Male Gametophyte
The development of the male gametophyte mostly follows a uniform pattern,
except in Welwitschia and Gnetum (see Chaps. 19, 20).
The initial step in the pollen grain is the formation of one (cycads) or
two prothallial cells which are usually inconspicuous and ephemeral. The
prothallial cells are absent in Cupressaceae, Taxodiaceae, Cephalotaxaceae
and Taxales. The microspore functions directly as the antheridial initial.
After the formation of prothallial cells, the antheridial initial divides to
form a small antheridial cell and a large tube cell. The latter is usually
vacuolate and shows a large nucleus, while the small antheridial cell remains
attached to the intine at the site of prothallial cell(s). The antheridial cell
generally divides periclinally to form the stalk cell towards the pollen wall,
and body cell. Initially, the stalk cell is surrounded by a distinct wall which
eventually breaks down, and its cytoplasm merges with that of the tube
cell. In later stages the stalk and tube nuclei are indistinguishable from
each other. The body cell enlarges, has dense cytoplasm, and a large nucleus.
It divides and gives rise to male gametes.
The formation of male gametes is schematized below:
Male
gametel
{
~e~y
Antheridial Male
cell gamete 2
Antheridial { Stalk
initial cell
Central Tube
cell cell
Prothallial
cell 2
Prothallial
cell 1
14 The Gymnosperms
The male gametes may be flagellate spermatozoids, as in cycads

(Fig. 8.14 E, F) and Ginkgo (Fig. 12.15 E), or unflagellate male cells (equal/
unequal), as in all the other taxa. The male cells are common in the members
of Cupressaceae, Taxodiaceae, Araucariaceae and Gnetum spp., while male
nuclei (needs confirmation) have been reported in taxa where the archegonium
is placed individually, as in Pinaeae. Reports of male nuclei are erroneous,
and all male gametes have a cytoplasmic sheath even if there is no cellulosic
wall. In a few species of Cupressus, and occasionally in Juniperus, the body
cell divides several times to produce multiple male gametes.
Winged pollen grains occur in several taxa, e.g. Podocarpus (Fig. 15.4.7 K),
Microcachrys, Pherosphera, and members of Pinaceae. The shape and
structure of the wings vary.
The number of cells in mature pollen, at shedding, differs in various
taxa. The pollen grain is one-celled in Taxus, Chaemaecyparis, Callitris,
Cryptomeria and Cupressus; two-celled in Cephalotaxus, Athrotaxis, Torreya
and Taxodium; three-celled in most cycads, Welwitschia and Gnetum; four-
celled in Pinus; five-celled in Cedrus and Ephedra; muticelled (due to a
large number of prothallial cells) in podocarps and araucarias.
Ovule
The young ovule has a central nucellus covered by a single integument.
Ephedra has two coverings and Welwitschia and Gnetum three. All along
the central region of integument up to the nucellus is a narrow passage, the
micropyle. There is a conspicuous chalaza, the funicule is not recognizable.
The ovule is mostly orthotropous in gymnosperms, both extinct and extant. In
the family Podocarpaceae the ovule is anatropous (Konar and Oberoi 1969 b).
Megasporogenesis
The young nucellus has one to several hypodermal archesporia! cells. They
divide periclinally to form the primary parietal and primary sporogenous
cells. The parietal cell and its derivatives divide periclinally and give rise
to a massive parietal tissue capping the primary sporogenous cells. The
latter may also divide once or twice, or one or more cells may function
directly as megaspore mother cell. The latter are elongated, have prominent
nuclei, dense cytoplasm, and a thick wall. They undergo meiosis and produce
triads (due to undivided upper dyad cell) or linear tetrads, generally the
lowermost megaspore functions. The division takes place in a layer of
callose, indicating that meiosis takes place under controlled conditions,
comparable to the division of the microspore mother cell giving rise to
microspores (see Konar and Moitra 1980). One to several layers of densely
cytoplasmic cells differentiate around the sporogenous cells, and become
progressively conspicuous during subsequent stages. This is designated the
spongy tissue or tapetum. Eventually, it degenerates and becomes compressed
between the female gametophyte and outer tissues of the ovule.
Seed Development 15
Megaspore Wall. Pettitt ( 1966) has reinterpreted the structure of the

megaspore wall in gymnosperms and the following generalizations have
been put forward: (a) The megaspore wall is usually a multiple structure,
and various parts are laid down in an orderly and controlled manner. They
can be linked to other formative events in the morphogenesis of the ovule~
There is considerable variation between the species. (b) For the construction
of wall, the material is supplied by both the sporophyte and gametophyte.
From a very early stage of development, polymer sporopollenin begins to
accrue in the wall.
Pettitt (1977) investigated the ontogeny of megaspore wall (preferring to
call it wall instead of membrane), and demonstrated features comparable
with that of pollen wall.
The foundation for the megaspore wall, laid on the free surface of the
coenocytic megaspore, consists of fibrillar matrix with radially aligned
hexagonal chambers. The fibrillar coating is interpreted as primexine, and
serves as the template of the sculptured sexine of the mature megaspore
wall (this role of sexine is similar in both pollen and megaspore wall). In
the beginning, a finely granular material (the probacula) accumulates in the
chambers in the proximal region, and later extends radially to form a series
of interconnecting columns (homologous to the bacula of sexine of pollen
wall).
Simultaneously, a continuous granular basement (homologous to nexine
I or foot layer) is deposited within the undersurface of the fibrillar matrix.
As development proceeds, the columns and basement layer become electron-
opaque and amorphous. At the same time, they become resistant to acetolysis.
These are regarded as evidence for the introduction of the polymer
sporopollenin into the wall, and the changes in appearance are visible
consequences thereof. Three separate and distinct substrata are subsequently
formed below the patterned layer. The first layer (which appears to be
absent in pollen wall) is rich in polysaccharide, and the material for its
development possibly originates in the dictyosomes of the megaspore
cytoplasm. The second substratum (equivalent to nexine II of pollen wall)
is acetolysis-resistant. It is formed when electron-opaque globules are extruded
across the plasma membrane from large, membrane-bound vesicles of
unknown origin. The third layer (interpreted as equivalent to intine of the
pollen wall), as the first, contains polysaccharides in vesicles, possibly of
dictyosomal origin. This layer appears when the female gametophyte enters
the cellular phase, and is structurally continuous with the radial walls of the
alveoli.
The principle features of construction and structure of the megaspore
wall in a number of cycad genera are comparable but differ from each
other, and from Ginkgo in several details (Pettitt 1977).
The nucellar tissue degenerates progressively as the megaspore and its
wall develop. This results in the formation of a thick covering of cellular
16 The Gymnosperms
detritus (containing sporopollenin and lipids) which accumulates against

the surface of the female gametophyte.
Various investigations have revealed that a megaspore wall occurs in all
extant gymnosperms. However, it has a more elaborate structure in the
most primitive members (cycads and Ginkgo) where fertilization is affected
by motile male gametes (Pettitt 1977).
Female Gametophyte
In monosporic development, the chalaza! megaspore undergoes several free-
nuclear divisions; the number of nuclei varies from ca. 256 in Taxus baccata
and Torreya nucifera, to ca. 8000 in Ginkgo biloba. The number of free-
nuclear divisions is usually constant for a particular species. These nuclei
become arranged in a thin peripheral layer of cytoplasm enclosing a central
large vacuole. The free-nuclear gametophyte enters the cellular phase through
the formation of alveoli followed by anticlinal walls laid down centripetally.
In the beginning, the alveoli have no walls on the inner side facing the
central vacuole, and the persistent spindles appear to guide the laying down
of wall material. Different alveoli close at various distances from the centre
of the gametophyte. A large number of alveoli are present at the micropylar
and the chalaza! pole of the usually oval gametophyte. Repeated periclinal
divisions occur in the closed alveoli, except in those which function as
archegonial initials. Initially, the gametophyte shows rows of radiating cells,
but in older gametophytes this arrangement is lost due to the laying down
of irregular walls. Generally, the nuclei of the cells adjacent to the megaspore
membrane are larger than those in the middle of the gametophyte.
In Welwitschia and Gnetum, instead of one large central vacuole in the
female gametophyte, there are several vacuoles and the nuclei are dispersed
throughout the cytoplasm. Wall formation takes place by free-cell formation.
In all gymnosperms, the female gametophyte develops a central cavity,
the corrosion cavity, to contain and nourish the developing embryos. Reserve
food materials like fat, protein and starch accumulate in the gametophyte
during maturation of the seed. This is utilised at the germination of "the
seed. Thus, the female gametophyte (usually termed endosperm after
fertilization) serves a dual function of bearing the gametes as well as
nourishing the embryo.
Archegonia. The archegonia develop in all gymnosperms, except Gnetum

and Welwitschia, mostly at the micropylar pole; lateral and chalaza! archegonia
are rare. Apical archegonial complex with a varying number of archegonia
are reported in members of the Cupressaceae and Taxodiaceae (except
Sciadopitys where four or five archegonia occur singly).
The closed and undivided alveoli function as archegonial initials. The
nucleus divides, and a periclinal wall is laid down, forming a large central
cell and a smaller neck initial. The latter undergoes several divisions to
Seed Development 17
form a short neck. It helps in the entry of the male gamete into the egg cell,
at the time of fertilization. The central cell enlarges considerably, its nucleus
lies just below the neck, and the cytoplasm becomes highly vacuolate,
foam stage of the archegonium. The nucleus of the central cell divides to
form an ephemeral ventral canal cell/nucleus and egg nucleus. The latter
enlarges considerably, and the cytoplasm of the egg accumulates numerous
proteid vacuoles. A closer look with an electron microscope has shown that
the proteid vacuoles are only cytoplasmic formations of several types; large
inclusions, small inclusions, microbodies and vesicular bodies (Fig. 15.1.14).
Besides these inclusions, the cytoplasm of the egg also shows a conspicuous
ER, plastids, mitochondria, Golgi bodies and ribosomes. The egg
nucleus is usually large and filled with nucleoplasm (see Konar and Moitra
1980).
EM studies of egg of various species (Pinus nigra-Camefort 1962; Biota
orientalis-Chesnoy 1967) show that a large number of mitochondria, present
in the cytoplasm, organize in a circle around the egg nucleus and form the
perinuclear zone. The latter increases in width due to continued migration
and multiplication of mitochondria from the general cytoplasm. The egg
nucleus is Feulgen-negative and the perinuclear zone is Feulgen-positive.
One to three layers of densely cytoplasmic cells, the jacket layers, surround
the central cell/egg/proembryo. There is a thick wall between the jacket
cells and the central cell of the archegonium, due to the secondary deposition
of the wall material. This thick wall with simple pits forms the egg membrane
during post-fertilization stages. The jacket layer is concerned in the nutrition
of egg/proembryo. The food reserve (starch or protein) in the gametophytic
cells becomes available to the egg/proembryo through plasmodesmata on
the inner tangential walls of jacket cells. These cells also secrete the enzyme.
The gametophytic tissue circumjacent to the archegonia undergoes divisions
and the cells enlarge, resulting in an upward growth forming a depression,
the archegonial chamber; the archegonia appear sunken.
Pollination and Fertilization
Pollination. In different gymnosperms the ovules receive pollen at various

stages of development: (a) sporogenous cells or megaspore mother cell in
Ginkgo, conifers and taxads; (b) free-nuclear gametophyte in many cycads
and Gnetum; (c) yamg archegonia in Macrowmia, and (d) mature archegonia
in Ephedra.
The pollen is produced in large quantities, is dispersed by wind, and the
adjoining areas become covered by yellow dust, the sulphur shower. In
Ephedra aphylla (Bino et al. 1984), Gnetum sp. (Kato et al. 1995) and
Welwitschia (Kubitzki 1990) there is effective insect pollination (see Chaps.
18, 19, 20).
At the time of pollination, the gymnosperms investigated so far show a
18 The Gymnosperms
sugary exudate at the micropyle. The exceptions are: (a) Abies, Cedrus, Larix,
Pseudotsuga and Tsuga have a stigmatic micropyle; (b) Araucaria, Agaihis
and Tsuga dumosa, where the pollen grains do not land on the micropyle.
The pollen is received on a pollination drop secreted at the micropyle of
the ovule, a short distance away from the nucellus. The pollination drop is
sucked in int-o the micropyle after the pollen grains are caught in it. Finally,
the pollen lands on the nucellus, where it germinates.
The pollination drop, its secretion, chemical composition and the mode
of transport of pollen grains require fresh study using modem tools and
techniques.
The pollen tubes grow towards the female gametophyte. At its tip the
tube contains the body cell, and later the male gametes. The short and
unbranched tube is quite prominent. In cycads and Ginkgo (see Chaps. 8,
12) the pollen tube is non-siphonogamous. It arises from the distal (upper)
end of the pollen grain, elongates and grows laterally and intercellularly
into the apical region of nucellus. The tube branches in Cycas and Ginkgo
but remains unbranched in Zamia. These pollen tube branches are enucleate
and are not concerned with the transport of male gametes for fertilization.
Their main function is to absorb nutrition from the circumjacent tissue.
From the proximal end also a small tube is formed through which the
motile spermatozoids pass into the pollen chamber (see Friedman 1987,
Choi and Friedman 1991, Johri 1992, Norstog pers. comm).
Fertilization. The release· of male gametes varies in different taxa of

gyrimosperms. (a) In cycads and Ginkgo, the gametes are released in the
archegonial chamber which may contain a fluid. (b) In gymnosperms where
the archegonia occur singly (Pinus, Podocarpus, Cephalotaxus, Taxus,
Ephedra), the neck cells degenerate and the pollen tube penetrates the egg
cell and releases the male gametes. (c) In taxa where the archegonial
complexes are laterally placed as in Athrotaxis and Callitris, the pollen tubes
grow adpressed to the neck of several archegonia. The male gametes, while
still inside the pollen tube, become closely attached to the neck cells (~hich
form a passage), which eventually degenerate. The male gametes then pass
into the egg cell leaving behind its cytoplasmic sheath. Two male gametes
can enter two archegonia (Brennan and Doyle 1956, Baird 1953). On entering
the egg cytoplasm, the male gamete (along with some cytoplasm) comes in
contact with the egg nucleus. The non-functional male nuclei, once inside
the egg cytoplasm, usually persist for some time.
Studies with the light microscope, as well as EM, on the characteristics
of the male and female cytoplasm during fusion and subsequent development
(in a few conifers), have led to the concept of neocytoplasm. The nucleoplasm
of both male and female nuclei form the ground substance of neocytoplasm.
The neocytoplasm alone takes part in the formation of embryonal cytoplasm;
the remaining cytoplasm of zygote degenerates. Further studies are called
for.
Seed Development 19
Embryo
The development of embryo can be divided into three phases.
(a) Proembryogeny. Development beginning with the division of zygote
up to the stage prior to elongation of the suspensor. There is heterogenity
in the development and four types can be distinguished: (i) Cycad
and Ginkgo type, (ii) Conifer type, (iii) Ephedra and Sequoia type, and
(iv) Gnetum and Welwitschia type.
(b) Early embryogeny. Varies in different taxa; it includes elongation
and proliferation of suspensors and formation of young embryonal mass.
(c) Late embryogeny. Further development of the embryo, and
establishment of polar meristems, i.e. root and shoot.
Seed
Seed Coat. The seed coat may develop (a) mainly from the tissue derived
from the chalaza] portion of the ovule, e.g. cycads, members of Pinaceae,
and Cephalotaxus; (b) both from chalaza and integument, e.g. Ginkgo;
podocarps, taxads and Gnetum; (c) mainly from integument, e.g. araucarias,
taxoqioids etc. Whatever the mode of origin, the initial changes are (i) an
increase in the number of cell layers in the particular region followed by
(ii) differentiation of mucilage canals (cycads) and resin ducts (in conifers),
(iii) differentiation of xylem and phloem in the provascular strands (cycads,
Cephalotaxus, taxads, Ephedra, Gnetum and Welwitschia), (iv) deposition
of tannin in several cells distributed all over the young seed coat, and
finally (v) the differentiation of three specific layers in seed coat-outer
parenchymatous sarcotesta, middle sclerenchymatous sclerotesta, and
innermost thin-walled endotesta. In members of the Pinaceae, a few layers
of cells of the ovuliferous scale, adjacent to the ovule, give rise to a wing.
In Araucariaceae, the wing arises from the entire bract scale, and in
Welwitschia from ·the outer envelope, the perianth (see Chap. 19).
Temporal Considerations
The time lag between ovule initiation and seed maturation in angiosperms
may usually be only a month or so, while in gymnosperms it takes much
longer. There are 1-year, 2-year and 3-year types of reproductive cycle.
The year is counted by the number of winter rests, are followed by growth
in the spring, through which the ovule passes. In most temperate
gymnosperms, the course of development of the reproductive structures is
seasonal, i.e. not continuous, while in the tropical and subtropical
gymnosperms like cycad, Gnetum and Ephedra, the development of the ovule
is a continuous process. In Ephedra the reproductive cycle takes about
3 months to complete. In other gymnosperms, the development of the male
cone and the ovule on a plant or a stand is usually synchronous. The details
of pollination and fertilization in different ovules are more or less the same.
3. Classification
One of the objectives of systematists is to discover a phylogenetic system

of classification. The evolutionary history of gymnosperms extends far
back in geological time (Figs. 1.1, 1.2); its fossil record begins in the Upper
Devonian and spreads over nearly 350 my B.P. (million years Before Present)
(Beck 1985). Most branches of the phylogenetic tree are extinct or have left
only fragmentary palaeobotanical records. Several presumed gymnospermous
remains have been described from the Upper Devonian and Lower
Carboniferous. It is, therefore, natural that there will be diversity of opinion
on the phylogenetic relationships and classification of plants. However, it
is felt that classification schemes should be flexible, and should be
continuously revised in the light of growing knowledge (Arnold 1948). The
evolutionary relationships of fossil plants are, in most instances, not completely
known (Miller 1985). Until reproductive organs are discovered in organic
connection with the vegetative parts, their taxonomic. placement is entirely
optional (Sporne 1965).
In classification, both vegetative and reproductive organs are important.
A knowledge of anatomy, embryology, cytology, palynology and
phytochemistry should be used in determining the validity of the system of
classification (see H. Singh 1978).
Sahni (1920) recognied two main phyletic lines in gymnosperms:
1 Phyllosperms. Leaves large and much divided, ovules borne on
leaves or organs regarded as such: (a) Pteridospermales, (b) Cycadales,
and (c) Bennettitales.
2 Stachysperms. Leaves simple, ovules borne on stems:
(a) Cordaitales, (b) Ginkgoales, (c) Coniferales, and (d) Taxales
including Taxus, Torreya and Cephalotaxus; separated from
Coniferales.
Florin (1948) upheld the separation of Taxales as an Order, coordinate
in rank with the Cordaitales, Ginkgoales and Coniferales (restricted). However,
he included only Taxus, Torreya, Nothotaxus; Amentotaxus, Austrotaxus
and their fossil relatives, and retained Cephalotaxus within Coniferales.
Chamberlain ( 1920, 1935)-almost simultaneously with Sahni-recognized
two main groups amongst gymnosperms:
Classification 21
1 Cycadophytes. (a) Cycadofilicales, (b) Bennettitales, and

(c) Cycadales.
2 Coniferophytes. (a) Cordaitales, (b) Ginkgoales, (c) Coniferales,
and (d) Gnetales.
The main difference between Sahni's and Chamberlain's classification
is the emphasis on the primary characters. Sahni's main basis is the
morphological nature of the ovule-bearing organs. This, according to Scott
(1923), is mainly a theoretical distinction for groups like Cordaites,
Bennettitales and Coniferales. Chamberlain emphasizes mainly the differences
in habit, stem anatomy and leaves.
Arnold (1948) gave a palaeobotanical foundation to Chamberlain's (1935)
system. He interpreted the class Gymnospermae as an artificial polyphyletic
group where the seed habit arose independently in cycadophytes and
coniferophytes. Arnold, therefore, recommended that the class Gymnospermae
should be dropped from the systems of classification. He divided
Gymnosperms into three phyla.
1 Cycadophyta. (a) Pteridospermales, (b) Cycadeoidales, and (c)
Cycadales.
2 Coniferophyta. (a) Cordaitales, (b) Ginkgoales, (c) Taxales, and (d)
Coniferales.
3 Chlamydospermophyta. (a) Ephedrales, and (b) Gnetales.
According to Arnold, the Cycadophytes and Coniferophytes represented
natural groups and had separate origin.
Pant (1957) modified Arnold's classification:
Division 1. Cycadophyta: Class Pteridospermopsida: (a) Lyginopteridales,
(b) Medullosales, (c) Glossopteridales, (d) Peltaspermales, (e)
Corystospermales, and (f) Caytoniales. Class Cycadopsida: (g) Cycadales.
Class Pentoxylopsida: (h) Pentoxylales. Class Bennettitopsida
(Cycadeoideopsida): (i) Bennettitales (Cycadeoidales).
Division 2. Chlamydospermophyta: Class Gnetopsida: (a) Gnetales,
(b) Welwitschiales.
Division 3. Coniferophyta: Class Coniferopsida: (a) Cordaitales,
(b) Coniferales, (c) Ginkgoales. Class Ephedropsida: (d) Ephedrales. Class
Czekanowskiopsida: (e) Czekanowskiales. Class Taxopsida: (f) Taxales.
Bierhorst (1971) presented a slightly different classification:
1 Cycadopsida: (a) Pteridospermales, (b) Cycadales, (c) Cycadeoidales,
and (d) Caytoniales.
2 Coniferopsida: (a) Cordaitales, (b) Coniferales, (c) Taxales, and (d)
Ginkgoales.
3 Gnetopsida: (a) Ephedrales, (b) Gnetales, and (c) Welwitschiales.
Most classifications of gymnosperms are based on criteria proposed by
Chamberlain (1935) and Arnold (1948). Of the two major lineages recognizyd
22 The Gymnosperms
by them, the Cycadophyta was presumed to have evolved from ferns, while
the origin of the Coniferophyta was unknown. Pitys and Callixylon, petrified
tree trunks with coniferophytic secondary xylem, were regarded to be the
oldest representatives. It was later discovered that Pitys is a lyginopterid
pteridosperm (Long 1963, 1979), Callixylon is a progymnosperm (Beck 1960),
and the Mesozoic seed ferns are much more diverse and abundant than was
formerly realized (Miller 1985). These discoveries conflict with Arnold's
scheme of classification.
Beck (1981) and Rothwell (1982) proposed schemes to account for the
role of progymnosperms. The progymnosperms show a combination of
pteridophytic (spore-producing reproductive structure) and gymnospermic
(arborescent, abundant secondary xylem, bordered pits on xylem tracheids,
and leaf traces without leaf gaps) characters (see Beck 1970). This group
is considered to be the progenitor of gymnosperms, i.e. the Cycadophyte
and Coniferophyte lines converging at the base.
Stewart (1983) proposed a classification which is a synthesis of several
different .natural systems. He used evidence from the fossil record which
has contributed extensively to establishing the natural relationships within
the major groups of vascular plants:
1 Progymnospermopsida (ancestors of gymnosperms): (a) Aneurophy-
tales, (b) Archaeopteridales, and (c) Protopityales.
2 Gymnospermopsida "Cycadophytes": (a) Pteridospermales,
(b) Cycadales, (c) Cycadeoidales; "Gymnosperms of uncertain
relationship": (d) Caytoniales, (e) Glossopteridales, (f) Pentoxylales,
(g) Czekanowskiales, (h) Ginkgoales; "Coniferophytes": (i) Cordaitales,
(j) Voltziales, (k) Coniferales, (1) Taxales.
3 Gnetopsida Gnetum, Ephedra, Welwitschia.
Meyen (1984, 1986) proposed a classification for all gymnosperms, past
and present. He erects three classes, namely, Ginkgoopsida, Cycadopsida
and Pinopsida, which agree with the currently available palaeobotanical
data. The classes are based on the analysis of the structural variation in
gymnosperms and interpretations of homologies of various organs. The
Ginkgoopsida is established as a distinct Class and a major phylogenetic
line coordinate with the Cycadopsida and Pinopsida. This is a significant
departure from the preceding classification. It comprises Calamopityaceae,
Callistophytales, Glossopteridales (Arberiales), Peltaspermales, Caytoniales,
and Ginkgoales, among others. The Cycadopsida and Pinopsida are relatively
natural groups. Meyen treats Gnetales and Welwitschiales in the Cycadopsida
(the reason for this treatment is not stated), and Ephedrales in the
Ginkgoopsida (because of its platyspermic seed and striated pollen). According
to Meyen (1986), the gymnosperms are a monophyletic taxon originating
from one of the progymnospermic Orders, probably Archaeopteridales.
During the past (nearly) 50 years, significant progress has been made in
Classification 23
understanding the characteristic features of the established taxa. However,

new taxa have not been described, and the interrelationships between the
existing 'taxa have also not changed significantly (Rothwell 1985).
In this book the classification (given below) is based on Stewart's (1983)
suggestions:
1 Progymnospermopsida: (a) Aneurophytales, (b) Archaeopteridales,
and (c) Protopityales.
2 Gymnospermopsida: (Cycadophytes): (a) Pteridospermales,
(b) Cycadeoidales, (c) Cycadales; (Gymnosperms of uncertain
relationships): (d) Caytoniales, (e) Glossopteridales, (f) Pentoxylales,
(g) Czekanowskiales, (h) Ginkgoales; (Coniferophytes): (i) Cordaitales,
G) Voltziales, (k) Coniferales, (1) Taxales.
3 Gnetopsida: (a) Ephedrales, (b) Welwitschiales, (c) Gnetales.
For a more convincing scheme of classification of gymnosperms, data
from all disciplines (mentioned earlier) should be critically analysed to
frame a phylogenetic tree - as "natural" as possible.
4. Progymnospermopsida
Beck's (1960) discovery of the organic connection between the leaf of a

free-sporing fern, Archaeopteris, and a stem, Callixylon, with gymnospermous
characters led to the establishment of the Class Progymnospermopsida, the
probable progenitors of gymnosperms (Namboodiri and Beck 1968). This
finding of "plants having gymnospermic secondary wood and pteridophytic
reproduction" had a profound effect on the interpretation of the origin of
gymnosperms and their systematics.
The above specimen, from the Upper Devonian (ca. 395 my B.P.) beds
in New York State, is 80 em long. It had compressed fertile and sterile
foliage of Archaeopteris macilenta type attached to partially permineralized
stem and rachis axes of Callixylon zalesskyi. Beck (1962) reconstructed the
plant as a tree up to 18 m tall with a crown of spirally arranged bipinnate
fronds (Fig. 4.1). He used the name Archaeopteris for the entire plant,
according to nomenclatural priority.
Before 1960, Archaeopteris was presumed to be the foliage of a Late
Devonian fern-like plant with large compound leaves. Nearly 15 spp. are
now known. The bipinnate frond with opposite or subopposite fertile and
sterile pinnae are the penultimate branches of the frond. The fertile pinnules
bear one or two rows of adaxial fusiform sporangia which dehisced
longitudinally. Callixylon is also a Late Devonian/Lower Carboniferous
taxon which often occurs in the same bed as Archaeopteris. Permineralized
specimens range from small twigs and branches to trunks with a diameter
of 1.5 m and height of nearly 20 m. In transection, many of them show a
conspicuous pith with a ring of mesarch primary xylem strands at the
periphery in contact with the secondary xylem. The latter is compact, and
the wood is typically pycnoxylic (Fig. 4.2 A) as in modern conifers. Most
of the trunk is made up of secondary wood and the tracheids are pitted in
a characteristic manner (typical Cordaitean type). On the radial walls, the
pits are arranged in groups separated by unpitted regions, and this pattern
is repeated on adjacent tracheids. In a longitudinal radial section, continuous
horizontal bands of pits are separated by unpitted bands (Fig. 4.2 B). This
feature makes the genus immediately recognizable.
Progymnospermopsida 25
Fig. 4.1. Archaeopteris sp., reconstruction (After Beck 1962)
Phylogenetic Considerations of Archaeopteris

The arborescent habit, megaphyllous leaves, secondary growth, circular
bordered pits, ray tracheids, collateral vascular bundles and probable
heterosporous reproduction suggest Archaeopteris to be intermediate between
the Psilophyte and the seed plant (Beck 1962). It had, however, advanced
beyond the Lower and Middle Devonian psilophytic level and approached
the Carboniferous gymnosperms. Archaeopteris is also similar to the
pteridosperms in foliage and fructifications, and in its primary body to both
pteridosperms and some coniferophytes. The secondary wood is remarkably
similar to the coniferophytes. There is, thus, considerable evidence in support
of its being a progymnosperm (one of the advanced genera). It had either
evolved concurrently with (and in some ways parallel to) both primitive
pteridosperms and coniferophytes, or was directly ancestral to some
pteridosperms or coniferophytes. Further information is, however, necessary
for its final determination (Beck 1962).
26 The Gymnosperms
Fig. 4.2 A, B. Callixylon newberryi. A Transection secondary wood shows ray tracheids
(rt). B Radial section, secondary wood with grouped pitting. (After Beck 1970)
5. Gymnospermopsida
According to Beck (1976), the Progymnospermopsida represents a network

of Devonian age from which Cycadophytes and Coniferophytes evolved
(Chap. 4). The class Gymnospermopsida (excluding Gnetopsida) forms a
natural group of vascular plants comprising at least two divergent phyletic
lines (Cycadophytes and Coniferophytes), which evolved certain distinctive
characteristics (Stewart 1981).
In Cycadophytes (Chaps. 6-8) including Pteridospermales, Cycadeoidales
and Cycadales, the plants have (a) large pinnate frond-like leaves, (b) stem
with a well-developed pith and cortex, (c) manoxylic secondary xylem with
very long tracheids of large diameter, several rows of bordered pits mostly
on the radial walls, and high multiseriate wood rays, (d) the ovules and
pollen organs are borne on modified leaves, and (e) a dicotyledonous embryo
(wherever known) in a seed.
In Coniferophytes (Chaps. 13-16), including Cordaitales, Voltziales,
Coniferales and Taxales, the plants generally have (a) spirally arranged
simple leaves, (b) compact woody stems with pycnoxylic secondary xylem
of tracheids and narrow vascular rays, a gymnosperm-type eustele where
leaf traces diverge from sympodia and (c) heterospory in most species.
Besides these two groups, there are other Mesozoic seed plants whose
relationship are unknown/uncertain (Chaps. 9-12).
6. Pteridospermales
The discovery of this group of gymnosperms from the Palaeozoic is one of

the notable achievements of the palaeobotanists, and of enormous importance
to phylogeny. It includes fern-like assemblage of seed plants' which first
appeared in the Upper Devonian. It extended to the Mesozoic through the
Carboniferous and Permian (Fig. 1.1) and became extinct millions of years
ago. Originally, they were presumed to be ferns and the name age of ferns
given to the Carboniferous period was largely due to this misconception.
Williamson (1887) first recognized, in the carboniferous flora, plants
combining in their anatomical structures the characters of both ferns and
cycads. Keeping this in view, H. Pot6nie (1899) named this group
Cycadofilices to indicate its composite character. It was later discovered
that most of these fern-like leaves were associated with true seed structures.
Because of their similarities to the pteridophytes on the one hand, and
spermatophytes on the other, the name Pteridospermae was proposed by
Oliver and Scott (1904). It also included Cycadofilices (plants containing
seeds).
A criterion for the separation of true ferns from the seed plants has been
recognized (see Thomas 1955). The fossil leaves of Cycadales, Cycadeoidales
(Bennettitales), Ginkgoales and Coniferales have cuticles which resist the
action of strong acidic oxidizing agents. They could be separated and, after
suitable treatment, examined for the shape of epidermal cells and structure
of stomata (see Chap. 7). The fragments of fossil fern leaves dissolve
completely when treated in a similar manner. Some of the Palaeozoic and
most of the supposed Mesozoic pteridosperm fronds have cuticles similar
to gymnosperms. The same treatment reveals the cutinized membranes in
fossil seeds, pollen-bearing structures, and pollen grains. By comparing the
form, size, appearance of epidermal cells, and the stomatal apparatus of
reproductive organs with those of associated leaves; it could be determined
from which leaves the fertile organs were derived. Cuticular studies thus
helped in the reconstruction of original plants.
The fossil remains of most of these plants are, however, of fragmentary
nature and only a few are completely known. The description is, therefore,
based on isolated parts which are assigned to a large number of form
genera.
Pteridospermales 29
Some of the features that generally characterize pteridosperms are: (a)

Plants rather large with slender stems. There is a solid or a medullated
protostele with mesarch (rarely exarch) primary xylem, occasionally polystelic.
The bifacial vascular cambium produced secondary phloem and a limited
amount of manoxylic secondary wood composed of tracheids with multiseriate
pitting, especially on the radial walls. There is a prominent and characteristic
outer cortex of longitudinally aligned fiber strands. (b) The leaves are
mostly large, fern-like, and often multipinnate. (c) The ovules and
microsporangia are borne on unmodified or only slightly modified foliage.
The seeds are borne on the fronds in a variety of ways. In some, they are
partially enclosed in a distinct cupule. The microsporangiate organs are
terminally clustered sporangia which usually form large complex synangial
fructifications (Andrews 1961 ).
The Pteridospermales are more diverse in their reproductive biology
than any other group of vascular plants. They have been studied intensively,
as they are central to our understanding of gymnosperm evolution.
The Palaeozoic Pteridospermales are generally classified into four families:
Calamopityaceae, Lyginopteridaceae, Medullosaceae, and Callistophytaceae.
It is presumed that the seed ferns originated from protostelic Aneurophytales
(Middle Devonian) through the Calamopityaceae (Upper Devonian and Lower
Carboniferous) where the other families diverged. The Callistophytaceae
and Lyginopteridaceae are abundant in the upper Carboniferous along with
Medullosaceae which continued into Permian (Fig. 1.1).
Lyginopteridaceae
Among the Palaeozoic pteridosperms, the Lyginopteridaceae represent some
of the oldest (stratigraphically), simplest and most primitive forms (Pigg
et al. 1986). The members occur in the Carboniferous (see Fig. 1.1). Several
stem genera, ovules and different types of pollen organs have been discovered
from permineralized remains alone (Taylor and Millay 1981 ). However,
many questions still remain unanswered, concerning the morphology, affinities
of disarticulated organs and reconstruction of whole plants.
Lyginopteris oldhamia (Calymmatotheca hoeninghausi)

This is the most familiar and best known of the pteridosperms. Different
plant parts have been discovered at various times and given separate names.
It illustrates a classic case of fossil plant reconstruction from 1828-1929.
L. oldhamia is Upper Carboniferous plant discovered in England, Europe
and North America, and is extremely common in coal balls from the coal
mines of Lancashire and Yorkshire.
Habit. The frequently branched stem (L. oldhamia) is long, slender,

ca. 3-4 em in diameter, and soft-textured. It bore ca. 0.5m long spirally
arranged leaves. The radial symmetry of the stem indicates that the plant
30 The Gymnosperms
grew erect, although it seems impossible that such a small stem could have
supported a large crown of leaves without additional support. It is assumed
that the plant reclined somewhat against other plants, or against steep cliffs,
and grew in thickets or jungle-like associations, or that they were small
scrambling lianas, or possibly shrubs.
Root. The roots (Kaloxylon) are adventitious and grow from the leaf-
bearing portion of the stem, as well as from the older parts from where the
leaves had already fallen. The larger roots have a diameter of about 7 mm
and produce numerous lateral roots. It has exarch protostele with considerable
secondary growth. The diarch lateral roots arise exactly opposite to the
protoxyleni.
Stern. Well preserved petrifactions of the stem show a large central pith
of parenchymatous cells and clusters of thick-walled cells (sclerotic nests)
with dark contents. The size of the pith, and the amount of the two types
of tissues, with respect to each other, vary. Several mesarch (nearly exarch)
primary xylem strands are present around the pith. There is a continuous
ring of secondary wood (Fig. 6.1A) of large tracheids with numerous angular
bordered pits irregularly arranged on the radial walls. The rays are 1-12
cells wide and four or five cells high. Occasionally, the amount of internal
secondary wood varied. In well-preserved stems, the secondary phloem-can
be distinguished immediately outside the secondary wood, followed by a
narrow band of radially aligned cells referred to as a periderm. The outer
cortex consists of radially elongated fibre bands which form a regular
anastomosing network (Dictyoxylon cortex) in their longitudinal course
through the stem (Fig. 6.1 B). The windows of this network are filled with
parenchymatous cells. The fibrous outer cortex makes Lyginopteris one of
the easiest fossils to identify and must have given rigidity to the stem. In
transection, the disposition of fibres gives the general impression of Roman
numerals on a clock face (Fig. 6.1 A).
Leaf. The leaves were borne in a 2/5 phyllotaxy. The frond (Sphenopteris
hoeninghausii) forked 10 em from its point of junction with the stem, and
each of the two divisions was three-times pinnate (Andrews 1961). The
petiole was slightly swollen at the base, and forked about half way up the
frond into two arms. The pinnae were laterally arranged on the rachis
below the fork as well as above it. Secondary pinnae bear two alternate
rows of small-lobed pinnules (Fig. 6.2 D). The epidermis is cutinized. The
lamina showed a mesophyll of palisade and spongy parenchyma. Stomata
occur on the lower side. Figure 6.2 C shows a compression-impression
fossil of Sphenopteris-type foliage terminating in clusters of cupules (with
characteristics of Stamnostoma; see Stewart 1983).
Pteridospermales 31
Fig. 6.1 A, B. Lyginopteris oldhamia stem. A Transection. It leaf trace. oc outer fibrous
cortex. B. Outer cortex in tangential longisection. (After Andrews 1961)
32 The Gymnosperms
Fig. 6.2 A-F. A Lyginopteris oldhamia, capitate gland. B Lyginorachis sp., transection
petiole. C Compression-impression fossil of Sphenopteris type foliage terminating in
Stamnostoma cupules. D Sphenopteris pinnules. E Lagenostoma ovoides in cupule
of Calymmatotheca. F Lagenostoma ovoides, longisection ovule in cupule (cu ), capitate
glands (cg ) occur on cupule. cc central column. i integument. f lagenostome. 'mg
megagametophyte shows archegonia. n nucellus. (A After Scott 1909, B, C after Long
1963, D after Arnold 1947, E after Oliver and Scott 1904, F after Long 1944)
The base of the petiole shows two strands. A leaf trace originates by
tangential division of a primary wood strand, passes out through the secondary
xylem (by way of a large ray.) and is associated with a small strip of
secondary tissue on its outer side. The trace divides in the inner cortex of
the stem; the two resultant traces traverse the cortex and ultimately unite
again at the base of the petiole to form a V-, Y- or W-shaped bundle with
Pteridospermales 33
the concavity towards the upper side (Fig. 6.2 B). The xylem is completely
surrounded by phloem, and there are sev.eral protoxylem groups on the
lower side. The hypodermis consists of thick-walled cells, and the inner
cortex contains plates of stone cells. The isolated preserved fragments of
petioles show "butterfly" traces and are usually placed in the form genus
Lyginorachis.
Except the roots, all other parts of the plant have capitate glands which
provide the clue to the connection between the seed and the vegetative
organs. The glands are stalked or sessile. The spherical head contained a
mass of cells which probably had a secretory function (Fig. 6.2 A). The
glands are considered as emergences, formed by the epidermis and outer
part of cortex.
Sporangia. Pollen-bearing organs have not been observed in organic

connection with the stems or seeds of Lyginopteridaceae. However, most
palaeobotanists now presume that Crossotheca is the likely microsporangiate
fructification of Lyginopteris. The fertile organs attached to foliage are
classified as Sphenopteris. In Crossotheca, the fertile branch tip is slightly
expanded into a circular or oval limb, described as hair brush or epaulettes.
On the margins of the lower surface these bear a fringe of suspended
bilocular sporangia, 3 mm by 1.5 mm (Fig. 6.3). The sporangia are apparently
devoid of annuli and contain spores with triradiate markings. Crossotheca-
type microsporangia produce trilete prepollen of the dispersed type observed
frequently in Lagenostoma pollen chambers (Stewart 1983).
Several authors have suggested that biotic pollination may have occurred
in some Carboniferous pteridosperms. To attract pollinators (insects and
animals), there could be some ea~ly type of nectary as glandular hairs, e.g.
capitate hairs on the cupule lobes of Lagenostoma (Taylor and Millay 1979).
Seed The seed (Lagenostoma lomaxi) is usually without a cupule. However,

during early development, it was surrounded by a lobed cupule covered by
capitate glands (Fig. 6.2 E). The cupules terminate the ultimate naked
branches of the frond, which also bears sterile pinnules. The cupules are
tulip-shaped or like the husk of a hazel nut. The envelope formed by the
cupule has eight to ten lobes in the distal half. Reconstructions show that
the lobes apparently open outward, which allowed the ovule to shed. The
cupule enclosed a single erect ovule attached to it by its base.
The seeds are barrel-shaped, radially symmetrical and 5.5 mm long, 4.4
mm in diameter. There is a single vascularized. integument which adheres
to the nucellus, except in the apical region.
The nucellus shows a conical prolongation at its apex and is surrounded
by a moat-shaped pollen chamber (Fig. 6.2 F). In well-preserved specimens,
numerous pollen grains have been observed in this space. The integument
is quite stout, there are nine vascular bundles, and there are no free lobes
34 The Gymnosperms
Fig. 6.3 Crossotheca sp., male fructification, fertile pinnule shows pendent sporangia.
(After Andrews 1961)
at the apex. There is a simple hole through which the lagenostome (a

pollen-capturing device) protruded slightly (Fig. 6.2 F). Long (1944) described
a specimen of a closely related species, Lagenostoma ovoides Williamson,
where the female gametophyte was preserved and showed several archegonia
near the micropylar end. The central apical portion of the megagametophyte
elongated into a "tent-pole" apparatus. It may have ruptured the megaspore
membrane and floor of the pollen chamber, and exposed the archegonia to
the microgametophytes. After pollination, it may have pushed the central
column into the distal end of the micropyle and plugged it.
Most of the seeds discovered in English petrifactions are isolated. It is
presumed that they were released from the cupule by a basal abscission
mechanism (Andrews 1961 ).
Phylogenetic Considerations
For several years, the pteridosperms were presumed to be an evolutionary
link between the ferns and cycads. There is, however, evidence that the
Pteridospermales 35
origin of Filicopsida is independent of the Cycadophytes. Thus, any similarity

between the two groups must be interpreted as parallel evolution (Stewart
1983).
The Palaeozoic pteridosperms display several evolutionary adaptations
and variations which, in later geological time, became established in the
vascular plants that evolved from them (Stewart 1983). Some major trends
in evolution are: (a) The planated bilaterally symmetrical frond evolved
from three-dimensional radially arranged telome trusses. (b) The evolution
of (gymnospermous) eustele from protostelic ancestors; and manoxylic wood.
(c) Cyclic synangia evolved from clusters of microsporangia which terminated
fertile telome trusses. (d) A variety of pollen and prepollen types evolved
from trilete spores. (e) The protective layers, around the megasporangium,
evolved from adjacent telomes to form integumentary layers and cupules.
(f) Special mechanisms such as the lagenostome and pollination drops evolved
to direct pollen to the pollen chambers. Pollen tubes appeared for the first
time.
7. Cycadeoidales
Foliage generally attributed to Cycadophytes (cycads and cycadeoids) has

been observed from the Permian. In the Mesozoic there are several remains
of foliage considered to be definitely of cycads and cycadeoids (they cannot
be separated by morphological features alone). The leaves had characteristic
leathery texture due to heavy cutinization of the epidermis, and abundant
supporting tissue within the lamina and around the veins. This is because
the foliage was exposed to arid and semi-arid climates for 200 or more
million years and, therefore, could survive the adverse conditions of the
Permian much better than the Pteridosperms (Arnold 1953). These tissues
are able to resist decay, the compressed foliage often retains part of the
cutinized epidermis which can be separated from the rock and studied
(microscopically).
Extensive studies of the epidermal structure of cycadophytes have been
made to understand the relations between living and fossil forms. Nathorst
( 1902, 1909), a pioneer in the investigation of cutinized material, developed
a technique where the pattern of epidermal cells could be shown by colloidion
films. Thomas and Bancroft (1913) studied and compared the epidermal
characters of recent cycads with several Mesozoic forms. Florin (1931)
observed two types of stomata in gymnosperms. These differ in the mode
of origin of the guard and sub~idiary cells: (a) Haplocheilic-the stomatal
mother cell divides once, and the daughter cells form guard cells. The
adjoining cells are the subsidiary cells and they often encircle it. (b)
Syndetocheilic-the mother cell divides twice to form two lateral subsidiary
cells, the third division produces the guard cells. The haplocheilic stomata
are characteristic of Pteridosperms, Cycadales, Ginkgoales, Cordaitales,
Coniferales and Ephedrales; the syndetocheilic of Cycadeoidales,
Welwitschiales, Gnetales and some angiosperms. P. Maheshwari and V.
Vasil (1961 b) report haplocheilic stomata in Gnetum (see Chap. 20). Harris
(1932) showed that the epidermal cells of cycads vary in shape, and the
depth at which the stomata lie below the surface also varies. The cuticle
covers the entire outer surface of the epidermis, except the actual stomatal
opening (Fig. 7.1 A-E). In Cycadeoidales, the cuticle which covers the
guard cell extends backward under the subsidiary cell (Fig. 7.1 F,G), and
is thickest where the cuticle of the subsidiary cell joins it.
Cycadeoidales 37
Fig. 7.1 A-G. Stomatal apparatus in cycads (A-E) and cycadeoides (F, G). A, B
Zamia muricata, C Stangeria paradoxa, D Cycas revoluta, E Dioon edule, F Pterophyllum
rosenkrantzii, G Ptilophyllum pecten. (A-D, F, G After Harris 1932, E after Florin 1931)
The cycadophytic stomatal apparatus often presents a confusing picture,

in surface view' due to the outlines of the subsidiary and encircling cells
which rise above the guard cells. In living cycads, the structure can be
analyzed by sections and microchemical tests (Harris (1932). In fossils,
these techniques cannot be applied, as only the surface cuticule of the
guard cell is all that remains.
The cycadophytic leaf remains are placed in two Orders: (a) Nilssoniales
(cycads)-the general structure is similar to living cycads, the stomata are
haplocheilic, the guard cells have a thin deposit of cuticle, the epidermal
cells are not oriented in rows and have straight walls, e.g. Nilssonia. (b)
Bennettitales (cycadeoids) -the stomata are syndetocheilic, the guard cells
are heavily cutinized on their outer and dorsal walls, and the stomata tend
38 The Gymnosperms
to be oriented at right angles to the veins. The epidermal cells are arranged
in distinct rows and have walls with wavy outline. Some common frond
genera are Pterophyllum (abundant), Zamites and Otozamites.
The Cycadeoidales, which coexisted with Cycadales during the Mesozoic,
became extinct by the end of the Cretaceous. Some of the well-known taxa
in this Order are: Cycadeoidea (Cycadeoidaceae), Williamsonia,
Williamsoniella and Wielandiella (Williamsoniaceae).
Cycadeoidaceae
Cycadeoidea. Cycadeoidea with more than 30 spp. flourished from the

middle Triassic throughout the Jurassic to Early or Middle Cretaceous.
They had a wide geographical distribution: USA, Mexico, Italy, Belgium,
France, Germany, Austria, Poland and India, The first specimens were
collected from the Isle of Wight and Isle of Portland. In America, most of
the specimens are from the Black Hills of South Dakota. These are petrified
trunks and the specimens are heavy, bulky and hard, and resembled beehives
so much that the collectors were misled to interpret them as beehives,
wasp's nests, corals, etc. The trunks are stout, globular, armoured with
spirally arranged leaf bases embedded in a ramen tum of flat tongue-shaped
scales. Some of them .were unbranched, while others branched near the
ground level into a cluster of short trunks which resembled a bunch of
pineapples (Fig. 7.2).
Stem. A transection of the stele of Cycadeoidea shows a large pith in the

centre surrounded by a ring of endarch primary xylem (Fig. 7.3 B), and a
broad zone of manoxylic secondary xylem and phloem in nearly equal
proportion. The wood comprises scalariform tracheids with circular bordered
pits on radial walls. The ring is dissected by wide rays. In addition, several
uni-or biseriate rays are also present. Each leaf trace is C-shaped at the
point of origin. As it passes through the cortex, it splits into a number of
mesarch bundles which form closed cylinder at the base of the leaf. There
is no indication of girdling and the leaf traces directly supply a leaf base.
Delevoryas (1959, 1960) studied th~ formation of the cone vascular
trace and its subtending leaf trace. In C. wyomingensis, the cone trace is
derived from the fusion of four leaf traces deep in the cortex. In its outward
course the large cone trace gives rise to four leaf traces, and one of these
supplies the leaf subtending the cone.
Leaves. Specimens with attached mature leaves have not been discovered,
but it is assumed that they were spirally arranged and borne in a crown at
the apex of the trunk (Fig. 7.3 A). The leaves were simple and pinnate, as
in a bud containing young unexpanded leaves. They were assigned to
Ptilophyllum or a Dictyozamites type.
Cycadeoidales 39
Fig. 7.2 Cycadeoidea marshiana trunk with profuse branches. (After Wieland 1906)
Reproduction
The reproductive organs are bisporangiate cones (Crepet 1972, 1974) borne
among persistent leaf bases (Fig. 7.4 A). More than 500 cones, one in the
axil of every leaf, have been observed on a single trunk of Cycadeoidea
dartonii and appear as rosette-like bodies. Figure 7.3 C is a tangential
section of the trunk of C. jenneyana which shows leaves and reproductive
branches in cross section. In C. dacotensis the cones are 6 em long and
5-10 em in diameter, borne on a peduncle of comparable length. In the
upper portion of the peduncle there are 100-150 elongated hairy bracts
which enclose the young reproductive organs.
The ovulate receptacle is subtended by a whorl of microsporophylls. A
reexamination of the massive pollen-bearing region led Delevoryas (1963)
to suggest that its structure may be fundamentally different from the frequently
illustrated flower-like reconstruction by Wieland (1906). Presumably, it
comprises a whorl of about 20 sporophylls, each with a pinnate structure
and each pinnule bearing two rows of synangia. They appear around the
40 The Gymnosperms
Fig. 7.3 A-C. A, B Cycadeoidea sp. A Reconstruction of plant. B Transection of trunk

with a large pith (p), poorly developed secondary xylem (sx), and thick ramentum (r)
with embedded cone (c) and leaf bases (lb). C C. jenneyana, a tangential section of
trunk shows leaves and reproductive branches in cross sections. (A After Delevoryas
1971, B after Crepet 1974, C after Bierhorst 1971)
Cycadeoidales 41
Fig. 7.4 A-C. Cycadeoidea sp. A Longisection cone shows ovulate receptacle (or) ,
microsporophylls bear synangia (sy) and investing bracts (b). 8 Longisection ovulate
receptacle bear stalked ovules separated by interseminal scales. C Stalked ovules and
interseminal scales. (After Crepet 1974)
apical meristem of the cone (Fig. 7.4A; 7.5A). Crepet (1974) examined
well-preserved permineralized specimens, and observed that each pinnate
microsporophyll became revolute as it matured, with the synangium-bearing
pinnae folded inward parallel to the plane of the pinna-rachis (Fig. 7.4 A).
The recurved microsporophyll rachises were fused with one another above
the ovulate receptacle, and at their tip. The latter was occasionally fused to
its base. A synangium is a kidney-shaped structure containing 8-20 tubular
sporangia (within the periphery of the synangium). Monocolpate grains
have been observed within the sporangium. Thus, wind pollination is
eliminated in Cycadeoidea, and selfing appears to be the chief mechanism;
pollination by boring insects is also a possibility (Crepet 1972, 1974). The
imposition of self-pollination was an important factor in the extinction of
the group, which did not continue beyond Cretaceous (Crepet 1974).
The ovuliferous receptacle is centrally located, conical or dome-shaped
and at the tip of the cone axis (Fig. 7.4 A, B). It comprised upright stalked
ovules enveloped by club-shaped vascularized interseminal scales with
expanded tips (Fig. 7.4 B, C) . The orthotropous ovules are small (up to
3 mm long), have a tubular unvascularized integument which extended and
surrounded a long micropyle. The ovules are enveloped except at the tip of
42 The Gymnosperms
the micropyle (Fig. 7. 5 B). Crepet and Delevoryas (1972) observed ovules
with remnants of a linear megaspore tetrad. However, stages in
megagametophyte development have not been observed, which is quite
unusual, as well-preserved cycadeoid seeds with dicot embryos have been
discovered.
B
Fig. 7. 5 A, B. A Cycadeoidea fructification in median longisection. B C. morieri, ovule
and adjacent scales in longisection. m micropyle, i integument, is interseminal scale.
(A After Seward 1969, B after Crepet 1974)
The origin and relationship of cycadeoids have aroused considerable interest.
Several vegetative characters are cycad-like, while the reproductive bodies
are unique-the ovules are borne on long stalks interspersed with interseminal
scales.
In their general morphology, the fronds of cycadeoids are remarkably
similar to pinnate leaves of the cycads (except their stomatal structure).
Their stem anatomy also resembles the cycads except in the absence of leaf
girdles, and presence of abundant and compact secondary xylem with
scalariform tracheids.
The cycadeoids have bisporangiate cones, while in cycads they are
monosporangiate. The morphological nature of the stalked ovules and
interseminal scales in cycadeoids is difficult to interpret. The whorls of
microsporophylls with pinnate synangium-bearing branches are also different
from spirally arranged microsporophylls with abaxial sporangia in cycads.
However, both groups have monocolpate pollen.
Cycadeoidales 43
Consequently, when vegetative characters alone are considered, a close

relationship between cycads and cycadeoids seems likely but, when
reproductive structures are also considered, the relationship appears remote.
Therefore, this may be interpreted as two distinct groups with a possible
common ancestor among the Palaeozoic pteridosperms (Chamberlain 1920).
The presence of syndetocheilic stomata in pteridosperms is in favour of this
interpretation.
8. Cycadales
The plants have woody, columnar, unbranched stems with a crown of large,
pinnately compound leaves, which make it look like a palm tree. Mucilage
canals are present in the pith and cortex, and the wood is of manoxylic
type. The leaf trace is diploxylic. The plants are dioecious, the male and
female cones are mostly terminal. The megasporophyll has two to eight
large orthotropous ovules. The microsporophylls have microsporangia on
the abaxial surface. The motile male gamete (spermatozoid) has a spiral
band of flagella.
Fossil Cycads
The beginning of Cycadales is not known. The fossil remains provisionally
assigned to Cycadales are meagre, in the Upper Palaeozoic. From the Triassic
have been reported undoubted cycads which spread rapidly over the earth.
Tertiary cycads mostly belonged to the modem genera, or were closely
related to them (Arnold 1953). The most complete and well-preserved remains
are from the Upper Triassic and Jurassic.
Gould (1971) described a well-preserved permineralized specimen of
Lyssoxylon, a stem genus, from the Upper Triassic of Arizona. The stem
has a large central pith, a cylinder of compact secondary wood, and an
outer cortical region surrounded by persistent spirally arranged leaf bases.
The stele has well-defined "growth rings", rays of two types, and tracheids
with multiseriate alternate circular bordered pits of the araucaroid type, on
the radial walls (cf. extant Dioon spinulosum). These characteristics indicate
that the affinities of Lyssoxylon are with the Cycadales.
Delevoryas and Hope (1971), from the Upper Triassic beds of North
Carolina, described a remarkably complete compression-impression fossil
of a cycad stem with leaves and pollen cone attached. The stomata are
haplocheilic, and other characteristic features are also cycadean. The attached
fronds are comparable to the isolated cycad frond type Pseudoctenis. A new
genus Leptocycas (Fig. 8.1 A) has been established. The reconstruction
depicts a slender stem, ca.1.5 m tall, bearing a crown of leaves, some
exceed 30 em in length. Persistent leaf bases on the stem are absent. The
general morphology of Leptocycas suggests that primitive cycads had slender
stems; the bulky, fleshy stem of a typical extant cycad is derived.
Cycadales 45
Fig. 8.1 A-D. A, B, D Reconstruction of extinct Cycad plant. A Leptocycas gracilis.

B Bijuvia simplex. C Palaeoycas, an ovule-bearing organ. D Cycad bearing Beania
ovulate cones and Nils sonia type foliage . (A After Delevoryas and Hope 1971,
B,C after Florin 1933 a, D after Harris 1961)
By making use of imaginary trunks, Florin ( 1933a) and Harris (1961)

also prepared reconstructions of extinct cycads. After an exhaustive study
of epidermal and stomatal structure of isolated compression-impression
fossils of leaves and reproductive structures, Florin (1933a) reconstructed
a plant of an Upper Triassic cycad and named it Bijuvia simplex (Fig. 8.1 B).
46 The Gymnosperms
It showed Doratophyllum leaves around a cluster of spirally arranged

Palaeoycas megasporophylls (Fig. 8.1 C; their stomatal arrangement and
structure are comparable) which crowned the apex of an imaginary unbranched
stout trunk ca. 3 m high.
Harris (1961) placed the reproductive organs and foliage on an imaginary
branched stem which had an armour of leaf bases (Fig. 8.1D), and
reconstructed a Jurassic cycad. It had crown of Nilssonia tenuinervis leaves
(Fig. 8.1D), ovulate cones of Baenia sp. (described later), and pollen cones
of Androstrobus sp. (described later). All the genera are from the Jurassic
Cayton Bay beds of Yorkshire.
Nils sonia. This is one of the most common and widely distributed Mesozoic
cycadean leaf types-slender, 60 em long, 10 em wide. The lamina is
entire or dissected into pinnule-like segments and is supported on a strong
mid-rib from which simple veins pass obliquely or at right angles to the
margin (Fig. 8.2A). The veins are rarely forked. There is considerable
Fig. 8.2 A-E. Fossil cycads. A Nilssonia cornpta, leaf. B, C Beania gracilis. B Seed
"cone". C Megasporophyll with one seed in longisection. D, E Androstrobus manis.
D Microsporangiate cone. E Microsorophyll. (A After Andrews 1961, B-E after Harris
1941, 1964)
Cycadales 47
variation within a species. The stomatal characters display cycadean affinity.

The genus is nearly worldwide in distribution from Triassic to Cretaceous.
Baenia. Two species are known. In B. gracilis, peltate sporophylls (1.5-

2.5 em) are arranged spirally in a loose cone (10 em long) around a central
axis, each bearing two ovules (Fig. 8.2 B,C) with their micropyles towards
the cone axis. The mature seeds (detached seeds are also called Baenia) are
7 x 7 mm to 16 x 16 mm. On maceration, four cutinized layers have been
observed. Their distribution indicates that the integument is fused to the
nucellus throughout the lower two-thirds of the seed, and is free at the apex
(Fig. 8.2 C). There is an outer fleshy, middle stony, and an inner fleshy
layer. There is evidence of vascular bundles running longitudinally up to
the level where the integument became free from the nucellus. The remains
of pollen grains (identical with those of Androstrobus manis) have been
observed inside the micropyle.
Androstrobus manis. The cones are compact, ca. 5 em long and 2 em

broad (Fig. 8.2 D). Each cone has a large number of spirally arranged
peltate microsporophylls, each is terminated by a rhomboidal scale and has
numerous finger-like pollen sacs on the abaxial side (Fig. 8.2 D,E). They
are arranged in sori with dehiscence slits along their inward-facing walls.
The pollen isolated from the rnicrosporangia are small, ovoid and monocolpate,
as in extant cycads.
Living Cycads
The cycads first appeared in the Upper Triassic, reached the peak of
diversification and development during Mid-Mesozoic, and have been on
the decline since then. At present there are 11 extant taxa with more than
100 spp. They have a disconnected distribution (see Chap. 1) and form an
inconspicuous member of the vegetation. Most cycads grow in exposed
habitats, and are considered xerophytes. A few plants (Zamia pumila,
Ceratozamia) grow in shady and moist woods. One species of Zamia is an
epiphyte (Arnold 1953). The plants are slow-growing. A plant of Macrozamia
takes ca. 100 years to reach a height of 1m (Schuster 1932); Encephalartos
takes 200-300 years to grow 1.5-1.75 m tall.
Root. The root system of cycads consists of a tuberou's, contractile taproot

with narrow racemosely branched lateral roots, and swollen dichotomously
branched coralloid roots. The latter have been variously referred to as
aerial roots, apogeotropic roots, nodules, tubercles, or pneumatophores.
Most of these terms are unsuitable, as th~ features they describe are either
absent or inconsistent (Staff and Ahem 1993). The term coralloid is accepted
as appropriate for mature invaded (by Cyanobacteria) roots. A new term,
precoralloid, has been introduced to describe root which are uninvaded but
48 The Gymnosperms
have the potential to be so and develop into coralloid roots (Staff and
Ahern 1993).
Coralloid roots represent the only known, naturally occurring symbiosis
between plant roots and nitrogen-fixing cyanobacteria (Nostoc, Anabaena
and rarely Calothrix-see Ahern and Staff 1994). Early claims of bacterial
initiation have been dispelled by successful sterile culture experiments (see
Ahern and Staff 1994). Reinke {1872) discovered coralloid roots in Cycas
revoluta. Since then it has been recorded in all taxa and all species examined,
including the new genus Chigua (Stevenson 1990b). These roots can reach
up to 10 em in diameter and 500 gm weight and contribute significantly to
the nitrogen metabolism of the plant as well as nitrogen economy of the
ecosystem (Halliday and Pate 1976).
Ahem and Staff (1994) investigated the origin and development of coralloid
roots in Macrozamia communis and described, for the first time, the sequence
of precoralloid initiation, maturation, cyanobacterial invasion and coralloid
development at the morphological level. The different stages can be identified
by specific characters. The earliest stage is initiation of young apogeotropic,
papillose roots (a distinct papillose sheath covers the root)-the precoralloids.
The active apices continue to produce papillose tissue, which is gradually
replaced (as it matures) by a thin dermal layer with scattered lenticels.
Cyanobacterial invasion has been observed at different stages of precoralloid
maturation and stimulates the irreversible development of precoralloids into
coralloids. Although continuous, these two coralloid regions may be
recognized by their external morphology. New coralloid growth involves
cessation of formation of papjllose sheath, change in gravitropic response,
proliferation of distinctive apicallenticels depicting transitional stage and
differentiation of a cyanobacterial zone.
Stem. Cycads are pachycaulous plants with stems either aerial and columnar
(arborescent) or subterranean (geophilous), tuberous, and fleshy. In Bowenia,
Stangeria, Zamia, and some species of Macrozamia and Encephalartos, the
stem is nearly spherical. In Zamia pumila and Stange ria eriopsis, the vertical
shoots are subterranean, the shoot apex remains underground even though
new shoots are added as a result of leaf development. The external surface
of these stems have a wrinkled appearance in the distal regions. The height
of the leaf scars decrease basipetally because both the fleshy taproot and
the stem contract, pulling the shoot underground. This contraction results
from the collapse of horizontal rows of cells in both pith and cortex. As
this collapse of cells occurs (Stevenson 1981), the vascular cylinder (which
appears straight in longitudinal section) becomes sinuously distorted.
Most cycads appear palm-like due to their columnar trunk and apical
clusters of large pinnate leaves. The tallest cycad is Macrozamia hopei, which
is ca. 20 m high, while the smallest Zamia pygmaea has an underground
stem 2 or 3 em in diameter and is ca. 25 em high. The stem apex of cycads
Cycadales 49
is often several mm across, and is the most massive among all vascular
plants.
The phloem in cycads has not been studied in detail (see Paliwall992).
The available data are based mainly on Cycas circinalis (Strasburger 1891)
and Microcycas calocoma (Chrysler 1926).
The axial system of the secondary system consists of sieve elements,
parenchyma cells and fibers. The sieve elements are long with inclined end
walls, their faces merge with the radial lateral walls. The sieve areas have
variable shape. They occur on end walls and the radial face of lateral walls.
Parenchyma cells are of two types: those containing starch (occasionally
druses of calcium oxalate), and the albuminous cells. In the non-functional
phloem the starch-containing cells remain intact, while the albuminous
cells collapse. Pits develop on the walls between adjacent parenchyma
cells. According to Chrysler (1926), the pits are identical in s-hape and
distribution to the sieve areas in the sieve elements.
In the cycads, the primary phloem of the leaf appears to be devoid of
fibres. In the older petioles and stems, the protophloem is crushed.
Metaphloem tissue contains wide sieve· elements and narrow parenchyma
cells. The tracheids are separated from the sieve elements by parenchyma.
Within the transfusion tissue (located on the flanks of the bundle) the
parenchyma cells, next to the phloem, ar~ rich in cytoplasmic contents and
have rather large nuclei. According to Strasburger (1891), they serve as
intermediate cells comparable to the albuminous cells.
The ultrastructure of sieve elements of Zamia pseudoparasitica reveals
that the ontogeny is identical to that of the Pinaceae· (Parthasarthy 1975).
However, unlike the necrotic nuclei reported by various authors, in Zamia
nuclei have been observed at various stages of degradation. P-protein is
absent (as in other gymnosperms). The o_ther cell organelles also exhibit a
similar behaviour during differentiation. Aggregates of ER normally occur
on both sides of sieve areas in the mature sieve cells and appear to be
interconnected by elements of ER traversing the sieve pores.
Leaf. The leaf length varies from 5 or 6 em (Zamia pygmaea) to about

3 m (Cycas circinalis), and is tough, fibrous and pinnate, bipinnate in Bowenia.
The leaflet in Chigua has a prominent mid rib, the lateral veins depart at
an acute angle and branch dichotomously. Circinate vernation has been
observed in Bowenia (young rachis and pinnae), Cycas (only pinnae), and
Stangeria and Zamia (only rachis). All cycads produce periodic crowns of
foliage and scale leaves. In plants with columnar trunks, the' lear' bases
persist and form a characteristic armour. In geophilous stems, the ·leaf
abscisses from the very base so that the stem surface is covered by a
smooth corky layer. The stomata in all the cycads are haplocheilic.
50 The Gymnosperms
Reproduction
The cycads are dioecious, the male and female sporophylls are organized
into strobili, except in Cycas, where the megasporophylls remain loose and
do not form a cone. Meagasporophylls fn Chigua are peltate, hexagonal.
Raised area or a mound is present at each angle of the hexagon. The length
of the cone varies from 2 em (Zamia pygmaea) to 80-100 em (Macrozamia
peroffskyana). A ripe female cone of Dioon spinulosum is nearly 61 em
long, and weighs up to 28 kg (see Bierhorst 1971).
Thermogenesis is a widespread phenomenon in the group (Tang 1987a),
occurring in the strobili in 42 species of cycads. Heat production generally
follows a circadian rhythm; the temperature and its duration can be correlated
with the size of the strobili. The male strobili have a high concentration of
starch that may supply the energy for thermogenesis.
The strobili of all the species examined release odours that are sweet,
resinous or musty (Tang 1987a). Heat production may help volatilize odours
that attract insects for pollination. The cone odours of some of the taxa
(Cycas, Dioon) are very strong and can be detected outdoors several metres
away. The odours may be released throughout the day (Cycas, Stangeria),
or may be noticeable only during heat production (Bowenia, Ceratozamia,
Zamia). The odours may be sweet and fruity (Bowenia, Ceratozamia,
Stangeria, Zamia), musty (Cycas, Dioon, Microcyas), or resinous, often fruity
(Encephalartos, Lepidozamia, Macrozamia).
Tang (1987b) conducted wind·and insect exclusion pollination experiment
in a wild population of Zamia pumila in Florida. Cones from which insects
(but not wind) were excluded did not produce viable seeds. Cones from
which wind (but not insects) were excluded produced abundant viable seeds.
Two beetle species (Pharaxonotha zamiae, Rhopalotria slossoni) have been
identified which may be affecting pollination. Abundant adults and larvae
of both beetles are present on the male cones. On .the female cones adults
and larvae are present occasionally. Z. pumila produces sugar and amino
acid-rich micropyle droplets which serve as pollinator reward.
Pant and Singh (1990) observed (in cycads growing in Allahabad) that
the male cones of Cycas circinalis alone are invaded by pollenivorous small
beetles, and other insects including ants while the compact crowns of
megasporophylls of C. rumphii and C. revoluta are ignored. The beetles
inside the male cone continue to be active and produce subsequent generations.
These insects do not play any part in the pollination of the plant. This is
further confirmed by the abortive nature of seeds in the neighbouring female
plants of Cycas.
The microsporophylls are flat with a large number of sporangia arranged
in sori on the under surface. In Zamia the microsporophylls are peltate
(cf. Equisetum).
The structure and development of male and female gametophytes are
more or less uniform through the group; Microcycas is the only exception.
Cycadales 51
In the development of the male gametophyte, the stalk cell divides several
times and forms a row of body cells which results in the formation of as
many as 16 (Norstog 1990) or/and 22 (Caldwell 1907, Downie 1928)
spermatozoids from a single male gametophyte.
Spermatozoid
The spermatozoids exhibit a developmental gradient in complexity and
mobility; first-formed spermatozoids are better organized and more actively
motile as compared to the last-formed spermatozoids (Norstog 1990).
All cycads have motile ciliated spermatozoids, first discovered by Ikeno
(1896) in Cycas, and are the largest ciliate male gametes known in plants.
They measure 180-210 Jlm in length in C. revoluta, 230 Jlm in length and
300 Jlm in diameter in Dioon, 332 J1m in length and 180 Jlm in diameter
in Zamia floridana, 180-300 J1m in size in Z. integrifolia and 400 Jlm in
diameter in Z.. chigua.
Norstog (1967a, 1968, 1974, 1975, 1977) studied the ultrastructure of
the male gamete of Zamia. The d}vision of the body cell produces two
spermatids, each with one blepharoplast (diameter ca. 10 Jlrn) and surrounded
by cytoplasmic asters. The bulk of the organelle consists of closely packed
procentriolar bodies, each with a hub-and-spoke arrangement, and nine-fold
symmetry (Mizukami and Gall1966). During maturation, the blepharoplast
forms an attenuated spiral band, and the procentrioles contribute directly to
the formation of the flagella of the mature spermatozoid. The spermatozoids
are oriented "back to back". The greater part of the spermatozoid in Zamia
(and other cycads) is the nucleus surrounded by a shallow zone (ca. 10 Jlm)
of cytoplasm. A flagellated spiral band occupies the anterior half of the
cell. About 10 000-12 000 flagella, each ca. 60 Jlm long, are attached to
this spiral band.
The spiral band (excluding flagella) is composed of three regions: (a) an
outermost electron-dense layer (DL), (b) an intermediate lightly staining
granular region, and (c) a four-layered complex of electron-dense tubules,
fins and compartments termed Vierergruppe (VG). The VG of Zamia is
similar to spermatozoids of Marchantia (Carothers and Kreitner 1967),
Pteridium (Tourte and Hurel-Py 1967, Duckett and Bell 1971), Equisetum
(Duckett and Bell1969, Duckett 1973) and to the three-layered structures,
Driergruppe, present in spermatozoids of Polytrichum (Heitz 1960, Paolillo
1965, Paolillo et al. 1968) and Bryum (Bonnot 1967).
An extensive system of microtubules is associated with the VG in Zamia.
It is similar to a microtubule system termed spline (Carothers and Kreitner
1968) which occurs in Marchantia spermatozoids. However, microttibules
are much more numerous in Zamia (ca. 60 000) than in Marchantia (only 17).
The flagella are subtended by a relatively long (ca. 4 Jlrn) basal body.
Much of this l~mgth is occupied by an internal system of fibres with a
stellate pattern similar to the flagella of algae (Manton 1964), bryophytes
52 The Gymnosperms
(Paolillo 1965, Carothers and Krietner 1967) and pteridophytes (Duckett

and Bell1969, 1971, Duckett 1973). The basal bodies terminate in the DL
of the band.
The cytoplasm is characterized by a network of a granular ER.
Mitochondria dispersed in the cytoplasm and are relatively small with fewer
cristae [comparatively, the spermatozoid of Pteridium has a conspicuous
localized system of rnitrochondria with well-developed cristae associated
with the flagellated band (Bell et al. 1971)].
Ovule
The general structure of the ovule in all cycads is essentially similar.· The
egg is perh~ps the largest in the plant kingdom (P. Maheshwari and Sanwal
1963). A typical feature of the archegonium is its massive nucleus. In
Dioon the egg nucleus is ca. 600 J.Lm long (Chamberlain 1906), and in Zamia
umbrosa 710 J.Lm x 490 J..Lm, 1330-J.Lm-long archegonia (Bryan and Evans
1956).
Pollen Tube
The process of fertilization, embryogenesis and seed germination is more
or less similar. The pollen tube in cycads is not involved in the conduction
of sperm to the egg (unlike the pollen tubes of conifers, gnetophytes and
angiosperms). It functions primarily to obtain nutrients (cf. a haustoria!
fungus) for the growth and development of the male gametophyte. The
cycad pollen tube penetrates the nucellar cells by enzymatically destroying
them. Choi and Friedman (1991) studied pollen tube growth and its relation
to the nucellus in Zamia fuifuracea using light and transmission electron
microscopy (TEM). Following germination of pollen grain at the distal
end, the pollen tube grows intercellularly through the subepidermal layers
of the micropylar apex of the nucellus. Additional localized outgrowths of
the pollen tubes, penetrating the walls of the individual nucellar cells, have
been observed. Intracellular haustoria! growth ultimately leads to a complete
destruction of each penetrated cell, and also appears to induce degeneration
of proximal unpenetrated nucellar cells. This pattern of intracellular
penetration of the sporophyte by the male gametophyte in Z. furfuracea
suggests that the heterotrophic and tissue-specific relationships that male
gametophytes of seed plants have with their host sporophytes are much
more diverse than had previously been known (Choi and Friedman 1991).
According to Johri (1992) 1 the investigation on Z. furfuracea leave no doubt
about the haustoria! role of the pollen tube (so far there is no such study
on any angiosperm). The pollen tube growing in the nucellar tissue is
interpreted as a vegetative structure since it is not concerned in the
reproductive processes (Choi and Friedman 1991).
Professor Knut J. Norstog (Miami, Florida, USA) has devoted considerable
attention to the reproductive biology of the cycads. According to him (in
Cycadales 53
a personal discussion with Professor B. M. Johri;· see Johri 1992), in

Microcycas and Zamia the proximal (lower) pole of the pollen grain does
produce a pollen tube ca. 1.5-2 mm wide and 3-5 mm long. This proximal
extension of the microgametophyte expands very rapidly in about 3-4 days,
breaking through the inner nucellar epidermis into the space of the archegonial
chamber. This growth occurs at the expense of nucellar fluids. As the
pollen tube elogates, the nucellus becomes dry and papery. During this
period, the spermatids swim about in the pollen tube for a day.
Chamberlain (1906) showed pollen tubes quite clearly during fertilization
in Dioon edule (see Cycas described later). According to Johri (1992), a
type of siphonogamy has been initiated in the cycads and they should no
longer be regarded strictly as non-siphonogamous.
The spermatozoids of cycads swim in a fluid medium (enclosed within
the developing ovule) whose solute concentration is approximately 0.6 M
(Norstog 1975). The spermatozoids remain motile for a relatively short
time interval (15 min or less) when collected in pollen tube liquid or in
20% sucrose solution. Zamia spermatozoids in sucrose solution exhibit an
initial burst of rapid swimming for ca. 5·min, followed by flexing movements
of the anterior portion of the cells accompanied by slow flagellar beating.
Finally, only vibration of the flagella is observed, which ceases after
approximately 30 min. This short-lived activity of the spermatozoid is possibly
due to the low volume of cytoplasm compared to the nuclear volume and
to the absence of a direct mitochondrial association with the flagellated
band of this massive cell. It may also be due to the unnatural conditions
(such as sucrose solution) under which spermatozoids were observed (Norstog
1975).
The spermatozoid swims with a rotating motion, with its flagellated
band directed forward. There is a wave-like beating of the flagella and a
rapid flexing of the anterior of the cell and occasional contractions and
extensions of this region. Such movement, more appropriately termed
euglenoid (Norstog 1965a) than amoeboid (Webber 1901), may contribute
to the passage of the spermatozoid through the neck canal of the archegonium.
In Zamia integrifolia, at the time of fertilization, the four neck cells becQme
turgid and separate and a passage (ca. 50 J.LM in diameter) is formed. The
neck cells shrivel soon after fertilization. Actual passage of spermatozoids
through the archegonial neck canal has not been observed. Spermatozoids
of Zamia and other cycads (Lawson 1926) assume a special form after
passage through the neck. The flagellated band remains in contact and
several spermatozoids are seen within the egg cytoplasm of a single
archegonium (Norstog 1975).
Embryo
The embryogeny (in one or two species in_.a few taxa) has been investigated
adequately (see Dogra 1992). Information is available on embryogeny for
54 The Gymnosperms
Bowenia (Lawson 1926), Ceratozamia (Chamberlain 1912), Cycas (!keno

1898, Treub 1884, Chamberlain 1919, 1935, Swamy 1948, De Silva and
Tambiah 1952, Rao 1963), Dioon (Chamberlain 1910), Encephalartos (Saxton
1910, Sedgwick 1924, De Sloover 1963), Macrozamia (Brough and Taylor
1940, Chamberlain 1913, Baird 1939), Stangeria (Chamberlain 1916) and
Zamia (Coulter and Chamberlain 1903, Bryan 1952).
Early divisions in the zygote occur in situ, are synchronous and in quick
succession. Later divisions are not synchronous, some of the nuclei may even
fail to divide, so that the final number of free nuclei is less than expected.
The nuclei are evenly distributed in the proembryo. Ten free mitoses occur
(highest in gymnosperms), before walls appear in Dioon edule, Stangeria
paradoxa, Encephalartos friderici-guilielmi, Cycas circinalis and Cycas
species (embryogeny of Cycas described later in this Chapter). More than
eight free-nuclear divisions in Zamiafloridana, and six free-nuclear divisions
(lowest in cycads) occur in Bowenia serrulata. A decrease in the number of
free-nuclear divisions (from ten to six mitoses) in the cycads has been observed.
An "advanced" trend (over proembryo with evenly distributed nuclei)
has been observed when the free nuclei migrate from scattered position to
the base of the proembryo (cf. conifers; see Dogra 1992). Further division
mostly occur at the base. Thus the proembryo has two physiologically
different region~-the upper inactive and the lower active.
Wall formation takes place in the proembryo after the last free-nuclear
division. Depending on the position of the free nuclei at wall formation, the
segmentation may be complete or partial.
In Encephalartos friderici-guilielmi, E. villosus and Macrozamia spiralis,
the segmentation of nearly the entire egg cell results in the forination of a
primary proembryo which has a dense, compact, active basal region. In
Macrozamia reidlei wall formation occurs throughout the proembryo except
in a small central region which contains free nuc;:lei and cytoplasm. This
region breaks down in later stages so that there is a hollow cavity in the
centre. In Cycas the cellular proembryo formation is restricted to the periphery
and the base. In Stangeria, Zamia and Bowenia, the proembryo cells are
formed only at the base (as in conifers; see Dogra 1992).
The primary proembryo develops directly into late, embryo without
elongation of the suspensor. The internal division and the U.S.E.(U-upper
tier, S-suspensor tier, ~mbryonal tier) pattern of conifers (see Chap. 15)
are absent.
The cellular primary proembryo gradually differentiates (morphologically)
into a large inactive primary upper region, and a very active primary embryonal
region. The primary upper region becomes gradually reduced to an inactive
region enclosing a central vacuole. Later, these cells degenerate. The primary
embryonal region consists of compact, densely staining, actively dividing
uniform cells concentrated at the (basal) tip. This is the meristematic cell
region of the embryo. The numerous proximal cells elongate to give rise to
Cycadales 55
tubular cells which (in all cycads) form a very long, elongated cord. This
carries the meristematic primary embryonal tip of the late embryo into the
prothallial tissue. In Zamia and Bowenia, there is a layer of elongated cells
around the compact terminal embryonal cells (cap cells). Cleavage
polyembryony is absent in cycads.
The embryo is mostly dicotyledonous, in Encephalartos there are three
cotyledons, and Ceratozamia has only one, which always develops on the
side of the seed facing the ground. If such a germinating seed is rotated in
a clinostat throughout the developmental period, it develops two cotyledons
(Dorety 1908). The size of the mature seed varies-Cycas spp. and
Macrozamia denisonii have the largest seed (ca. 6 or 7 em), and Zamia
pygmaea the smallest (ca. 5-7 mm). The seeds of all cycads germinate
without a resting period.
Chromosome Number
According to Khoshoo (1962), among the cycads the karyotype of Cycas
and Microcycas is most advanced. Vovides and Olivares (1996) observed
that Zamia loddigesii forms a morphologically variable complex on the
Yucaten Peninsula, Mexico. Several dipolid chromosome numbers have
been recorded in the species: 2n = 17, 24, 25, 26 and 27. The karyotypes
differ, in individuals of the same population, in the number of metacentric
and telocentric chromosomes present. Centromere fission as well as pencentric
inversions and unequal translocations are probable mechanism for the
karyotype variation. A correlation between higher chromosome number,
increasing dryness of the habitats along with asymmetrical karyotypes suggest
that karyotype evolution in Z. loddigesii is recent (Vovides and Olivares
1996).
The various taxa display well-marked affinity, yet it is difficult to arrive
at a satisfactory classification of the genera (within the group). The
classification, therefore, varies and the taxa are included in one, two or
three tribes/sub-families/families. According to Johnson (1959), there are
three families:
a) Cycadaceae - Cycas
b) Stangeriaceae - Stangeria
c) Zamiaceae - Lepidozamia, Macrozamia, Encephalartos, Dioon,
Microcycas, Ceratozamia, Zamia, Bowenia
Cycadaceae
Cycas
Cycas is the most widely distributed taxon (s.ee Chap. 1), and has 20 species
(see Willis 1966).
56 The Gymnosperms
Morphology
Cycas has a columnar stem with a cluster of pinnately compound large
leaves and gives the plant the appearance of a palm tree (Fig. 8.3A). Normally,
it is unbranched, but beyond a certain age, branching trunks are common;
branching can also be induced by injury. A bonsai (see Chap. 23) of Cycas
revoluta can be induced to branch more profusely than happens in nature.
Sometimes, dwarfs hundreds-of-years-old may have nearly 20 crowns (Pant
1973 ). The branches develop from adventitious buds called bulbils, arising
from the lower fleshy portion of old leaf bases. In the beginning there are
a number of scale leaves around a small stem; later, crowns of leaves are
produced. When they have grown larger, these bulbils develop branches on
the stem. Occasionally, they may have even adventitious roots at their base.
On separation they grow into new plants (vegetative propagation).
A B
Fig. 8.3 A, B. A Cycas revoluta, female plant. B C. circinalis, columnar trunk shows
alternate bands of large (foliage leaves) and small (megasporophyall) rhomboidal bases .
(A After Strasburger 1930, B after Pant 1973)
For the most part the stem is covered with a thick persistent armour of
regularly alternating bands of large and small rhomboidal leaf bases (Fig.
8.3B); the larger one is the base of the foliage leaf and the smaller of a
scale leaf as well as of the megasporophyll (in a female plant). The
approximate age of a plant can be calculated from the whorl of leaves and
Cycadales 51
megasporophylls produced every year. However, the leaf bases at the base
of the trunk may be shed off by abscission, leaving behind the corky
surface of the stem which now appears narrower than the apical portion .
There are two types of leaves. (a) A cluster of scale leaves which alternate
with green leaves and are often roughly felted. (b) Large unipinnate compound
foliage leaves. The leaflets are tough, leathery, have a mid-rib, and show
circinate vemation (Fig. 8.4 A,B). The rate of leaf development is slow but
uniform.
\18
Fig. 8.4 A-D. Cycas revoluta. A Adaxial view of foliage shows acropetal unrolling
of circinnate pinnae. B Transection rachis shows young pinnae in circinnate vemation .
C, D Transection pinna. C Shows revolute margins. (A, B After Foster and Gifford
1959, C, Dafter Pant 1973)
In the beginning, there is a normal taproot system. Later, it is replaced

by strong branched adventitious roots which develop a number of apogeotropic
(negatively geotropic) roots. These roots grow horizontally and vertically,
and have lenticels on their surface. There is an incursion of a blue-green
alga(Cyanobacteria) which multiplies rapidly and forms a conspicuous zone
in the cortex. The alga causes a distortion of the rootlets, giving rise to a
mass of tubercles which look like corals, hence these roots are called coralloid
(Fig. 8.5 C, D). They are greenish or brownish and are abundant in the
58 The Gymnosperms
seedling stage, especially in the nurseries. Two species of Nostoc and one
of Anabaena (family Nostocaceae), Oscillatoria (Oscillatoriaceae) and diatoms
have been reported (Fritsch 1945). The presence of numerous lenticels on
the surface of these roots, (also on normal roots), abundant air spaces in the
cortex, and the emergence of these roots above the ground suggest that they
also serve as aerating organs.
Fig. 8.5 A-F. Root. A, 8, D-F Cycas revoluta, C, C. rumphii . A Transection of diarch
normal root (diagrammatic). 8 Inset from A. ipd inner periderm, opd outer periderm,
ph phloem, sx secondary xylem, tanc tannin ceii.C, D Coralloid roots. E Transection
of triarch coralloid root, the algal zone (alz) is in the middle of the cortex. ic inner cortex .
oc outer cortex. F Portion of coralloid root with algal zone. (A-C After Pant 1973,
D-F after Wettstein 1935)
Cycadales 59
Anatomy
Root. A normal young root, at the point of attachment to the base of the
stem, has up to eight protoxylem groups. The roots become progressively
thinner, and the number of protoxylem groups becomes reduced to a diarch
condition. The branch roots are mostly diarch. A typical periderm is produced
and older roots show accessory cambia. The first cambium appears away
from the xylem so that in an old root the primary and secondary xylem
become separated (Fig. 8.5 A, B). The protoxylem shows characteristic
spiral thckenings (Greguss 1955). The secondary xylem is manoxylic with
abundant multiseriate rays. Pitting of the tracheids is similar to that in
secondary xylem elements of the stem. Some tracheids in the wood of the
root have peculiar delicate spirals (Greguss 1955). The vascular tissue is
surrounded by a pericycle of starch cells. The endodermis shows typical
casparian strips. Tannin-filled dark brown cells are interspersed all over.
Druses, or rhomboidal crystals, occur in the root, either in isolation or in
longitudinal series.
The anatomy of a coralloid root is comparable to that of a normal root
(Fig. 8.5 E), except that (a) the development of vascular tissue is poor, (b)
the secondary growth is absent, and (c) the cortex has a conspicuous greenish
zone of radially elongate thin-walled cells with large intercellular spaces.
These cells contain a blue-green alga (Fig. 8.5 E, F). Initially, these cells
are more richly cytoplasmic than the adjacent cells.
Grilli (1963) studied (with EM) the blue-green alga/e in the root nodules
of Cycas revoluta. The "infected" cells showed remarkable polymorphism,
in the organization of the chromatoplasm, characterized by either a lamellar
system or a reticulum, or both. The polymorphism in structure is probably
related to the light intensity, age, and the depth of the root nodules in the
soil. The ultrastructure of the heterocysts, and their abundance and
development in relation to various environmental factors have been studied.
Stem. A young stem of Cycas has an irregular outline on account of

persistent leaf bases (Fig. 8.6 A, B). A central broad pith, surrounded by
a ring of numerous vascular bundles, is connected to the cortex by medullary
rays. The stem always remains predominantly parenchymatous with scanty
development of xylem (manoxylic). Numerous canals (clear circular areas)
occur throughout the parenchymatous tissue (Fig. 8.6 C) and petiole, and
are interconnected through leaf gaps. These canals contain mucilage, which
is partly miscible with water and dries up into a hard, clear, somewhat
crystalline mass. The parenchymatous cells have abundant starch.
The poor development of xylem is balanced by the thick and rigid armour
of leaf base:. and periderm. The primary vascular cylinder is an endarch
siphonostele in adult plants, but mesarch in the seedling. The bundles are
conjoint, collateral and open. The primary cambium is short-lived, but Cycas
60 The Gymnosperms
.·.· ..
1 · · · · ·· · · .· ~- 0 ·.· ·:·o · ~ muc

@:· ~ -··· a · .. ··.· ~:
... o & .. . . · a &v ·. .;
:: __ .. : '. ~:.-%\~~~~;~:~;j_\r · · ,·:~. :'A!f
··.... ~ .,:.•)· ):. ·.\ .. ,.,'miU~Ull . . . ~ ..
~ ·..:.--•·> ..•,.. \~~1\\\\\}\W.::~ :~-l'l'
:. ·. · \:r:~~~{i\~~;\t~~~k~ .}
\l''..<i"'
..:'li
. fl!lf..
~~~~:"
.. ~'/~:'S~~-~~
·o .... ·. >'_·>:·.::·
..
• ••
.
•
. . ·. :.·:
·
• 0
Fig. 8.6 A-F. Stem. A, B, D-F Cycas revoluta, C Cycas sp. A Transection young.stem
(diagrammatic), note leaf bases, gtr girdle traces. 1b leaf base. vb vascular bundle.
B Transection of stem, one sector (diagrammatic) shows successive vascular rings and
persistent leaf bases. C Transection of stem, sector(diagrammatic) shows vascular bundles,
cortical strands and mucilage canal (muc) . D-F Xylem in radial (D, F) and tangential
(E) section. (A After de Bary 1884, B, C after Pant 1973, D-F after Greguss 1955)
is polyxylic (Fig. 8.6 B). As soon as the primary cambium has stopped
functioning, a second cambium differentiates in the cortex and develops a
stronger second ring of wood. Such rings are formed successively one after
the other, but the outer rings become gradually weaker. Annual rings are
absent. According to Terrazas ( 1991 ), observations of the vascular tissue of
Cycas shoots provide evidence that the first vascular cambium (originating
from procambium), as well as subsequent successive cambia, are
Cycadales 61
simultaneously active (several previous workers, namely, Coulter and

Chamberlain 1917, and Pant 1973, have stated that the original cambium is
short-lived). The second cambium is established during the seedling stage
and differentiates from the cortical cells. Cambial activity also occurs within
phloem parenchyma cells of the first vascular cylinder. In all cambial layers
conspicuous cell divisions occur and undifferentiated xylem and phloem cells
are present. Tracheids in the first and successive vascular cylinders are generally
of the same length. However, there is a trend towards increasing length within
the successive cylinders possibly because successive cambia are long-lived
(Terrazas 1991 ).
The metaxylem usually has scalariform, and occasionally multiseriate pitted
elements. In older plants, the growth is slow and the protoxylem may also
show scalariform thickening. The secondary xylem has bi- or tri- or multi-
seriate bordered pits on radial walls, and rarely on tangential walls. These pits
are in alternate rows, comparable to those in Araucariaceae (Fig. 8.6 D-F).
The pits are oval or round (Fig. 8.6 D,E). Ultrastructural studies have shown
that the torus is absent in the pit membranes of Cycas (Eicke and Metzner-
Kuster 1961). The presence of tertiary spirals (Fig. 8.6 D, E) in tracheids (as
in Taxus) has been reported. They are probably similar to spirals reported in
the wood of the root by Greguss (1955).
Leaf. A peculiar feature in Cycas (and allied genera) is the girdling of the
leaf traces (Fig. 8.6 A). A leaf trace, after arising from the stelar cylinder,
usually does not pass directly into the nearest leaf but turns round a semi-
circle or "girdles" the stem horizontally, and enters the leaf almost opposite
to its point of origin. It is joined at intervals by other traces, so that a number
of them enter each rachis. The rachis is cylindrical, and the pinnae are inserted
on it (Fig. 8.7 A). It has a thick-walled epidermis covered by a thick cuticle.
and stomata are irregularly distributed. There is a broad outer zone of fibrous
cells intermixed with short chlorenchyma, and an inner zone of large
parenchymatous ground tissue with mucilage canals. Several collateral open
vascular bundles are embedded in it. Usually, only two large and a variable
number of small strands enter the base of the leaf. Immediately on entering,
the two main bundles branch and form numerous bundles which become
arranged like the inverted Greek letter omega-n (Fig 8.7A). Towards the
tip of the rachis the number of bundles becomes reduced and they become
arranged in a C-shaped arc. The centripetal xylem appears like a wedge, with
the protoxylem at the apex facing the phloem (Fig. 8.7 B). An arc of cambium
lies on its outer side (towards the phloem). A few centrifugal xylem elements
are present on its inner side, usually in two o.r three separate groups or in a
continuous arc (Fig. 8.7 B). The centrifugal xylem is separated from the
protoxylem and adjacent centripetal xylem by a few parenchyma cells. On
its outer side, towards the primary phloem, a few layers of radially seriated
secondary phloem cells are present. Each vascular bundle is surrounded by
62 The Gymnosperms
Fig. 8.7 A, B. Cycas revoluta . A Transection rachis shows inverted omega arrangement
of bundles. B Enlargement of part of A. cfx centrifugal xylem. cpx centripetal xylem.
muc mucilage canal. ph phloem. px protoxylem. (After Pant 1973)
a fibrous sheath. This characteristic diploxylic foliar bundle is seen in the

rachis as well as in the pinnae (Fig. 8.4 C, D).
Leaf traces are endarch at the point of origin in the stem, become mesarch
in the cortex, and farther out in the pinnae many traces are completely
exarch. During their course, the centripetal xylem becomes progressively
large, while the centrifugal xylem becomes reduced.
In their longitudinal course, the bundles of the rachis frequently dichotomize
and anastomose. The bundles lying at the two open ends of the Q-shaped
arc, facing the grooves in the rachis, are the first to enter the pinnae. The
next two bundles of the arc either directly enter the next set of pinnae or
Cycadales 63
may branch, and the outer strand enters a pinna. This process continues till
all the pinnae in the leaf have become vascularized.
The leaf has xerophytic characteristics. The upper and lower epidermes
are heavily cutinized. There is a hypodermis or one or two layers of
sclerenchymatous cells, followed by a palisade tissue, and finally the lower
epidermis. The stomata are sunken and haplocheilic. Between the palisade
and the spongy tissue, there are several layers of transversely elongated
thin-walled colourless cells, the transfusion tissue, helpful in conduction.
The vascular bundle of the median vein has a mesarch xylem, the phloem
lies below the xylem, and there is a sclerenchymatous sheath around the
bundle (Fig. 8.4 C, D).
The stomata, 75 J.Lm x 34 J.Lm, of cycads are generally larger than those of
other gymnosperms. Pant and Mehra (1963) have studied the development
in Cycas circinalis, C. revoluta (Fig. 8.8 A-F) and C. rumphii. In all these
species, the stomata are haplocheilic. In a young stoma, the cells which surround
the guard cell or the guard mother cells divide tangentially, form the subsidiary
and encircling cells, and a radial division increases their number. In C. revoluta,
the stomata are deeply sunken and amphicyclic, i.e. surrounded by two rings
of cells (Fig. 8.8E). The first ring consists of four to eight subsidiary cells
which partly overlap the stoma. The second ring of cells has a thicker layer
of cuticle, and forms elongated finger-like processes overarching the deeply
sunken stoma, resulting in a conspicuous epistomatal dome. In C. circinalis,
C. rumphii and other species, the stomata have only an irregular ring of
subsidiary cells around a stoma which is not deeply sunken. In C. revoluta,
the guard cells show radiating striations on their surface.
A part of the guard cell wall is lignified (in lateral and polar lamellae),
the rest is chiefly cellulosic. The rigid lignin lamellae help in the opening
and closing of the stomatal aperture. When the guard cells become turgid,
the cellulosic region stretches readily, while the lignified lamellae direct
the stretching, resulting in the widening of the stomatal aperture. The cuticle
has no function in the opening mechanism, and the subsidiary and the
encircling cells are regarded as accessory to the stomatal apparatus.
Reproduction
All Cycas spp. are dioecious. The reproductive cycle starts after several
years of vegetative growth.
Male Cone
The short-stalked male strobilus is large and compact (Fig. 8.9A). The
length of a ripe cone is ca.40 em (C. revoluta) to 60-80 em, or even more
(C. circinalis). Normally, the cone is terminal, and the apex of the main
stem exhausts itself during its production. Then the growth of the trunk is
continued by a lateral bud which arises from the base of the peduncle,
pushes the cone to one side, and takes its place. This new shoot apex
64 The Gymnosperms
A B
Fig. 8.8 A-F. Cycas revoluta, development of stomata. A Guard cell mother cell, lower
epidermis of young leaflet. B Guard cell mother cells in division . C Young guard cells
(dotted outline) slightly sunk below the adjoining cells. D Two guard cells (dotted outline)
surrounded by subsidiary and encirciling cells. E Mature stoma in surface view.
F Vertical section of young stoma. (After Pant and Mehra 1963)
appears terminal, forms fresh crowns of scales, leaves, and later a cone.
This process is repeated with the production of each new cone.
A male cone is an oval or conical structure with numerous spirally-
Cycadales 65
B E
c D
Fig. 8.9 A-G. A Cycas revoluta, male cone. B-D C. circinalis. B Sporangia in sori,
longitudinal slits radiate from the center of each sorus. C, D Dorsal (C) and lateral (D)
views of microsporophyll. E-G Cycas sp. E Transection of microsporophyll. F, G
Tran- and longisection of microsporangium. (After Pant 1973)
66 The Gymnosperms
arranged microsporophylls which are almost perpendicularly attached to

the cone axis. Each microsporophyll is parenchymatQus, has a hairy epidermis,
stomata are present mainly on the lower surface, and numerous vascular
bundles and mucilage ducts traverse through it (Fig. 8.9 E).
A mature sporophyll is hard and woody. It has a wedge-shaped, distally
expanded, fertile portion which bears a large number of sporangia on its
entire lower surface (Fig. 8.9 C, D). The sterile end tapers and has a
pointed upcurved apex (Fig. 8.9 D). The sporangia are arranged in definite
groups or sori (cf. ferns like those of the family Marattiaceae). Each sorus
has three to six sporangia placed around an indusial papilla from which
they are originally formed. The mature sporangia can be easily distinguished
by the lines of dehiscence which radiate round the indusial papilla (Fig. 8 9 B).
A mature sporangium is an oval sac with a short, and stout stalk. The wall
consists of a thick-walled epidermis called exothecium (Fig. 8.9 F, G). A
cuticle is present on its outer surface. The middle zone consists of three or
four layers of thin-walled cells and an inner lining of tapetum, which mostly
consists of small cells with large nuclei and thick granular cytoplasm. The
sporangium is packed with pollen grains (Fig. 8.9 F, G).
Microsporangium
In Zamia and Stangeria the archesporium has a hypodermal origin (Pant
1973). One, sometimes several hypodermal cells undergo periclinal divisions,
resulting in the primary parietal cells on the outer side and primary
sporogenous cells on the inner side. These cells continue to divide and give
rise to a several-layered wall, central mass of sporogenous tissue, and a
tapetum between the two. In Cycas (Swamy 1948, De Silva and Tambiah
1952), the sporogenous cells repeatedly divide mitotically, enlarge, become
rounded and form microspore mother cells (mime). The tapetal cells with
large nuclei and thick granular cytoplasm breakdown, and the nucleated
cytoplasm is absorbed by the mime. Some of the inner cells of the sporangia!
wall may also be absorbed. Each mature mime has a large nucleus and a
number of starch grains (Fig. 8.10 A).
Microsporogenesis. The mime Wldergo reduction divisions (Fig. 8.10 B-E).

After meiosis I, a ring-like thickening appears on the callose wall of the
mother cell in the equatorial plate. Usually, the wall laid down after meiosis I
seems to be thicker than the one which developes after meiosis II. It is
likely that the abundance of starch, especially on the equatorial plate, obstructs
a clear view of the cell plate (Fig. 8.10 B-E).
The cytoplasm of each microspore contracts and secretes a membrane
which forms its independent wall (Fig. 8.10 F, G). Thus, each member of
a tetrad is contained within its own chamber. The original wall of the
mother cell, which appears to be made up of pectic substances, collapses
and no trace is left after a week of division. The pollen grain of cycads
Cycadales 67
A
B c D
F H
pr
J L
Fig. ~.10 A-L. A-J Cycas rumphii, development of micros pores and male gametophyte.
A, B Microspore mother cells, Meiosis I. C-E Meiosis II, starch grains (sgr) on equatorial
plate. F, G Micros pore tetrads. H-J Delimitation of prothallial cell (pr) and larger cell.
K, L Cycas sp. K Larger cell in division. L Cross section of pollen grain at shedding
shows prothallial cell, generative cell (g) and tube nucleus (tn). (A-J After De Silva and
Tambiah 1952, K, L after Pant 1973)
shows an oval germinal furrow .or sulcus on its distal face (Millay and
Taylor 1976). The furrow develops by a modification of the exine in the
region and can be differentiated into a peripheral and a central zone (Audran
1965, 1971). According to Gullvag (1966), in C. revoluta the exine is made
up of two layers. The outer one comprises orbicules and/or a sculptured
granular layer, while the inner layer is lamellated. The newly formed
microspores are uninucleate. Excluding the exodermis, the sporangia! wall
now consists of three or four layers of thin-walled polygonal cells; most of
the tapetum has already been consumed.
Male Gametophyte. The wall of the uninucleate pollen grain shows the
exine and intine. The latter is the last layer to be laid down and is of
variable thickness (Audran 1965), thinnest at the proximal or prothallial
cell end, and thickest at the distal end of the grain. Numerous Golgi vesicles
take part in its formation. The pollen contains abundant starch grains. The
nucleus divides assymetrically, forming a small lenticular prothallial cell,
and a large central cell (Fig. 8.10 H-J). The prothallial cell persists, unlike
in other gymnosperms. It remains attached to the lower region (proximal)
68 The Gymnosperms
of the pollen grain wall. The antheridial initial (the central cell acts as an
antheridial initial since there is only one prothallial cell), divides again to
form a small antheridial cell and a large tube cell (Fig. 8.10 K, L). The
former is attached to the intine at the site of the prothallial cell. The tube
cell is usually vacuolate and has a large nucleus. The pollen grains are
shed at the three-celled stage (prothallial cell, antheridial cell, and tube
cell).
The microsporangia dehisce by longitudinal slits. The lines of dehiscence
of various sporangia radiate out from the centre of the sorus towards the
tip of the sporangia, and are usually marked by unthickened cells of the
exothecium. After the sporangia open, the cone axis elongates and the
tightly pac~ed sporophylls separate from each other; this helps in the release
of pollen. In a strobilus, the microsporophylls mature progressively from
the apex to the base, and it takes several days for all the pollen in a
strobilus to be shed. The numerous light pollen grains are easily blown by
wind.
Megasporophyll
Instead of a female strobilus in Cycas, the megasporophylls form a loose
crown at the apex; the apical growing point continues further growth. Each
megasporophyll is a leaf-like structure, 15-30 em long, and pinnately dissected
in its upper region. In the lower portion, one to five pairs of ovules are
borne on either side of the sporophyll. The sporophylls and ovules are
covered with yellow hairs (ramenta) which fall off as the ovules ripen. The
seeds develop a soft orange colour. In various species there is a gradual
reduction in the expanded part of the sporophylls. In C. siamensis, C. pectinata
and C. revoluta, the megasporophyll is large with elongated pinnae (Fig.
8.11 A, B, D). In C. rumphii, C. beddomei and C. <;ircinalis, it is narrow
and the pinnae are reduced to almost mere serrations (Fig. 8.11 C, E, F).
In C. normanbyana, the sporophyll is highly reduced and bears only two
ovules (Fig. 8.11 G).
Ovule. The earlier stages of development of the ovule in cycads have not
been fully studied. This may be due to the difficulty in locating very young
ovules since they are deeply embedded in the crown of the leaves, and are
not easily visible (from outside). For the collection of such ovules, one
must dissect the entire crown. In these slow-growing plants, most of the
ovules (on a plant) are at the same stage of development. Therefore, by
"destroying" one plant, only one developmental stage can be obtained. The
megasporophylls emerge from the crown when the ovules are already in
the free-nuclear stage of the gametophyte. Owing to such difficulties, the
details of megasporogenesis have been worked out for only a few cycads
(P. Maheshwari and H. Singh 1967).
The large orthotropous unitegmic ovule may be up to 6 em long and
Cycadales 69
s'
'~·. c
\ ";~
~?1
.
~ E .'
G
Fig. 8.11 A-G. Megasporophylls. A Cycas siamensis. B C. pectinata. C C. rumphii.

DC. revoluta. E C. beddomei. F C. circinalis. G. C. normanbyana. (After Pant 1973)
4 em in diameter. The integument is fused with the nucellus, and is free

from it only at the apex. It can be differentiated into three layers: (a) the
outer fleshy layer which becomes variously coloured at maturity, (b) middle
stony layer, and (c) an inner fleshy layer which is resorbed and becomes
papery even before maturity. The vascular supply at the base of the ovule
divides into three strands, the middle one supplies the base of the sporangium,
70 The Gymnosperms
and the two laterals enter the integument and divide again, The thicker
strand supplies the outer fleshy layer, and the thinner ramifies into the
fleshy inner layer (Fig. 8.12 F). The integument is open at the tip to form
the micropyle. Due to disorganization of some of the nucellar cells at the
apex, a concavity (the pollen chamber) is formed. The epidermal cells of
the nucellus divide periclinally to form a nucellar cap (Pant 1973). A mature
nucellus is beak-shaped. Its epidermis is heavily cutinized and has stomata,
although it is enclosed by a thick integument.
A
8 c
Fig. 8.12 A-F. Megasporogenesis and female gametophyte. A-D C. rumphii .

A Megaspore mother ceii.B, C Dyad (B) and triad (C) . D Functional chalaza), and two
micropylar degenerated megaspores . E, F Cyms sp., Iongisection of ovule (diagrammatic).
E Cellular female gametophyte. F Mature ovule. (A-D After De Silva and Tambiah
1952, E, F after Pant 1973)
Cycadales 71
Megasporogenesis. Generally, the young nucellus has one to several

hypodermal archesporia! cells which divide periclinally to form the inner
primary sporogenous and the outer primary parietal layer. The latter and its
derivatives repeatedly divide periclinally to form a massive parietal tissue
above the primary sporogenous cells. The latter may divide so that the
sporogenous tissue increases. One of the sporogenous cells functions as the
megaspore mother cell. One or more megaspore mother cells (mgmc) become
distinct by their prominent nuclei and dense cytoplasm (Fig. 8.12A). The
elongated cells show a thick cell wall (under the light microscope). The
mgmc undergoes reduction divisions, but in C. rumphii the upper dyad fails
to divide, so that a row of three cells is formed (Fig. 8.12 B-D, De Silva
and Tambiah 1952).
Female Gametophyte. Repeated free-nuclear divisions occur in the

functional chalaza! megaspore, and the nuclei become distributed in the
peripheral layer of cytoplasm (around a large central vacuole). The actual
number of free nuclei before centripetal wall formation have not been
counted in Cycas (Pant 1973). Gradually, the entire female gametophyte
becomes cellular (Fig. 8.12 E). A further investigation will provide detailed
information on the development of the female gametophyte.
A nutritive tissue, the spongy tissue or tapetum, of densely cytoplasmic
cells differentiates around the sporogenous cells. The cell walls of the
spongy tissue are acetolysis-resistant, and perforated. The walls of the
innermost tapetal cells form a thin sheet of resistant material with the inner
surface covered with sudanophilic droplets, or merely an aggregation of
droplets. These droplets coalesce to form clusters on the tapetal cell wall.
According to Pettitt (1966), the droplets are responsible for the deposition
of material on the outer surface of the megaspore membrane. Eventually,
the spongy tissue degenerates and lies compressed between the female
gametophyte and the outer tissues of the ovule. Pettitt presumes that this
layer has been interpreted as the megaspore membrane in cycads (and
Ginkgo). When the female gametophyte has become cellular (after the
degeneration of the spongy tissue), an "endospermic" jacket of large and
multinucleate cells develops around the gametophyte.
Archegonium
At the micropylar tip of the gametophyte (prothallus), two to six archegonia
differentiate. The archegonial initial (Fig. 8.13 A) divides to form a primary
neck cell and a central cell. The former divides anticlinally, resulting in
two neck cells (Fig. 8.13 B, C). They divide once again, just before
fertilization, to form four neck cells. The second wall is much thinner than
the first one. This is a regular feature in Cycas (Norstog 1972). By the time
the nucleus of the central cell is ready to divide, the neck cells become
large and turgid and project into the archegonial chamber. The central cell
72 The Gymnosperm s
Fig. 8.13 A-F. Archegonium. A-C Cycas rumphii. A Archegonial initial (ari).
B Archegonium. C Two neck cells of archegonium in transection . D-F C. circinalis.
D Longisection (part of) archegonium. E Plugs (p[) of egg cytoplasm (ec). F Cells (devoid
of contents) show dark-staining bodies. (A-C After De Silva and Tambiah 1952, D-F
after Rao 1961)
continues to grow for several months before its nucleus divides. The ventral
canal nucieus moves up into the neck of the archegonium (Fig. 8.13 D),
where it soon disorganizes. Occasionally, however, it enlarges and simulates
the egg nucleus. The maturation of different archegonia in a gametophyte
is synchronous. The egg cytoplasm shows a number of proteid vacuoles
which increase after fertilization . In C. rumphii they are rich in proteins
(De Silva and Tambiah 1952).
The gametophytic region around the archegonia grows upwards and forms
a depression in the middle, the archegonial chamber.
The archegonia occur singly, and each has a well-developed single-
layered jacket. There is usually a thick wall with simple pits between the
jacket cells and the central cell of the archegonium. Stopes and Fujii (1906)
observed distinct plasmodesmata! connections between the walls of the
jacket cells and the egg. They refute any "communica tion" through simple
pores between the inner layer of the jacket and the egg cells, as claimed by
Ikeno (1898) in Cycas. According to Swamy (1948), the pores become
occluded by plug-like thickenings, and the jacket cells become depleted of
their contents (Fig. 8.13 E, F). The thick cell wall forms the so-called egg-
membrane during the post-fertilization stages. The egg membrane thickens
as the egg matures. It retains its form and remains connected with the
Cycadales 73
suspensor of the embryo when the latter is fairly well advanced. There are
prominent pits on the thick wall. Extension of the egg cytoplasm come in
contact with the cytoplasm of the jacket cells, through the plasmodesmata
present in the pit areas.
Pollination
The male cones mature when the young megasporophylls have emerged on
the female plant, and the ovules are at the free-nuclear gametophytic stage
and ready for pollination. The single integument is drawn out into a short
micropylar tube, which secretes a sugary exudate (pollination drop) close
to the nucellus of the ovule. The wingless pollen grains usually float into
the pollination drop, which retracts soon after. The pollen grains show a
germinal furrow, which closes in dry weather and is wide open in high
humidity. After landing on the pollination drop, the pollen grains imbibe
water and other substances from the exudate, become heavy and sink down
the micropyle to reach the nucellus, where they germinate.
The male cones of Cycas growing in warm climate emit a strong odour
(described earlier); several insects visit them when the pollen ripens (see
Chamberlain 1935, Pant 1973). Thus, these insects may also be carriers of
pollen in cycads, although this has to be confirmed by critical field observation.
Following pollination, the ovules enlarge and reach their full size. Unpollinated
ovules remain small and eventually dry up. In the female plants of C. rumphii
(growing in the Peradinya Botanic Garden, Sri Lanka), some of the ovules
could develop to full size even when no male plant of the same species was
growing in the vicinity. This is due to pollination by pollen of other cycads
like Encephalartos and Macrozamia. The pollen grains germinate in the
pollen chamber and the male gametophyte develops to an advanced stage
although sperms are not formed (Le Goc 1917). Such ovules do not form
embryos. Pant (1973) occasionally observed fully developed ovules in the
megasporophylls of Cycas revoluta and C. rumphii growing in the Botanical
Garden of the Allahabad University, where the nearest male plants of
C. circinalis were about 5 km away. The stimulus for the development of
these ovules was quite likely provided by the pollen grains of some other
plant, possibly an angiosperm or conifers, like Pinus, Thuja, Cupressus
(Pant 1973).
Niklas and Norstog (1984) studied the implication of aerodynamics on
reproduction, and the pattern of pollen deposition on megasporophylls of
Cycas (and megastrobili of Dioon and Zamia). A characteristic air-disturbance
pattern around megasporophylls influences the quality and arrangement of
wind-borne pollen grain deposition on the surface of megasporophylls and
ovules. The largest number of pollen grains adhere on the windward profiles.
The megasporophyll deflects the air flow passing over it leeward, distal
surfaces, where pollen grains accumulate. The preferential concentration of
pollen on the distal portions may help pollination. Water dislodges the
74 The Gymnosperms
adhering pollen grains, which flow along the glabrous ovule-bearing margins
of megasporophylls and collect near or on the micropyle. The authors suggest
two phases in cycad pollination: (a) the transport of wind-borne pollen
grains to megasporophyWmegastrobili (Cycas, Dioon, Zamia); (b) subsequent
transport of adhering pollen to ovules by water and/or passive sifting (Cycas)
or insect activity (Dioon and Zamia) .
After the pollen grains land in the pollen chamber (situated in the upper
beak-shaped portion of the nucellus), a group of cells lying below begin to
degenerate and extend the lower limit of the chamber. This post-pollination
extension is the lower pollen chamber or intermediary chamber.
The intine of the pollen grain ruptures the exine at the distal end, grows
into a tube in the region of the furrow, and the tube nucleus migrates into
it (Fig. 8.14 A). The pollen tube grows laterally and intercellularly into the
tissue of the nucellus, instead of towards the archegonia as in other
gymnosperms. During its course through the nucellus, the tube often branches
horizontally, derives nourishment as it digests its way, and accumulates
g
pr
B
Fig. 8.14 A-G. Development of antherozoids. A-D Cycas rumphii, pollen tube with
prothallial (pr) and generative (g) cell. B Generative cell divides to form stalk (sc) and
body (be) cells. C Prothallial cell extends into the stalk cell. D The body cell shows two
well-developed blepharoplasts (bl) . E, F C. revoluta. Ciliated antherozoids, before (E)
and after (F) free movement. G Cycas sp. Antherozoid with ciliated spiral band.
(A-D After De Silva and Tambiah 1952, E, F after Miyake 1906, G, after Pant 1973)
Cycadales 75
considerable amount of starch for the development of the male gametophyte.

The pollen tube does not transport the male gametes but establishes only
a haustoria! system. ,
In the beginning the grain-end of the pollen tube is enclosed in the exine
but, eventually, it enlarges and cannot be contained therein. This proximal
end of the pollen grain contains the prothallial, stalk and body cells (or
male gametes). The broad, short (Fig. 8.14 C-F) proximal end grows towards
the female gametophyte. The branched pollen tube continues invasion of
the nucellus and the haustoria! activity increases.
The antheridial cell divides periclinally; the cell adjacent to the prothallial
cell is sterile and designated the stalk cell, the other cell is the body cell
(Fig. 8.14 A-C). The stalk cell is surrounded by a distinct cell wall. It
enlarges considerably, and accumulates starch and other materials.
Simultaneously, the prothallial cell bulges and its upper end penetrates
through the stalk cell, which forms a life-belt-like ring around the prothallial
cell. The body cell (Fig. 8.14 C, D) begins to enlarge and elongate along
the longitudinal axis of the pollen grain. The nucleus of the body cell also
enlarges, and two small granular bodies (the blepharoplasts) appear, one on
each side of the prothallial cell (Fig. 8.14 D). Cytoplasmic rays (also called
astral rays) radiate from the blepharoplasts. The astral rays are the rnicrotubules
which extend from the matrix or the surface of the blepharoplasts. The
latter comes to lie on the two poles of the body cell along the longitudinal
axis of the pollen grain. The body cell enlarges, becomes spherical, and the
blepharoplasts move through 90° and come to lie at right angles to the long
axis of the pollen. The blepharoplasts increase in size and become very
prominent.
The mitotic spindle of the body cell lies at right angles to the long axis
of the pollen tube, so that the two sperms lie side by side (Fig. 8.14 E)
enclosed in the wall of the body cell till they mature. The blepharoplast
forms a part of clock-spring-band-bearing cilia, around the gradually or
abruptly tapering distal part of the sperm (Fig. 8.14 F, G). This band in
C. revoluta shows a right-hand coiling (as seen from the apex of the sperm).
The spiral band has six turns (gyres), and in a radial section (C. circinalis)
appear stair-like. The sperms start rotating movement while still enclosed
in the body cell. The sperms of C. revoluta are 180-210 Jlm in length, and
visible to the naked eye. They are released by the rupture/dissolution of the
outer walls of the sperm mother cells into the basal end of the pollen grain.
By their cilia they move freely in the fluid of the turgid pollen grain-end
and remain active for several hours.
The nucellar tissue between the pollen and archegonial chamber collapses,
and the proximal end of the pollen grain pushes down through this opening
and "hangs" freely in the dry archegonial chamber (Fig. 8.15 A). A bunch
of pollen grains grow downward and disorganize it completely between the
intermediary chamber and the female gametophyte. The proximal end of
76 The Gymnosperms
A
8
Fig. 8.15 A-F. Fertilization . A Cycas sp., longisection upper part of ovule
(semidiagrammatic). B-F C. revoluta, successive stages in the fusion of egg and sperm
nuclei. cilb ciliary band. en egg nucleus.sp spermatozoid. spc spermatozoid cytoplasm.
spn supernumerary spermatozoid. (A After Pant 1973, B-F after Ikeno 1898)
Cycadales 77
the pollen grain lies in a longitudinally continuous cavity in the middle of

the nucellus (Fig. 8.15 A).
Fertilization
The proximal end of numerous pollen grains hang into the archegonial
chamber (Fig. 8.15 A). Finally it bursts and forms a short pollen tube
through which it discharges the male gametes into the archegonial chamber.
The gametes swim for about 15 min with a forward and circular motion;
the band of flagella form the anterior end. The large spermatozoids enter
the egg through the opening between the neck cells within a few minutes
of their release. The male gametes often become distorted during the passage,
since the opening is narrow. On entering the egg cytoplasm, the flagellate
band of the male gamete is cast off, and is left at the tip of the egg
cytoplasm (Fig. 8.15 B), where it gradually dissolves and is eventually
represented by dark-staining granules. The naked sperm nucleus shrinks
as it moves downward to unite with the egg nucleus. The male nucleus
penetrates deep into the larger egg nucleus before its membrane disappears
(Fig. 8.15 B-F). In C. circinalis the two sets of chromatin can be easily
distinguished from each other by the different morphology of their threads.
The spiral band usually persists in the egg cytoplasm long after fertilization.
Occasionally, a sperm may get close to the egg nucleus with its ciliary
coat, which is cast off just before uniting with the egg nucleus. After a
sperm entered an archegonium (Fig. 8.15 C-F), it is sealed off by a dark-
staining bubstance which is probably derived from a degenerated sperm.
Often one or two, sometimes up to five, additional sperms penetrate the
cytoplasm of an egg cell; only the first sperm fertilizes the egg nucleus.
While still far from the fertilized nucleus, the supernumerary sperms
degenerate without casting off their spiral bands or sheaths. Rarely, they
may even penetrate close to the egg nucleus and cast their coats there.
Embryogeny
The zygote nucleus divides in situ followed by several free-nuclear divisions
(Fig. 8.16 A-E). The nuclei are distributed throughout the young proembryo.
In later stages, the free nuclei mostly concentrate at the base of the proembiyo
(Fig. 8.16 F) and only a few nuclei are present in the upper thin cytoplasm.
Subsequently, only the nuclei at the base divide while the upper nuclei
show signs of degeneration. At the time of wall formation, there are 512
free nuclei in C. circinalis (Rao 1963). Cells are formed only in the lower
portion of the archegonial cavity (Fig. 8.16 F-J), the nuclei present in the
upper area remain free. During later stages, the free nuclei, along with the
circumjacent cytoplasm, degenerate and form a plug. Following wall
formation, the cells at the base divide and function as embryonal cells. The
upper cells (just above the embryonal cells) differentiate into a suspensor
and the uppermost layer of cells as the buffer cells, the latter form a few
layers of tissue around a central vacuole (Pant 1973).
78 The Gymnosperms
A 8
D E
c
F
L M
Fig. 8.16 A-M. Embryogeny. A-E, J·M Cycas sp. A·E Zygote, 2-, 8-, 16- and 32-
nucleate proembryo. F, G, I, C. circinalis, H C. rumphii. F Young proembryo shows
segmentation. G-J Apical end of young proembryos. K Three embryos at different stages
of development. L Dicotyledonous embryo. M Longisection seed. (A-E After Swamy
1948, F, G, I after Treub 1884, H, K after De Silva and Tambiah 1952, J after Pant
1973, L after Schuster 1932, M after Richard, from Engler and Prantl 1926, see Pant
1973)
Cycadales 19
The thick inner tangential walls of the jacket cells persist till an advanced
stage of proembryo. 'The suspensor penetrates the chalazal end to the thick
wall which is very prominent and is called the egg membrane.
Differentiation of Embryo. After the proembryo becomes cellular, the

cells of the upper region elongate to form a very long suspensor. It grows
rapidly, and thrusts the basal embryonic region through the egg membrane
and archegonial jacket deep into the gametophytic cells, which have abundant
food reserve. This is due to enzymatic digestion of the gametophytic cells
when they come in contact with the embryonic cells. Due to rapid elongation,
the suspensor becomes much coiled and twisted (Fig. 8.16K). Usually,
more than one archegonium in an ovule is fertilized, and the development
of multiple zygotes leads to simple or archegonial polyembryony
(Fig. 8.16 K). At first, all the zygotes in an ovule appear to develop with
equal vigour, the embryos project into the gametophytic tissue, and their
suspensors become closely intertwined with each other. Due to competitive
growth, ultimately only one embryo develops while the rest abort at various
stages of development (Fig. 8.16 K, L). The suspensor of the aborted embryos
remain attached to the tough egg membrane, and persist for a long time.
The suspensor of the successful embryos is, therefore, a composite structure
of the coiled suspensors of the entire group of embryos in an ovule. When
stretched, the suspensor may be up to 10 em long.
Seed
The studies on late embryogenesis in cycads are inomplete. A number of
cells are formed in the embryonic region before the differentiation of various
organs. The maturation of the embryo in seed takes over a year after
fertilization. The seed is shed at any stage during this period, and the
development of the embryo is completed on the ground. To begin with, the
embryo only increases in size without differentiation into its organs. Then
the coleorhiza develops at the micropylar end of the embryo. The latter
shows internal differentiation into two polar meristems, epidermis, cortex,
procambium and pith; the hypocotyl is rather short in cycads. The number
of cotyledons varies from one to three, closely adpressed to one another
and appear as a single structure. The coleorhiza is partly derived from the
suspensor and becomes quite "hard" in a mature embryo. It is morphologically
equivalent to the root cap. At maturity several outer layers of the root cap
become especially thick-walled to form a distinctive caplike structure. There
are abundant mucilage cells in the tissues of the embryo.
The attractive, red or orange fleshy seeds contain abundant food reserve
(Fig. 8.16 M). Quite likely, they are dispersed by birds and rodents. The
cycad seeds do not have a resting period and the viability is short. Germination
of the seed is epigeal.
80 The Gymnosperms
Chromosome Number
Cycas circinalis, C. media var basaltica, C. revoluta var revol, C. revoluta
var taiwaniana and C. siamensis have similar karyotypes of 2n = 22. There
are 2 median-centromeric, 4 long submedian-centromeric, 4 short submedian-
centromeric and 12 terminal-centromeric chromosomes. Kokubugata and
Kondo (1996) compared the similar karyotypes with each other by using
(fluorescent staining method with double counter-staining reagents binding
for opposite base-pairs) chromomycin A3 (CMA) specifically for guanine-
cytosine and 4 ', 6-diamidino-2-phenylindole (DAPI) specifically for
adeninethymine. These techniques are useful to investigate the chromosome
variability and to mark the species-distinctive regions on chromosome for
cytotaxonomy. A CMA band has been observed at the terminal region of
each of the 4largest submedian-centromeric chromosomes. The 12 terminal-
centromeric chromosomes display CMA bands at the terminal and pericentral
region (2 of the chromosomes carry a CMA band in the interstitial region
of the long arm in different positions in each of the 4 species). All the
chromosomes exhibit DAPI at the centromeric region.
Similarities of the karyotypes, the CMA and the DAPI fluorescent patterns,
observed in the species of Cycas, suggest that speciation of the genus may
have occurred without any major karyotypic change. Cycas thus appears to
be a monophyletic group in morphological and anatomical (Stevenson 1990a),
and cytological characters (Kokubugata and Kondo 1996).
Temporal Consideration
As compared to the temperate gymnosperms, the reproductive cycle of
cycads occurs at different times of the year. The cones in the South Indian
C. circinalis are initiated probably in April, and show micro- and megaspore
mother cells during June-July (Rao 1963). The ovules have a free-nuclear
gametophyte at the time of pollination ( 8 months after initiation) in December.
Fertilization occurs in May-June (5-6 months after pollination), followed
by a very slow development of the embryo, in which the cotyledons appear
during November-December. The seeds with an immature embryo are shed
during May and June (1 year after fertilization). The embryo matures and
attains full size, and the seeds germinate during September-October
(4 months after shedding). Thus, this taxon takes 2 years and 5 months to
complete its life cycle, although there is no winter rest. In C. rumphii, the
ovules are pollinated in May (De Silva and Tambiah 1952), fertilization
occurs 13 months later (i.e. the following June), and the seeds are shed in
January but the embryo matures only by March. Consequently, the time
lapse between pollination and fertilization is much longer.
The general reproductive cycle of cycads is:
1. There is a long time gap from the time of ovule initiation to pollination
(the ovules are in an advanced stage of development at the time of pollination,
as compared to conifers).
Cycadales 81
2. There is a short time lapse between pollination and fertilization. Most

of this time seems to be taken up in the maturation of archegonia and
filling up of their cytoplasm.
3. The embryo takes a long time to mature, partly on the plant and partly
after shedding.
Phylogenitic Considerations
Morphological studies suggest that the immediate ancestors of the living
cycads are extinct. No extant taxa can be considered to be the forerunner,
and they cannot be arranged into a series considering all the characters
because every taxon shows primitive and advanced characters. Cycas is
regarded as a primitive form, because its megasporophylls are loosely arranged
and do not form a strobilus, and its circinate young leaves are fern-like.
The compact male cones, pitted secondary wood, and single-veined pinnules
display advancement. According to Gaussen (1950): (a) Cycas shows 12
primitive and 70 advanced features. (b) Zamia is the most advanced in the
group. However, the species with tuberous stem have scalariform tracheids
(filicinean feature). Gaussen (1950) lists 67 primitive and only 33 advanced
characters in Zamia. (c) Encephaltirtos has fern-like leaves (it was classified
with the ferns until the cycadean nature was revealed after observation of
the cones), the stomata are least specialized, the epidermal cells of the leaf
have undulate lateral walls (cf. ferns) and presence of scalariform wood,
and centripetal xylem in the cone axes. (d) The male and female gametohytes
of Microcycas are presumed to be the most primitive in seed plants because
of the largest number of male gametes (16 to 22) and archegonia (64 to
200). The vegetative anatomy, and the cones are considered to be advanced.
A genus which appears advanced in some respects may thus be primitive
in others. There is a similarity (in the group) apove the generic level. Each
of the ten (eleven) extant taxa of the Cycadales presents a terminal branchlet
of a phylogenetic tree where all connecting links have disappeaFed (see
Arnold 1953). Therefore, the interrelationships in modern cycads are diffic~lt
to visualize. According to Chamberlain (1920), the cycads did not give rise
to any other plant group, and will probably become extinct in the next
geological period.
9. Caytoniales
This is a Mesozoic Pteridosperm which is often described as a distinct

Order. In some classificatory systems, three families are included in this
Order: Caytoniaceae, Corystospermaceae and Peltaspermaceae (see Stewart
1983).
Caytoniaceae
Several spp. of leaves (Sagenopteris), pollen organs (Caytonanthus) and ovule-
bearing organs (Caytonia) are known in association, but not in organic
connection, from Greenland, Sardinia, western Canada, the eastern USSR,
England and Siheria. The leaves, known for over a century, are scattered
geographically, and stratigraphically from Upper Triassic to Lower Cretaceous
rocks (Fig. 1.1). The reproductive organs were first described by Thomas
(1925) from Mid-Jurassic rocks at Cayton Bay in Yorkshire. There is a
similarity between the epidermal cell structure (specially stomata) of
Sagenopteris with that of the fruit axes, and the presence of caytonanthus
type pollen in the nucellar beak cavity of Caytonia show that the three organs
are congeneric.
Leaf. Sagenopteris phillipsi leaves have a slender petiole which bears

four lanceolate, palmately compound, terminal leaflets (Fig. 9.1 A). Each
leaflet has a prominent mid-rib and reticulate venation (Fig. 9.1B).
Haplocheilic stomata (Fig. 9.1 C) are present on the lower surface. The
entire leaf was shed by an abscission layer (Harris 1951 ).
Pollen-Bearing Organs. The structure of Caytonanthus is unusual. It has

a dorsiventral rachis which bears opposite or sub-opposite, irregularly
branched pinnae. The ultimate branches bear terminal, pendant, tubular
synangia (Fig. 9.2 A). Each synangium has three or four sporangia which,
on dehiscence, separate from each other except at the tip (Fig. 9.2 B). The
synangium is radially symmetrical (Harris 1935) and differs from bilateral
symmetry in extant flowering plants. The pollen grains are winged
(Fig. 9.2 C).
Caytoniales 83
c
B
Fig. 9.1 A-C. Sagenopteris phillipsi. A Palmate leaf. B Leaflet shows reticulate venation.
C Stoma. (A, B After Thomas 1925, C after Harris 1964)
Raunsgaard and Fries (1986) studied by light microscopy, TEM and

SEM, the pollen grains extracted from Caytonanthus kockii (lowermost
Jurassic of Scoresby Sound, East Greenland) and C. arberi (Middle Jurassic
of Yorkshire, England). The pollen grains are small, ovate, bisaccate, and
have a nearly smooth surface except for the granular sulcus area.
In transmitted light the internal structure of sacci appears reticulate with
elongated meshes (as in several conifers). Ultrastructural studies with TEM
reveal: (a) A spongy infilling of the sacci formed by elongated, cylindrical,
solid elements that radiate from the central corpus, and branch irregularly.
A similar infilling of the sacci (this appears to be an unique feature) has
also been observed in other Mesozoic pteridosperm. (b) The outermost part
of the pollen wall is thin, homogenous and has a smooth surface, while the
innermost layer (endoexine) is thick, and has a laminate to thready structure.
The thick laminate endexine and the sacci distinguish the pollen of
Caytonanthus from the angiosperms and group them with gymnosperms.
84 The Gymnosperms
Fig. 9.2 A-G. A, B Caytonanthus kochi. A Microsporophyll. B Cross section synangium

shows four microsporangia. C Caytonanthus, bisaccate pollen grain. D, E Caytonia
nathorsti. D Reconstruction megasporophyll. E Young cupule shows lip and opening.
F C. thomasi, longisection cupule shows position of ovules. G Caytonanthus type pollen
grains in micropyle of Caytonia ovule. (A, B After Harris 1937, C, E after Harris 1964,
D after Thomas 1925, F after Harris 1933, G after Harris 1951)
However, the ultrastructure of the sacci of the pollen differs from coniferopsid
pollen.
Ovule-Bearing Organs. Caytonia consists of an axis ca. 4 em long, which

bears two rows of lateral ovule containing cupules (Fig. 9.2 D). A cupule
is ca. 4.5 mm in diameter, globose, recurved and has a lip-like projection
adjacent to the pedicel above an opening (Fig. 9.2 E); it closes at maturity.
The cupule encloses a U-shaped row of ovules which are in a single cavity
in C. sewardi or in separate ones, as in C. thomasi (Fig. 9.2 F). The number
of oyules varies; in C. sewardi there are 8 in a single row, in C. nathorstii
15, and in C. thomasii about 30 in a double row. Each ovule is elliptical,
orthotropous and has a short micropyle which points toward the opening
below. Some of the ovules and the wall on which they are borne are quite
fleshy . Pollen grains have been seen within the micropyle of the ovules
(Fig. 9.2 G). The opening must have been very small. It is likely that the
Caytoniales 85
pollen reached the micropyle by means of a "pollen drop mechanism", as

in many extant gymnosperms. The pollen grains trapped in the drop floated
up the channels to the seeds, and the two lateral bladders on the pollen
grains may have acted as flotation devices. It is also possible that at pollination
time it was a much more open structure. Later, the opening became small
by differential surface growth.
There is a single integument, which is free from the nucellus. The cuticle
covering the nucellus is exceptionally thick. There is no evidence of vascular
system in the integument.
The relationship of Caytoniaceae is not at all clear, and it appears to be
quite isolated. Few fossils have created such a stir among morphologists as
Caytoniaceae when it was first discovered by Thomas (1925). He emphasized
similarities with both pteridosperms and angiosperms. He assumed that it
provides a clue to the origin of flowering plants. He considered the flange
on the cupule as a stigma, the cupule itself as a kind of carpel and the
Caytonanthus synangium as the angiosperm stamen (despite its radial
symmetry, and lack of a filament and connective). However, the pollination
in Caytonia is still at the gymnosperm level (Harris 1964 ). Thus, a decision
on its phylogenetic relationships should be taken only after we know more
about the nature of its cupules (Sporne 1965).
10. Glossopteridales
During the Lower Carboniferous, the flora of the North and South
Hemispheres was more or less similar. By the Upper Carboniferous and
Lower Permian, there was a completely different flora in South America,
much of the southern part of Africa, Australia, the Indian Peninsula,
Antarctica, New Zealand and other smaller land masses. These regions
together formed the supposed continent of Gondwanaland, separated by the
Tethys Sea from the other continent of the Northern Hemisphere.
The Gondwana era started with a cold environment and was followed by
a warm moist climate which supported a new type of vegetation. This flora
is often called the glossopteris flora because of the abundance of the leaves
known as Glossopteris. They appear in the Upper Carboniferous, extend
into the Triassic, and decline thereafter (Fig. 1.1). Leaves with characteristics
of Glossopteris have also been reported from the Jurassic of Mexico
(Delevoryas 1969). The geology of the Gondwana system is best known in
India, where several studies of the flora have been conducted.
Glossopteridaceae
The name Glossopteris was proposed by Brongniart (1828), for the leaves
which were lanceolate/tongue-shaped, 3 to 40 em in length. For a long
time, it was presumed to be a fern, as the leaves resembled a modem taxon
such as Polypodium musaeifolium (Spome 1965).
Evidence accumulated by Gould and Delevoryas (1977) and Pant (1977)
clearly shows that Glossopteris was a large tree. Gould and Delevoryas
made a detailed reconstruction of the plant (Fig. 10.1A). The trunk is
nearly 6 m tall, with gymnospermous wood of araucarioxylon type. It is
supported by a root system of the vertebraria type, conspicuous growth
rings are present in roots, trunks and branches. The leaves are in spirals or
whorls, probably on short shoots (Pant 1977). The plants are deciduous
(Surange and Chandra 1976), as shown by the presence of a large number
of isolated leaves and very few reproductive organs.
Root. The permineralized plant remains from the Upper Permian contain
silicified remains of Vertebraria, the detached roots of a Glossopteris plant
Glossopteridales 87
8
Fig. 10.1 A-C. A Glossopteris tree, reconstruction. B Vertebraria, transection.
C Glossopteris sp., leaf (reconstruction) shows mid-rib and reticulate venation.
(A, C After Gould and Delevoryas 1977, B after Gould 1975)
88 The Gymnosperms
(Gould 1975). They can be easily identified by their characteristic wedge-

like sectors (Fig. 10.1A) which radiate from the centre of the axis. This is
the secondary xylem, arranged around a central polyarch protostele, with
the protoxylem strands alternating with the radiating arms (Fig. 10.1B).
The cavities between the radiating arms suggest that Vertebraria is the root
system of a plant which flourished in a semi-aquatic environment. The
roots branched commonly (indicated by the presence of branch root traces
and smaller roots in the rock specimens). The secondary xylem is identical
to the pycnoxylic wood of Araucarioxylon arberi trunks. Both have opposite/
alternate circular bordered pits in five or six vertical rows. They may be in
groups of two to seven, crowded on the radial walls so that the pits are
hexagonal. Uniseriate rays are 1-20 cells high. The various observations
undoubtedly show that silici.fied roots of Vertebraria and stems of
Araucarioxylon arberi are congeneric (Stewart 1983).
Leaf. Glossopteris and Gangamopteris are the most notable elements of

the flora. Studies on epidermal structure confirm the existence of numerous
species. According to Surange (1966), a fair degree of speciation can be
expected during the 50 million years of their existence on earth. Glossopteris
had a prominent mid-rib and reticulate venation (Fig. 10.1 C). In a few
species, a distinct petiole was present. Gangamopteris was similar, but lacked
the midrib. Glossopteris leaves are characteristically hypostomatic (stomata
on lower surface), the stomata are haplocheilic and irregularly placed between
the veins. Internally, the leaf is differentiated into an upper and lower
epidermis, and a mesophyll of palisade and spongy parenchyma. The mid-
rib is made up of closely spaced branching and anastomosing vascular
bundles. The lateral bundles which form the veins are associated with
sclerenchyma strands in the lamina.
There are different types of staminate and ovuliferous structures which
belong to this group of plants. They were borne separately. It is not known
whether they were borne on different parts of the same plant or on separate
plants. More than 30 genera of fructification are attributed to Glossopteris
and Gangamopteris.
Male Fructification. Pollen organs have been recovered from Permian

deposits of South Africa, India and Australia (Surange and H.K. Maheshwari
1970, Lacey van Dijk and Gordon-Gray 1974, 1975, Gould and Delevoryas
1977). Surange and H.K. Maheshwari reconstructed the microsporophyll
Eretmonia as a unit. It has a stalk which bears an expanded, nearly triangular,
distal lamina. Two branches arise, just beyond the mid-point, on the adaxial
surface of the stalk each bearing whorls of arberiella-type sporangia
(Fig. 10.2A,B). Venation pattern and epidermal features show their undoubted
glossopterid affinities. Abundant Arberiella sporangia and striatites type of
bisaccate pollen grains with transverse striations on the corpus (Fig. 10.2C)
Glossopteridales 89
A
c
Fig. 10.2 A-C. A Eretomonia sp., fertile leaf with two clusters of microsporangia. B
Arberiella, a glossopterid microsporangium. C Striatites type of pollen of Glossopteris.
(A After Surange and H.K. Maheshwari 1970, B, C after Pant 1977)
are dispersed among the permineralized specimens of Glossopteris (Gould

and Delevoryas 1977).
Glossotheca is another pollen organ which shows clusters of Arberiella
sporangia (Surange and H.K. Maheshwari 1970). The spatulate unit has a
single branch, which arises from near the mid-vein, and bears paired laterals
terminating in clusters of sporangia.
Ovulate Structures. The ovulate organ consists of a stalked fertile

head/capitulum. In a few specimens it is attached to modified leaf-like
bracts. Gould and Delevoryas (1977) report it to be attached (jn one specimen)
to true leaves, either in the axil of a bract or to its upper surface. The fertile
bracts may sometimes bear several capitula, as in Lidgettonia (Fig. 10.3A)
or Denkania (Fig. 10.3 B).
In Lidgettonia, the fertile leaves are shorter than the sterile leaves,
spathulate, and have a round apex. The veins spread from the petiole of the
leaf into the lamina, where they fork and anastomose. The petiole of the
leaf bears two rows, each containing three or four disc-like capitula on the
abaxial side (Fig. 10.3A). Each capitulum is peltate and stalked. There are
six or seven ovules in a row on the underside of the capitulum, near the
90 The Gymnosperms
Fig. 10.3 A-C. A Lidgettonia mucronata, fertile leaf shows disc-shaped capitula in
two rows. B Denkania indica, adaxial surface of glossopterid leaf with a row of pedicels.
Each has a terminal cupule which contains a single ovule. C Dictyopteridium sp.,
ovuliferous capitulum with subtending leaf (seen from underside). (A After Surange and
Chandra 1972, B after Surange and Chandra 1971 , C modified from Surange and Chandra
1975, and Gould and Delevoryas 1977, see Stewart 1983)
undulating margin in L. mucronata. In Denkania indica. the pedicels arise

in a row from the adaxial surface of the subtending glossopterid leaf
Glossopteridales 91
(Fig. 10.3 B). Each pedicel terminates in a cupule containing a single ovule
whose impression can be seen inside.
In Arberia, the more divided capitula appear to be axillary, while in the
entire forms such as Dictyopteridium they are surface-attached (Fig. 10.3 C;
10.4 B). Thus, these plants have leaf-borne reproductive organs, which is
unique among gymnosperms. It is likely that they originated in the axillary
position and were later incorporated to the leaf surface (Pant 1982).
Fig. 10.4 A-D. A Ottokaria bengalensis, capitulum and stalk adnate to subtending leaf.
B-D Glossopteris (Dictyopteridium). B Ovuliferous capitulum with subteJ,lding leaf.
C Transection ovule-bearing capitulum. D Single ovule from capitulum in longisection.
(A After Schopf 1976, B-D after Gould and Delevoryas 1977)
The capitulum could be entire or divided, and always bears the ovules
on the lower surface which may be enfolded (Fig. 10.3 C). There is a wide
range of variation in the number and arrangement of their ovules. In Scutum
there are up to 75 ovules which are spaced randomly, while in Senotheca
the capitulum is reduced and the ovules are spaced closely in two rows.
Denkania has only one ovule on each capitulum which appears cupule-like
(Surange and Chandra 1972). In Dictyopteridium, the edges curve over and
enclose the ovules in a mass of hairs (Fig. 10.4 C), while in Ottokaria the
space between the ovules is filled by a filamentous structure. The ovules
are small and numerous, sessile, ovoid to pyriform (Fig. 10.4 C), and point
92 The Gymnosperms
towards the centre of the enclosing capitulum. The integumynt is thicker

around, the micropyle. The nucellus is free from the integument except at
the chalazal end (Fig. 10.4 D). The pollen chamber occasionally contains
bisaccate pollen grains with transverse striations on the corpus (a characteristic
of Glossopteris pollen). A few ovules have been 'observed with
megagametophytes containing a single archegonium. The embryo has not
been observed in any ovule. In addition to the filaments which fill the
spaces between the ovules, there are conical webs of filaments which extend
from each micropyle. This suggests that pollen-drop liquid, which completely
filled the structure, directed the pollen towards the micropyles.
It is not certain whether the glossopterids are a single group of plants;
the nature- of the ovules and their mode of attachment are quite varied.
Surange and Chandra (1976}presumed there are two groups of glossopterids:
a) With strobiloid receptacle: in Dictyopteridium the strobiloid ovule-bearing
receptacle is borne in the axil of a stalked fertile bract that covers it like
a protective spathe (Fig. 10.3 C). The receptacle and its fertile bract are
subtended by and attached to the petiole of a Glossopteris leaf. In Scutum,
according to Surange and H.K. Maheshwari'(1970), the structure at the end
of the pedicel is a cone of spirally arranged ovules. Later, a Scutum sp.
was also observed to have a bilateral receptacle bearing the ovules, which
were covered on one side by a protective scale leaf with glossopteris type
of venation (Surange and Chandra 1972). Both receptacle and its subtending
covering scale are borne on the pedicel. Earlier, Plumstead (1952, 1956)
reported that the bivalved cupule of Scutum is attached by a pedicel to the
midrib of the leaf. The fructifications are bisexual, i.e. half of the cupule
nearest the adaxial surface be-ar carpels, while the other half is presumed
to bear microsporangia. The affinity of the genus with the angiosperms
(due to the presence of carpels by the reproductive structure of Glossopteris)
had been claimed, but was seriously questioned and ultimately rejected (see
Thomas and Spicer 1986). In Ottokaria, a Permian genus, the ovulate
fructification is attached to the foliage of Gangamopteris (Fig. 10.4 .A),
and the ovules are borne in cones.
b) With foliar receptacle: i.e. ovules borne on modified leaves, e.g. Lidgettonia
(Fig. 10.3 A) and Denkania (Fig. 10..3 B).
According to Schopf (1976), Ottokaria has a flattened capitulum with a
marginal frill bearing the ovules on the undersurface and not in a cone.
Gould and Delevoryas (1977) also present evidence, from permineralized
specimens from the Permian of Australia, to suggest that the ovule-bearing
structure of their Glossopteris specimens (including Scutum and Ottokaria)
is foliar.
It is difficult to recognize the ancestors of the glossopterids because of the
ovules and their different modes of attachment. There is also no clear
Glossopteridales 93
relationship with any geologically younger group. According to Schopf

(1976), they probably evolved from the Cordaitales; or from earlier Northern
Hemisphere pteridosperms (Gould and Delevoryas 1977) ..Surange and
Chandra (1975) place some of them in Pteridospermales and others in
Glossopteridales. Pant (1986) suggests that the glossopterids can be either
included in a broadly defined group of Pteridospermopsida, or kept altogether
isolated, since their ancestors or descendants are presently unknown. It is,
therefore, best to consider the glossopterids as a highly successful group of
gymnosperms which dominated large areas of vegetation in their time and
environment. They subsequently became extinct, due either to climatic change
or the migration of more vigorous plants into the habitat (Thomas and
Spicer 1986).
11. Pentoxylales
The Pentoxylales is a small group of plants of relatively recent discovery.

The stem genus Pentoxylon and ovulate cone Carnoconites have been
described by Srivastava (1946), leaf genus Nipaniophyllum by Sahni (1948),
and micro-sporangiate organs Sahnia by Vishnu-Mittre (1952). Sahni (1948)
reconstructed Pentoxylon (Fig. 11.1 A) and proposed the name Pentoxyleae
for the group.
Pentoxylon
Habit. The habit of the plant is unknown. It was probably a shrubby

plant which grew beside aquatic surroundings (Bose et al. 1984) and had
erect branched leafy shoots. The latter, after a few seasons of growth,
flopped to the ground or on to other stems and made a thicket.
Stem. The stem (Fig. 11.1 A) is dimorphic (cf. Ginkgoales, Coniferales)

and the long shoot is an axis ca.\ em in diameter and with 5-7.5-mm-thick
short shoots. Both long (Stewart 1983) and short shoots are covered with
an armour of closely aggregated, spirally arranged scale and foliage leaf
bases (Figs. 11.1 A; 11.2 A). P. sahnii varied from 2 to 3 mm across, and
frequently had five closely aggregated steles (which gives the name to the
genus and the group) arranged in a ring around a· central ground tissue (Fig.
11.2 B). The number of steles is mostly five, occasionally six. According
to Vishnu-Mittre (1957), the number of steles varied along the length of
stem. The variation in the number of steles (4-1 0) is due to branching and
the branches anastomose (Stewart 1983). Each stele has a tangentially
elongated mesarch primary xylem mass, and a cambium which persists
throughout the year in the young stem. In the older stem, the secondary
wood is eccentric due to excessive development towards the centre of the
stem (Fig. 11.2 B). The vascular steles in the long shoot present a polystelic
condition (cf. Medullosaceae). There are five additional, much smal~er steles,
probably of the short shoots (Sahni 1948). They are almost entirely of
secondary wood.
The wood is pycnoxylic made of compact tracheids which have uni- or
Pentoxylales 95
Fig. 11.1 A-C. Pentoxylon sahnii. A Reconstruction shows long and short shoot with
spirally arranged leaf bases, and leaves on short shoot. B, C Camoconites. B Reconstruction,
short shoot bearing cones. C Longisection ovule. (After Sahni 1948)
hi-seriate circular bordered pits crowded on the radial walls. The wood rays
are uniseriate and one to five cells high. The growth rings are well defined.
Leaf. Nipaniophyllum raoi is strap-shaped, 7 em long and ca. 1 em wide,

with a prominent mid-rib (Fig. 11.1 A). The lateral veins are mostly
unbranched and without anastomoses. The slightly sunken syndetocheilic
stomata (cf. Cycadeoidales) are confined to the lower surface. The petiole
is winged. There are about six leaf traces; arranged in an arc, which enter
the base of each leaf. Traces to a leaf arise in pairs, one from each protoxylem
96 The Gymnosperms
Fig. 11.2 A, B. Pentoxylon sahnii. A Long shoot shows spirally arranged leaf bases.
B Transection long shoot shows five steles. It leaf trace. (A after Stewart 1983, B after
Sahni 1948)
strand of adjacent steles (Stewart 1976). The paired traces divide in their
upward and outward course through the cortex to a leaf base, and form six
to nine bundles arranged in a tangential row (cf. Cycads; Stewart 1976).
Microsporangiate Organs. Sahnia nipaniensis were borne terminally on

short shoots which resemble Pentoxylon. It has ca. 24 microsporophylls
arranged in a single whorl round a conical receptacle, and are fused at the
base into a disc. Each microsporophyll is filiform at its free end and bears
short, spirally arranged branches which terminate in unilocular sporangia
borne singly or in groups. The pollen grains are monocolpate (Cycadophyte
type).
Ovulate Cones. Carnoconites compactum and C. laxum have been

described. The peduncle usually dichotomizes or may remain unbranched.
Each branch terminates in a mulberry-like single cone (Fig. 11.1 B). The
latter varies in length from ca. 1.8 em (C. compactum) to ca. 3 em (C. laxum).
Each cone has ca. 20 sessile erect ovules attached to a central receptacle.
Each ovule has a single integument, which is free from the nucellus, and
the micropyle is directed outward (Fig. 11.1 C). The integument has a thick
sarcotesta and an inner sclerotesta (appearing platyspermic in a cross section).
Each ovule is supplied by a trace at its chalaza) end, which arises from a
ring of vascular bundles in the cone axis.
Pentoxylales 97
This enigmatic group displays a unique, combination of characters. The
polystelic stems resemble some of the Palaeozoic Medullosaceae, yet the
secondary wood is pycnoxylic (coniferous). The leaves show both cycadean
and cycadeoidean features in the leaf trace anatomy and stomata. Both the
pollen- and seed-bearing organs are stachyosporous (borne on stems). The
microsporangiate organs morphologically resemble cycadeoids, and the pollen
is cycadophytic. The ovulate cones are unlike any gymnosperm and their
structure is peculiar to themselves. Most palaeobotanists agree that this
group should be given a status similar to Cycadeoidales and Cycadales.
Investigations during the last 30 (or more) years have not clarified the
exact affinities of this group. According to Rao (1981), at the present level
of our knowledge it cannot be referred to any known group of plants,
exclusively. It can only be regarded as an isolated synthetic group which
shows a mixture of features in common with Pteridospermales, Cycadales,
Cycadeoidales and Coniferales. The correct phylogenetic placement must
await further information (or arguments).
12. Ginkgoales
Ginkgoales is an ancient group which appeared in the Permian, and was

well-represented and nearly worldwide in distribution in the Mesozoic.
Their maximal diversity occurred during the Middle Jurassic, and remains
have been collected from many countries: Alaska, Greenland, Scandinavia,
Siberia, Mongolia, England, Europe, China, Japan, Australia, New Zealand,
India, tip of South Africa and South and North America. These plants
flourished for over 150 million years. By the beginning of the Oligocene
(Tertiary), only 2 out of ca. 19 genera survived; Ginkgo adiantoides is one
of these. This species became extinct by the Pliestocene (Tertiary), and
Ginkgoales has been represented by the extant Ginkgo biloba-the "living
fossil". It appeared in the Jurassic, and became extensively distributed in
the Tertiary. In later periods, it was abundant in the more northern latitudes.
At present, its natural occurrence is restricted to a small inaccessible region
in southeastern China (see Chap. 1).
Fossil Taxa
Abundant compression-impression leaf remains have been collected, the
reproductive organs are meagre. There are about seven or eight Mesozoic
leaf genera included in Ginkgoales (Harris 1935, 1974). Ginkgoites and Baiera
have the maximal number of species. These two taxa are indistinguishable
in their cuticular and stomatal characteristics. However, Baiera lacks a petiole
and its lamina is comparatively more wedge-shaped, than that of Ginkgoites.
The latter has a distinct petiole with two traces which diverge into the basal
edge of a bilobed lamina. The fossil leaves, which cannot be distinguished
from those of the extant Ginkgo, are included in this taxon, but the species
are different. Those leaves which can be distinguished by their morphological
and anatomical characteristices (size and shape of epidermal cells, distribution
of stomata, structure of subsidiary cells, mesophyll and distribution of resin
bodies) are placed in the genus Ginkgoites.
Generally, the primitive leaves were linear and deeply dissected. The
leaves of Arctobaiera show a deeply dissected to entire margins
(Fig. 12.1 A, B). Sphenobaiera (Lower Permian to Lower Cretaceous) has
a characteristic dissected lamina (Fig. 12.1 C) with several dichotomised
Ginkgoales 99
T'I
··:
..'·
~
i.•
1,'
' t• t
ii .,
B
A c
Fig. 12.1 A-K. Leaves and stomata. A, B Arctobaeira. C Sphenobaiera . D, E Baiera

spectabilis. F Ginkgoites minuta. G G. taeniata. H Ginkgoidium. I Ginkgo digitata.
J G. lamariensis. K Ginkgoites lunzensis, stomatal apparatus . (A·C after Florin 1951,
D, F-H after Harris 1935, E after Schenk 1867, I, J after Brown 1943, K after Andrews
1961)
veins. Baiera (Middle Jurassic to Lower Cretaceous) had wedge-shaped

leaf with an indistinct petiole (Fig. 12.1 D, E). The extent of lobing of
lamina and variations in venation pattern in Ginkgoites .spp. (Late Triassic)
can be arranged in a series (Harris 1935). The leaf in G. minuta is highly
dissected by equal dichotomies (Fig. 12.1 F), in G. taeniata the unequal
dichotomies make each half of the leaf appear three-lobed (Fig. 12.1 G),
and in Ginkgoidium the lamina is entire except for a median notch, and
prominent marginal and unbranched secondary veins (Fig. 12.1 H). Thus,
100 The Gymnosperms
there is a general trend from deeply dissected Jurassic leaves to those of the
Tertiary, which tend to be entire except for a median sinus (see Stewart
1983). This series may have an evolutionary significance, as shown by
Mesozoic species of Ginkgo: G. digitata (Early Mesozoic) has a deeply
dissected wedge-shaped petiolate leaf (Fig. 12.1 1), G. lamariensis
(Cretaceous) has undissected wedge-shaped leaves (Fig. 12.1 J), and in
G. adiantoides (Tertiary) the leaves are indistinguishable from those of
extant G.' biloba.
The stomata are haplocheilic. There are four to six subsidiary cells each
with a blunt papilla which tends to overarch the guard cells as in Ginkgoites
lunzensis (Fig. 12.1 K).
There are very few verified reports of reproductive structures. Numerous
dispersed ovules of Allicospermum (Harris 1935) -similar to those of cycads
and G. biloba-are associated with Jurassic and Cretaceous Ginkgo-like
remains. Kp,rkenia is an ovulate fructification from the Cretaceous of Argentina
(Archangelsky 1965). It is a short stalk crowded with 100 or more ovules
associated with Ginkgoites tigrensis. Paired ovules joined by a pad of tissue
with Ginkgo-like stomata have been reported from Yorkshire Jurassic beds
(Harris 1976), where abundant Ginkgoites huttoni occurs. In the same beds,
small pollen-bearing catkins are present. Each consists of a stalk (ca. 5 mm
long) to which are attached rather lax stalks with pairs of microsporangia
at the tips. The pollen grains are monocolpate like those of G. biloba.
The Ginkgo line can be trac~d to the Lower Permian Trichopitys (often
placed in its own order/family). Florin (1949) investigated T. heteromorpha,
known since 1875 from the Lower Permian of southern France, and interpreted
it to be the earliest member of the group (see Phylogenetic Considerations).
It has spirally arranged dichotomously branched leaves without lamina
(Fig. 12.2 A). There are small branched ovuliferous trusses (branch system,
Fig. 12.2 B) in the axil or on the upper surface of the leaf base. Each
ultimate branch bears a terminal, recurved ovule (Fig. 12.2 C), unlike Ginkgo.
The male organs of Trichopitys are not known. However, Sphenobaiera
furcata (Triassic) bears clusters of microsporangia at the branch tips of a
bifurcating axis, which are borne in tum on short shoots, along with leaves
similar to Trichopitys.
Ginkgoaceae
Ginkgoaceae is a monotypic family. The deciduous leaves are fan-shaped
with parallel veins. The tree is dioecious; male flowers are catkin-like
while the female is long-stalked with (usually) two ovules. The male gametes
are motile, and the fruit is drupaceous.
Ginkgo
Morphology
Ginkgo biloba is a tree more than 30 m high and exceeds 1.5 m in diameter.
Ginkgoales 101
8
c
Fig. 12.2 A-C. Trichopitys. A Portion of shoot with sterile telome trusses (leaves) and
axillary ovule-bearing shoot. B Ovule-bearing shoot. C Ovule. (After Florin 1949, 1951)
It resembles a conifer in its general habit. The crown becomes broad,

irregular with age, and shows a variable pattern of branching. The main
axis branches profusely. There are two kinds of shoots: (a) long shoots
which elongate rapidly and bear scattered leaves, and (b) dwarf shoots
which grow slowly and bear a terminal cluster of leaves; the older portion
is covered with leaf scars of previous years (Fig. 12.3 A, D, F). Bud scales
cover young spur (dwarf) shoots. Frequently, the dwarf shoots become
more active and tum into a long shoot, while (more rarely) the terminal
growth of a long shoot may be retarded and resemble a lateral spur shoot
(Gunckel and Wetmore 1946). This suggests that there is not much
fundamental difference between them. The apical meristems of the two
types of shoots are also essentially similar; the difference between them
depends on the duration of cell division and cell elongation in the stem
tissues derived from the shoot apex (Foster 1938). There is experimental
evidence to show that the dwarf shoots are formed due to the inhibitory
effects of auxins produced by the tissues of the long shoots (Gunckel et al.
1949).
The shape and venation of the deciduous foliage leaves are unique. The
102 The Gymnosperms
Fig. 12.3 A-F. Ginkgo biloba. A Dwarf shoot bearing male strobilus. B, C
Microsporophylls. D, F Dwarf shoot bearing young strobili (D) and seeds (F).
E Longisection female strobilus, (A·E After Ganguli and Kar 1982, F after Andrews
1961).
petiole is long, smooth, black and slender, traversed by two collateral vascular
bundles. The lamina is broadly wedge-or fan-shaped, variously lobed, and
the venation is conspicuously dichotomous-(Fig. 12.4 D). The leaves resemble
those of Adiantum (maidenhair fern) in form and venation, hence the popular
name maidenhair tree. The old leaves are shed in autumn, when they change
colour to a golden yellow; the new leaves appear in spring. There is
considerable difference in the lobing of the leaves on the same tree. They
may be nearly entire or two-lobed due to a conspicuous, often deep, apical
notch. They are mostly bilobed on long shoot and entire on short shoot. In
seedlings the leaves have several notches which give a palmatifid appearance
(as in the extinct taxa).
Ginkgoales 103
Fig. 12.4 A-E. Ginkgo biloba. Stem anatomy. A, B Transection long (A) and dwarf
shoot (B). C radial longisection, secondary wood shows circular bordered pits, bars of
sanio, and ray cells with cross-field pits. D, E Leaf. D Single leaf shows venation.
E Vertical section (A, B, E After Ganguli and Kar 1982, C, D after Stewart 1983)
104 The Gymnosperms
Anatomy
Root. The young roots are usually diarch. The endodermis has conspicuous
thickenings on its radial walls and there is a broad pericycle. Older roots
may be tetrach or hexarch. A radiallongisection shows that the spiral elements
of the protoxylem are followed successively by traceheids with (a) reticulate
pitting, (b) transversely elongated simple pits and (c) bordered pits.
Secondary growth occurs, but annual rings are not pronounced. The
tracheids have thinner walls. Some xylem parenchyma cells include crystals,
and thick-walled fibres are abundant in phloem. The medullary rays are
one to several cells high and often show crystals.
Stem. The shoot apex consists of superficial apical initials, a prominent

subapical zone of central mother cells, and a zone of rib meristem around
it on its proximal side (Foster 1938).
A young stem comprises an epidermis, a parenchymatous cortex and
pith, and vascular cylinder. The stele is an endarch siphonostele. A long
shoot has a thick zone of wood and comparatively narrow pith and cortex
(Fig. 12.4 A). In the dwarf shoots, the ring of wood is narrow and there is
a broad cortex and pith (Fig. 12.4 B). The primary xylem consists of a
number of separate strands. The phloem forms a broad zone. The double
leaf traces are very conspicuous.
A radial section of the wood .shows tracheids with numerous round ai:l.d
opposite pits separated by bars of Sanio (Fig. 12.4 C). The uniseriate rays
are one to three cells deep ip. the long shoots, and much deeper in dwarf
shoots. Occasionally, a few xylem parenchyma cells contain calcium oxalate
crystals. In secondary wood the tracheidal overlap is not extensive, and
several tracheids end at the same level. This makes the wood very brittle,
and in high wind storms the trees suffer heavily. When a tree is felled. the
log frequently shatters as it strikes the ground (see Bierhorst 1971.
The secondary phloem is composed of sieve elements, parenchyma strands
and fibres in the axial system and rays in the radial system (Srivastava
1963a). The albuminous cells associated with the sieve cells lack starch
and are crushed in the old phloem. They are connected to the sieve cells
by one-sided sieve areas. Lateral connections between the sieve elements
and albuminous cells could not be traced, as callose was not present in
sufficient quantity. These cells also have plastids but do not store normal,
detectable starch. The fibers are elongated, tapering, tangentially flattened
and non-septate. They have narrow lumen, the wall is thick, lamellated and
appears to be cellulosic. They do not stain positive with phloroglucinol and
HCl and are strongly birefringent under polarized light '(see Paliwal 1992).
Outside the phloem there is a ring of sclereids, probably pericyclic, and
an inner ring of thickened cells, whose walls appear to be gelatinized.
Throughout the plant numerous mucilage canals occur in the pith and the
cortex.
Ginkgoales 105
Leaf. The leaf has a double trace. A transection of the petiole shows two
endarch vascular bundles. The primary xylem of the stem branches
sympodially when the leaf traces are given off. The two traces to any leaf
therefore arise ind~pendently from two different primary strands. They
divide at the base of the blade and the resultant four strands fork repeatedly
to form the dichotomous system of veins which occasionally anastomose in
the lamina (see Stewart 1983). The venation of each of the two halves of
the leaf is completely independent. Mucilage canals are present even between
the veins of the leaf.
A vertical section of the lamina shows: (a) a thick cuticle, (b) stomata
mostly on the lower surface of the leaf, (c) a distinct palisade only in the
leaves on the long shoots (Fig. 12.4 E), (d) mucilage canals and (e) \!lsually
endarch vascular bundles with traces of centripetal xylem represented by
one or two tracheids. The bundles are surrounded by a sheath of thick-
walle<;l cells.
On the lower epidermis, stomata occur irregularly scattered between the
veins. They are haplocheilic, surrounded by four to six ·subsidiary cells,
each with a blunt papilla which projects over the guard cells (see Stewart
1983). The characteristic accessory cells of the stomata are also recognizable
in the extinct taxa.
Reproduction
Ginkgo is dioecious. 1 The male cones are pendant and catkin-like, borne on
short shoots in the axil of normal leaves or scale leaves (Fig. 12.3 A).
The ovulate cones are borne in groups at the apex of the dwarf shoot
(Fig. 12.3 D, F). They are reduced, and each shoot bears two ovules on a
long peduncle in the axil of a scale leaf (Fig. 12.3 D, E).
Male Cone
A male cone comprises 40-50 microsporophylls (Fig. 12.3 A). Each
microsporophyll has a terminal knob, which contains a mucilage sac
(Fig. 12.3 B, C), and there are two (occasionally three to seven) pendulous
microsporangia which dehisce by longitudinal slits (Fig. 12.3 C).
Microsporangium. The strobili are initiated in summer, and appear as

small papilae in the axils of bracts. Wolniak (1976) examined more than
300 cones, and observed: (a) There is no acropetal or basipetal progression
in sporangia! development; (b) There is no correlation between the size of
the sporangium and its development. There is, however, variation in
1ln the Botanical garden at lnsbuck (Austria), on a female tree of Ginkgo, a branch from
a male tree was grafted. The male cones developed and produced fertile pollen grains,
pollination and fertilization occurred normally, and numerous (apricot-coloured) ripe
seeds developed on the female tree (BMJ, pers. observ. 1957).
106 The Gymnosperms
development, and in a sporangia! pair the rnicrosporocytes are ontogenetically

similar. Earlier stages have not been observed but there is evidence of a
single hypodermal cell which divides by anticlinal and periclinal divisions.
The outer cells form a wall of four to seven layers, and the inner cells give
rise to a large group of sporogenous cells. A tapetum surrounds the
sporogenous tissue, and a peritapetal membrane has been reported (Pettitt
1966).
An· endothecium differentiates, which is an exception, since in
gymnosperms only an exothecium is reported. The endothecium develops
from one to three layers of subepidermal cells, becomes thick-walled and
develops fibrous thickenings (Fig. 12.5).
Fig. 12.5 Ginkgo biloba. Longisection microsporangium and a part of microsporophyll,

several layers of wall show endothecial thickenings . (After Jeffrey and Torrey 1916)
Ginkgoales 107
Microsporogenesis. Meiosis in microspore mother cells coincides with

the opening of the bud scales of the spur shoot. The distribution of starch
in the microsporangium is specific, and first appears at the archesporia!
stage. Prior to meiosis, starch reappears in the sporogenous cells and then
in tapetal cells. During meiosis I starch grains accumulate at the equatorial
plate at metaphase, move to the opposite poles during anaphase and telophase,
and become equally distributed after the completion of meiosis. After
telophase I, besides starch, entire plastids and mitochondria shift to the
equatorial region of the microspore until nuclear divisions cease. Microtubules
and ER proliferations appear to stabilize.the organelle distribution through
meiosis II. Tetrads are formed by centripetal wall formation, and the organelles
become distributed equally (Wolniak 1976). A callose wall has been observed
during microspore mitosis (Gorska-Brylass 1968).
Male Gametophyte. The rnicrospore nucleus cuts off two prothallial cells
(Fig. 12.6 A, B); the first cell towards the wall is ephemeral while the
second persists. The antheridial initial divides and forms a smaller antheridial
cell, which remains attached to the intine, and a larger tube cell, which
becomes vacuolate and has a conspicuous nucleus (Fig. 12.6 C, D). The
antheridial cell divides periclinally to form the stalk cell (toward the pollen
wall) with a distinct wall, and the body cell (Fig. 12.6 E). The stalk and
body cells persist in situ. The persistent prothallial cell remains active and
grows into the stalk cell which lies next to it. The stalk cell thus appears
to form a jacket around the protruding prothallial cell (Fig. 12.6 D-F).
The microsporangium dehisces by a longitudinal slit along the inner
face. The pollen is shed at the four-celled stage: two prothallials, one
antheridial and a tube cell.
Ovule
The peduncle bifurcates and bears on each branch a single sessile ovule
with a fleshy collar around its base (Fig. 12.3 E). The morphology of the
collar has been variously interpreted (see Chamberlain 1935); it does not
grow after pollination.
Usually there are only two ovules on each peduncle, occasionally three,
four or more. Whatever the number of ovules, the peduncle always has
twice the number of vascular bundles.
The morphology of the meristem which gives rise to the ovule needs a
critical reinvestigation. The ovule is orthotropous with a beaked nucellus
which has a heavily cutinized epidermis. The nucellus has a well-differentiated
strand of elongated cells and extends almost to its entire length (Fig. 12.7 A).
Its degeneration forms a narrow, deep pollen chamber (De Sloover-Colinet
1963). The inner cells degenerate first followed by the epidermis
(Fig. 12.7 B, C). There is a single integument, which is free from the nucellus
at the apex. Two unbranched vascular strands supply the base of the
integument.
108 The Gymnosperms
A 8
Fig. 12.6 A·F. Ginkgo biloba. Male gametophyte. A·F Development of male
gametophyte. al antheridial cell, be body cell, pr prothallial cell, sc stalk cell, tn tube
nucleus. (A·D After Chamberlain 1935, E. F after Favre-Duchartre 1956)
Megasporogenesis. One or more megaspore mother celVs become distinct

by their prominent nuclei and dense cytoplasm. Due to a considerable
thickening of the middle lamella (Fig. 12.8 A), and the development of a
double-layered wall, the wall of the mother cell becomes thick and two-
layered (Stewart and Gifford 1967). The latter has densely staining outer
layer which resembles the circumjacent nutritive tissue, and an inner layer
which is similar to the middle lamella except for a tighter arrangement of
the fibrillar structure. The young megaspore mother cell is spherical, and
has a large nucleus in the centre. Its cytoplasm occasionally shows a small
vacuole, relatively scanty endoplasmic reticulum (Stewart and Gifford 1967),
Ginkgoales 109
8
c
Fig. 12.7 A·C. Ginkgo biloba. A·C Longisection nucelli show successive stages of
development of pollen chamber. (After De Sloover-Colinet 1963)
and randomly placed starch-bearing plastids, mitochondria and dictyosomes.

The mother cell elongates as it matures. The cytoplasm in the micropylar
half shows a large and complex system of ER (Fig. 12.8 B), while it is
relatively meagre and there is no definite pattern in other parts of the cell.
However, all over the cell, ER is only sparsely associated with ribosomes.
In the mature mother cell the micropylar ER becomes reticulated with
several "loops" and "circles" (Fig. 12.8 C). A vacuole appears below the
nucleus of a mature mother cell, and several small vacuoles in the micropylar
part. From the micropylar half, the plastids and mitochondria shift laterally
to the chalazal end of the maturing mother cell (Fig. 12.8 D); the micropylar
ER may have a role in the migration of these organelles. In a mature cell
the plastids with a few mitochondria are restricted to the region below the
nucleus, the mitochondria lie just below the plastids (Figs. 12.8 D; 12.9).
Other organelles, like dictyosomes, lipid droplets, dense bodies bounded by
unit membrane, and an occasional multivesicular body, do not show polarity
in their distribution.
The nuclear envelope of the mother cell shows a large interruption in the
lateral wall of the cell (Fig. 12.9), and remains identifiable until the onset
of meiosis I. Starch accumulates only in the chalazal region of the megaspore
mother cell. Following meiosis I, plastids and mitochondria become restricted
to the chalaza} dyad. Generally, a linear tetrad of megaspores is formed. In
a triad-upper undivided dyad and two megaspores-it is the functional
chalaza} megaspore which has most of the plastids. The starch grains are
consumed during the enlargement of the megaspore. It appears that starch
begins to accumulate, much in advance, at the site of the functional megaspore.
Female Gametophyte. The female gametophyte develops from the haploid

110 The Gymnosperms
Fig. 12.8 A-D. Ginkgo biloba. Electron micrographs, megaspore mother cell. A Portion
of megaspore mother cell (mgmc), outer (cw) and inner (i/) layer of mgmc wall, middle
lamella (mdl), and spongy cells (sg). B, C Part of micropylar half of mother cell.
B Extensive ER. C Reticulate ER. D Part of chalaza! region of mother cell; note starch
(sgr)- bearing plastids (p), near the nucleus (nu), and mitochondria (m) lower down. (After
Stewart and Gifford 1967)
Ginkgoales 111
Fig. 12.9. Ginkgo biloba. Megaspore mother cell (diagrammatic) . er endoplasmic

reticulum.go golgi apparatus. m mitochondria. nu nucleus. p plastid. v vacuole. (After
Stewart and Gifford 1967)
chalaza) megaspore. The gametophytic tissue, however, is not uniformly

haploid, as shown by cytological studies and cytophotometric measurements
of the DNA content (A vanzi and Cionini 1971 ). At the beginning of the
cellular stage, 5% nuclei of the gametophyte show 1C DNA content, 50%
2C, more than 40% 4C, and the remaining nuclei 8C or higher DNA content.
This variation in the DNA content is attributed to endopolyploidy. The
cells with 4C content are mostly located in the outer region of the
gametophyte. The nuclei with higher DNA content (8C or more) degenerate
in the young gametophyte. In older stages, most cells contain 2C DNA.
Free-nuclear divisions (Fig. 12.10 A-C) occur in the megaspore for about
4 weeks. According to Favre-Duchartre (1958), there are 13 successive
112 The Gymnosperms
ljij !
I
@! . • I
•
OQ··i
A ·:
\CF.; ~
I
·/;;) I
._, :
·"""·'""'• I
®··:
\.;
.r-r'.. - ~
A B 0
Fig. 12.10 A-H. Ginkgo biloba. Longisection of peripheral region of female gametophyte.
A, B Cytoplasm with free nuclei. C Nuclear divisions D-G Formation of alveoli and
initiation of walls. H Cellular gametophyte, note the thick wall. (After Favre-Duchartre
1956, 1958)
mitotic cycles, so that more than 8000 nuclei are formed. The divisions are
initiated at the chalaza! end, and proceed towards the micropyle. The
prothallial cytoplasm, throughout cenocytic phase, adheres to the megaspore
membrane. Walls are laid down at the end of the 14th mitotic wave. The
gametophyte remains colourless throughout the free-nuclear stage. Typical
alveoli are formed (Fig. 12.10 D-G), followed by cellularization (Fig. 12.10 H).
The gametophyte becomes green (due to the presence of chlorophyll) and
starch is synthesized. The female gametophyte of Ginkgo biloba is the only
seed plant with a chlorophyllous gametophyte. The relative transluscence
of the integumentary tissues of the ovule permits sufficient light to induce
the synthesis of chlorophyll (Friedman and Goliber 1986). The plastids do
not contain an organized thylakoid membrane system (Pettitt 1977). When
cell formation begins, the female gametophyte has a light green colour,
attributed to chlorophyll. EM of chloroplasts demonstrate stacking of thylakoid
Ginkgoales 113
membrane in the grana. Plastids located deeper in the gametophyte have

fewer thylakoid membranes and may also show prolamellar bodies.
The presence of chlorophyll has been confirmed by Burgerstein (1900).
He showed that an alcoholic extract of the gametophyte fluoresces red;
Carothers (1907) suggested it may be capable of photosynthesis. A
measurement of photosynthetic active radiation (PAR) indicates that a
gametophyte (growing within an ovule) can receive significant quantities
of light, i.e 70 f.1 mol photons m· 2 s· 1 (Friedman and Goliber 1986). This
unique ability to produce chlorophyll and perform photosynthesis results
from its·exposure to sufficient levels of light, and an inclination to react to
this stimulus by the development of functional photosynthetic apparatus.
The entire gametophytes were dissected free from the ovules. They were
capable of gross photosynthesis (not net photosynthesis) under experimental
conditions. On a dry weight basis, the maximum rate of carbon fixation, in
near-saturating light intensities, was 3.64 x 10· 3 ).1 mol C02 g·'s·' (Friedman
and Goliber 1986).
A mature prothallus contains water, nearly 60% of the whole mass (Favre-
Duchartre 1956). Reserve food accumulates in both the fertile and sterile
prothalli, and is therefore independent of fertilization. In addition to starch,
a mature prothallus also contains lipids, and its density decreases sharply
from the epidermis inwards (Fig. 12.11 A). The concentration of lipoptoteins
increases during the maturation of the prothallus. The starch grains vary
from 4-5 )1m in diameter in the epidermal cells to 15-20 )1m in the inner
cells. They are simple, irregular in outline, and have a central hilum of
indefinite shape (Fig. 12.11 B-F).
The apical part of the gametophyte regularly forms an unusual structure,
the "tent pole" (Fig. 12.11 A). The central portion of the micropylar region
grows by active cell divisions to form a column (the tent pole) which grows
into the nucellus, or may even reach the pollen chamber.
At the micropylar end of the cellular gametophyte, one or two cells
enlarge and function as archegonial initials. Each initial divides transversely
and forms a large central cell and small neck cell of the archegonium
(Fig. 12.12A). At maturity there are four swollen neck cells (Fig. 12.12 B),
which project upward and may have secretory role (Lee 1955). In the
beginning, the four neck cells are in one tier but later they may show a two-
tiered arrangement (Favre-Duchartre 1956). In a female gametophyte the
maturation of different archegonia is synchronous. The central cell nucleus
divides to form a small ventral canal cell and the egg cell (Fig. 12.12 A, B).
The mature egg cell is spherical, and measures ca. 500 )1m in diameter with
its nucleus ca. 100 )1m across (Camefort l965a). The chromatin is confined
to small globule so that the nucleus appears almost colourless with Feulgen
reaction (Fig. 12.12 B).
The single-layered archegonial jacket comprises uninucleate cells. Several
simple pits occur on the wall between the jacket cells and the central cell
114 The Gymnosperms
~'
' ~
~'
i .
~
Fig. 12.11 A-F. Ginkgo biloba. A Longisection mature female gametophyte, note two
archegonia and tent pole at the micropylar end, and distribution of starch (open circles),
lipids (black dots) and lipoproteins (radial lines). B Three cells from the margin, outer
layer is sudanophilic. C-F Cells from different layers of the gametophyte, at various
stages of development, show accumulation of starch , lipids and lipoproteins. (After
Favre-Duchartre 1958)
(Hirase 1895). Occasionally, lobes of the cytoplasm of the central cell may
project into these cells. Plasmodesmata are also present in these pits (Stopes
and Fujii 1906), and have been confirmed by ultrastructural studies (Maugini
and Fiordi 1970). Probably, the nutrients are absorbed through the jacket.
Avanzi and Cionini (1971) measured the DNA content of the jacket cells
cytophotometrically. The large uninucleotate nuclei had DNA content
corresponding to 2C, 2C-4C, 4C or 4C-8C, due to endoduplication. Cionini
( 1971) characterized the DNA in jacket cells by HCl hydrolysis curves and
observed two types of DNA complexes: (a) Feulgen-stainable after 5 min.
and (b) after 7 min. of hydrolysis. This is probably related to the functional
activity of jacket cells or formation of nascent DNA during endoduplication.
The passage of materials from the jacket into the egg, and to the coenocytic
(later cellular) proembryo has been studied using electron microscopy
Ginkgoales 115
Fig. 12.12 A, B. Ginkgo biloba. development of archegonium. (After Favre-Duchartre

1956)
(Maugini and Fiordi 1970). The wall between the jacket and central cell of
the archegonium is fairly thick, and has prominent pits (Fig. 12.13 A). The
cytoplasm of the central cell forms short and blunt projections at the site
of the pits (Fig. 12.13 B, C). The cytoplasm is separated only by the thin
pit membrane, and they remain in contact through plasmodesmata. Soluble
materials move across this contact. Granular material, which lies outside
the plasma membrane of the egg cell, is deposited around the cytoplasmic
projection of the egg (Fig. 12.13 C). According to Maugini and Fiordi
(1970), the granular material represents the temporarily stored nutritive
116 The Gymnosperms
Fig. 12.13 A-D. Ginkgo biloba. Central and jacket cell. A Longisection shows thick
and pitted wall between central (cl) and jacket layer U). B Pit region shows plasma
membrane (pm) in contact with the pit region of the cell wall. C Later stage, central cell
cytoplasm (d) extends into the pit, laterally disposed granular material (gm), endoplasmic
reticulum (er) in the cytoplasmic appendage. D Degenerated jacket cell, the contents
pass into the proembryo through open pit. (After Maughini and Fiordi 1970)
Ginkgoales 117
material on its way from the jacket cell to the egg cell. The plasma membrane
of the coenocytic proembryo shows small invaginations and short and irregular
microvilli. The increased surface provides a greater absorptive area for
nutritive materials. Starch and-protein granules, stored in the gametophyte,
are translocated to archegonium (in a soluble form) through the pits. When
the proembryo is free-nuclear, or immediately after cell formation, the pit
mambrane breaks down at places. Through these passages, the mitochondria,
plastids, dictyosomes, portions of endoplasmic reticulum and nuclei (whole
or in parts) flow (from degenerated jacket cells) into the egg cytoplasm
(Fig. 12.13 D). This is perhaps due to a sudden lowering of pressure inside
the egg. The contents of the jacket cells partially accumulate between the
cell wall and the plasma membrane of the proembryo.
The egg cytoplasm has the usual organelles like ER•. plastids, Golgi bodies,
ribosomes and a large number of mitochondria. Observations with an electron
microscope (Camefort 1965a) have revealed the following ·cytoplasmic
formations (so-called proteid vacuoles): (a) The morphology of small
inclusions is somewhat different from others. They are completely enclosed
in the double membrane of ER whose components stay together. An
enveloping vacuole is thus absent. (b) Microbodies are present in abundance,
have dense contents enclosed by a single membrane of ER. Their morphology
is similar to certain lysosomes in animal cells. (c) Vesicular bodies occur
seldom and comprise a mass of vescicles enclosed by a single membrane.
The amyloplasts in a mature egg are distributed at the periphery. They
are enclosed in a layer of endoplasmic reticulum, in addition to their own
membranes (Camefort 1965a). The amyloplasts continue to fragment until
the egg is mature.
Pollination
The ovules are pollinated soon after megaspore formation. A pollination
drop is secreted at the micropyle of the ovule. The wind-borne pollen, after
landing on the pollination drop, imbibes nutrients from the fluid, becomes
heavy, sinks down the micropyle, and reaches the nucellar tip. Unpollinated
ovules drop from the tree about 4 weeks after anthesis.
Post-Pollination Development of Male Gametophyte. On the return of

favourable weather after the winter, the pollen grains germinate in the
pollen chamber (Favre-Duchartre 1956, De Sloover-Colinet 1'963), The exine
splits along the germinal furrow, the pollen tube emerges at the distal end,
and the vegetative (tube) nucleus may pass into it. The tube grows laterally
and horizontally into the massive nucellus and branches profusely. The
pollen tube grows in between the nucellar cells, in a hypha-like manner
(Fig. 12.14 A). The branched pollen tube in the nucellar tissue has been
studied from sections and dissections (Friedman 1987), and undoubtedly
has a haustoria! function (see Chap. 2).
118 The Gymnosperms
The proximal end of the pollen grain contains the prothallial, the stalk
and body cells/male gametes. In the beginning it is enclosed in the exine,
but later enlarges and bursts out of the exine. The short, broad end of the
"tube" is usually unbranched (Fig. 12.14 B-F), and a bunch of them grow
into the nucellus towards the female gametophyte. Consequently, the nucellus
becomes completely disorganized between the intermediary chamber
and the female gametophyte. At the grain end, the pollen tubes appear to
~ ~
q:.:: ·:&.?iL'7'§_.: . ~
\·/·.¥·::·:.~~!~?::..·
~ . .. . =: ·:.¥:f
. . .. B.
...·.....· ~ ·..... :·... :' :.: ·... ·...
· . · . ·.·. -: 0
-
c
E F
Fig. 12.14 A-F. Ginkgo biloba. A Haustoria! ramifications of pollen tube between
nucellar cells. be body cell. tn tube nucleus. B-F Diagrammatic representation of upper
portion of nucellus and female gametophyte, longisections. B-D Pollen tube and
degenerated inner portion of nucellus. E Pollen tube close to the archegonium. fd
fertilization fluid, tp tent pole. F Post-fertilized shrivelled nucellus. (A After De Sloover-
Colinet 1963, B-F after Favre-Duchartre 1958)
lie in a longitudinally continuous cavity in the middle of the nucellus

(Fig. 12.14 D-F). Thus, the pollen tube has: (a) a vegetative portion which
Ginkgoales 119
grows laterally into the nucellus and is haustoria! in function, and (b) a
fertile portion which carries the motile gametes.
The occurrence of motile/ciliate sperms in Ginkgo was discovered by
Hirase (1895). 2 This was a landmark in the history of (embryological)
study of gymnoserms.
The ultrastructure of male gametes has been investigated by Gifford and
Lin (1975). The body cell remains spherical during maturation (Fig. 12.15
A, B). Two blepharoplasts, from which cytoplasmic/astral rays radiate,
appear on one side of the body cell (Fig. 12.15 A, B). These rays are the
microtubules, which extend from the matrix or the surface of the
blepharoplasts. The latter move through 90° and come to lie at a right angle
to the long axis of the pollen tube (from the very begnning). The blepharoplasts
bl
c
Fig. 12.15 A-E. Ginkgo biloba. Body cell and male gamete. A Body cell with irregular
nucleus; two blepharoplasts (bJ) at oppO>ite poles. B Body cell with a lens-shaped nucleus
and two osmiophilic globules ~og), one on either side. C Division of body cell. D Two
male gametes, one gamete shows a beak-like extension. E Gamete with three gyres of
ciliate band (top view). (A, B After De Sloover-Colinet 1963, C-E after Favre-Duchartre
1956)
2This tree, in the Botanical Garden, Tokyo, is still in a healthy state.

120 The Gymnosperms
increase in size and become prominent. The entire surface of the matrix is
covered by a single layer of probasal bodies (Fig. 12.16 A, B). The total
number of probasal bodies on one blepharoplast is 1000. Each probasal
body shows a hub-and-spoke arrangement in the centre and nine-fold
symmetry. A central tubule is present along the entire length of the probasal
body. Microtubules/astral rays/cytoplasmic rays originate from the interior
of the blepharoplast, pass between probasal bodies, and extend into the
cytoplasm. The probasal bodies separate from each other and form the
basal bodies of flagella. The matrix of the blepharoplast comprises dense
and less dense regions, the latter appearing to be infiltrated by a network
of tubules (Fig. 12.16 C, D).
Fig. 12.16 A-D. Ginkgo biloba. electron micrographs. A Whole-mount (portion) of

grain-end of pollen tube; the body cell shows lens-shaped nucleus and two prominent
osmiophilic globules ( og) on either side of nucleus. B Osmiophilic globule and
blepharoplast (bl) in ( a part of) body cell. C Numerous probasal bodies; microtubules
in less dense regions (ldr-arrows) . D Peripheral portion of blepharoplast; microtubules
in groups extend from probasal body. (After Gifford and Lin 1975)
Ginkgoales 121
During its maturation, the nucleus of the body cell becomes lens-shaped
(Figs. 12.15 B, 12.16 A). A vacuole, which is an osmiophilic globule (it has
no membrane around it), appears on either side of.the nucleus (Figs 12.15 B;
12.16 A) and is attached to the blepharoplasts (Lee 1955). A dumbell-
shaped lipoprotein granule arises in the cytoplasm close to the nuclear
membrane. In addition, the cytoplasm of the body cell shows other organelles,
like protoplastids (numerous, electron-dense, showing some lamellar
developments), mitochondria, small vacuoles, lipid bodies, abundant
ribosomes, and relatively sparse ER and dictyosomes (Fig. 12.16 B). The
body cell divides, the lipoprotein granule splits into two, and each half is
incorporated into a spermatozoid. The mitotic spindle of the body cell lies
at right angles to the long axis of the pollen tube, so that the two sperms,
enclosed in the wall of the body cell, lie side by side (Fig. 12.15 C, D) until
they are mature. The vacuole (osmiophillic globule), earlier attached to the
blepharoplasts, shifts to the opposite pole of the cell and becomes attached
to the sperm nucleus. Later, it acquires granular contents due to diffusion
of chromatin from the nucleus (Lee 1955). The blepharoplast forms a part
of the tapering distal part of the sperm. The spiral band has three turns/
gyres (Fig. 12.15 E). There is simultaneous division of the nucleus of the
body cell in the pollen tube and of the central cell in the archegonium.
Fertilization
In zooidogamous gynmosperms, autolysis of the nucellus, megaspore wall,
and the gametophytic tissue around the archegonia form an archegonial
chamber (Pettitt 1977). The turgid nucellar cells abruptly release their vacuolar
contents, a fluid is produced which forms a pool and floods the archegonial
chamber and the space above it (Fig. 12.14 E). The male gametes are
released in this fluid (Lee 1955). The spermatozoids, with the band of
flagella at their posterior end, swim about in the chamber with a forward
and circular motion. The four neck cells open out as the egg cell pushes
through the disintegrated ventral canal cell to form a beak. The egg nucleus
may elongate and extend into the beak. As soon as the spermatozoid becomes·
attached to the projection of the egg, the elongated nucleus withdraws·
toward the centre and the beak of the egg retreats. According to Lee (1955),
only the head of the sperm (made of a vacuole-like structure and cytoplasm)
flows through; most of its body is left behind, outside the archegonium,
and disorganizes immediately. However, Favre-Duchartre (1956) observed
portions of the ciliate band of the sperm inside the egg. Extra sperms are
prevented from entering 'into the egg cytoplasm by a thickening of the
plasma membrane of the egg (Lee 1955).
During karyogamy, the paternal chromosomes (12) become short. They
can be stained clearly by Feulgen reaction as soon as the male pronucleus
penetrates the female pronucleus. They mix with the maternal chromosomes
(12) during the first somatic prophase (Favre-Duchartre 1958).
122 The Gymnosperms
Several pollen grains may germinate in a pollen chamber. However, as

soon as one pollen tube has discharged its spermatozoids, the body cells in
the other tubes frequently fail to divide and degenerate. When two
spermatozoids are discharged, two eggs can be fertilized, but a second
embryo rarely develops.
Both fertilized and unfertilized ovules are shed from the tree at irregular
intervals. The ovules may be sterile or may enclose one or two embryos
ranging from the coenocytic to the dicotyledonous stage. Unfertilized ovules
shed from the tree can undergo fertilization and subsequent embryonic
development.
Embryogeny
The zygote nucleus divides in situ followed by several free-nuclear divisions
(Fig. 12.17 A). The nuclei become distributed throughout the young
proembryo. Sometimes, evanescent walls (Fig. 12.17 B) appear during the
free-nuclear period. During later stages, the nuclei become distributed almost
evenly in the cytoplasm. Wall formation takes place when there are 256
free nuclei; the newly formed cells fill the entire proembryo (Fig. 12.17 C).
Within the female prothallus, the cellular proembryo develops continuously;
it is considerably influenced by the prevailing temperature. The cells at the
base divide and function as embryonal cells, while the upper cells elongate
to form a massive suspensor (Fig. 12.170). There is, however, no well-
defined suspensor; it is a micropylar region of elongated cells.
Differentiation of Embryo. There is a cap-like structure formed by the

upper cells (towards the neck of the archegonium) which is pierced through
by the radicle at the time of germination (Favre-Duchartre 1958). The
embryo develops (slowly) after the seeds have been shed.
The mature embryo is dicotyledonous, occasionally three cotyledons are
present. The embryo may reach its maximum size in 3 months after
fertilization, if the development is accelerated in an incubator. In nature,
the embryos may remain healthy for 12 months if the ovules are preserved
in a damp place. This is because the embryo is surrounded by the prothallial
cells, which provide necessary humidity for its survival (Favre-Duchartre
1958). There is no after-ripening requirement, and the seeds germinate
whenever a suitable substrate is available.
The most conspicuous change in a female gametophyte is the deposition
of reserve food like fat, starch and protein. Unlike other gymnosperms, the
accumulation of food reserves in Ginkgo occurs before fertilization (cf. cycads;
Favre-Duchartre 1958).
Abundant starch as reserve food accumulates in the gametophyte even
when the archegonia are still immature. Lipids occur as variously-shaped
globules which decrease sharply from epidermal cells inwards. Lipoproteins
appear nearly 2 months before fertilization, at first in the inner layers and
Ginkgoales 123
............···-·-····· ....................
D
Fig.12.17 A-D. Ginkgo biloba. Proembryogeny. A Longisection free-nuclear proembryo
with the jacket layer and part of gametophyte. B Proembryo in non-median section,
formation of evanescent walls. C Cellular proembryo. D Proembryo, the upper cells
elongate to from the suspensor. em egg membrane. (A, C, D After Lyon 1904, B after
Favre-Duchartre 1956)
later towards the periphery. In a mature gametophyte, there are four zones
of storage cells: (a) The peripheral zone of three or four layers of isodiametric,
vacuolate cells containing numerous round and densely aggregated lipid
124 The Gymnosperms
droplets in the cytoplasm. Where the density of the lipid droplets is low,
the cytoplasm shows dictyosomes, a few mitochondria with short cristae,
ER and chloroplasts. ER comprises rounded or irregular vescicles. Small
starch grains occur embedded in the chloroplasts (Dexheimer 1973).- (b)
The middle zone contains lipoproteins and starch. The cells are large and
vacuolate. The lipoprotein inclusions are irregular (diameter 5-50 f.Lm). They
are either enclosed in the central vacuole or are limited by a fine membrane
in the periphery of the cpll. Amyloplasts are abundant in these cells while
other cytoplasmic organelles like ER, dictyosomes, mitochondria and
ribosomes are rather sparse. (c) The inner zone contains starch. these cells
are very large and show a thin parietal layer of cytoplasm. (d) The central
zone cells do not have any reserve food material (Dexheimer 1973).
Seed. The seed coat is contributed both by the chalaza and integument.
The integument differentiates into three zones: (a) outer (parenchymatous)
sarcotesta, (b) middle (sclerenchymatous) sclerotesta and (c) innermost (thin-
walled) endotesta. The sarcotesta is 5-6 mm thick in the equatorial region,
and 2-3 mm in the micropylar and chalaza! region; it is the only "live"
portion of the integument. The epidermis has a ca. 15-f.Lm-thick cuticle which
is interrupted above the stomata. The cells contain chloroplasts, some of
them also have druses of monohydrate calcium oxalate crystals. The bulk
of the integument is formed of large turgid cells, the latter further enlarging
towards the sclerotesta, while the number of chloroplasts and the size of
starch grains decrease (Favre-Duchartre 1958). The stomata comprise two
kidney-shaped cells filled with starch grains; the frequency is about eight
stomata per mm2 •
The sclerotesta is hard and lignified. It forms the shell, which is slightly
flattened laterally, and usually has two ribs (facing each other) which represent
the suture. It is unlignified in the micropylar half, and forms as many
valves as there are ribs.
a
The endotesta has withered appearance. In the micropylar half it adheres
to the sclerotesta-a~d is free from the nucellus. It is parchment-like, has a
golden-brown c.olq~r, and is thin and translucent. In the chalaza! half, the
endotesta unites :with the nucellus, adheres to the prothallus, but is always
separate from sclerotesta (Favre-Duchartre 1958).
The seed coat is vascularized. Several unbranched vascular bundles are
arranged in a ring, the traces enter the chalaza, pass through the basal plate
of thick-walled cells (see Schnarf 1937).
The mature seed is the size of a small apricot. The seed coat has an outer
orange-coloured fleshy portion rich in butyric acid, and emits an odour like
rancid butter. Inner to the fleshy layer is the stony layer, followed by the
innermost papery layer. The dicotyledonous embryo is in the centre of the
gametophyte, the so-called endosperm. The two cotyledons are normally
equal and hypogeal, and have stomata mainly on the adaxial surface.
Ginkgoales 125
Germination. During germination, the plumule is pushed out of the testa

by elongation and arching of the cotyledonary base. A portion of the cotyledon
remains inside the seed, enlarges and persists through the first season, and
functions as a haustoria! organ. The first two or three leaves on the seedling
are small and scale-like. The young stem stops· its rapid elongation
during the first winter after it has formed a close crown of leaves and large
terminal bud.
Chromosome Number
In Ginkgo biloba, the haploid number of chromosomes is 12. There is
evidence of karyotypic changes within the species. The basikaryo type is
more or less constant; there are differences in the exact location of centromeres
and in number and position of the satellites (see Khoshoo 1962).
The duration of the life cycle in G. biloba is 14 months (Favre-Duchartre
1958).
The ovules are pollinated (in Paris) at the megaspore stage in the second
half of April (first year). The pollen grains begin to germinate 3 weeks
after landing in the pollen chamber.
In the female prothallus, the coenocytic phase continues until the middle
of June, and archegonial initials can be observed by the end of the month.
Fertilization begins early in September. Both fertilized and unfertilized
ovules are shed from the tree at irregular intervals from October to April
(second year).
The cellular proembryo develops within the prothallus. As soon as the
embryo matures, it germinates in June (second year), in adequate water and
at suitable temperature, without a period of dormancy.
The fossil record provides only a few clues as to when the Ginkgoales
appeared first, and the ancestral group from which they have been derived
(see Stewart 1983).
Trichopitys is one of the Palaeozoic genera which may have affinities
with the Ginkgoales. However, there is no evidence of differentiation of
Trichopitys axes into short shoots (a feature of Ginkgo biloba). Florin (1949)
contemplates that as short shoots evolved, the ovulate branches present on
the long branches of Trichopitys were transferred to the short shoots, and
the number of ovules on a branch became reduced to one. or two. Some
support for this hypothesis can be derived from abnormal specimens of
G. biloba, where several ovules are formed on an axillary branch system
quite similar to that of the fertile truss of Trichopitys. If Trichopitys represents
an early stage in the evolution of Ginkgophytes, then the relationship of the
group appears- to be with the conifer type. This is because the fertile shoot
126 The Gymnosperms
of Trichopitys is in the axil of a foliar unit (cf. Cordaitales) and is not a

megasporophyll with ovules (Pteridospermales). On the other hand, evidence
from the abnormal production of ovules on leaves of G. biloba does not
support this view. Embryologically, too, they are similar to cycads (pollen
grains, male gametophyte, motile sperms, development of female
gametophyte). This implies that the origin of Ginkgoales from the conifer
type is far from settled. According to Stewart (1983), it is best to presume
that the Ginkgoales originated from a Palaeozoic ancestor, until there is
more evidence from the fossil record.
While we know a good deal about various aspect of biology of Ginkgo
biloba, priority attention should be paid to the gaps in our knowledge. A
detailed and critical investigation should be made by applying newer
techniques. A fascinating area of research would be the examination of
living material through cine films. Microcinematography will bring out the
sequenees of development (such as in the development of motile male
gametes) which are otherwise difficult to follow. The scope in this method
of study is endless.
13. Cordaitales
For a long time, the Cordaitales have been regarded as the dominant
gymnosperm group of the Palaeozoic era. After the discovery of
Pteridospermales, it became apparent that both the groups were present in
the Palaeozoic. Although recorded from the Lower Pennsylvanian deposit
(Good and Taylors 1970), cordaites became an important element of the
Carboniferous in the middle Pennsylvanian dunes, and flourished into the
Permian. The leaves of Cordaites are reported from the Permo-Carboniferous
of Siberia, China, India, Australia and South America (see Stewart 1983).
Leaves, stems, roots, cones and seeds of these plants are particularly abundant
in the Mid-Pennsylvanian location of Iowa and Kansas in the U.S.A. The
order includes a single family, Cordaitaceae.
Cordaitaceae
Cordaites is an organ genus for detached foliage of the Cordaitaceae.
According to Arnold (1967), the name may also be employed, without
typification, for the entire cordaitean plant.
Habit. Cordaites is a tree which attained a height of nearly 30 m and a

diameter of more than 1 m at its expanded base. It had a crown of branches
near the apex, which bore spirally arranged sessile leaves (Fig. 13.1A).
Cridland (1964) reconstructed a cordaitean plant as a small tree ca. 5 m
high (Fig. 13.1B). According to him, it had stilt roots (comparable to those
of mangroves), and it inhabited swamps, estuaries and along seashores.
Root. The roots, known as Amyelon, are profusely branched and shallow,
and form a pad of stilt roots which support the stem. A transection shows
a central exarch actinostele with protoxylem strands at the tip of each arm
of the stele. It is surrounded by a thick layer of secondary xylem. A well-
developed periderm is present. As it grows it produces an outer layer of
phellem of empty radially arranged cells (Fig. 13.2); lenticels are also
present. A broad zone of aerenchymatous phelloderm is formed around the
stele. As the root elongates, the protostele gives way to a siphonostele by
medullation of the metaxylem to form a pith. The protoxylem points persist
128 The Gymnosperms
A 8
Fig. 13.1 A, B. Cordaitean plant, reconstruction from Carboniferous (A) and

Pennsylvanian (B) era. (A After Scott 1909, B after Cridland 1964)
at the margin of pith and lie adjacent to the secondary xylem (Cridland
1964).
Stem. A transection of Pennsylvanioxylon (Figs. 13.3 A; 13.4 A) and

Mesoxylon (Fig. 13.4 C) shows a large pith surrounded by pycnoxylic
secondary wood. In the former, the distinction between primary and innermost
secondary wood is apparent only in radial section. The latter also shows the
innermost narrow protoxylem elements with spiral thickenings, followed
by larger spiral and reticulate elements, and finally by tracheids with
scalariform bars (Fig. 13.3 B). The compact secondary wood of
Pennsylvanioxylon and Mesoxylon is similar in structure and comparable to
Araucarioxylon. Bordered pits are present on radial walls in two or more
rows. They are alternate and densely crowded, so that their borders appear
Cordaitales 129
Fig. 13.2. Ameylon radicans. Transection root. mx metaxylem, pre primary cortex,
ph phloem, px protoxylem, src secondary cortex, sx secondary xylem. (After Scott 1909)
Fig. 13.3A, B. Pennsylvanioxylon. A Transection young branch. as axial sympodium.

It leaf trace. sx secondary xylem. B Wood, radial section . (After Takhtajan 1956)
hexagonal (Fig. 13.3 B). The pore of the pit is a transverse or inclined
elliptical slit. The rays in the secondary wood are uniseriate and vary in
height.
A well-preserved specimen of Mesoxylon shows a layer of secondary
phloem (Fig. 13.4 C) which consists of radially arranged sieve cells,
130 The Gymnosperms
parenchyma and fibres. In a young stem, the cortex shows a hypodermal

system of vertical strands of anastomosing fibres and secretory sacs. In
older stems, an early formation of periderm in the cortex is evident. It
apparently replaces the cortical tissues as the stem increases in diameter.
Fig. 13.4A-C. A, B Pennsylvanioxylon, oblique (A) and longisection (B) stem shows
septate pith . C Mesoxylon, stem transection. bt branch trace. c cortex, pi pith , sp secondary
phloem . sx secondary xylem. (Courtesy Dr. G .W. Rothwell)
The large parenchymatous pith , in a well-developed stem, shows

conspicuous transverse lens-shaped cracks which alternate with the diaphragms
Cordaitales 131
of parenchyma cells (Fig. 13.4 A, B). Such septate pith casts are named
Artisia.
Leaf. The leaves, Cordaites, are spirally arranged, generally 15-20 em

long; unusually, ca. 1 m long and 15 em broad. These are linear, lanceolate
to spatulate, and may have a blunt or pointed tip. The leaves are tough,
leathery, thick at the region of attachment, taper proximally, and have
parallel venation (Fig. 13.5A). Several species have been described, which
differ from each other in their epidermal and internal structure. Transfer
specimens of thick cuticles of compression fossils show rectangular cells in
rows. The cell wall is thick and straight, and the cells are parallel to the
long axis of leaf. The haplocheilic stomata are arranged in longitudinal
rows between the ribs formed by the vein in the lower epidermis. In a few
species they are scattered on the upper epidermis (Stewart 1983). The
stomata are elongated parallel to the veins, guard cells are in a pit surrounded
by four to six subsidiary cells, of which two are polar.
The palisade layer is poorly differentiated and the spongy mesophyll
shows large lacunae between the veins. Hypodermal strands of sclerotic
tissue, and their extent of development and distribution, are diagnostic to
distinguish the species. In some species the fibrous strands occur only
above and below a vein, while in others they extend from the upper to
lower epidermis, enclose the vein, and form !-shaped girders. The centripetal
xylem in the veins is surrounded by a two-layered vein sheath. Most species
also show centrifugal xylem (Fig. 13.5 B).
Several authors have made a detailed study of the leaf and branch trace
of Pennsylvanioxylon and Mesoxylon (see Stewart 1983). The branching is
axillary, and the leaf trace appears double at its point of inception at the
periphery of the pith. The sympodia from which the leaf trace arises is
mesarch in Mesoxylon and endarch in Pennsylvanioxylon. Each double leaf
trace is flanked by two branch traces. As the leaf traces traverse the cortex
of the stem and enter a leaf base, they dichotomize repeatedly to form
8-16 bundles. In Mesoxylon the leaf trace (within the leaf) shows both
centripetal and centrifugal xylem, in Pennsylvanixylon exclusively centripetal
xylem.
The two branch traces fuse after reaching the cortex, and form a single
(branch) trace.
Fructifications
Because of morphological similarity, both pollen and ovulate fructifications
are designated as Cordaianthus. The compound fructifications consist of a
primary axis with secondary shoots/cones in the axils of modified leaves
called bracts. The cones bear spirally arranged modified sterile (mostly)
and fertile leaves, the scales. Close to the apex, a few fertile scales terminate
in pollen sacs or ovules. The cones are monosporangiate. Figure 13.5A
132 The Gymnosperms
Fig. 13.SA, B. A Cordaitean branch bearing leaves, vegetative bud and fertile shoots.
B Cordaites sp ., vertical section leaf. ep epidermis. hyp hypodermis. lc transversely
elongated cells connecting bundles, mx metaxylem . px protoxylem. (A After Grand 'Eury
1877, B after Renault 1879)
Cordaitales 133
shows a branch bearing a vegetative bud (upper right), leaves and scattered
fertile shoots.
Cordaianthus concinus. Besides compression-impression fossils several

permineralized specimens have been discovered from the Middle
Pennsylvanian coal ball material (Delevoryas 1953). A cone has, on a
secondary shoot, ca. 25-40 scales, and only 5-10 distal scales are fertile
(Fig. 13.6 A, B). Each fertile scale usually terminates in six microsporangial
pollen sacs fused at the base (Fig. 13.6 C, D). They contain pollen grains
of the Florinites type, monosaccate with reticulate ornamentation on the
inner wall of the saccus. The latter is attached to the body of the pollen
grain (corpus) on its distal and proximal surfaces (Millay and Taylor 1974).
Cordianthus pseudofluitans. Most of the species are compression-

impression fossils from the Upper Carboniferous of England and Europe;
very few permineralized specimens have been discovered. According to
Florin (1939, 1950a, 1951), the morphology and anatomy of the ovulate
fructification is similar to that of the male fructification. A cone and its
subtending bracts are distichous on the primary axis. Each cone has ca.
16-20 spirally-arranged scales, and the distal four to six scales are fertile.
Each scale may have a single reflexed ovule at the tip, or it may dichotomize
so that there are two or more pendulous ovules at the tip of the branched
scale (Fig. 13.7 A, B). The cordate ovules are platyspermic. In compression-
impression fossils, the ovules are of the samaropsis type, while the structurally
preserved permineralized ovules are assigned to Miterospermum (Lower-
Upper Pennsylvanian) and Cardiocarpus (Middle Pennsylvanian to Permian).
In transection, the ovules are biconvex, the primary plane of the ovule
being parallel to the long axis, and the secondary plane at right angles to
it. In C. spicatus the conspicuous testa consists of a sarcotesta; its outer
zone comprises large thin-walled cells, and the inner !>mall cells. The
sclerotesta has thick-walled sclerotic cells and spine-like projections into
the sarcotesta. The endotesta cells line the sclerotesta. In the primary plane,
the sarcotesta of Cardiocarpus and Miterospermum expands into a flattened
"wing" in some species (Fig. 13.7 C).
The nucellus is free from the integument, except at the base. Its distal
portion, below the inner opening of the micropyle, is differentiated into a
beak and well-developed pollen chamber (Fig. 13.7 D). Several workers
(see Stewart 1983) have observed the florinites dispersed type of pollen in
this chamber. A few specimens showed well-preserved gametophytes with
at least two micropylar archegonia flanking a "tent pole' (as in some
Pteridosperms and Ginkgo). In the mature seeds the nucellus appears thin
and inconspicuous.
A single vascular strand enters the base of the ovule. In Cardiocarpus,
it terminates in a plate of tracheids at the base of the nucellus. Two vascular
134 The Gymnosperms
I
c
\ 0
Fig. 13.6 A-D. A Cordaianthus concinnus, portion of primary axis of fertile shoot
shows two male secondary shoots with spirally arranged sterile and fertile scales. B, C
C. penjoni, D C. saportanus, B Longisection secondary shoot shows sterile and fertile
scales with terminal pollen sacs. C, D Fertile scales. (A After Delevoryas 1953, B after
Renault 1879, C, Dafter Florin 1951)
strands arise from the central vascular supply and extend laterally, in the
primary plane, into the sarcotesta, and terminate almost at the apex of the
ovule. In Miterospermum compressum, the two lateral strands pass through
the sclerotesta into the sarcotesta where each strand divides (in the primary
plane) to form several bundles (Fig. 13.7 D).
Cordaitales 135
Fig. 13.7 A-D. A Cordaianthus pseudofluitans. B Cardiocarpus cordei, ovulate

fructification. C, D Mitrospermum compressum, trans- (C) and longisection (D) of ovule.
mic micropyle. pc pollen chamber. sa sarcotesta. sci sclerotesta. vb vascular bundle.
w wing. (A After Florin 1951, B after Zimmermann 1959, C, Dafter Taylor and Stewart
1964) .
Cordaitales is the oldest truly coniferophytic group of plants (Miller 1977).
We do not precisely know how or from what ancestral group the Cordaitales
evolved. An appraisal of the ontogeny, structure and reproductive biology
of the most ancient seed ferns , Upper Carboniferous seed ferns, cordaites,
and conifers shows differences between the last two groups and their supposed
ancestors. The analysis is based principally on the investigations of
Cordaixylon dumusum, Mesoxylon priapi (Cordaitales) and Lebachia and
Ortiseia (Coniferales). The data indicate that the evolution of Cordaitales
from the Lower Carboniferous seed ferns included several significant steps
(Rothwell 1986) such as:
1. The transition from hydrasperman reproduction (the most primitive
type of reproduction in gymnosperms), where the ovules had a complex
pollen chamber which sealed the microgametophytes with radial, trilete
exine structure, i.e. prepollen, to more modem type, where the integument
plays an active role in pollination and post-pollination sealing of the pollen
chambers.
2. The vegetative leaves changed from pinnately compound to strap-
shaped simple leaves with entire margins.
136 The Gymnosperms
3. Reduced number of ovules and microsporangia per microsporophyll.

4. Aggregation of sporophylls on compound shoots.
It is uncertain wh~ther changes in anatomical features and cuticular anatomy
(important in the identification of Cordaitales) were also instrumental in
the evolution of Cordaitales. The evolution of conifers probably resulted
from changes in vegetative (the conifer type of leaves along the entire
branch system) and reproductive morphology (aggregation of sporophylls
on compound shoots with derived appendage arrangement on secondary
axes) among plants that already had the more modem type of gymnospermous
ovules, as in Cordaitales and some Upper Carboniferous seed ferns (Rothwell
1986).
14. Voltziales
In the history of the plant world, the transition from the Carboniferous to
the Permian has attracted much attention. Coexisting with the Cordaitales
in the Late Paleozoic and persisting into the Mesozoic ar~ the Voltziales,
often called the transition conifers. They exhibit the redpction series and
changes in symmetry of a cordaianthus-like fertile dwarf shoot (see Chap. 13)
that lead to the ovuliferous scale of extant conifers. Florin (1939, 1944,
1945, 1950a, b) made an intensive study of the Voltziales. He compared
the fertile dwarf shoot of Cordaianthus with the ovuliferous scale of modem
conifers, and cited several intermediate stages within the Voltziales (Florin
1951 ). However, the V oltziales do not fill the ·many gaps between the
cordaites and conifers (Stewart 1983).
Morphology
Externally, the Voltziales resemble extant taxa such as Araucaria excelsa,
the Norfolk Island pine (Florin 1951). Older plants show monopodia! stems
with a regular arrangement of their (five or six) branches in a whorl
(Fig. 14.1A). The leaves on the vegetative branches are acicular,. while
tl:iose on fertile branches have a bifurcate tip.
The best-known genera are Lebachia and Ernestiodendron. They are based
on vegetative remains which were formerly included in Walchia, and are
distinguishable from one another primarily by differences in their cuticle.
They appear to lack calcium oxalate crystals in the cuticular layers. Lebachia
and Ernestiodendron have simple unicelled hairs particularly on the underside
of the leaves (unlike contemporary Cordaites or Voltziaceae). The haplocheilic
stomata are amphistomatic (on both surfaces of the leaf) and have four to
ten subsidiary cells. The leaves and bracts are supplied by a single vein.
Lebachia leaves are "entire", except those ori penultimate shoots and bracts
of ovulate cones, which have bifurcate tips. They are decurrent along the
branch (Fig. 14.1 B) and slightly spreading. The stomata are longitudinally
oriented in two bands. The leaves of Ernestiodendron are borne at right
angles to the branch (Fig. 14.1 C). The stomata are in isolated longitudinal
rows. Walchia represents fossils which lack sufficient preservation of the
cuticle for identification, either in Lebachia or Ernestiodendron.
138 The Gymnosperms
/)
c
Fig. 14.1A-E. A, D, E Lebachia piniformis, B Lebachia sp., C Ernestiodendron.
A Whorled branches at the apex of shoot. B, C Leafy shoot, spirally-arranged leaves
(B), and leaves at right angels to the axis (C). D, E Branches with pendulous pollen
cones (D) and erect ovulate cones (E). (After Florin 1951)
Anatomy
The wood is not widely known in the Late Palaeozoic forms (Miller 1977).
Those on record show a eustele with endarch primary xylem (Florin 1951 ).
Secondary xylem has tracheids with araucaria type of pitting (one to three
rows of closely arranged alternating bordered pits). Occasionally, xylem
parenchyma is also present. The rays are uniseriate and resin ducts are
absent. Early or older Voltziales (see later) had fewer rows of pits on the
radial tracheid walls than Cordaites. A relatively large and irregularly ruptured
pith is present (Florin 1951). In addition to the above characteristics, Rothwell
(1982) observed in Lebachia twigs (from the Middle Pennsylvania of
Voltziales 139
Oklahoma) the origin of leaf traces from the eustele, and apparent resin
ducts without epithelial cells in the pith and cortex of twigs and mesophyll
of leaves.
Reproductive Organs
There is considerable variation in the structure of the seed cone within the
Voltziales. The changes begin in the Palaeozoic and continue into the
Mesozoic. On the basis of relative modification of the ovuliferous shoot
and its subtending bract, Florin (1951) recognized two groups of Voltziales.
These are placed in two families: (a) Lebachiaceae (Pennsylvanian and Early
Perrnian)-the older Voltziales. (b) Voltziaceae (Late Permian and mainly
Mesozoic)-the younger Voltziales.
Lebachiaceae
The Lebachiaceae have inflorescences with strobiloid nature and show
relatively much less modification than that of the Cordaianthus (the strobiloid
nature of the dwarf shoot is evident).
The plants are monoecious. Morphologically the fructifications appear
like cones. They are monosporangiate, ellipsoidal to cylindrical borne at
the tip of leafy branches. The male cones are pendulous and the female
cones upright (Fig. 14.1 D, E).
The male cone of lebachias is radially symmetrical to ellipsoidal. There
is a primary axis around which are attached spiral dorsiventral scales or
microsporophylls. In these cones there are no bracts subtending a secondary
shoot. Florin (1951) and others (see Stewart 1983) interpret these cones as
simple, and each microsporophyll has a narrow stalk-like base and a distal
upturned leaf-like portion (Fig. 14.2 A, B). There are two microsporangia
on the abaxial side (Fig. 14.2 A), and the pollen grains are monosaccate or
bisaccate (Fig. 14.2 C).
The compact ovulate cone has a primary axis which bears several spirally
arranged bracts with bifurcate tips (Fig. 14.2 D, E). In the axil of each bract
is a tangentially flattened secondary shoot with scales attached assymetrically
(Mapes 1982, Rothwell 1982). In Lebachia, there is only one fertile scale,
terminated by a single erect bilaterally symmetrical ovule (Fig. 14.2 E). In
Ernestiodendron and Walchiostrobus, there are several ovule-bearing scales
(unlike Lebachia ). In the former, all the scales on the secondary shoot are
fertile (Fig. 14.2 F); in Walchiostrobus, sterile scales are present on the
proximal and fertile scales on the distal portion of the secondary shoot
(Fig. 14.2 G). The ovules may be erect or reflexed (Fig. 14.2 F, G). Florin
(1951) regarded the lebachia type of cone as primitive, and Ernestiodendron
as more advanced.
The ovulate fructifications in Lebachia and Cordaianthus are compound,
this is the most important character which indicates their relationship.
The primary difference is that in Cordaianthus the secondary shoot-bract
140 The Gymnosperms
Fig. 14.2A-G. A, B Lebachia hypnoides, microsporphyll. A Two microsporangia.

B Lateral view. C Lebachia sp., pollen grains; proximal (above) and distal (below)
views; optical Iongisection in centre. D, E L. piniformis, secondary shoot. D Abaxial
view, bract with bifurcate tip. E Adaxial view, spirally arranged scales and erect ovule.
F Ernestiodendron filiciforne, bract and secondary shoot complex; all scales fertile .
G Walchiostrobus sp., secondary shoot shows fertile and sterile scales and recurved
ovules. (After Florin 1951)
complex is lateral and distichous on the primary axis, and spirally arranged
in Lebachia.
Voltziaceae
Voltziaceae includes plants where the seed scale of the ovulate cone shows
significant modification from the strobiloid condition of the Lebachiaceae,
yet with parts sufficiently free from one another and from the bract to
indicate derivation from a strobilus. They show reduction and fusion of
fertile dwarf shoot elements, change in their symmetry from radial to bilateral
(dorsiventral), and also fusion between the fertile shoot and its subtending
bract. These changes begin in the Palaeozoic and continue into the Mesozoic
(Miller 1977).
Several types of modifications of the bract-scale complex are known.
Some of the genera may possibly represent intermediate evolutionary stages
between the Lebachiaceae and modern conifers, while others show interesting
variations, which probably became extinct without giving rise to an extensive
lineage (Miller 1977).
In Pseudovoltzia (Permian) five sterile lobes, their bases fused to one
another, occur axillary to each bract (Fig. 14.3 A, B). The middle and two
marginal lobes are larger than the remaining two, and each bears a recurved
ovule. This lobed unit is fused to the subtending bract for a short distance.
Florin (1951) pointed out that the five lobes and three ovules represent
eight distinct parts. Schweitzer (1963) studied petrified material of
Pseudovoltzia, where the original three-dimensional arrangement is preserved.
It shows a greater fusion of parts than was formerly known, and also
Voltziales 141
OS
D E
c
Fig. 14.3A-F. A-D Pseudovoltzia liebeana. A Ovuliferous scale shows three recurved
ovules (o). B Abaxial view of bract-scale complex. b bract, os ovuliferous scale. C, D
Vascularization of bract-scale complex (C) and ovuliferous scale (D). E Ullmania bronii,
ovuliferous scale, adaxial surface shows single ovule. F Glytolepis longibracteata, fertile
ovuliferous scale, note five sterile lobes and two recurved ovules. (A-D After Schweitzer
1963, E, F after Florin 1951)
supports Walton's ( 1928) view that the three larger lobes and their associated
ovules represent megasporophylls and there are only five distinct parts to
the unit. The vascular bundles to the bract and scale are not fused, the lobes
and ovules are each vascularized by a single strand, five of which arise
from the end of the scale bundle (Fig. 14.3 C, D). The three larger lobes
and their associated ovules represent megasporophylls. Pseudovoltzia is, thus,
reinterpreted as too advanced to be an intermediate between the Lebachiaceae
and the Voltziaceae (Miller 1985). Voltzia also has five sterile elements in
its cone scales, but there are only three ovules. The middle and two marginal
lobes are larger than the remaining two lobes, as in Pseudovoltzia. The seed
stalks are fused to their subtending sterile lobes and are free only at their tip.
142 The Gymnosperms
Ullmania (Permian) has a single recurved ovule adaxial to a large orbicular

scale (Fig. 14.3 E) subtended by a bract. In the structurally preserved
material of U. frementaria, it is still uncertain if the seed has a separate
attachment or is fused to the scale (Schweitzer 1963). The bract-scale complex
is similar to that of Araucaria in general morphology and orientation of
the single adaxial ovule.
Glyptolepis (Permian and Triassic) has elongate lax cones apparently borne
in groups (Schweitzer 1963). The secondary shoot is planated and bilaterally
symmetrical. It consists of five or six sterile lobes and two recurved ovules
in lateral position on either side of the sterile scales (Fig. 14.3 F). The unit
is axillary to a bract. In older species the bract is shorter than the sterile
scales, while in younger species, it is longer (Florin 1951). The bracts and
their secondary shoots are spirally arranged on the lax primary axis.
Voltziopsis (Early Triassic and possibly Permian) is known only from the
Southern Hemisphere. Cylindrical seed cones terminate short branches and
have ca. 25 bract-scale complexes. The narrow bracts have bifurcate apices
and are nearly as long as the scale. The ovuliferous scale consists of a thick
but flattened stalk which expands apically into five lobes (rarely six) that
separate from the stalk at different levels. Each lobe bears a single recurved
ovule which is free from the lobe but adnate to the seed stalk. Both complete
cones and isolated bract-scale complexes are known, which suggests that
the cones may have dissociated when mature (Townrow 1967).
The pollen-bearing structures in Cordaianthus are produced in sporangia
which occur at the tip ofmicrosporophylls (see Chap. 13). In Lebachiaceae
and most other conifers, the sporangia are in the basal region of a laminar
microsporophyll. These sporophylls (in Cordaianthus and L~bachiaceae) are
arranged in a helix around an axis to form a strobilus, which is further
organized to form compound strobili in Cordaianthus. The presence of simple
or compound strobili is important to distinguish Cordaitaceae from
Lebachiaceae and, according to Miller (1985), makes it difficult to accept
Florin's (1951) hypothesis that the Lebachiaceae evolved from Cordaitaceae.
lnsteaq, it is more likely that the two groups share a common ancestor
which was contemporary with or earlier than the earliest Cordaites. This
may also explain the diversity seen in Pennsylvanian Lebachiaceae (Miller
1985).
As there is no precise information on how or from which ancestral group
Cordaitales evolved, the major changes that occur in the evolution of
compound female cones ar~ interpreted from the existing information of
the ovulate fructification of the upper Carboniferous-Permian Cordaitales
and Voltziales.
Voltziales 143
The secondary shoot in the axil of a bract of Cordaianthus (see Chap. 13)
is homologous with the bract and axillary ovuliferous scale of a conifer
seed cone. Ernestiodendron and Walchiostrobus are like Cordaianthus in
the radially symmetrical secondary shoots composed of spirally arranged
sterile and fertile scales. The evolutionary trends apparent in Lebachiaceae,
without implying an evolutionary series (Stewart 1983), which lead to the
compound ovulate cone, are: a reduction in the number of ovules on the
secondary shoot (Lebachia); the erect ovules become inverted with their
micropyles directed towards the primary axis of the fructification
(Ernestiodendron sp. and Walchiostrobus sp.), and a tendency towards bilateral
symmetry by flattening of the secondary shoot and ovule (Lebachia).
In Voltziaceae the secondary shoot is planated and bilaterally symmetrical.
In Glyptolepis and Voltziopsis, the fertile and sterile scales lie in a single
plane and there is hardly any tangential fusion. The number of ovules is
reduced to two. Ullmannia shows maximum tangential fusion of sterile
scales to form a ovuliferous scale (cf. several conifers), and the single
ovule is inverted. Permineralized specimens of Lebachia (Mapes 1982,
Rothwell 1982) indicate how the shift occurred from the radial symmetry
of the secondary shoot (Cordaitales and Lebachiaceae) to bilateral
symmetry (Voltziaceae) followed by tangential fusion of scales to form an
ovuliferous scale with adaxial ovules and their micropyle directed towards
the primary axis, as in compound or ovulate cones of extant conifers~
According to Stewart (1983), this is undoubted evidence that the ovuliferous
scale of a conifer cone is a highly modified secondary shoot in the axil of
a bract.
Late Permian conifers combine features of various Mesozoic taxa, and
the Mesozoic and Cenozoic conifers those of many modem taxa. According
to Miller (1982), the extant families may not have a common origin among
the Voltziaceae. Some of them may· have· diverged directly from the
Lebachiaceae, without intermediates from the Voltziaceae.
Certain characters of the conifers can be traced back to the Late Devonian.
The growth habit appears to have already evolved (as seen in Callixylon ),
with 9-m-tall trunks and stumps 1.52 m in diameter. All features of
coniferophytic secondary xylem, except the torus, had evolved by the end
of the Devonian and are common to the progymnosperms.
The Coniferophytes were much more widespread and diverse during the
Mesozoic than they are now. Also, modem conifer families appear to have
originated somewhat earlier than was formerly presumed. This makes it
difficult to visualize their evolution from the known Voltziales, where petrified
seed cones show greater modification than was formerly known. According
to Miller (1977), it would be better to search for precursors of modem
families in Early Triassic and Palaeozoic sediments. While Florin's (1951)
hypothesis remains valid, it may not be t~.e only route that conifers followed
in their evolution (see Miller 1985).
15. Coniferales
The conifers constitute more than three-fourths of the living gymnosperm

flora, and cover vast areas of the temperate regions of both Northern and
Southern Hemispheres.
The largest continuous, dense forest in the world is coniferous, and it
encircles the earth just below the northern tundra of the Arctic and extends
from Canada, Russia, Northern Europe to northern China and Japan (see
Chap. 1). It began developing (ca. 10 000 years ago) when the glaciers
retreated, and most of it still exists today. It extended southward along
mountain ranges and merged. with the deciduous forests of the south. The
coniferous trees are ideally suited for the cold and snowy climate of the
north. The dense stands deflect the wind, the snow slips off the down-
sweeping branches of the trees, and the thick bark and waxy evergreen
needles retain moisture for photosynthesis even in winter. The thick,
rough bark of many conifers helps the tree to survive forest fires. There are
52 genera and ca. 566 species, which are classified into six families:
Pinaceae, Taxodiaceae, Cupressaceae, Podocarpaceae, Araucariaceae and
Cephalotaxaceae.
The plants are mostly trees or shrubs. Most trees are pyramidal or conical
when young, round- or flat-topped when old. Many have columnar trunks
free of branches to a considerable height. The tallest tree in the world is the
Californian redwood-Sequoia sempervirens (Taxodiaceae), over a 100m
in height. The redwoods prefer the humid conditions of the Pacifi~ coast,
where fog is prevalent and the deep soil enables their roots to grow.
Sequoiadendron giganteum (Taxodiaceae) is known as the giant sequoia, and
grows on the western slopes of Sierra Nevada in California. Some of the
trees are over 76 min height and 9 min diameter. The 'General SQ.erman'
tree, the largest living e~ample, measures 90 min height and has a diameter
of 33.5 m. The giant sequoias are more massive and heavier than the coastal
redwood. Each tree (redwood or giant sequoia) yields a great deal of high-
quality ·timber, and at one time it was felt that t3e trees would become
extinct due to overcutting. Now, however, they are protected, and many of
the areas where they grow have been declared National Parks. There are
other giants like Dacrydium cupressinum (Podocarpaceae), which reaches
Coniferales 145
a height of 55 m. Taxodium mucronatum is the "big tree of Tule" from

southern Mexico and· has a diameter of 17 m.
Massive buttresses support a few conifers, such as the Taxodium distichum
(bald cypress) and some of the larger species of Chaemaecyparis and
Cryptomeria. Such buttressed trees may have a circumference three times
as great at ground level as at 2 or 3 m higher. Others may have no apparent
swelling at the base, but have radiating roots which anchor the tree against
strong winds. Taxodium distichum likes damp soil when growing in swampy
habitats, and develops hollow vertical pneumataphores ca. 1 m high and 30
em across.
Young trees of Chaemaecyparis-lawsoniana (lawson cypress) and several
other species have downwardly sweeping branches, the lower ones often
rooting in the ground at their tips. Dacrydium laxifolium is the smallest conifer
known. Mature specimens with cones are ca. 8 em tall. It is a native of
mountains of New Zealand. Juniperous horizontalis (Cupressaceae) is a
completely prostrate form. Some trailing or creeping junipers, less than 30
em tall, have been horticulturally developed as ground covers (see Chap.
23). Podocarpus ustus (Podocarpaceae), confined to New Caledonia, is the
only conifer suspected to be a partial parasite on the roots. of Dacrydium
taxoides. The leaves are described as reddish bronze or purple (see Sporne
1965).
Some of the conifers have an exceptionally long life. In the Inyo National
Forest of California, USA, there is a tree of Pinus aristata which is more
than 4600 years old, and which even produces cones occasionally
(P. Maheshwari and Konar 1971). The giant sequoias live perhaps as long
as 3000 years, while the redwoods live for 2000 years. A cross section
from the base of a Cedrus deodara (Pinaceae) tree-displayed in the timber
museum of Forest Research Institute, Debra Dun (India)-shows that this
tree was .a sapling in the 12th century and died early in the 20th century
(P. Maheshwari and Biswas 1970).
Most conifers have needle-like leaves and are evergreen. In Araucaria
the leaves may persist and remain green for as long as 15 years; the leaf
base expands as the branch bearing it enlarges. A few conifers are deciduous,
shedding their leaves or branches in autumn: Larix decidua, Pseudolarix,
Metasequoia and Taxodium distichum. In Phyllocladus (Podocarpaceae) the
foliage leaves are scaly and succeeded by phylloclades, which arise in their
axils. The strobili are borne at the edge of the phylloclades near the base.
Anatomically, transfusion tissue is associated with veins in the leaves of
all investigated conifers. The prominence and arrangement of this tissue
varies in the genera. It may surround the veins (Pinus), form a cap adaxial
to the xylem (Araucaria), occur as wing-like projections lateral to the vascular
bundles (Cephalotaxus, Podocarpus), or be abundant lateral to the phloem.
Transfusion tissue may consist entirely of tracheids, or have parenchyma
interspersed with the tracheids (see Griffith 1971).
146 The Gymnosperms
The roots of conifers have ectotrophic mycorrhiza which occurs naturally.

The mycorrhiza is a symbiotic association between the root cells and fungi.
It has been established that the symbiosis increases the uptake of phosphorous
by the host plants for the mycelium. In Araucariaceae, the mycorrhiza is
endotrophic, and Podocarpaceae also have root nodules (cf. Leguminaceae).
15.1 PINACEAE
Pinaceae is the largest and the most recent of the modem conifer families.
The exact time of origin is not clear, but the family is evident by the Early
Cretaceous. Seed cones of this Cretaceous assemblage show considerable
diversity but have more features characteristic of Pinus than of any other
modem taxa (Miller 1977). Anatomically also, the cones are distinctly
Pinus-centred (which occurs only in the Early Cretaceous). Pseudolarix, and
possibly Picea, may have evolved by the end of the Cretaceous, but reliable
evidence of other modem taxa is absent during the Mesozoic.
The characteristic features of the family are: spirally arranged parts; two
pollen sacs in a microsporophyll, pollen mostly bisaccate; ovuliferous scale
free or slightly fused at the base of the bract scale, two ovules per scale,
and seeds generally winged. There are over 200 spp. included in ten taxa:
Abies, Cathaya, Cedrus, Keteleeria, Larix, Picea, Pinus, Pseudolarix,
Pseudotsuga and Tsuga. All are trees, and the family is economically very
important (see Chap. 23).
Pinus
The genus has ca. 80 valid species (Dallimore and Jackson 1966). They are
divided into two natural subgenera with distinct characters:
(a) Haploxylon or soft pines-the scaly shoot at the base of the short shoot
is deciduous, ray tracheids in secondary wood of stem and root have smooth
wall; the needle (leaf) has a single vascular bundle.
(b) Diploxylon or hard pines-the scaly shoot present at the base of the
short shoot is persistent; the ray tracheids have a corrugated wall, and the
needle has two vascular bundles.
Morphology
Young pine trees are pyramidal with horizontal branches at regular intervales
(Fig. 15.1.1). This symmetry is lost as the tree matures and the crown
becomes round, flat or spreading.
There is a primary tap root with a large number of laterals called long
roots. The primary root soon becomes arrested while the long roots continue
to grow and bear clusters of dwarf roots. Some of these roots branch
dichotomously, form coralloid masses, have an ectotrophic mycorrhiza
(Fig. 15.1.2 A,B) and are termed mycorrhizal roots.
Coniferales 147
Fig. 15.1.1. Pinus roxburghii, plantation at Ranikhet (India). (After P. Maheshwari

and Konar 1971).
The majority of the fungi forming ectotrophic mycorrhiza belong to the

families Boletaceae and Agaricaceae (seeP. Maheshwari and Konar 1971).
In the soil these fungi occur as loose hyphal strands and can infect directly.
A seedling with mycorrhiza acts as an inoculum for the spread of infection
to the adjoining seedlings. Excessive mortality has been recorded when
good-quality seedlings have been grown in soil other than forest soil. In
P. nigra var. calabrica, stocks grown in nurseries without proper mycorrhizal
conditions suffer on an average 45% mortality, while only 7% has been
recorded in those raised in a soil with a mycorrhizal fungus like Boletus
bovinus (Raymer 1947).
The intensity of mycorrhizal infection in soil depends on strong sunlight,
adequate soil moisture, aerated and acid soils, and low levels of soil fertility
(Bakshi and Kumar 1968). There is a positive correlation between the
intensity of infection and increase in the mobilization of soil nitrogen. It
has been suggested that the internal nutrient status, especially N2, phosphorus
and potassium is responsible for determining the intensity of infection.
When their concentration is optimum in the roots, there is resistance to
mycorrhizal infection.
The effect of ectomycorrhizal fungi on growth and phosphorus uptake of
P. sylvestris seedlings at increasing phosphorus levels has been studied.
Even a low intensity of infection by LLlccaria laccata greatly stimulates growth
of the seedlings, which seems to be more related to its capacity to produce
growth substances than to its capacity to stimulate phosphorus uptake. The
poor efficiency of Hebeloma crystuliniforme compared with LLlccaria laccata,
148 The Gymnosperms
c D
Fig. 15.1.2A-E. A-C Pinus sp., mycorrhiza. C Transection ectotrophic mycorrhiza shows
mantle (rna) and Hartig's net ( hn). D E P. roxburghii foliar spur. D Young, and
E unfolded needles. cat cataphyll. ne needle. pro prophyll. (A, B After Hatch 1937,
C after Hatch and Doak 1933, D, E after P. Maheshwari and Konar 1971)
at any level of phosphorus, could result from differences in diversion of

carbohydrates from the host to the fungal structure.
Two types of branches are present: long shoot with unlimited growth;
dwaif shoot with limited growth. The long shoots occur on the main stem
as lateral buds in the axils of scale leaves, and terminate in an apical bud.
The latter is enclosed by a number of bud scales closely surrounded by a
thick mat of hairs. The lateral buds grow, nearly horizontally, to a certain
length termed nodal growth. In uninodal pines a single internode, while in
multinodal pines two or more internodes are formed in a year.
The dwarf shoot or foliar spur develops on a long shoot in the axil of
a scale leaf (Fig. 15.1.2 E) and lacks a terminal bud. Each dwarf shoot
initially has two opposite scales-prophylls followed by 5-13 cataphylls
Coniferales 149
(Fig. 15.1.2 D). Finally, needle-like foliage leaves (Fig. 15.1.2 E) develop
on the spur shoot, in fascicles of one (P. monophylla), two (P. sylvestris,
P. merkusi), three (P. insularis, P. roxburghii, P. gerardiana), four
(P. quadrifolia) and five (P. griffithii, P. wallichiana, P. armandi). The
number of needles present is constant for a species and is used for
identification of different pines.
The plants are monoecious. The male and female cones are borne on
different branches of the same tree. The male cones are modified dwarf
shoots and appear in clusters (Fig. 15.1.3A) on the lower branches of the
tree. The number of cones in a cluster varies from ca. 15 (P. griffithii) to
140 (P. roxburghii). Each cone arises in the axil of a scale leaf, replaces
a dwarf shoot and is surrounded by several bracts. The female cones replace
the terminal buds of the long shoot and are a modified long shoot
(Fig. 15.1.3 B). In the beginning, the female cones are protected by an
involucre of bracts. The cone has an average length of 1.5-2 em and a
diameter of 0.8-1 em. The cone axis elongates (at the time of pollination)
and the cone protrudes beyond the envelop of scales and is open to receive
pollen grains. After pollination the cone closes. Originally, the cones are
pale green but about the time of pollination the colour changes to reddish
purple, and finally to glaucous green or purplish (P. wallichiana). The seeds
are shed nearly 27 months from the time of initiation of the cone. The
cones (at this time) are ca. 20-24 em in length and 5.5-6 em in diameter.
In most species the hard and woody mature female cone opens to release
the seeds (P. roxburghii, P. wallichiana), in others the seeds are released
only after the cones fall to the ground and rot. In a few species (P. jlexilis)
fc
8
Fig. 15.1.3A, B. A Pinus roxburghii, long shoot bearing a cluster of male cones (me).
B P. wallichiana, long shoot bearing second year female cones (/c). (After Konar 1960)
150 The Gymnosperms
the cones remain on the tree for several years and open only when scorched
by forest fire.
Anatomy
Root. In a mycorrhizal root the rootlet is enclosed by a fungal sheath, and
the fungus is restricted to its cortex in a net of hyphae called Hartig's net
(Fig. 15.1.2 C). The piliferous layer and the root hairs are replaced and
covered completely by a mantle of mycelium. In P. radiata the invading
fungus lyses the middle lamellae of the cortical cells, separates them by
mechanical action, and occupies the intercellular spaces (Foster and Marks
1966). In the early stages of symbiosis, deep intracellular hyphae occur in
the cortical cells, which are later digested by the host tissue due to its
increasing resistance, and the fungus remains primarily in the intercellular
regions.
The young long roots (in transection) have a circular outline (Fig. 15.1.4 A).
The epidermis consists of isodiametric cells, and several cells become filled
with tannin. The cortex is broad, parenchymatous and the cells frequently
contain starch. The endodermis consists of suberized cells, usually impregnated
with tannin which gives a brownish-orange colour. Casparian strips are
indistinct. There is a six- or seven-layered pericycle, and many cells contain
tannin. The stele is generally diarch or tetrarch, occasionally pentarch.
A resin duct is associated with each protoxylem (Fig. 15.1.4 A). The
protoxylem consists of mostly scalariform or scalariform-pitted tracheids,
the metaxylem of only pitted tracheids. The phloem has parenchyma, sieve
cells and tannin cells, and abundant starch (occasionally tannin) is present
in the pith cells.
Secondary growth sets in even before the primary tissues are fully
differentiated. The cambium differentiates from the parenchymatous cells
beneath the phloem (Konar 1963a). It forms (by repeated periclinal divisions)
secondary xylem towards the pith and secondary phloem towards the cortex.
In the region of the resin ducts, the cambium cuts off only parenchymatous
cells, which results in broad xylem rays (Fig. 15.1.4 B). Later, with the
development of more secondary wood, the rays become reduced to a single
cell. Secondary xylem tracheids have bordered pits on their tangential and
lateral walls. In older roots tyloses block the tracheids. The rays are uni-
or multiseriate, the latter associated with a resin duct. The secondary phloem
consists of radially oriented rows of cells.
Simultaneously with the differentiation of the vascular cambium, a cork
cambium is formed in the outer region of the pericycle. It cuts off cork
cells externally and parenchyma internally. The cork cells may be highly
suberized and cutinized (P. roxburghii), and some of them may be filled
with tannin.
Stem. Shoot Apex. The apex of the long shoot remains dormant from
Coniferales 151
Fig. 15.1.4 A, B. Pinus roxburghii, transection root. A Young tetrarch root. BOld diarch
root shows secondary growth. rd resin duct. sp secondary phloem. sx secondary xylem.
(After Konar 1963a)
152 The Gymnosperms
October to March. In P. ponderosa, P. lambertiana and P. roxburghii, the

shoot apex is distinguishable into: (a) the region of apical initials, (b) the
central mother cells, (c) the peripheral tissue zone and (d) rib meristem.
During the dormant phase, the apex has a low parabolic dome averaging
72 J.Lm in height and 277 J.Lm in diameter. It resumes its activity in April,
and by May attains a tall parabolic form. At this stage the needles produced
in the axils of the scale leaves project beyond them. An active shoot apex
is 175-300 Jlm in height and 305-500 Jlm in diameter. By the end of August,
the phase of rapid growth ceases. The dwarf shoot apex is dome-shaped
during the active period of growth and appears elongated and cone-like at
maturity. There are four zones, highly telescoped, corresponding to those
of the long shoot. The apical initials (by anticlinal divisions) contribute to
the outer layer of the peripheral tissue zone, and by periclinal and oblique
divisions to the rib meristem and the inner layers of the peripheral tissue
zone.
A transection of a young stem, slightly below the apex, shows superficial
ridges and furrows due to adpressing of the surrounding leaves. The epidermis
is followed by a broad parenchymatous cortex, the vascular bundles are
arranged in a ring, separated from one another by broad medullary rays.
The resin ducts arise schizogenously, have an inner lining of epithelial cells
and are arranged in a ring in the cortex. The pith is parenchymatous. Several
cells of the cortex and pith contain tannin.
The interfascicular cambium arises in the region of the medullary rays
and connects with its intrafascicular counterpart to complete the cambial
zone, which is the entire area of undifferentiated dividing cells. The cambium
forms two types of cells: (a) fusiform initials-these are elongated cells
with tapering ends, and give rise to longitudinal or vertical systems of
xylem and phloem; (b) ray initials-consist of isodiametric cells of relatively
small size and give rise to the ray cells.
The vascular bundles increase in width due to tangential extension of the
fascicular cambium. Further activity of the cambium produces a continuous
zone of secondary xylem and phloem (Fig. 15.1.5. A). At places, the cambium
cuts off parenchymatous cells which form the secondary medullary rays.
These are mostly uniseriate, rich in cytoplasmic contents and have thick
walls.
The phloem initial transforms directly into the sieve element (Srivastava
1963b). Occasionally, it may divide once transversely, the resultant cells
develop into short sieve cells. The cambial derivative (destined to become
a sieve cell) increases in cell volume accompanied by radial expansion.
The secondary wall thickenings (characteristic of the family) are lamellate.
The older cells have more lamellae in their walls than the younger ones. At
this time, the cytoplasm has the usual complements of organelles. Rough
ER is evident in the form of cisternae and vesicles and the ground substance
is rich in free ribosomes. Occasionally, fine fibrillar material can be detected
Coniferales 153
Fig. 15.1.5 A-D. Pinus roxburghii, stem. A Old stem in transection to show growth
rings (gr) and resin ducts (rd). B, C Radial longisection. B Secondary phloem with lateral
compound sieve plates. C Vascular ray parenchyma shows simple pits on radial walls.
D Tangential longisection with uni- and multiseriate rays. (A, C, D After Konar 1963a,
B after P. Maheshwari and Konar 1971)
in the vesicles of the rough ER. Mitochondria and dictyosomes are abundant
and the cortex of the cell contains microtubules, which are usually parallel
to the long axis of the cell.
The plastids also undergo changes. They accumulate a few electron-
dense bodies in the stroma, some of these are 1.2 Jlm in older sieve elements
(Srivastava and O'Brien 1966, Barnett 1974). These crystals, presumed to
be proteinaceous, show weak refrigence under polarized light. Their origin
and significance is not known, but they may develop from the osmiophilic
intralamellar inclusions in the plastids of cambial initials. In Pinus pinea,
Wooding (1966) has described, in the ray plastids, accumulation of granules
comparable in staining to the starch grains.
154 The Gymnosperms
During further differentiation, significant changes occur in the organelles

and membrane system of the cell. The cisternae of rough ER vesiculate
further, lose their ribosomes, and subsequently the vesicles disappear.
Simultaneously, a new reticulum is formed, composed of large cisternae
which mostly occur near the wall. Bouck and Cranshaw (1965) refer it as
the sieve tube reticulum (STR) while Srivastava and O'Brien (1966) termed
it sieve element reticulum (SER). While these changes take place in the
ER, the ribosomes and dictyosomes (in the cytoplasm) disappear.
Evert and Alferi (1965) report necrotic nuclei in the mature (coniferous)
sieve cells. Comparable stages of nuclei in the mature sieve elements have
been identified in Pinus strobus (Muramanis and Evert 1966, Srivastava
and O'Brien 1966), P. pinea (Wooding 1966, 1968) and P. resinosa. In the
elements immediately adjacent to the crushed phloem, the vesicular masses,
nuclear remnants, and mitochondria associated with longitudinally stranded
material persist along the wall, and the lumen of sieve elements appears
empty. The inclusion of the sieve element plastids disappears by the time
it is mature, only granules, resembling starch in their staining behaviour,
persist (Wooding 1966, Wooding and Northcote 1965). In P. radiata and
P. strobus the crystalline inclusions change to fibrillar form.
A mature sieve element of pine is approximately of the same size as the
cambial initial from which it originates. It has a thick lamellated secondary
wall, separated from the cytoplasm by a persistent plasmalemma. The
peripheral cytoplasm is composed of an elaborate system of smooth ER in
the form of long cisternae. Somewhat abnormal mitochondria are often
enmeshed between these cisternae. Plastids are present in various stages of
breakdown, alongwith the starch grains and protein crystalloids. The ground
substance is composed of a fine fibrillar material derived either from the
stroma of degenerated plastids, or from the rough ER, or both. Degenerated
nuclei are mostly present, but the vacuolar membranes, dictyosomes, and
ribosomes are absent. Microtubules may be present at maturity but they are
difficult. to distinguish with certainty. Plasmalemma persists even in the
advanced stages of sieve-element differentiation. Barnett (1974) has recorded
stacks of hexagonally packed tubules in the lumen of sieve elements of
P. radiata.
Initiation of the sieve area has been noticed soon after the cell wall
formation has begun. The formation of sieve pores needs to be explained,
whether the sieve areas arise from the pit fields in the fusion initials from
which the sieve cells have been derived, or whether their origin is de novo.
According to Srivastava and O'Brien (1966), some boring of the wall occurs
because a cavity median nodule is formed where none existed before.
Furthermore, strands from the two sides may join. The "pores" in the walls
of dead sieve elements are much wider than the plasmodesmata. Barnett
(1974) did not observe simple pits in the radial wall of the cambium in
P. radiata. Gradually, the wall thickens in these regions. In an almost
Coniferales 155
mature sieve area, the difference in the structure between the cell wall and
the material of which the cell plate is made is more obvious. Usually, a
sieve plate is homogeneous, ·stains evenly, while the remaining cell wall is
lamellar in structure. This is the stage when communication channels begin
to develop between the cells. Later, they widen enough for entire organelles
to pass through them. The pores in the wall are lined by plasmalemma
(Srivastava and O'Brien 1966), Wooding (1966) observed these as lined by
callose (as revealed by fluroscence microscopy).
The sieve elements have compound sieve plates with either individual or
small groups of (two or more, very closely placed) sieve areas (Fig. 15.1.5 B)
on the radial walls and consist of numerous narrow channels (0.1-0.5 pm
in diameter) lined with callose. These channels widen and coalesce to fonn
a central lacuna in the middle of the wall. The sieve elements are in close
contact with albuminous cells through a rather specialized compound pore.
The sieve element towards the pore is similar to a half-sieve area, but the
channels connecting the central lacuna with the albuminous cells are much
narrower.
The xylem comprises tracheids which have bordered pits on their radial
and tangential walls. Frequently, bars or rims of sanio are present between
the pits. Every year, the activity of the cambium reaches its peak during
spring. The xylem elements produced in this season are broader, thin-
walled and somewhat polygonal. With the onset of summer, the activity
slows down and the diameter of elements becomes appreciably smaller,
they are thick-walled, narrow, squarish and compact. The summer wood
passes rather abruptly into the spring wood, which results in the formation
of growth rings (Fig. 15.1.5 A). In P. halepensis (seeP. Maheshwari and
Konar 1971) the number of circles in one ring of spring or autumn wood
reflects the climatic conditions of a given year much better than the anatomical
features of the tracheids. The constant presence of water table under the
roots also leads to a very low increment in wood thickness.
Resin ducts are distributed all along the wood (Fig. 15.1.5 A) and reaches
maximal number just before the onset of summer.
In the secondary xylem, the rays are differentiated into two distinct
zones. In the central zone the cells are similar and have only simple pits
on their walls (Fig. 15.1.5 C). On either side, one or two layers of cells
form the marginal ray tracheids. They are thick-walled and rectangular
with bordered pits on their radial and tangential walls. In a tangential
section the medullary rays can be distinguished into two types: (a) u~iseriate
rays which are single-cell wide, the number (of cells) vary from 1-12; (b)
multiseriate or fusiform rays which are always more than one cell wide and
several cells in height, and have a resin duct in the centre (Fig. 15.1.5 D).
Simultaneously with the differentiation of the vascular cambium in the
stelar region, a cork cambium arises in .the first or second layer of the
cortex. It divides periclinally and gives rise to the cork or bark on the
156 The Gymnosperms
periphery, and secondary cortex towards the centre. By intermittent anticlinal

divisions it increases the number of radial rows and enables the periderm
to keep pace with the increase in circumference of the stem. The scaly bark
thickens gradually, cracks externally and ultimately wears away.
Compression Wood. It is formed along the under-surface of branches,

on the trunks of leaning or fallen living trees. It apparently forms in response
to geotropic stimuli and may also be due to the concentration of auxin, as
shown in P. strobus by Wershing and Bailey (1942).
Dwarf Shoot. Anatomically, the dwarf shoot resembles a long shoot except
for its narrow diameter. It has a small cortex, few resin ducts, and the
vascular bundles are open and collateral. The meduallary rays are broad
and parenchymatous. The tracheids are mostly scalariform, occasionally
pitted. A large parenchymatous pith has many cells filled with tannin.
Leaf. A cross section of needle (Fig. 15.1.6 A) shows an epidermis of

isodiametric cells with lignified walls. It is discontinuous in the region of
the stomata, which are sunken (Fig. 15.1.6 B) and arranged in axial rows
all along the surface of the leaves.
The hypodermis is two- or three-layered, of polygonal or rounded cells
of uniform thickness.
The mesophyll, the bulk of the leaf, is delimited on its inner side by the
endodermis (Fig. 15.1.6 A). It consists of closely fitting chlorenchymatous
cells with a number of plate-like infoldings projecting into the cell cavity
(Fig. 15.1.6 C). Two or more resin ducts are interspersed in the mesophyll
tissue. Depending on their position, the resin ducts are classified into:
external (touching the hypodermis), internal (touching the endodermis), medial
(touching neither hypodermis nor epidermis), and septal (touching both
hypodermis and endodermis to form a septum).
The stele occupies the centre of the leaf. The prominent endodermis
(Fig. 15.1.6 D) consists of a single layer of barrel-shaped cells, followed
by the transfusion tissue present all round the bundle with a larger amount
on the abaxial side (Hu and Yao 1981 ). There are several tracheidal cells
(Fig. 15.1.6 E) interspersed with parenchyma and albuminous cells.
The number of vascular bundles in a needle may be one (P. sylvestris
var. monophylla, P. wallichiana), two or more (P. roxburghii, P. contorta).
When two bundles are present, they are disposed at an angle to each other
(Fig. 15.1.6 D). The mature leaf has a partly crushed protoxylem. The
metaxylem has helically thickened tracheids with bordered pits. The radial
rows of xylem elements are interspersed with rows of parenchymatous
cells. The phloem parenchyma is more abundant than xylem parenchyma.
Many of its cells contain starch, or proteins, or crystals.
Structure and ontogeny of the stomatal complex has been studied in
Coniferales 157
Fig. 15.1.6 A-E. A Pinus gerardiana , needle in transection. rd resin duct. B-E
P. roxburghii. B Epidermis shows sunken stoma (st). C Mesophyll cell with infoldings
of the wall. D Vascular bundles. end endodermis. tr transfusion t1acheid. E Transfusion
tracheid. (After Konar 1963a)
P. strobus and P. banksiana (Johnson and Riding 1981). The stomatal complex
is eight-celled: two guard, two polar, two lateral and two hypodermal
subsidiary cells. The stomatal complex (in these two pines), therefore, cannot
be readily classified as haplocheilic because a polar subsidiary cell arises
from the same protodermal cell as the guard cell mother cell. A modification
of the classical concept of stomatal development is necessary to describe
stomata as eumesoperigynous (Johnson and Riding 1981 ).
Reproduction
Male Cone
A male cone consists of a number of spirally arranged microsporophylls
with upturned scaly apices (Fig. 15.1.7 A-C). Two microsporangia are
158 The Gymnosperms
borne on the abaxial side (Fig. 15.1.7 B-C) and dehisce by a longitudinal
slit.
Microsporangium. A transection of young microsporangium shows a mass

of meristematic tissue surrounded by an epidermis. One, two or more cells
of the hypodermal region, by repeated divisions, give rise to an archesporia!
tissue which has dense cytoplasm and prominent nuclei (Fig. 15.1.7 D).
The peripheral cells of the archesporium undergo periclinal divisions and
cut off the primary parietal layer towards the periphery (Fig. 15.1.7 E) and
sporogenous cells on the inside. Further periclinal divisions in the parietal
layer forms an outer three- or four-layered wall and the inner mass of
sporogenous tissue (Fig. 15.1.7 F). The innermost layer of the wall cells
develop into the tapetum (Fig. 15.1.7 G). Simultaneously, the epidermal
cells undergo anticlinal divisions; most of the cells are filled with tannin
except two rows of smaller cells, which form the line of dehiscence
(Fig. 15.1.7 G). At the time of dehiscence of the microsporangium, the
epidermis attains its maximal development with fibrous thickenings, and
functions as the exothecium (Fig. 15.1.7 H). As the microspore mother
cells initiate divisions, the epidermal cells lose their nuclei, become filled
with a homogeneously staining substance which may extend (P. strobus) to
the subepidermal layer of the wall. The outer wall of the epidermal cells
gradually becomes cutinized.
The tapetum is of the secretory type in P. sylvestris (Willemse 1971b},
P. banksiana (Dickinson and Bell1972, 1976a) and P. roxburghii (see Moitra
and Bhatnagar 1982). Young tapetal cells are richly cytoplasmic and
multinucleate. They become very conspicuous during meiosis in the
microspore mother cells, and degenerate soon after the spores are released
from the tetrads. The tapetum is involved in the nourishing of the sporogenous
cells/young microspores, and in the formation of exine on the spores
(Dickinson 1970, 1971, Dickinson and Bell1972, 1976a, b, Willemse 1971d).
The development of tapetum and sporogenous cells takes place
simultaneously. There appears to be a correlation between the stage of the
microspore mother cell (meiocyte) and the structure of tapetal cytoplasm.
Accordingly, three phases (with subphases) of development of tapetal cell
can be distinguished corresponding to pre-meiotic, meiotic and post-meiotic
phases of microsporogenesis.
Pre-meiotic Changes. In P. banksiana the development of tapetum has

been studied in detail (Dickinson and Bell 1976a). In a very young
microsporangium, ultrastructurally the tapetal and sporogenous cells are
alike. The membrane investing the plastids is difficult to distinguish and
the contents of the organelles appear in continuity with the ground cytoplasm.
Later, the envelope on the plastids becomes distinct, the number of
dictyosomes and associated vesicles increases, and most of the starch is
~
.."~ ·~:· ·
' .
.I ' ,
\ .
B
' ~c
/~
~~··~· · ~
;:s
fl.] 1r::~"' G S;
~
~
Fig. 15.1.7 A-H. A-C, H Pinus wallichiana. A Male cone. B, C Microsporophyll, dorsal (B) and lateral (C) views. D-G P. roxburghii, (portions
of) longisection of microsporangium shows archesporium (D), sporogenous tissue (E), microspore mother cells (F) and microspore tetrahedral tetrads, ~
and 2-nucleate tapetum (G). H Microsporangium (transection) shows fibrous thickenings in the epidermis. and 4-celled (2 prothallial cells have .....
til
degenerated) pollen grains. (A-C, H After Konar and Ramchandani 1958, D, E after Konar 1960, F, G after P. Maheshwari and Konar 1971). IC
160 The Gymnosperms
lost from the plastids (Fig. 15.1.8 A). These vesicles, which look "coated",
may also surround a mass of fibrogranular, electron-opaque material. Small
portions of rough endoplasmic reticulum (RER) appear. The protoplast of
tapetal cells begins to contract from the fibrous PAS positive cell walls.
Small quantities of a fibrous material accumulates between the contracting
protoplasm and the original cell walls. From this stage onward, the tapetal
and the sporogenous cells become distinct from each other. A large
accumulation of RER and other associated coated vesicles become evident
in the tapetal cells. The mitochondria in tapetal and sporogenous cells
increase in size and frequency, and become more conspicuous (Fig. 15.1.8 A).
The tapetal cells become radially compressed, and gather considerable RER
at the end of this phase.
Meiotic Changes. The tapetal cells increase in volume at the onset of

meiosis. The RER continues to accumulate until the periphery of'the tapetal
cells is loaded with these membranes. The ribosomes are abundant in the
form ofpolysomes only. Numerous large sudanophilic globules ca. 250 }.tm
in diameter are visible. The initially strongly PAS-positive tapetal cell walls
begin to loosen, with a concomitant increase in thickness. Osmiophilic
globules arise outside the tapetal protoplast on middle lamellae between the
outer tangential wall of the tapetal cells and the inner tangential wall of the
innermost wall layer. These globules increase gradually from 0.1 J.Lm to
1.0 }.tm in diameter. Occasionally, they adhere to the middle lamella between
the radial walls of the tapetal cells. The globules continue to be deposited
until a thin layer surrounds the pollen loculus. The tapetal cell walls show
no change during the second half of meiosis.
Post-Meiotic Changes. During the early tetrad stages, the electron density
of the cytoplasm decreases. The tapetal cells become active again as the
pollen wall formation begins. The Golgi bodies produce numerous vesicles,
about 60% of each tapetal cell is filled with vesicles/inflated cisternae. The
accumulation of RER continues. It is possible that sporopollenin precursors
are stored or synthesized in these inflated cisternae and later passed on to
the thecal fluid through these vesicles. The deposition of sporopollenin
starts on the peritapetal membrane (described later) and orbicules. The
peritapetal membrane completely invests the tapetum and developing pollen
grains in P. roxburghii (see Moitra and Bhatnagar 1982). It is acetolysis-
resistant and can be dissected as a bag containing pollen grains. During
pollen wall formation, the tapetal cytoplasm again becomes osmiophilic
and highly vacuolate. In P. sylvestris, Golgi bodies stop vesicle production
(Willemse 1971 b). At the tetrad stage, radial walls between some of the
tapetal cells break down to form coenocytic cells. Tapetal cells degenerate
soon after the release of microspores from the tetrad (Willemse 1971 d,
Dickinson and Bell 1976b), but the deposition of sporopollenin continues.
Coniferales 161
Fig. 15.1.8 A, B. Pinus sp., tapetum electron micrographs. A Tapetal protoplast in pre-
meiotic stage shows conspicuous dictyosomes (d) and a starch grain in one of the
plastids (p) . m mitochondrion. 8 Tapetal protoplast during meiosis, note the deposition
of sporopollenin on the lipid cores of orbicules and membrane-bound vesicles (arrows)
discharged from the tapetal cytoplasm. (After Dickinson and Bell 197 6a, b)
162 The Gymnosperms
The cell organelles become swollen and show electron-transparent contents.

After the young microspore stage, the tapetal cells disappear completely
and the orbicules adhere to the peritapetal membrane.
A detailed developmental sequence of peritapetal membrane, at
ultrastructural level, has been investigated in P. banksiana (Dickinson 1970,
Dickinson and Belll972). The membrane develops between the innermost
layer of the anther wall and the tapetum. Globules of saturated lipids (as
shown by staining reaction) accumulate (at this site) and fuse to form a
continuous outer electron-luscent layer, which coats the entire periphery of
the loculus. A second layer of sporopollenin is deposited on the lipid
substratum after the release of microspores from the tetrads (Fig. 15.1.8 B).
This double-layered structure is called a peritapetal membrane. Deposition
of sporopollenin is first detected as an accumulation of electron-opaque
material upon the earlier-formed membrane. A large number of vesicles
between the peritapetal membrane and tapetal protoplast persist even after
the release of the tetrads from the callose wall.
The tapetal nuclei in younger sporangia show a higher Feulgen stainability
than in older sporangia. In many tapetal cells the nuclei undergo regular
mitotic divisions followed by inhibited cytokinesis which forms binucleate
(rarely multinucleate) cells.
The tapetum generally has a higher RNA content than the microspore
mother cell. There is a gradual increase of RNA synthesis in its cells from
the time they are formed till the onset of meiosis in microspore mother
cells. Thereafter, it declines sharply until final dissolution of tissue. In
P. banksiana (Dickinson and Bell 1976a) the accumulation of proteins in
the tapetal cells occurs from its initiation to the meiotic divisions in mother
·cells. This is associated with an increase in ribosomal population during the
same phase.
In young microsporangia of P. roxburghii, there is a difference in staining
pattern of the cell walls of sporogenous and tapetal cells and of the other
wa,lllayers. The cell walls of the wall layers are thicker and stain a brighter
magenta by periodic acid-Schiff' s (PAS) reaction, than the cell walls of
tapetum and sporogenous cells. The epidermal and middle layer cell walls
are largely cellulosic, 'while those of tapetal and sporogenous cells are
mostly pectinaceous (see Moitra and Bhatnagar 1982).
There is a definite pattern of starch distribution in the microsporangium.
Minute starch grains appear at the archesporia! cell and are absorbed. They
reappear in the sporogenous cells immediately prior to meiosis. In tapetal
cells starch appears only after it appears in the sporogenous cells. During
meiosis starch shows a very regulated pattern of distribution. At metaphase
I the grains accumulate at the equatorial plate, and move to the oppbsite
poles during anaphase I and telophase I, and are equally distributed at the
end of meiosis.
Throughout their development, the sporogenous cells are rich in RNA
Coniferales 163
which becomes maximal in meiocyte prior to meiosis. The young microspore

is poor in RNA but the level rises again at the time of formation of the
antheridial cell (see Moitra and Bhatnagar 1982). The archesporia! and,
later, the sporogenous cells become rich in proteins. The level is high in
meiocytes but low during meiosis and, subsequently, rises during microspore
maturation (P. banksiana-Dickinson and Bell 1976a, b).
Microsporogenesis. The differentiation and maturation of the sporogenous

tissue in Pinus starts in the centre of the sporangium and proceeds
centrifugally. Electron microscopic studies have revealed many more details
of the basic course of development than is shown by light microscopy.
The meiocytes undergo a gradual stretching, thinning, and loss of PAS
stainability before meiosis. With the dissolution of their walls, a thin layer
of callose develops between the cell wall and the plasma membrane. It
gradually thickens with the advance of meiosis.
Meiosis is asynchronous, and prophase I to telophase II are seen in
adjoining meiocytes.
During meiosis, several changes occur in the cell organelles. EM studies
reveal that the structure of nuclear pore in microspore mother cells
(P. sylvestris) is very complex and remains constant throughout
microsporogenesis. The annulus (made up of eight circular regions) and the
centre of the nuclear pore are electron-dense (Willemse 1971 a, b, c). At the
beginning of meiosis, the evaginations (formed during premeiotic stages)
of the nuclear membrane disappear; the large nucleus shows homogeneous
nucleoli and the chromatin begins to shrink. The c)lromosom.es comprise
typical electron-transparent chromatin fibrils, covered by a membrane-like
boundary. The nucleus is homogeneous till diplotene, but later it seems to
produce granules which disperse in karyoplasm and mix with the cytoplasm
after telophase I . The granules are the precursors of ribosomes and polysomes.
The nucleolus disappears during diakinesis. The nuclear membrane shows
invaginations during the young tetrad stages (Dickinson and Bell 1970b,
Willemse 1971c), and transports chromatin-like and other materials into
the cytoplasm. This peculiar phenomenon is connected with "primexine"
formation (described later) in the spore tetrads.
During anaphase and telophase II, the Golgi bodies become active and
produce a variety of vesicles which become arranged in the region of the
future wall, and fuse with each other. Microtubules appear perpendicular to
the fusing vesicles (Willemse 1971 b). A very thin line appears in the direction
of the cell plate in the centre of the fused vesicles. It is replaced by a fine
fibrillar material, which is eventually taken over by the electron-transparent
callose.
Following meiosis, wall formation is _simultaneous, callose grows
centripetally, separating the four nuclei_, and tetrahedral and isobilateral
tetrads are formed (Fig. 15.1.9 A-F). There are no plasmodesmatal connections
164 The Gymnosperms
A 8 c D
E F
pr -----:ll~i@!i·~~~
ac----~~~~~ ~\~--
tn .....,...........,=;;::oo,;<'+.-"="'
(Fig. 15.1.9 A-1. Pinus roxburghii. A-H Development of microspore and male
gametophyte. I Pollen grain at shedding. ac antheridial cell, pr prothallial cell, tn tube
nucleus. (After P. Maheshwari and Konar 1971).
either between the dividing mother cells, or the microspores in a tetrad.

Callose has low permeability, isolates the mother cells and their products
from the circumjacent influences, and the meiocytes follow an independent
course of development (during meiosis).
The development of the pollen grain wall has been intensively investigated,
mainly by using EM, and described in P. sylvestris (Willemse 197lc.) and
P. banksiana (Dickinson 1971). The outermost layer, sexine, is laid down
while the spores are still within the tetrad and callose covering. The Golgi
bodies excrete small unit membranal vesicles (ca. 0.1 fJm) with granular
electron-dense contents, and coalesce during transportation towards the
plasmalemma. The unit membrane of the vesicles fuses with the plasmalemma,
secretes its cellulosic contents outside, but within the callose layer. It is the
laying down of primexine. In P. sylvestris the granular content of these
vesicles changes into fine fibrillar network. The sexine pattern is determined
by placement of the vesicle content on the plasmalemma. A space forms
between the plasmalemma and the callose wall, and is much enlarged in the
region where subsequently air sacs develop. The plasma membrane shows
Coniferales 165
many protrusions. Tectum and baculae are absent in the region between
sacci where the germinal pore of the pollen develops later. In P. banksiana
also sexine in not formed in the germinal pore region.
The next layer laid down is nexine I, formed by the deposition of
sporopollenin on trilaminar tapes (at least five layers) present in the inner
part of the primexine during the tetrad phase. In P. banksiana it is formed
by the thickening of the plasma membrane and the appearance of a new
membrane under it. This process is repeated until several layers develop.
In P. sylvestris electron-dense tapes have been observed lying along the
plasma membrane, and extending deep into the cytoplasm. Lipid granules
form the primary source for nexine I and the tapes probably originate from
plasma membrane or endoplasmic reticulum. The nexine surrounds the
cytoplasm except at the regions where wings develop and is thicker on the
side of germinal pore than on the other side.
The callose degradation begins after the formation of nexine I. The
wings enlarge in volume due to an increase in the sexine layer. The
sporopollenin in the thecal fluid and the tapetum are continuously deposited
on the sexine.
The development of air sacs has been traced in P. banksiana using light,
phase contrast, electron microscopy and histochemical techniques (Dickinson
and Bell 1970a). They begin to form within the tetrad as the primexine is
laid down between the plasma membrane and the callose layer. The sacci
develop in an electron-transparent space containing PAS-positive fibrillar
material. The callosic covering is conspicuous at this stage. The second
phase of sacci development begins when microspores are released from the
tetrad. The PAS-positive materials between sexine and nexine of the early
pollen wall expands several times. The mechanism of expansion is not well
understood. Lipid appears to be the main structural component of the wings
as they c.ollapse after lipid extraction. The wings are autofluorescent in
P. roxburghii (see Moitra and Bhatnagar 1982).
The intine is the last layer to be laid down in P. banksiana, P. sylvestris
and P. roxburghii. It is thinnest at the proximal end (the region of prothallial
cells) and thickest at the distal end of the grain. In P. sylvestris the in tine
is lamellar. In P. roxburghii, a thin layer of callose (callosic outer intine)
starts developing at the two sites where (later) the wings are formed. It
spreads and covers the proximal (prothallial) end of the pollen grain. 'the
inner intine is made up of cellulose and pectin (see Moitra and Bhatnagar
1982).
The nuclear envelope in a mature pollen grain consists of two membranes
with occasional pores (ca. 0.1 Jlm in diameter). In P. banksiana and
P. sylvestris finger-like invaginations (involving both layers of the nuclear
envelope) extend into the nucleoplasm. They appear when the microspores
are still within the tetrad. At the distal end of these invaginations, structures
similar to nuclear pores have been observed. Fibrillar material, interpreted
166 The Gymnosperms
as chromatin, fills up the invaginations, which are discharged into cytoplasm,

and the nuclear envelope becomes regular once again. In addition, in the
outer nuclear membrane, evaginations are present which extend into the
microspore cytoplasm. The latter becomes filled with vesicles (ca. 1 p.m in
diameter) surrounded by unit membrane. Nuclear pores occur at unvaginated
portions only. Chromatin-like material is associated with invaginations in
the nucleoplasm, and not with the evaginations in the cytoplasm. The
invaginations, evaginations, and the formation of Golgi complexes are
correlated with the production of the sexine layer.
Male Gametopbyte. The uninucleate microspore/pollen grain is the first

cell of the rnale gametophyte (Fig. 15.1.9 G). In all its divisions inside the
microspore, the nucleus divides equally, but the cytoplasm becomes unequally
distributed.
As the intine is being laid down, the divisions of microspore nucleus
lead to the formation of two prothallial cells. Both the prothallial cells are
ephemeral, and their remnants become embedded in the intine (Fig. 15.1.9 H, I).
After the formation of prothallial cells, the antheridial initial divides, giving
rise to a small antheridial cell, which remains attached to the intine at the
site of prothallial cells, and a large vacuolate tube cell with a conspicuous
nucleus (Fig. 15.1.9 1). The electron density and organelle distribution of
the antheridial and tube cells are similar (Dexheimer 1969, 1970), but there
is a cytochemical difference in the amount of DNA, RNA and proteins (see
Moitra and Bhatnagar 1982).
From the end of April (at Simla, Mussoorie, Chak:rata-Northem Himalaya,
India, 2000-3000 m) to the beginning of June, P. wallichiana pollen grains
are shed at the four-celled stage, i.e. two prothallials, antheridial and tube cell
(Fig. 15.1.9 I). The pollen germination is arrested and further development
takes place on the nucellar apex.
Female Cone
A young cone is small, elongated to spherical, and green in P. roxburghii
and maroon in P. wallichiana. Each cone consists of 80-90 ovuliferous
scales in the axil of bract scale, two tog~ther are termed seed-scale complex.
They are arranged spirally on the cone axis (Fig . .15.1.10 A-C). Independent
vascular traces supply the ovuliferous and bract scale (Fig. 15.1.10 C); in
the former it is inversely oriented. The number of seed-scale complex
varies with the species. Those present at the base and apex of the cone are
sterile. An ovuliferous scale arises in the axil of a bract scale, which encloses
it till the time of pollination. It bears two ovules on its dorsal surface, their
micropyles face the cone axis. Later, the ovuliferous scale outgrows the
bract scale (Fig. 15.1.10 C-E).
Ovule. The ovule is unitegmic. The integument is free from the nucellus
Coniferales 167
Fig. 15.1.10 A·H. A, B, D, E Pinus wallichiana. A, B Young female cone, A before

and B at the time of pollination. C P. roxburghii, longisection young female cone.
D, E Adaxial (D) and lateral view (E) of megasporophyll bearing ovules. brs bract scale.
ovs ovuliferous scale. w wing. F-H Ovules in adaxial (F), abaxial (G) and lateral (H)
views of cone scales. The micropylar canal is two-pronged, note pollen grains on the
stigmatic prongs. (A·E After P. Maheshwari and Konar 1971, F-H after Doyle and
O'leary 1935)
except at the chalazal end, and forms a symmetrical tube well beyond the
level of the nucellus (Fig. 15.1.10 F-H). Adaxially, the edges of the integument
extend into two long arms, which curve inwards before pollination, outwards
168 The Gymnosperms
during and curve back after pollination and finally dry up. There is no
vascular supply to the ovule (Konar and Ramchandani 1958).
Megasporogenesis. The archesporia! cell becomes distinguishable while

the female cone is still covered by the scale leaves. It differentiates at the
broad apical end of the nucellus (Fig. 15.1.11A), and divides transversely
to give rise to primary parietal and primary sporogenous cell. The former
undergoes both vertical and transverse divisions, so that the sporogenous
cell is pushed deep into the nucellus, and later functions as the megaspore
mother cell (Fig. 15.1.11 B, C). Starch grains accumulate at the chalaza!
end of the megaspore mother cell (P. sylvestris). The latter undergoes meiosis I
and produces a dyad (Fig. 15.1.110, E). Only a triad (Fig. 15.1.11F) is
formed if the upper dyad cell does not undergo meiosis II, or a tetrad if
both the dyads undergo meiosis II. In a triad and linear tetrad, the upper
dyad cell and the adjoining megaspores or the upper three megaspores in
a tetrad usually degenerate. The chalazal megaspore functions so that the
development of the gametophyte is monosporic.
Three to five layers of cells round the functional megaspore become
densely cytoplasmic with prominent nuclei. This is the spongy or nutritive
tissue. In the ovules, if the megaspore mother cell, or functional megaspore,
fails to develop, the adjoining cells of the spongy tissue enlarge simulating
a cellular gametophyte. The spongy tissue comprises a definite zone of
physiologically active cells which are concerned in the nutrition of the
young gametophyte, especially at the resting stage. The cells of this zone
contain abundant starch.
Female Gametophyte (First Period of Growth). The functional megaspore

enlarges and shows a large vacuole even before the nuclear divisions
commence. It forms only a few free nuclei and apparently remains inactive
for 8-9 months (first period of rest).
Pollination. Each tree produces an enormous number of pollen grains

dispersed by wind. The surrounding area becomes clouded by the yellow
powder (also known as sulphur shower). However, only a few pollen grains
reach the pollen chamber and develop further.
A pollination drop is secreted (possibly from the nucellus), at the flared-
out tip of the integument. The fluid contains sucrose, glucose and fructose
(McWilliam 1958). The secretion starts a few days after the female cones
emerge out of the scale leaves, and the ovuliferous scales separate sufficiently
to permit the free entry of pollen. Under high humidity and cell turgor, the
secretion begins around midnight, and by early morning the micropyle
dries up. The arrival of the wind-borne pollen at the micropyle is purely
a chance phenomenon. The pollen is caught in the pollination drop, grains
stick to the two-pronged "stigmatic" micropylar canal (Fig. 15.1.1 0 F, H),
and "migrate" to the nucellar tip.
Coniferales 169
Fig. 15.1.11A-F. Pinus roxburghii. A, B Nucellus shows a primary parietal and a

sporogenous cell in A and three parietal and three sporogenous cells in B. C Deep-seated
megaspore mother cell. D Megaspore mother cell at metaphase. E A dyad. F Linear
triad. (After P. Maheshwari and Konar 1971)
The pollen grain has a germinal furrow which closes in dry weather, but
remains wide open in high humidity, or when in contact with the pollination
drop. The pollen tube emerges through this furrow .
In Pinus the ovuliferous scale is "inverted" and the pollen grains are
170 The Gymnosperms
winged. Doyle and O'Leary (1935) and Doyle (1945) have studied the
mechanism of pollination. According to Doyle, the wings orient the grain
on the hanging pollination drop in such a way that the germinal pore/
furrow of the grain faces the surface of the drop. This orientation is particularly
necessary since the ovules are inverted. The pollen then floats upward (due
to buoyancy caused by the air sacs) and reaches the nucellus with the
germinal furrow faceing the nucellus. The pollen tube enters the nucellus
without curving or twisting. McWilliam (1958) emphasizes that in P. elliottii,
P. nigra and P. wallichiana, the stickiness of the neck and arm of the
micropyle may be due to either a local secretion, or sugar residue resulting
from the retreating micropylar fluid. This is an effective method for retaining
the pollen at the site, and the pdme mover of the grain is the active absorption
of the fluid. McWilliams did not observe any preferred orientation of pollt!n
on the nucellus.
At the time of pollination. the integument is four-to-five-layered, and
soon after the pollen grains reach the nucellus, the micropyle closes due to
a rapid division and enlargement of the cells of the inner layer of the
integument. In P. wallichiana pollination takes place in May when the female
cone contains a functional megaspore (Konar and Ramchandani 1958).
Post-Pollination Development of Male Gametophyte. The pollen

deposited on the nucellar apex germinates immediately and produces a
short tube. lnitially, the growth of the pollen tube is very slow as compared
to the rapid. development of the ovule. The apical part of the nucellus
(penetrated by pollen tubes during the previous year) becomes thickened,
and the cells lose their contents. The underlying cells are meristematic and
rich in starch. Due to their growth, the tip of the nucellus with the pollen
tubes is lifted above and away from the developing female gametophyte.
Towards the end of March or later, depending on the species
(P. Maheshwari and Konar 1971), the pollen tube resumes active growth.
In P. sylvestris the nucellar cells adjoining the pollen tube degenerate; the
hemicellulose and pectin are affected first (Willemse 1968, Willemse and
Linskens 1969). During this period, the antheridial cell in the pollen grain
divides to form the stalk and body (spermatogenous) cell. However, there
is much variation in the time of division, not only in different species but
also in the same species. Willemse and Linskens (1969) observed that,
following the division of antheridial cell (called generative cell by them),
the stalk cell has fewer organelles as compared to the body (spermatogenous)
cell. Also, the cytoplasm of the stalk cell is vacuolate, but that of the body
cell is dense. The stalk and body cell migrate into the pollen tube. Soon
after, the body cell divides into two male gametes. To begin with, both the
male gametes are equal, but later, one of them enlarges considerably. These
two unequal male gametes have a distinct cytoplasmic sheath around them.
The male gametes are Feulgen-negative, stain faintly for RNA, and are rich
Coniferales 171
in proteins (P. roxburghii). In P. sylvestris the cytoplasmic sheath contains

plastids, mitochondria and ribosomes, and the ER remains coiled up like a
watch spring (Willemse and Linskens 1969).
Female Gametophyte (Second Period of Growth). When the growth is

resumed in the following year in spring, the free-nuclear divisions in the
female gametophyte continue (Fig. 15.1.12 A-C). In P. roxburghii and
P. wallichiana, when ca. 2500 nuclei have been formed, wall formation
begins by alveoli (see Chap. 2) which grow centripetally (Fig. 15.1.12 D, E).
Fig. 15.1.12 A-E . Pinus wallichiana, development of female gametophyte. A-C Free-
nuclear. D Gametophyte (longisection), note wall formation by alveoli. E Inset from D.
(After Konar and Ramchandani 1958)
The additional tissues formed in the ovules after the resumption of growth
are green. Both ovuliferous and bract scale become much enlarged and the
end part of the ovuliferous scale is pushed up.
As the ovule increases in size, the gametophyte also enlarges and occupies
the entire basal and central part of the nucellus. The densely cytoplasmic
172 The Gymnosperms
two- or three-celled thick spongy tissue surrounds the gametophyte. Due to

its continuous growth, the nucellar cells become flattened. When wall
formation is initiated in the free-nuclear gametophyte, the spongy tissue
begins to disintegrate and is absorbed by the time the archegonia are ready
for fertilization.
After the formation of the cellular gametophyte, the archegonia differentiate
at the micropylar end. Occasionally, lateral and chalaza! archegonia have
also been observed. The archegonia occur singly, 1-7 in a gametophyte,
the number varies with the species. A superficial cell of the gametophyte
functions as the archegonial initial. It is larger and has a prominent nucleus.
Its development (see Chap. 2) corresponds to the pattern in other gymnosperms
(Fig. 15.1..13 A-F):
A mature archegonium has a multicellular neck, an egg cell, and a small
ephemeral ventral canal cell. In P. nigra, the egg cell is 600 J.Lm long and
300 Jlm wide, its nucleus is ca. 150 Jlm in diameter, and migrates to the
centre of the cell.
Each archegonium has a single-layered jacket derived from the uninucleate
cells with dense cytoplasm, adjoining the archegonial initial. In P. palustris,
P. gerardiana and P. pinea the thick jacket wall has numerous pits on its
inner wall, while in P. wallichiana and P. roxburghii the pits are poorly
developed. The archegonium maintains contact with the circumjacent tissue
through these pits. As the egg cell matures, the pit membrane between the
egg and jacket cells breaksdown. Through this passage, the mitochondria,
plastids, dictyosomes, portions of endoplasmic reticulum and even entire
nuclei pass into the egg cytoplasm (Corti and Maugini 1964). The contents
of the jacket cells are incorporated in the egg cell.
In P. nigra the ultrastructure of the egg cytoplasm has been investigated
(Camefort 1959, 1960, 1962, 1965b, 1968). This study has invalidated the
classical interpretation of the origin and nature of its inclusions. There are
two types of inclusions:
a) Small inclusions, called granules by earlier workers. They are ca. 4 Jlm
in diameter, and surrounded by a small ring or crescent form of vacuole.
b) Large inclusions, variously called paranuclei, vitellus, granules and proteid
vacuoles. They are ca. 40 Jlm in diameter and structurally more complex
(Fig. 15.1.14).
The small inclusions comprise cytoplasm and are partly enclosed in a
simple membrane which is a portion of the envelope of the vacuole that
caps the cytoplasmic nodule. These inclusions usually remain connected
with the general cytoplasm by means of short peduncles. The enclosed
cytoplasm is fairly granular with a scattered mass of osmiophilic granules,
dictyosomes and mitochondria. The included cytoplasmic islet is situated
within an electron-transparent ground substance which itself is separated
from the general cytoplasm by double membrane.
The large inclusions arise by transformation of pre-existing plastids in
Coniferales 173
~
~
A~
~ B
E F
Fig. 15.1.13 A-F. Pinus walliehiana, development of archegonium. vee .Yentral canal
cell. (After Konar and Ramchandani 1958)
the young central cell. These are elongated and contain only a few lamellae.
They become deformed while invaginating, and assume the form of a ring
(in section) which encloses the islet of cytoplasm. This deformation is
followed by others that completely change the structure of plastids to give
them finally the general appearance of large inclusions. The plastid origin
of these hypertrophied formations can always be distinguished by the presence
of two layers around the various compartments. These layers separate the
inclusions from the general cytoplasm.
Camefort (1962, 1965b) investigated the development, growth and
maturation of the cytoplasm of the central celUprogamete in P. nigra. The
174 The Gymnosperms
Fig. 15.1.14. Pinus sp. Mature archegonium with circumjacent gametophytic eel
(longisection). The dense egg cytoplasm contains numerous large (hi) and small (s
inclusions, while the contents of the egg nucleus (en) are relatively thin. j jacket eel
(After Camefort 1968)
Coniferales 175
"young" transparent cytoplasm contains mitochondria, plastids and

dictyosomes, which are grouped at the periphery of the cell or near its
nucleus. Numerous small vesicles tend to come together. Under the light
microscope, the central cell shows a large number of vacuoles (foam stage;
Fig. 15.1.14). The EM shows that these vacuoles are bounded by a simple
membrane, which is often open, and the transparent content is continuous
with the general cytoplasm and can enclose cytoplasmic organelles. During
the growth of the cell, the vacuoles increase in number. The simple bounding
membrane disappears and the contents become as dense as that of the
general cytoplasm.
The cytoplasmic vesicles of the young central cell fuse and form layers
of endoplasmic reticulum which caps the small inclusions. The plastids
also become deformed before the vacuoles disappear. The general cytoplasm
becomes increasingly dense. A mature central cell has fully differentiated
protoplasm with abundant inclusions and a concentric layer of endoplasmic
reticulum (as seen in cross section). The mitochondria are dispersed throughout
the cell. They appear to have undergone structural regression (as shown by
a reduction or disappearance of the tubules). The young central cell shows
only a few mitochondria with short tubules. As the cell grows, the
mitochondria become arranged in long, sinuous and branched chondrioconts
(see Chesnoy and Thomas 1971). They appear constricted at this time,
possibly fragment, which accounts for their increased number in the mature
egg.
The nucleus of the egg cell (P. sylvestris, P. uncinata, P. nigra) shows
chromosomes which consist of bundles of fibrils 100 A in diameter (each
30 A thick) made up of a light central line between two dark ones. These
fibrils are parallel to one another and lie along the length of the chromosome.
This particular arrangement appears to suit the generally inert female
gametophyte before fertilization (Camefort 1964).
The nucleoli are formed of 50 A thick fibrils. They may be distributed,
tightly packed, in the dark zones of the nucleolus, or dispersed in the light
zone. At the periphery of the nucleolus, the nucleolar substance is scattered
either -in the form of fibrils separating from each other, or as small muriform
masses (0.1-0.03 J..Lm in diameter) consisting of several granules of 170 to
200 A. The latter indicates the beginning of a change from a fibrillar to a
granular structure (Camefort 1964).
Fertilization
The pollen tube, just before fertilization, grows rapidly and reaches the
neck of the archegonium (Fig. 15.1.15A, B). The tip has a dense cytoplasm,
a large number of starch grains, tube nucleus, stalk cell and the two male
gametes enveloped in a common cytoplasmic sheath. The pollen tube
penetrates into the neck of the archegonium, and discharges its contents at
the tip of the cytoplasm of the female gamete.
176 The Gymnosperms
Fig. 15.1.15 A-F. Pinus wallichiana. A Longisection ovule shows pollen tube in the
neck of the archegonium. B Pollen tube from A. mgi> mg 2 male gametes. sc stalk cell.
tn tube nucleus. C-F Stages in the fusion of one of the male gametes with egg. E, F
Portion of archegonium from C, D. (After Konar and Ramchandani 1958)
One of the two male gametes moves towards the female gamete. The
second male gamete, along with the stalk cell and tube nucleus, and. the
original male cytoplasmic sheath, persist at the apex of the egg and eventually
degenerate. The functional male gamete is devoid of the cytoplasmic sheath
as it moves down the archegonium to fuse with the female gamete (Willemse
and Linskens 1969). The mitochondria and plastids brought by the pollen
Coniferales 177
tube are morphologically different from those of the female gamete. They
remain grouped in the upper part of the egg cytoplasm during the division
of the zygote (Camefort 1965b, 1967).
The functional male gamete lodges in a depression on the female gamete
(Fig. 15.1.15 C-F). As soon as contact is established, junctions between the
nuclear membranes of the two gametic nuclei are established at several
points. These areas of communication between the nucleoplasm gradually
A B
Fig. 15.1.16 A-E. Pinus lambertiana. A Male and female chromatin in separate groups
in egg, each surrounded by spindle fibres to form a multipolar spindle. B, C Two
chromosome groups on multipolar and diarch (B) and bipolar (C) spindles. D Zygote
nucleus in mitosis, note separate male and female chromatin. E Polar view of transection
through late metaphase shows diploid (24) chromosomes. (After Haupt 1941)
enlarge to form islets of cytoplasm surrounded by nuclear membrane. The

latter slowly breaks down at the region of contact between the two nuclei
and the two groups of chromosomes [which are initially quite distinct
(Fig. 15.1.16 A)] converge and lie on a common spindle (Fig. 15.1.16 B).
Finally, the chromosomes of the two gametes lose their identity
(Fig. 15.1.16 C).
178 The Gymnosperms
Embryogeny
Proembryo. At the time uf mitosis of the zygote, the nuclear membrane

separates the nucleoplasm from the surrounding cytoplasm (Fig. 15.1.17 A, B).
Here and there the nuclear membrane is deeply invaginated into the
nucleoplasm and collapses by the end of first mitosis. The nuclei of the
coenocytic proembryo are formed within the nucleoplasm of the zygote
(Camefort 1967) which progressively organizes into the cytoplasmic area
of the embryo. It is essentially of nuclear origin and distinct from the
cytoplasm of the egg (Fig. 15.1.17A-C) and is termed the neocytoplasm.
During the first embryonal mitotic division (P. banksiana), the two groups
of chromosomes come to lie in one group at the equator of the spindle. In
Fig. 15.1.16 D (P. lambertiana) the first embryonal mitosis shows separate
male and female chromosomes. Each chromosome splits longitudinally, the
daughter halves move to the poles. A transection through mitotic late
metaphase shows (Fig. 15.1.16 E) polar view of diploid complement of 24
chromosomes (Haupt 1941). The two proembryonal nuclei divide once
more and the four daughter nuclei migrate to the base of the proembryo and
become arranged in one layer (Fig. 15.1.18 A, B). Initially, they are very
small, as after every division there is a decrease in the size of the proembryonic
nuclei; as they migrate to the base they enlarge considerably. All these
nuclei divide with vertical spindles, resulting in eight nuclei which organize
into eight cells arranged in two tiers of four cells each (Fig. 15.1.18 C-F).
They are designated the primary embryonal tier (p£) and primary (pu) upper
tier (Fig. 15.1.18 E, F). The cells of both the tiers divide transversely by
internal division (nuclear division inside proembryo cells), so that four tiers
of four cells each are formed. The lower two tiers comprise the embryonal
tier (E) folllowed by disfunctional suspensor (ds) previously termed the rosette
tier. The uppermost tier, U, is the upper derivative of pu (Fig. 15.1.18 G, H).
After the proembryonal nuclei migrate to the base of the archegonium,
the cytoplasm of the female gamete degenerates. The degeneration begins
close to the neocytoplasm, extends gradually throughout the egg cell, and
is completed by the time the proembryo has become cellular. A cytochemical
study of the cytoplasm shows that there is a non-specific acid phosphatase
activity in the endoplasmic reticulum formations enclosing the inclusions.
This activity becomes intense in all the inclusions after the migration of the
proembryonal nuclei to the basal pole of the archegonium (Camefort 1966,
1967).
Differentiation of Embryo. The upper four cells of the £-tier elongate and
function as the embryonal suspensor (Es), and the lower four cells form the
embryonal mass (Fig. 15.1.18 I). Subsequently, there are several layers of
Es, which are termed Es~> Es 2 , Es 3 ••• ,formed by divisions in the £-tier. The
Es cells elongate rapidly and thrust the terminal cells (embryonal cells)
forward into the corrosion cavity (see Chap. 2). The latter enlarges and
Coniferales 179
Fig. 15.1.17 A·C. Neocytoplasm differentiation in Pinus. A Zygote nucleus in telophase

surrounded by archegonial cytoplasm . The nucleoplasm appears to form the ground
substance of neocytoplasm (neo). B, C Progressive stages in the differentiation of
neocytoplasm at two-, four- and eight- nucleate proen:bryo. The archegonial cytoplasm
does not seem to be involved in the process. (After Camefort 1969)
180 The Gymnosperms
A
./
\ ltiJ c 0
~)" E
"
pE G H
Fig. 15.1.18 A-0. Embryogeny . A-D, I Pinus sp., diagrammatic representation of the
development of proembryo. A, B Four-nucleate proembryo, the nuclei are in the middle
(A) and at the base (B) of the archegonium. C, D Wall formation at eight-nucleate stage.
E -H P. wallichiana, lower portion of proembryo. E Eight-celled proembryo shows primary
upper (pU) and primary embryonal (pE) tier. F, G Internal division in pU (F) and pE
(G). ds disfunctional suspensor, U upper tier. H 16-celled proembryo. E Embryonal tier.
I Formation of embryonal suspensor es 1• J-0 P. roxburghii, development of embryo.
(A, B, I After Buchholz 1929, C, Dafter Dogra 1967, E-H after Konar and Ramchandani
1958, J-0 after P. Maheshwari and Konar 1971)
facilitates elongation of the Es. Later, as the elongation of Es exceeds the

rate of enlargment of the corrosion cavity, the Es becomes coiled and twisted
(Fig. 15.1.18 M). The elongation of the Es system pushes the growing
embryonal mass into the central portion of the female gametophyte. As
development proceeds further, the earlier suspensor system collapses, while
newer Es are formed.
Coniferales 181
Polyembryony is common in Pinus. Additional embryos originate by simple

polyembryony, i.e. from multiple zygotes, as well as due to cleavage
polyembryony, i.e. a single zygote gives rise to multiple embryos by cleavage
or splitting of the embryonal tier E into several embryonal units
(Fig. 15.1.18 J-M). The separation into embryonal units occurs at Es 2
formation (see H. Singh 1978). These units remain unchanged during the
earlier stages of Es elongation, but later they divide and form a multicellular
mass. There is a period of embryonal selection due to competition between
the four embryos from each zygote, and between the embryos from different
zygotes. This phase of competition lasts for about 6 weeks from the time
of fertilization (P. Maheshwari and Konar 1971). Generally, the deep-seated
terminal embryo succeeds and develops further. The remaining embryos
become arrested at different stages of development.
The embryonal mass develops from a single cell (Fig. 15.1.18 N, 0). An
apical cell with three cutting faces is formed during the earlier development,
but ii becomes indistinguishable soon afterwards.
The fully formed embryonal mass is a smooth paraboloid structure. It
has a hemispherical apex at its distal end (Fig. 15.1.19 A, B), and a suspensor
which is continuous with it at the proximal end. Further development at the
proximal end gives rise to a well-developed root cap. It differentiates
independently into a column and a peripheral region; the former does not
contribute to the development of the latter (P. Maheshwari and Konar 1971).
As the root cap and root initials organize at the basal end, an incipient
pith is recognizable in the hypocotyl between the epicotyl and the root
initials. It is evident by the predominant transverse divisions, cell enlargement
and vacuoliation. In P. lambertiana (Berlyn 1967) a pro-cambium
differentiates at a lower level, following the formation of the pith. The
cortex becomes demarcated soon after. The formation of pro-cambium is
associated with the early phases of cotyledon formation though it is well
developed before distinct cotyledonary primordia originate. Simultaneously,
several long, uninucleate and multinucleolar secretory cells become
distinguishable in the cortical region. Finally, 3-18 cotyledons and the
shoot apex differentiate (Fig. 15.1.19 C, D). The cotyledons are traversed
by the procambial strands, and show development of mesophyll. Generally,
only one embryo matures, occasionally even two may reach maturity. The
mature embryo has distinct epicotyl, root axis with remnants of the suspensors,
and a hypocotyl-shoot axis bearing several cotyledons (Fig. 15.1.19 D).
The cells of the ds tier occasionally elongate and resemble suspensor
cells. In some species it may undergo divisions to form a few to a mass of
cells; they do not complete development, and very often abort.
Seed. The seed ripens within a few weeks without much change in the
size of the embryo.
In the young ovule the integument is three-layered. It becomes six- or
182 The Gymnosperms
c 0
Fig. 15.1.19 A-D. Pinus roxburghii, progressive development of embryo. (After
P. Maheshwari and Konar 1971)
Coniferales 183
seven-layered at the time of pollination (Fig. 15.1.20 A). There is an epidermis

and hypodermis, the cells of the inner layers are richly cytoplasmic, have
prominent nuclei and some of them are tanniniferous. The cells divide
anticlinally and periclinally to form a 10-12-layered integument which
differentiates into three distinct zones: outer fleshy, middle stony, and inner
fleshy (Fig. 15.1.20 B, C).
Fig. 15.1.20 A-E. Pinus wallichiana, seed coat. A-C Longisection of integument at
the time of pollination (A), free-nuclear female gametophyte (B) and archegoinal stage
(C) . D, E Mature seed coat shows outer fleshy (of>, middle stony (stl), and inner fleshy
(if) layers. (After Konar and Ramchandani 1958)
The outer fleshy zone finally becomes seven-or eight-layered, the outer
two or three layers contain a shiny granular material. At the time of seed-
shedding, these cells become round with vacuolated cytoplasm and the
nucleus shrinks (Fig. 15.1.20 E).
The middle stony layer differentiates when the ovule is at the archegonial
stage. At maturity it comprises ca. 18-20 layers of pitted cells, the outermost
layer has refractive granules. Lignification of the cells begins in the innermost
layer and proceeds outward (Fig. 15.1.20 D, E).
The inner fleshy layer is seven or eight cells wide. The cells are thin-
walled and elongated with scanty cytoplasm. At maturity only two or three
184 The Gymnosperms
layers persist and the rest are absorbed (Fig. 15.1.20 D, E).
In some species of Pinus (P. roxburghii, P. insularis, P. balfouriana,
P. strobus), the seed is winged and the development of the wing is closely
linked with the development of the seed coat. The latter extends into a
wing which cannot be detached without injury. The wing is thin and papery
or thick at the base.
Embryo development and the type of food reserve in endosperm has
been studied by Hakansson (1956). Starch appears first around the apex of
embryo when suspensor cells begin to elongate. There is no starch in the
upper region around the empty achegonia while other parts of the gametophyte
have scanty starch. Later, three zones can be demarcated in the endosperm
(P. sylvestris): (a) the cells bordering the central cavity have no starch or
only simple grains, (b) the middle zone has abundant starch, (c) the cells of
the peripheral zone have dense contents (but no solid storage material).
Later, the entire endosperm becomes packed uniformly with starch. The
accumulated food reserves in the gametophyte are utilized at seed germination.
An ultrastructural study of the dry seed of P. sylvestris shows that the
endosperm and embryo cells have the same components. Their different
physiological and morphological role in the development of young seedlings
needs further study (Simola 1974).
There is a cytochemical difference in the distribution of proteins and
amyloplasts in shoot apex of the embryo in a dry seed of P. banksiana (Mia
and Durzan 1974). The size and frequency of protein b()dies and amyloplasts
increase from the apical zone towards the flanks and subapical region.
Free sugars such as sucrose, stachyose, raffinose and 19 amino acids and
2 amides have been reported from the embryo and endosperm of P. banksiana
(Durzan and Chalupa 1968). In the seeds of P. thunbergii Katsuta (1961)
reported globulin, albumin and glutalin.
Hatano (1957) observed that pyruvic acid and alpha-keto acids are
consumed during the first step of conversion of organic acid to amino acid.
The fat content in the seed of P. roxburghii is 31% of the total weight
(Konar 1958). In P. sylvestris linoleic, oleic, palmitic and stearic acids and
an unidentified component C have been reported (Nyman 1966).
Germination. The food stored in the seed breaks down during germination
and is the only source of energy for the germinating embryo. Several changes
take place after the seed has imbibed water. Mitochondria, dictyosomes
and ER-at first absent (or present in small quantities)-appear and gradually
increase (Durzan et al. 1971, Simola 1974). Small microbodies, interpreted
as glyoxysomes, have been observed in P. sylvestris (Simola 1974),
P. ponderosa (Ching 1970) and P. pinea (Lopez-Perez et al. 1974). These
are the sites of fatty acid oxidation in oil-containing seeds.
Stachyose, rafinose and sucrose are the main sugar reserves in the seeds
of P. thunbergii andP. sylvestris (see Konar and Moitra 1980). In the former,
Coniferales . 185
as germination proceeds, raffinose and stachyose are digested and sucrose,

including glucose and fructose, appear. In P. sylvestris, small quantities _of
glucose and fructose, already present in dry seeds, rapidly increase during
germination. In P. strobus the increase in the above sugars in the seedlings
is due to photosynthesis. By using radioactive methods, it has been observed
that of the total sugars formed sucrose is maximal followed by glucose and
fructose together, while raffinose is minimal.
Almost all the seeds contain protein reserves as a source of N2 for the
young seedlings before they are able to absorb nitrogen by their roots. The
breakdown of protein reserves in the endosperm and embryo and appearance
of new proteins in other parts of the seedling takes place during germination.
Durzan et al. (1971) and Hatano and Asakawa (1964) observed that reserve
protein is high in basic amino acids. In the shoot apex of P. radiata (Riding
and Gifford 1973) there is an initial breakdown and subsequent increase in
protein content following seedling establishement. Fats and proteins are the
major reserves in the mature seeds of P. thunbergii (Goo and Negisi 1952).
During germination, most of the protein reserve moves from the endosperm
to the embryo, which shows a continuous increase in total nitrogen. With
the germination of embryo in the seed, a sustained synthesis of proteins is
indispensible and is accompanied by increased synthesis of RNA, which is
triggered off within a few hours of imbibition of water.
For further details and information on seed germination, readers are
referred to the excellent review by Konar and Moitra (1980).
The seed germinates 20-25 days after sowing. The radicle emerges and
penetrates into the soil (Fig. 15.1.21A-D). The hypocotyl elongates,
straightens, and carries above the ground the remnant of the seed alongwith
the cotyledons. The cotyledons absorb nutrition from the gametophyte till
it is exhausted, and then the seed coat drops off (Fig. 15.1.21E, F). The
epicotyl, deep within the whorls of cotyledons, gives rise to the stem and
leaves (Fig. 15.1.21 G, H). The cotyledons persist for a long time and are
shed only after the juvenile leaves and the long shoot have grown considerably.
Chromosome Number
The karyotype analysis shows that Pinus has uniformly n = 12 and 2n = 24
chromosomes (Mehra 1988).
The chromosomes are numbered I-XII according to their decreasing size.
Chromosome pair XII is heterobrachial in P. gerardiana, P. grifithii,
P. roxburghii and P. kesya. In P. kesya, chromosome XI is also heterobrachial.
In P. gerardiana there is a secondary constriction in the proximal arm
of the heterobrachial pair, as well as in isobrachial chromosome pair X,
where it is located a little away from the centromere in one of the arms.
In P. grifithii and P. roxburghii (Fig. 15.1.22 B, C) in the haploid
complement each of the six isobrachial chromosomes have a secondary
constriction. In P. roxburghii the six homologous pairs with secondary
186 The Gymnosperms
A
B
c
D
H
E F
Fig. 15.1.21 A-H. Pinus strobus. A-F Germination of seed. G, H Seedling with
cotyledonary and juvenile leaves. (After P. Maheshwari and Konar 1971)
constrictions are reported in the diploid complement (Fig. 15.1.22 A). In

P. kesiya, each of the two isobrachial pairs have secondary constrictions.
Meiosis has been studied in detail in P. roxburghii and P. patula (Mehra
1988). In both the species, meiosis is normal. However, not infrequently
the chromosome segments are left behind at the equatorial region as laggards
(P. roxburgliii), or are extruded (P. patula). Such irregularities can affect
the chromosomal constitution as well as the morphology of pollen grains.
Polyploidy is not known in the wild population of any species.
In P. roxburghii (growing at an altitude between 500 and 2500 m in the
NW Himalayas, India) the male cones are initiated in September. Pollination
takes place (pollen grains at the four-celled stage) in March. Pollen germinates
Coniferales 187
·:· ....
~--~··--··
B
Fig. 15.1.22 A-C. Pinus roxburghii. A Root tip (squasn) shows 2n = 24 chromosomes.
B, C Endosperm (squash) shows n = 12. h heterobrachial chromosome; arrows indicate
position of secondary constriction. (After Mehra 1988)
188 The Gymnosperms
on the nucellus immediately after shedding. There is a period of rest for

approximately 10 months (from May to February-2nd year). Growth is
resumed in March (2nd year) and fertilization occurs in April (2nd year).
The female cones are initiated in February. Soon after pollination in March,
the cone undergoes a period of rest from April to January for ca. 10 months.
The female gametophyte is at the free-nuclear stage at this time. Growth is
resum~d from February (2nd year), and fertilization occurs in April. The
embryo developes and matures by December (2nd year). The cones open
and shed their seeds in April-May (3rd year). Usually, the cones dehisce in
the third year; the undehisced cones may also contain mature seeds.
There are three species of Pinus-P. pinea, P. leiophylla and
P. torreyana-which have a 3-year-type reproductive cycle (Dallimore and
Jackson 1966, Francini 1958). These taxa undergo three winter rests, and
the interval between pollination and fertilization is 2 years. The cones are
initiated during autumn and the ovules overwinter (1st rest period).
Pollination occurs during spring when the ovules show sporogenous cells.
Megasporogenesis occurs during autumn and the ovule with a slightly enlarged
megaspore overwinters (2nd rest period). The megaspore matures through
the coming spring and summer. It undergoes free-nuclear divisions during
autumn. A massive spongy tissue develops around the megaspore, and the
young female gametophyte. The ovule overwinters again (3rd rest period)
at the free-nuclear gametophytic stage. In the coming. spring, the gametophyte
develops more rapidly and fertilization takes piace by June. The interval
between pollination and fertilization is 2 years. The embryogeny is completed
and the mature seeds are shed in autumn or early winter.
The reproductive cycle in tropical and temperate pines is conditioned by
the environmental factors and, therefore, the 2- and 3-year cycle. The cyto-
and histochemical changes during the rest period deserve further attention.
15.2 TAXODIACEAE
The characteristic features of the family Taxodiaceae are: monoecious trees;

leaves narrow and linear (occasionally apparently two-ranked); vegetative
buds without scales; young bract and pvuliferous scale nearly free from
each other but fuse in the mature cone; each ovuliferous scale has 2-9
ovules; microsporophyll bears 2-9 pollen sacs; pollen grains without wings.
There are 10 genera and 15 spp.: Athrotaxis (3 spp), Taxodium (3 spp),
Cunninghamia (2 spp.) Cryptomeria, Glyptostrobus,Metasequoia, Sdadopitys,
Sequoia, Sequoiadendron and Taiwania are all monotypic.
There are several genera of seed cones, and vegetative remains are reported
from the Jurassic. The Taxodiaceae were then a distinct line of evolution.
All modern taxa of the Taxodiaceae, except Sequoiadendron, are known from
the Mesozoic. The family shows increased diversity in the Cretaceous, a
number of new organ genera as well as many more of the modern ge.nera
Coniferales 189
are represented. The living species probably existed as persistent relics of

lineages that evolved nearly 100m y ago, and may be regarded as modem
representatives of lines of specialization (see Miller 1977).
Cryptomeria japonica
This monotypic genus includes ca. 25 varieties (Dallimore and Jackson
1966). It is a native of China and Japan. Japan owes much of the beauty
of its groves to this taxon. The seeds were first brought to India in 1844
(see Troupe 1921), and cultivation was started in Darjeeling, which now
has extensive forests of C. japonica. In the moist climate of the Eastern
Himalayas, it grows very rapidly and yields excellent planted f~rests. It has
also naturalized in the Western Himalayas and grows up to an altitude of
1800-2400 m.
Morphology
The plant is a tall, conical, much-branched evergreen tree, reaching a height
of 50 m or more and a girth of 7-8 m (Mehra 1988). The bark is fibrous,
reddish-brown, and peels off in long strips. The leaves are linear, awl-
shaped and spirally arranged (Fig. 15.2.1 A, B). They are keeled on both
the surfaces and have a somewhat spinous tip. The leaf bases are fused
with the stem and form its outer covering. The branches are in whorls. The
branchlets with the awl-shaped leaves are deciduous.
The plant is monoecious. Male and female cones are usually borne on
different branchlets (Fig. 15.2.1 A), occasionally on the same branchlet
(Fig. 15.2.1. B). The male cones arise in clusters near the tip of the young
shoots, while the female cones are terminal on the branchlets (Fig. 15.2.1 A).
Anatomy
Stem. The wood is reddish brown and fragrant. A cross section shows
clearly defined growth rings, variable in width and slightly wavy. Early and
late wood are well differentiated, the tracheids are square, occasionally
with smaller tracheids occurring between the larger ones. There are ca.
3600 tracheids/mm2 • The thickness of the wall is 1.5-3 }lm in early wood
and 4-6 Jlm in the late wood. The rays are relatively few, wide apart and
traverse several growth rings. Their walls are horizontal and sparingly pitted.
There is abundant wood parenchyma, chiefly in the late wood and at various
distances from the growth ring limits. The parenchyma cells are thin-walled
and can be readily distinguished from the thick-walled late tracheids. The
resin ducts are absent (Greguss 1955).
Numerous pits occur on the tangential walls of tracheids (especially
abundant in the late wood). The pits are 9-11 Jlm in diameter, commonly
in a row, circular with poorly defined borders. Apertures in the early wood
are circular or elliptic, in the late wood eye-shaped or slit-like. The tracheids
190 The Gymnosperms
Fig. 15.2.1 A·K. Cryptomeria japonica, external morphology. A, B Male (me) and
female cones borne on different branchlets (A) or on the same branchlet (B). C Male
cone with bract. D, E F Microsporophyll, abaxial, lateral and adaxial views.
G Microsporophyll with dehisced sporangia. H-K Female cones. H Young cones. I
Abnormal cone bearing a cluster of male cones at the tip. J Open cone scales at the time
of pollination. K Bract and ovuliferous scales show the orientation of ovules (ov), and
processes (ps) on the ovules. (After H. Singh and Chatterjee 1963)
are pointed. The rays are uniseriate with variable height (commonly 3-6
cells); tangential ray walls are thin and occasionally delicately verrucose.
The wood parenchyma cells are fairly abundant with nodular cross-walls.
The cells communicate with each other through simple pits, and with adjoining
tracheids through half-bordered pits. The latter are smaller than the tangential
pits, and their apertures differ in size and shape in the early and late wood.
There are about 40-45 rays and 150-160 ray cells per mm2.
A radiallongisection shows uniseriate (occasionally two- to three-seriate)
bordered pits. The pits are circular, and the apertures circular, elliptic,
linear or slit-like. In slits, the pits are obliquely inclined or nearly vertical.
The horizontal walls of the ray cells are relatively thick (1.5-3 Jlm) and smooth
except for sporadic local thinnings; the tangential walls are smooth and
thin (1-1.5 J.lm), while the radial walls show large, round simple pits with
half-bordered complementary pits in the tracheids. The apertures vary in
size and shape. Taxodioid cross-field pits are also present.
Coniferales 191
Leaf. The leaf is awl-shaped and amphistomatic. The stomata are

haplocheilic. A vertical section (Fig. 15.2.2 A-C) shows: stratified cuticle
of variable thickness; thick-walled epidermal cells with a much-reduced
lumen containing tannin or other inclusions; well-developed hypodermis of
thick-walled sclerenchymatous cells on one or both leaf surfaces.
The mesophyll is undifferentiated. It shows many elongated cells (especially
on the adaxial side) which appear like chains/filaments of long narrow cells
with large spaces between the rows. The outermost layer on the abaxial side
is somewhat palisade-like, with radially elongate cells (Fig. 15.2.2 B, C).
There is a single large resin duct with a two-layered wall beneath the
vascular bundle (Fig. 15.2.2 A). A bundle sheath is not very distinct. In
vertical section, the transfusion tissue appears as an inverted U-shaped arc
clasping the vascular bundle with the tracheids and albuminous cells situated
A 8
Fig. 15.2.2 A-D. Cryptomeriajaponica, leaf. A-D Transection. A Outline diagram for
Band C. B, C Internal structure. D Vascular bundle and transfusion tissue. alb albuminous
cells. tr transfusion tracheid. (After Kausik and Bhattacharya 1977)
laterally to the xylem and phloem, respectively (Fig. 15.2.2 D). The transfusion
parenchyma is distributed at random in the arc, especially on the adaxial
side (Kausik and Bhattacharya 1977).
Reproduction
Male Cone
A male cone cluster comprises 10-18 cones, each borne in the axil of a
192 The Gymnosperms
bract (Fig. 15.2.1 C). Each cone bears 18-25 spirally arranged
microsporophylls, the two lowermost microsporophylls are usually sterile.
The stalk of each sporophyll is almost at right angles to the axis of the
cone. It is a peltate structure which extends upward as a membranous flap
and bears three or four microsporangia on its lower (abaxial) surface
(Fig. 15.2.1 D-G).
Microsporangium. The hypodermal archesporia! cells differentiate on

the abaxial surface of the microsporophyll. They have dense cytoplasm and
large nuclei, and are arranged in two layers (Fig. 15.2.3 A, B). The epidermal
cells undergo anticlinal divisions; the archesporia! cells divide in all planes
to form a mass of cells.
The outermost layer of the archesporium divides periclinally to form an
outer primary parietal layer, which divides again periclinally and gives rise
to a three-layered wall. The innermost layer differentiates into a tapetum
(Fig. 15.2.3 B, C, E). The tapetal cells become binucleate (Fig. 15.2.3 F)
during reduction divisions of the microspore mother cells, and eventually
degenerate with the maturation of microspores.
The two-layered hypodermal cells undergo rapid anticlinal divisions
followed by tangential elongation to keep pace with the increasing
sporogenous tissue. Finally, the hypodermal layers degenerate. The epidermis
develops annular thickenings during the maturation of microspores
(Fig. 15.2.3 E, G, H). In vertical section, the sporangia appear sessile and
oval (Fig. 15.2.3 D). They dehisce by vertical slits which appear at the
base, extend upward, and face the cone axis.
Microsporogenesis. Only some of the sporogenous cells function as

microspore mother cells while the others degenerate. Cytokinesis follows
meiotic divisions and tetrahedral, isobilateral and decussate tetrads are formed
(Fig. 15.2.3 1-M). The callose wall around the microspores is consumed
and the original wall of the mother cells breaks down, releasing them into
the cavity of the microsporangium. The exine differentiates outwards to the
intine, except where the latter projects in the form of a papilla (Fig. 15.2.3 I,
N, 0). The latter is occasionally balloon-like; sometimes an additional sac-
like structure may occur beside the normal papilla (Fig. 15.2.3 P). The
exine and the intine are of the same thickness; later the exine becomes
thicker. Starch grains accumulate in the pollen grains, which show a high
percentage of sterility. The pollen is shed at the uninucleate stage, which
appears rare among the conifers; this condition is known only in Cupressus
(Mehra and Malhotra 1947, Konar and Banerjee 1963) and Taxus (Dupler
1917, Sterling 1948a).
Male Gametophyte. After pollination, the pollen grains imbibe moisture

and swell. The exine may persist in fragments or is entirely cast off. The
Coniferales 193
J
mi
rd
•
.
..
.
D ..
Fig. 15.2.3 A-P. Cryptomeria japonica, microsporangium and microsporogenesis.

A, D Longisection of male cone. mi microsporophyll. rd resin duct. B, C, E, G, H
Longisections of microsporophylls. B Archesporia! cells (arc). C Sporogenous cells and
wall layers. tanc tannin cell. E Later stage. G Epidermis, degenerated tapetum and
microspore tetrads. H Exodermis with annular thickenings and microspores. F, 1-M
Whole-mounts. F Binucleate tapetal cell. I, J Microspore mother cells, meiosis I and II.
K-M Tetrahedral, decussate and isobilateral microspore tetrads. N, 0 Uninucleate pollen
grain . P Abnormal pollen grain, note starch grains in N-P. (After H. Singh and
Chatterjee 1963)
194 The Gymnosperms
nucleus moves to one side, and functions directly as the antheridial initial
(prothallial cells are not formed in Taxodiaceae). The nucleus divides and
forms a lenticular antheridial cell and a large tube cell (Fig. 15.2.4 A). At
this stage, the starch grains are no longer discernible, the cytoplasm becomes
less dense, and the pollen grains germinate. As the pollen tub~ enters and
grows further, the nucellar cells disorganize (Fig. 15.2.4 B, C). Two to four
pollen tubes have been observed in a nucellus; occasionally, the pollen tube
branches (Fig. 15.2.4 D-F). The tube nucleus moves into pollen tube. The
antheridial cell gives rise to the stalk and body cells. The nucleus of the
body cell divides when the pollen tube has reached the archegonial complex.
The two male gametes are equal and disc-like with dense cytoplasm and
prominent nuclei (Fig. 15.2.4 G, H). The contents of the pollen tube are
discharged in the archegonial chamber (H. Singh and Chatterjee 1963).
Female cone
The apex of the branchlets is generally used up during the differentiation
of the terminal female cones (Fig. 15.2.1 H). Rarely, the axis of the cone
may grow into a vegetative shoot which bears a cluster of male cones
(Fig. 15.2.1 I).
The young female cone (ca. 4 mm in length and diame.ter) has a curved
stalk and emerges from the rosette of leaves at the time of pollination. At
this time, it has a flattened apex (Fig. 15.2.1 J), but ·subsequently becomes
almost spherical. A mature cone (19 mm in length and 21 mm in diameter)
is yellowish~brown, .and comprises 26-30 spirally arranged scales; the upper
nine or ten small scales are sterile. A mature fertile scale (ca. 9mm in
length and 11 mm in width) is concavo-convex, and bears on the upper side
three to five spinous processes at the distal end. The bract and the ovuliferous
scales are fused except at the tip, where the bract app~s as a recurved
process. Each scale bears three or four adaxial ovules with the micropyle
pointing away from the cone axis (Fig. 15.2.1 K).
Ovule. After the demarcation of the integument, one or two hypodermal

archesporia! cells can be distinguished by their longer size and prominent
nuclei (Fig. 15.2.5 A). These cells divide periclinally and form the outer
primary parietal and the inner primary sporogenous cells. The epidermal as
well as the parietal cells divide further and give rise to a massive nucellus,.
which thus has a dual origin (Fig. 15.2.5 B). The primary sporogenous cells
divide and form a large group of sporogenous cells, some of which
degenerate. At this stage the nucellar cells around the sprogenous tissue
contain prominent nuclei (Fig. 15.2.5 C), and later contribute to the spongy
tissue.
At its base, the integument is free from the nucellus (Fig. 15.2.5 E, G).
The micropyle becomes wide open before pollination, and points upward.
The lower part of the nucellus consists of smaller cells, while the upper has
Coniferales 195
B
A
H
Fig. 15.2.4. A-H. Cryptomeria japonica, male gametophyte. A Two-celled pollen grain
lying on the nucellus. a/ antheridial cell. tn tube nucleus. B, C, F Development of pollen
tube in the nucellus. be body cell. D Branched pollen tube. E Longisection of ovule
with pollen tubes in the nucellus. pi plugging tissue. G Outline diagram for H.
ace archegonial complex . H Pollen tube with male gametes (mg) in the archegonial
chamber (ac). (After H. Singh and Chatterjee 1963)
larger cells. At the free-nuclear stage of the gametophyte, a constriction

appears in the upper part of the nucellus;. the undei'lying cells contain
tannin. One of the sporogenous cells enlarges and functions as the megaspore
mother cell (Fig. 15.2.5 D). Pollination takes place at this stage. Soon after,
196 The Gymnosperms
Fig. 15.2.5 A·K. Cryptomeria japonica, megasporangium and female gametophyte.

A Longisection of ovule, initiation of integument and archesporium. B-D Megaspore mother
cell. E, G Longisection of cone scale before (E) and after (G) pollination. bs bract scale.
ovs ovuliferous scale. F, H Spongy tissue (sg) and functional megaspore. 1-K Free-nuclear
gametophyte. K inset x in J shows projection in the megaspore membrane. (After
H. Singh and Chatterjee 1963)
Coniferales 197
the epidermal and subepidermal cells lining the micropylar canal become
richly cytoplasmic and elongate inward. They divide irregularly, the walls
thicken at maturity, and the micropyle becomes closed (Fig. 15.2.5 G). One
or two ovules (on a scale) may degenerate just before or after pollination.
Megasporogenesis. Meiosis I results in two dyads, of which only the

lower undergoes meiosis II and a triad is formed. The chalaza! megaspore
functions, while the other megaspore and dyad degenerate (Fig. 15.2.5 F).
With the enlargement of the megaspore, the circumjacent sporogenous and
nucellar cells form the spongy tissue or tapetum. It is one to two cells thick
laterally but six to eight cells thick above and below the megaspore
(Fig. 15.2.5 C, D, F, H). The spongy tissue is very prominent and the
densely cytoplasmic cells remain uninucleate. The nucellar cells disorganize
in the immediate vicinity of the tapetum. The latter also collapses during
the enlargement of the free-nuclear gametophyte.
Female Gametophyte. The functional megaspore enlarges, the cytoplasm

becomes less dense, the nucleus also enlarges, migrates to one side of the
cell, and undergoes division (Fig. 15.2.5 H). The gametophyte enlarges and
a vacuole appears in the centre. The freee-nuclear divisions are synchronous
and the nuclei lie in a peripheral layer of dense cytoplasm. The early
gametophyte is ovate (Fig. 15.2.5 I). Later, it elongates, and at maturity
shows a broad slightly convex micropylar and a tapering chalazal end. The
megaspore membrane often has an uneven outline (Fig. 15.2.5 J, K).
A mature gametophyte shows rows of cells radiating from the centre to
the periphery, which indicates centripetal wall formation. These uni- or
binucleate cells are thin-walled and have scanty cytoplasm. With the
differentiation of the micropylar archegonial complex, the adjoining
gametophytic tissue grows upward and forms a prominent archegonial
chamber (Fig. 15.2.4 G, H). The development of the embryo is accompanied
by the deposition of starch grains in the gametophytic tissue.
A mature archegonial complex comprises 15-37 archegonia in a common
jacket (Fig. 15.2.6 E). or the archegonia may be individually separated by
the jacket cells and the gametophytic tissue. Usually, the archegonia develop
side by side; occasionally they are superimposed. Lateral archegonia are
frequent; they rarely differentiate at the chalaza! end (Fig. 15.2.6 F, G).
Sometimes, a gametophyte may show a chalaza} archegonial complex in
addition to the micropylar complex (Fig. 15.2.6 H, 1). In the chalaza! region,
probably due to fusion, the archegonia assume an irregular shape. These
archegonia do not become fertilized, their nuclei may divide but do not
give rise to embryos (H. Singh and Chatterjee 1963).
The development of the archegonium follows the usual conifer plan
(Fig. 15.2.6 A-C). A mature archegonium shows an ephemeral ventral canal
nucleus and a large egg nucleus. Due to rapid elongation of the ·egg cell,
198 The Gymnosperms
cl f•
. . ::i
.~\
-:> . ~·
mac
Ia
cha
Fig. 15.2.6 A·J. Cryptomeria japonica, archegonial complex and fertilization.

A-CDevelopment of archegonium. cl central cell. ec egg cell. ni neck initial. nk neck.
D Archegonial complex in tangential section; central archegonium shows a kinoplasmic
body at the base. j jaclet cell. E Archegonial complex in transection. ag archegonium.
F Longisection of gametophyte (outline), to show micropylar (mac), lateral (la) and
chalaza! (cha) archegonia. G Two archegonia (chalaza! portion of F). H Outline diagram
for I. I Chalaza! archegoinal complex from H. J Fertilization. mg male gamete. en egg
nucleus. (After H. Singh and Chatterjee 1963)
Coniferales 199
the cytoplasm becomes less dense and a large vacuole appears in the lower
part; the egg nucleus is pushed up to the upper part of the archegonium
(Fig. 15.2.6 C).
Electron microscopic studies show that in Cryptomeria japonica the egg
cell is smaller than in the Abietaceae. It contains nodules of dense cytoplasm
capped with a crescent-shaped vacuole, and is comparable with the "small
inclusions" of the Abietaceae. The plastids undergo no appreciable change
up to the maturation of the egg. These leucoplasts have few lamellae, and
the ground substance is opaque to electrons. Their membrane is invariably
supplemented with layers of endoplasmic reticulum, which lie parallel to it
(see Chesnoy and Thomas 1971).
The jacket is usually one-layered laterally and two-layered at the base of
the archegonial complex. The jacket cells contain dense cytoplasm and one
to three prominent nuclei (Fig. 15.2.6 D). Some of the cells may contain
rather large nuclei, probably due to the fusion of smaller nuclei.
Fertilization
The neck of the archegonium begins to disorganize before fertilization. The
contents of the pollen tube are discharged into the pollen chamber, one or
both of the male gametes enters the egg cell, and fertilization takes place
in its centre. A depression appears in the upper part of the egg nucleus
(Fig. 15.2.6 J), and the dense cytoplasm around the fusing nuclei contains
starch grains. Vacuoles appear in the upper part of the zygote and the
vacuoles in the lower part are no longer visible.
Embryogeny
Five to seven archegonia in a complex become fertilized, and the division
of the zygote nucleus occurs at the base of the egg cell. Two further
divisions follow, the resultant eight nuclei become embedded in dense
cytoplasm ~nd starch grains, and organize into the primary embryonal tier
(pE) and primary upper tier (pU). The latter divides into a middle suspensor
(S) and an upper open tier (U, Fig. 15.2.7 A-D). A mature proembryo has
13 or 14 cells arranged in three distinct tiers (E, S, U) and occupies nearly
one-third of the length of the archegonium. The U tier eventually degenerates.
The cytoplasm in the upper part of the proembryo appears less dense
and can be distinguished from denser cytoplasm in the lower part.
Occasionally, one or two nuclei (probably derivatives of the second male
gamete) have been observed in the upper part of the zygote; they may
rarely persist up to the cellular stage of the proembryo (Fig. 15.2.7 D).
The suspensor tier S elongates and pushes the attached embryonal cells
E beyond the archegonium. At the same time, the suspensor cells separate
from each other and lead to cleavage polyembryony (H. Singh and Chatterjee
1963). Some of the separated suspensor cells do not bear any embryonal
cell (the latter are fewer). The tip of such a suspensor cell becomes densely
200 The Gymnosperms
. •'"\ ...
..-~ ..-~
(~& .
'·.
'\ - ~--
~~··
..- .. .....
.·
. • 0
Fig.l5.2.7A-M. Cryptomeriajaponica, embryogeny. A·D Development of proembryo.

E Embryonal tier. pE primary embryonal tier. pU primary upper tier. S suspensor tier.
U upper tier. E Elongated suspensor tier. F Young proembryo system, suspensor cells
with or without an embryonal cell at their tip. G Septate suspensor with embryonal cells
at the tip. H Outline diagram for I. I Secondary suspensor (ss) from H. J Mature embryo
system. K, L Longisection gametophyte, development of embryo. emb embryo.fg female
gametophyte. M Transection of mature seed. cot cotyledon. mid middle layer. oz outer
zone. (After H. Singh and Chatterjee 1963)
Coniferales 201
cytoplasmic, its nucleus enlarges and a small "embryonal cell" is cut

off. Occasionally, a suspensor cell may become transversely septate
(Fig. 15.2.7 E-G). According to Sugihara (1969), the suspensor cells elongate,
but remain fused parallel to one another. Cleavage polyembryony takes
place by elongation of embryonal tubes at the tip of the suspensor. Meanwhile,
the cells of the gemetophyte adjoining the archegonial complex disorganize,
and form a loose cavity. The elongation of the suspensors pushes the
developing embryos (with the suspensors coiled around each other) into
this cavity.
The embryonal cells divide to form an embryonal mass, whose upper
cells elongate and produce secondary suspensors. The latter, eventually,
become massive. The embryonal masses attached to the secondary suspensors
(lying in the cavity of the gametophyte) show various stages of development.
The embryonal mass lying deep in the cavity continues to grow; the other
remains arrested (Fig. 15.2.7 H-J).
Differentiation of Embryo. The embryonal mass grows predominantly

by transverse divisions. The differentiation of root initials is followed by
the stem tip. The three cotyledonary primordia appear around the stem tip,
and elongate towards the chalazal end of the seed. The gametophytic tissue
below the shoot apex is not completely used up during the maturation of
the embryo, and persists between the developing cotyledons. A provascular
strand differentiates in each cotyledon. With the growth of the embryo, the
suspensor is gradually absorbed. A mature embryo comprises three cotyledons,
a well-organized root and shoot tip, and is surrounded by four or five
layers of gametophytic cells filled with starch and fatty food reserves
(Fig. 15.2.7 K-M).
Seed. The seed coat has the usual three zones : (a) an outer zone of thin-
walled sarcotesta, (b) a middle zone of thick-walled sclerotesta, and (c) an
inner zone of thin-walled endotesta (Fig. 15.2.7 M).
Germination. The seeds do not have a period of dormancy. The germination

(%) is rather low, and is epigeal as in conifers. The three cotyledonary
leaves are green, thick and linear, and show two resin canals, one near each
margin of the leaf. The vascular bundles are collateral and tangentially
elongated. The juvenile leaves are arranged in an opposite and decussate
manner, and differ from the mature leaves.
Chromosome Number
The diploid chromosome number is 22 (Fig. 15.2.8 A, B); six chromosemes
are heterobrachial and the rest isobrachial. Each of the two isobrachial
chromosomes has a secondary constriction (Mehra 1988).
202 The Gymnosperms
Fig. 15.2.8 A, B. Cryptomeria japonica, cytology. A, B Root tip mitoses, 2n = 22;

arrow shows secondary constriction. h heterobrachial chromosome. (After Mehra 1988)
In Mussoorie (Western Himalayas), the life cycle is completed in 1.5 years
(H. Singh and Chatterjee 1963 ). The male cones are initiated in June. The
pollen is shed in February and male gametes are formed by May. The
female cones appear in July--ca. 1 month after the male cones. It undergoes
a rest period from September to January. Growth is resumed in the beginning
of February of the following year and fertilization occurs in the middle of
May. The embryo develops and seeds mature by October. The female cones
dehisce from the beginning of December to the beginning of January.
15.3 CUPRESSACEAE
The trees or shrubs are characterized by shoots arranged in an opposite and

decussate manner or in whorls; there is no differentiation into long and
short shoots. The plants are monoecious. The scales in the female cone are
arranged cross-wise. There is a pronounced fusion between ovuliferous and
the bract scale. In some taxa, the cone scales overlap (Thuja), or are valvate
(Cupressus) and may even be fleshy (Juniperus). The ovules are erect and
3-20 (rarely 1 or 2) per scale. The number of pollen sacs also varies from
3-6 (rarely 2).
The family comprises 19 genera; about 9 are confined to the Northern
Hemisphere and the rest to the Southern Hemisphere. Eight taxa are
monotypic. Juniperus is distributed in a continuous broad belt round the
northern hemisphere, possibly due to its berry-like female cones which are
Coniferales 203
relished and thereby distributed by birds. This taxon includes 70 spp. and
is the second largest conifer genus.
The northern taxa are (species in parenthesis): Arceuthos (1), Biota (1),
Calocedrus (3), Chamaecyparis (6), Cupressus (20), Fokienia (2), Juniperus
(70), Thuja (4) and Thujopsis (1).
The southern taxa are: Actinostrobus (2 ), Austrocedrus (1 ), Callitris
(15), Diselma (1), Fitzroya (1), Libocedrus (5), Neocallitropsis (1),
Papuacedrus (3), Pilgerodendron (1), and Widdringtonia (5).
The Cupressaceae have existed for a considerable time. Their remains
have been recognized in sediments of the Late Triassic (Lemoigne 1967),
and again in those of the Lower to Middle Jurassic (Cha1oner and Lorch 1960).
Accordingly, most of the genera of the family can be viewed as relicts in
that they include only a few species with limited geographic distribution
(see Miller 1977).
Biota orienta/is
For a long time, the taxon Biota was included under Thuja. On the basis
of morphological characters, Endlicher (1847) raised Thuja orienta/is to a
new genus, Biota. His views were accepted by Buchholz (1929) and Martin
(1950), but opposed by Lindley (1853), Pilger (1926) and Pilger and Melchior
(1954).
Biota orienta/is is a monotypic genus. It is a native of the eastern part
of Central Asia, and is cultivated as an ornamental in many parts of the
world. This species has numerous cultivars.
Morphology
The plant is an evergreen tree reaching a height of ca. 15 m. It has an
entirely different shape when young and displays considerable variations
under cultivation. Wilson (1926) observed some plants in a park in China,
that have been growing undisturbed for centuries. These were large trees
with horizontal lateral branches, in contrast to the plants under cultivation
with vertical lateral branches. It appears that in younger stages the lateral
branches are vertical, but later become horizontal. Due to the slow-growing
habit of the plant, and constant trimming of its branches to give it the
desired shape, it almost never attains its natural appearance.
The bark is smooth, brownish, and separates from the older branches as
papery scales. The terminal shoots are divided into a spray of branchlets
(Fig. 15.3.1 A) covered by dark green, closely appressed, acute and decussate
leaves. Each leaf is fused with the stem along one-third of its length. It has
a long groove in the middle of its abaxial surface (Fig. 15.3.1B).
The plants are monoecious. The male and female cones occur on separate
branch systems. However, both types of cones may sometimes be present
on the same branch system (H. Singh and Oberoi 1962). The plants are
either predominantly male with a few female cones, or vice versa. The sex
204 The Gymnosperms
Fig. 15.3.1 A-K. Biota orientalis. A Twig with branchlets. B Leaf, abaxial surface
>hows longitudinal groove . C Young male cone enclosed by leaves. D Dehisced male
cone. E Microsporophyll, lateral view. F Branch bearing male cones. G, H Female cones
on curved branches. I Erect mature cone. J, K Lateral (K) and abaxial (J) views of scale-
bearing ovules. (After H. Singh and Oberoi 1962)
Coniferales 205
expression (in this species) is quantitative, perhaps controlled by a multigenic

factor (Mehra 1988).
Reproduction
Male Cone
The male cones are borne on the ultimate branchlets (Fig. 15.3.1 F); the
young cones are completely covered by leaves (Fig. 15.3.1 C). Later, they
emerge by the elongation of the basal portion of the cone axis. A male cone
is 2-3 mm in diameter, and bears four to six pairs of microsporophylls
arranged decussately (Fig. 15.3.1 D). At maturity the microsporophyll is
small, round, leaf-like, and attached to the cone axis by a short stalk. It is
slightly broad at the base, convex on its abaxial surface and has an inwardly
curved crenate margin (Fig. 15.3.1 E). Three to five microsporangia are
present on the abaxial surface near the base of each sporophyll. Young
cones are yellowish green but turn pale yellow at maturity. The
microsporophylls separate from each other and expose the sporangia
(Fig. 15.3.1 D). After the pollen is shed, the cones tum brownish-yellow,
dry up and fall off from the plant.
Microsporangium. A young sporophyll (in longisection) shows two to

three hypodermal archesporia! cells near the base of the abaxial surface.
The epidermis and the archesporia! cells divide anticlinally and their number
increases (Fig. 15.3.2 A-C). The archesporia! cells divide periclinally and
form a primary parietal layer and a primary sporogenous layer. The cells
of the latter give rise to a mass of sporogenous cells, by repeated divisions
in all planes (Fig. 15.3.2 D). The primary parietal layer divides periclinally
to form two layers, the layer in contact with the sporogenous tissue
differentiate into the tapetum. A young sporangium consists of the epidermis,
a single middle layer, a tapetum, and a mass of sporogenous cells
(Fig. 15.3.2 F). The epidermis and the wall layer continue to divide anticlinally
for a while, the sporangium becomes round or slightly elongated
(Fig. 15.3.2 E). Later, with an increase in the sporogenous tissue, the anticlinal
divisions become fewer. A mature sporangium shows a tangentially elongated
epidermis with cutinized outer walls, and develops annular thickenings
(Fig. 15.3.2 G). The cells of the middle layer degenerate as the mother cells
prepare for reduction divisions. The tapetal cells become binucleate and
collapse during maturation of the microspores.
Micro<;porogenesis. The microspore mother cells (mime) undergo reduction

divisions (Fig. 15.3.2 H-K). At anaphase I the cytoplasm of the mother cell
retracts slightly and a callose wall is secreted around it. A cell plate may
(Fig. 15.3.2 J) or may not (Fig. 15.3.2 I) be laid down between the two
dyad nuclei, so that cytokinesis is of the successive or simultaneous type.
Both tetrahedral and isobilateral tetrads are formed (Fig. 15.3.2 L-N).
206 The Gymnosperms
Fig. 15.3.2 A-Q. Biota orienta/is. A Longisection very young male cone.
B, C Longisection microsporophylls shows hypodermal archesporium. D Longisection
young microsporangium with primary sporogenous and primary parietal layers.
E Longisection male cone at microspore mother cell. F Young sporangium with wall
layers and sporogenous tissue; some tapetal (tap) cells are binucleate. G Sporangia! wall
at microspore stage; epidermis shows thickenings. H-N meiosis I and II; the microspore
tetrads are tetrahedral and isobilateral. 0 Microspore. P Microspore nucleus in division.
Q Pollen grain at shedding stage. at antheridial cell. tn tube cell. (After H. Singh and
Oberoi 1962)
Male Gametophyte. A microspore is spherical, and shows a thin exine,

a thick intine and a large nucleus (Fig. 15.3.2 0). Starch accumulates in the
cytoplasm, and may even mask the nucleus. The latter shifts laterally and
divides (Fig. 15.3.2 P) to form a small antheridial and a large tube cell
(Fig. 15.3.2 Q). With further enlargement of the pollen grains, the intine
Coniferales 207
becomes thinner and the starch grains begin to be digested. The pollen is
shed at the two-celled stage. Some of the microsporangia, and sometimes
all the sporangia on a plant, produce only sterile pollen (H. Singh and
Oberoi 1962).
Female Cone
The female cones are borne on the terminal branchlets, the male cones
slightly lower on the same branchlet. The young cones covered by leaves
are borne on branches that curve downwards (Fig. 15.3.1 G). These branches
become nearly upright when the cones emerge at the time of pollination.
Each cone consists of three or four pairs of decussate scales, and bears a
curved spine (Fig. 15.3.1 H, I, K). The scale is a fusion product of the bract
and the so-called ovuliferous scale. It bears one to three orthotropous ovules
near the base of its adaxial surface (Fig. 15.3.1 J, K). The uppermost pair
of scales is usually sterile.
At the time of pollination, the tip of each scale curves backward to
expose the ovules. With further growth, the scales become fleshy, come
close to each other and the cone becomes compact (Fig. 15.3.1 1), and
woody. A mature cone measures ca. 20-22 mm in length and ca. 18 mm
in diameter. The cone scales separate from each other along the original
margins and expose the seeds for dispersal. A seed is ca. 5 mm long and
is covered by a three-angled stony seed coat.
Ovule. The young cone shows well-differentiated ovules on the adaxial

surface of the scale (H. Singh and Oberoi 1962). The integument is three
or four cells thick, tapers to one layer at the tip, and is slightly longer than
the nucellus. One or two primary sporogenous cells differentiate in the
nucellus at the level of insertion of the integuments (Fig. 15.3.3 A). The
nucellar epidermis undergoes periclinal as well as anticlinal divisions. The
ovule continues to increase in size, and the primary sporogenous cells
undergo mitotic divisions to form the sporogenous tissue (Fig. 15.3.3 B).
Megasporogenesis. A single, more or less centrally placed mother cell

(Fig. 15.3.3 D) undergoes reduction divisions to form a row of three cells.
The uppermost cell is the undivided dyad; Tarchi (1969) observed callose
around it. The chalazal megaspore functions (Fig. 15.3.3 E, F).
Female Gametophyte. The functional megaspore enlarges and a large

central vacuole shifts the nucleus to one side. Repeated synchronous free-
nuclear divisions occur, and due to the formation of a large central vacuole
the nuclei become embedded in a peripheral layer of cytoplasm (Fig. 15.3.3 K).
With an increase in the nurrber of nuclei, they become smaller (Fig. 15.3.3H-J).
When the number of free-nuclei exceeds ca. 4000, centripetally advancing
walls (Fig. 15.3.3 L, M) bring about cellularization of the gametophyte.
208 The Gymnosperms
) 0)
Fig. 15.3.3 A-M. Biota orienta/is. A, B Longisection ovules with deep-seated sporogenous
cells . C Longisection upper part of ovule after pollination; micropylar canal closed.
pt pollen tube. D Megaspore mother cell surrounded by sporogenous tissue. E, F Megaspore
triads, chalaza! megaspore functional. G Two-nucleate gametophyte (jg) with well-
developed spongy tissue (sg) . H, I Free-nuclear gametophytes. J Inset from I.
K Longisection ovule with deep-seated gametophyte. L Cellular gametophyte. M Inset
from L. (After H. Singh and Oberoi 1962)
Coniferales 209
The megaspore membrane also becomes very conspicuous. The mature

gametophyte elongates, tapers slightly at the chalazal end, and is flat or
somewhat convex at the micropylar end.
A free-nuclear gametophyte is enclosed by two or three layers of a
conspicuous spongy tissue derived from the non-functional sporogenous
cells (Fig. 15.3.3 G). As the gametophyte enlarges, the nucellar cells outside
the spongy tissue collapse. By the time walls are initiated in the gametophyte,
the spongy tissue degenerates.
Archegonial Complex. A newly formed central cell along with a neck

initial shows the megaspore membrane flush with the neck initial; the jacket
cells are uninucleate (Fig. 15.3.4 A).
The neck initial divides twice vertically with the walls oriented at right
angles to each other, and forms a neck of four cells lying in one plane
(H. Singh and Oberoi 1962). Meanwhile, the central cell and its nucleus
enlarge considerably and a vacuole appears just below the nucleus. About
the time the pollen tube reaches the archegonial chamber, the nucleus of
the central cell divides to form a small disc-shaped ventral canal nucleus
(vcn) and a large egg nucleus (Fig. 15.3.4 C, D). Before the nucleus of the
central cell divides, the organelles of the cytoplasm, particularly the
mitochondria, collect below the nucleus and at the centre of the basal
cytoplasm below the vacuole. During early stages, the cytoplasm around
the vcn is vacuolar and distinct from that below it. As the body cell in the
pollen tube divides to form two male gametes, the vcn begins to degenerate
and is no longer recognizable by the time the male gamete enter the
archegonium.
The egg is ca. 300 J.lm long and 50 J.lm across. The nuclear diameter is
35-40 J.lm and is situated in the upper part of the cytoplasm. The egg cell
shows a vacuole representing the remnants of the vacuole earlier occupied
by the greater part of the central cell.
The cytoplasm of the central cell has a large vacuole in the middle of
the cell. The cytoplasmic organelles (Fig. 15.3.4 B) aggregate in its micropylar
portion (below the nucleus}, and the chalaza! portion (below the central
vacuole). The organelles comprise mitochondria and leucoplasts arranged
radially around a mass of ribosomes and microtubules. It has been suggested
that this area of organelle aggregation may also be the centre of organelle
multiplication (Chesnoy 1971).
The filling up of the vacuole below the nucleus occurs by the breakdown
of its tonoplast and the penetration of cytolasmic vesicles into the vacuole
(Chesnoy 1971).
Electron microscopic observations have been made on the egg cytoplasm
(Chesnoy 1971). The large number of mitochondria (present in the mature
egg) are spherical and dispersed. In section, some of these appear as a ring
enclosing a small core of cytoplasm. Only rarely do they reveal tubules,
210 The Gymnosperms
Fig.15.3.4 A-E. Biota orienta/is. A Longisection young central cell (d). ni neck initial.
j jacket cell. mm megaspore membrane. B immature archegonium; two asteroids lie in
the cytoplasm of the large cell. m mitochondria. nk neck cell. p plastid. v vacuole.
vc cytoplasmic vesicles. C, D Upper part of central cell. C Central cell nucleus in division.
D Ventral canal nucleus (vcn) and egg nucleus (en). E Nearly mature archegonium with
large egg nucleus, scattered mitochondria, the jacket cell is binucleate. si small inclusion.
(A, B, E After Chesnoy 1971 , C, D after H. Singh and Oberoi 1962)
which are very short and lie in a transparent matrix (Chesnoy 1969a, b).
The leucoplasts are narrow, very long and often grouped into bundles
(Fig. 15.3.4 E).
The archegonia are arranged in a single group or complex at the micropylar
end of the gametophyte (Fig. 15.3.5 A). Each complex has 15-28 (usually
22) archegonia (Fig. 15.3.5 C). Occasionally, one or two supernumerary
archegonia develop below the archegonial complex. In such an archegonium
Coniferales 211
mac
:::~ l.Ll_liiTITI
~}
____ .,..J B
Fig. 15.3.5 A-C. Biota orientalis. A, B Longisection female gametophyte with micropylar
(mac) and chalaza! (cha) archegonial complex. C Archegonial complex in transection.
(After H. Singh and Oberoi 1962)
the initial functions as the central cell and the neck cells are not formed.
These complexes degenerate, probably due to lack of fertilization. Rarely,
there is a chalazal archegonial complex with fewer archegrnia (Fig. 15.3.5 B).
With the development of the archegonia, the adjacent tissue grows upward,
which results in a depression, the archegonial chamber. The archegonial
complex is surrounded by a common jacket layer. After fertilization, most
of the jacket cells become binucleate; large polyploid nuclei may be formed
by nuclear fusions.
212 The Gymnosperms
Pollination
Pollination occurs by wind. At the time of pollination the tip of each scale
curves backward so that the ovules are exposed. There is a sugary exudation
at the micropyle (Fig. 15.3.6 A, B). The pollen grains swell when caught
in the pollination drop (H. Singh and Oberoi 1962).
I<'ig. 15.3.6 A, B. Biota orienta/is. A, B Female cones at pollination. (After H. Singh

and Oberoi 1962)
After pollination, the cells of the integument (lining the micropyle) elongate
radially and close the canal (Fig. 15.3.3 C), periclinal divisions may also
take place. These cells become thick-walled and show simple pits during
the maturation of the ovule.
Post-Pollination Development of Male Gametophyte. After the pollen

grains come to lie on the nucellus, the exine bursts and the intine extends
and forms a small pollen tube containing the tube nucleus (Fig. 15.3.7 A, B).
Meanwhile, the antheridial cell becomes free from the wall of the microspore,
moves into the pollen tube (Fig. 15 .3.7 B), and divides to form the stalk
and body cells (on entering the nucellus). Later, a wall cannot be made out
around the stalk cell and it persists as a nucleus.
Generally, three to five pollen tubes enter an ovule. They elongate and
Coniferales 213
enlarge so that the adjoining nucellar cells are crushed; their contents always
persist at the tip. Occasionally, a pollen tube may branch (Fig. 15.3.7 C).
The growth of the pollen tube is accelerated by the differentiation of the
archegonial initials.
The tube and stalk nuclei cannot be distinguished from each other and
always lie close to the body cell in dense cytoplasm (Fig. 15.3.7 C). The
spermatogenous (body) cell enlarges considerably (Fig. 15.3.7 E). On reaching
the archegonial chamber, the tip of the pollen tube swells and brings about
disintegration of the cells lining the archegonial chamber (Fig. 15.3.7 D, E).
The spermatogenous (body) cell divides to form two equal male cells (Fig.
15.3.7 F-H). The stalk and tube nuclei begin to degenerate.
A mature male gamete is ca. 50 pm in diameter, its spherical nucleus is
ca. 30-35 pm and shows a distinct nucleolus. It is separated from the
cytoplasm of the pollen tube only by a plasmalemma. Electron microscope
studies reveal that the cytoplasm is organized into three consecutive zones,
which do not show uniform structure. The mitochondria, amyloplasts, groups
of vesicles and abundant ribosomes collect in a deep zone ca. 3-15 pm in
thickness (Fig. 15.3.8), separated from the nucleus by a narrow perinuclear,
and from the plasmalemma by a marginal zone. The perinuclear zone is ca.
0.5 to 1 pm thick, free from organelles, and shows only fragments of reticulum
arranged parallel to the nuclear zone. The marginal zone is 2-3 pm thick
and contains only a few Golgi vesicles and dilated fragments of reticulum
(Chesnoy 1969 a, b).
The EM studies of the male gametes have shown the ultrastructural
characters of the organelles of the male cytoplasm, and have helped to
locate these organelles after the union of the gemetes. The matrix of the
mitochondria of the sperm cells is very dark and the cristae well developed.
The amyloplasts contain several starch grains (Chesnoy 1969 a, b).
Fertilization
The pollen tube tip breaks down and discharges the male cells into the
archegonial chamber. In a single chamber as many as 12 male cells have
been observed. There is simultaneous fertilization of two adjoining egg
cells by the two gametes from the same pollen tube. With the entrance of
the pollen tube into the chamber, the neck cells degenerate, making a
passage for the male cells to enter the archegonium. The male gamete (cell)
penetrates into the cytoplasm of the egg; its organization is only slightly
modified. The marginal cytoplasmic zone of the male gamete merges with
the cytoplasm of the female gamete during penetration of the sperm cell
into the egg. The male nucleus moves in the maternal cytoplasm surrounded
by the perinuclear, deep zone of its own cytoplasm. It partially frees itself
from the perinuclear, deep zone as it approaches the female nucleus. The
latter has a depression in its upper part in which the male nucleus establishes
itself. The nuclear membranes join together while the two nuclei turn at
214 The Gymnosperms
a l --+-o~
tn --""""'"'~:>E=o.,....t::J
E
Fig. 15.3.7 A-H. Biota orientalis. A Longisection ovule shows germinated pollen on
the nucellus. pt pollen tube. B Pollen tube from A. al antheridial cell. tn tube nucleus .
C Branched pollen tube in nucellus shows tube and stalk nuclei and the body cell (be).
D Longisection (upper part) female gametophyte shows pollen tubes. E Part of pollen
tube and female gametophyte from D. F Division in body cell. G, H Pollen tube with
two equal male cells. (After H. Singh and Oberoi 1962)
Coniferales 215
Fig. 15.3.8 Biota orientalis. Portions of two male cells, middle zone contains most of
the organelles. (Courtesy Chesnoy, see H. Singh 1978).
180° (see Chesnoy and Thomas 1971). At the commencement of the nuclear
encounter, the cytoplasm of the deep zone of the male gamete caps the two
nuclei. It gradually surrounds them and isolates from the maternal cytoplasm.
EM shows that the male cytoplasm retains all its cohesion during its descent
through the egg cell.
Embryogeny
The division of the zygote takes place within the nucleoplasm, which is
still bordered by the nuclear membrane that disappears only after the telophase.
The mitotic figure is intranuclear and slightly oblique with respect to the
long axis of the archegonium (Fig. 15.3.9 A). After the completion of
mitosis, the cytoplasm of the male gamete, along with its organelles, penetrates
into the original nucleoplasm and surrounds the two proembryonal nuclei.
The two nuclei with the cytoplasm originating primarily from the deep
zone of the male gamete migrate to the base of the archegonium and constitute
216 The Gymnosperms
--
,' ... . ~ .
s
A
sht
es
Fig. 15.3.9 A-H. Biota orienta/is. A Intranuclear division of the zygote nucleus .
B, C Two-and four-nucleate proembryos. ml male nucleus. D Three-tiered proembryo.
E Embryonal tier. S suspensor tier. U upper tier. E Young embryo. F Elongation of
embryonal suspensor (es) . e embryonal cell. G Whole mount embryo shows mass of
suspensor, embryonal suspensor and embryonal cells at the tip. H Longisection female
gametophyte shows mature embryo. cot cotyledon.fg female gametophyte . hp hypocotyl.
rtp root tip. sht shoot tip. (After H. Singh and Oberoi 1962)
Coniferales 217
the embryonic pole. The mitochondria, amyloplasts and cytoplasmic RNA

are transmitted to the embryo. A few mitochondria of maternal origin (carried
along by the male cytoplasm during its course through the egg cell) appear
in the proembryonal cytoplasm. The plastids of maternal origin have not
been obsrved in the proembryonic cytoplasm. EM studies confirm that the
cytoplasm of the proembryo and its organelles are essentially of paternal
origin (Chesnoy 1969 a, b).
Wall formation in the proembryo takes place at the eight nucleate stage.
The lower one-quarter of the zygote contains the embryonal nuclei and is
densely cytoplasmic (Fig. 15.3.9 B, C); the upper portion has scanty
cytoplasm, which seems to lose contact with the cytoplasm of the lower
portion.
Occasionally, one or two nuclei (probably derivatives of the second
male nucleus) have been observed in the upper region of the proembryo
(Fig. 15.3.9 B). These nuclei usually degenerate, but may persist up to the
cellular stage of the proembryo (Fig. 15.3.9 E).
Wall formation in the proembryo results in the upper pU and the lower
pE tier of four cells each. Two embryonal and five upper cells are not
uncommon. The cells of the pU tier divide and give rise to a proembryo of
12-16 cells arranged in three distinct tiers of (a) embryonal (E), (b) suspensor
(S) and (c) open tier ( U), and occupies about one-third the space of the
proembryo (Fig. 15.3.9 D).
Differentiation of Embryo. The suspensor tier elongates and pushes the

embryonal tier beyond the archegonial complex (Fig. 15.3.9 E). As soon as
the embryonal tier comes in contact with the gametophytic tissue, each of
its cells divides to form the embryonal suspensor (Fig. 15.3.9 F), which
elongates only after the primary suspensor tier ceases to function.
Cleavage occurs at this stage and the embryonal suspensors, carrying the
small embryonal cell at its tip elongate (into the corrosion cavity). The
suspensor system develops rapidly and becomes tightly coiled round
(Fig. 15.3.9 G), in the limited corrosion cavity. The degenerated suspensor
cells are pushed back by the developing suspensors. Occasionally, the
suspensor cells become separated and bulge outward with prominent nuclei
in the swollen portions. Rarely, the nucleus divides and as many as eight
free nuclei may be formed in the bulge. Sometimes, these cells elongate
considerably and simufate the suspensor. These can be distinguished by
their swollen appearance and absence of an embryonal cell at the tip.
The embryonal cell remains inactive throughout the period of elongation
of the suspensor cell. The first two divisions in the embryonal cells are
vertical and at a right angle to one another. Further divisions take place in
all planes, forming a small globular mass located at various levels in the
cavity of the gametophyte. The leading embryo (lying farthest in the
gametophyte) continues to grow, while the remaining embryos become
218 The Gymnosperms
arrested and gradually collapse. The proximal cells of the embryonal mass
divide and elongate forming a massive secondary suspensor. The embryonal
cells also increase by further divisions.
The root initials differentiate behind the apex of the embryonal mass,
and this is followed by the organization of the stem tip. Simultaneously,
two cotyledonary primordia appear and begin to elongate. The region between
the shoot apex and the root initials also elongates considerably to form the
hypocotyl. Each cotyledon is supplied by a provascular strand. As the
embryo elongates, the suspensor collapses and its remnants persist until
maturity (Fig. 15.3.9 H).
The mature embryo has two (rarely three) cotyledons with well-organized
root and shoot tip. The embryo is surrounded by five or six layers of
gametophytic tissue rich in starch.
Seed
Initially the integument comprises four or five layers of cells. These cells
enlarge prior to pollination and undergo only anticlinal divisions. The cells
of the outer epidermis and hypodermis contain tannin.
When the ovule contains a free-nuclear gametophyte, the integument is
10-12 layers thick. The cells lining the micropylar canal (after pollination)
become thick-walled and show simple pits. Such thickening of the cells
also extends downward (Fig. 15.3.10 A-C).
At maturity, the seed coat is 20-25 cells thick, comprising three distinct
regions-an· outer and inner zone of thin-walled cells, and an intervening
zone of thick-walled cells (Fig. 15.3.10 G, H). The outer, thin-walled zone
of the integument consists of three or four layers of elongated cells, the
outer two layers are tanniniferous. In the mature seed this zone becomes
disorganized and is no longer distinguishable when the cone "opens" to
shed the seeds. The middle thick-walled zone consists of six or seven
layers which are comparatively large and contain shrunken cytoplasm and
small nuclei. The walls show numerous pits (Fig. 15.3.10 D, E). The inner
zone consists of seven to ten layers of long, narrow cells which remain
thin-walled. The cells contain small nuclei and scanty cytoplasm (Fig.
15.3.10 F). The cells of the inner epidermis contain tannin. The mature
seed (in a cross section) is triangular (Fig. 15.3.10 H).
Germination. The seed germinates soon after shedding. The epigeal primary
root branches only at a late stage. The juvenile leaves look very different
from the mature leaves, and are arranged spirally on the stem. They are
elongated and lanceolate, and are not fused with the stem for any appreciable
length.
The epidermis of the cotyledons is covered by a poorly dveloped cuticle.
The mesophyll consists of an undifferentiated mass of parenchymatous
cells with abundant intercellular spaces. Each cotyledon has a single
Coniferales 219
a.,_ . _
G
- fg
-M
F.~ . ::.
tf; .: . ;;
~H
Fig. 15.3.10 A-H. Biota orientalis. A-C Longisection ovule at v,arious stages of
develoment. The thickening in the middle part of integument extends from micropylar
to chalaza! region. il inner zone. mid middle zone. ol outer zone. D, E, F Portions marked
A, B, C in C. Three regions of integument. E Cell walls of middle zone have become
thick. F All the cells are thin-walled. G, H Longi-and transection of mature seed. emb
embryo. fg female gametophyte. (After H. Singh and Oberoi 1962)
(somewhat tangentially elongated) collateral and endarch vascular bundle.

Resin ducts are not present in the cotyledons, although they are common
in the foliage leaves.
Chromosome Number
The chromosome number, as determined from root tip squashes, is 2n = 22
220 The Gymnosperms
(Fig. 15.3.11 A). Karyotypic analysis reveals three pairs heterobrachial

and the remaining eight pairs isobrachial. One isobrachial pair showed a
secondary constriction in one arm (Fig. 15.3.11 A, arrow). Two-chromosome
configuration at mixed anaphase in endosperm squash is shown in
Fig. 15 .3.11 B, C (Mehra 1988).
'
Fig. 15.3.11 A-C. Biota orienta/is. A Root-tip mitosis, 2n = 22. h B, C Endosperm

squashes show chromosomes at mixed anaphase I. (After Mehra 1988)
Coniferales 221
In Delhi, the male cones are initiated at the end of May/begining of June,
remain covered by foliage and emerge only after November; the growth of
the cone is slow during this period. By early January, the microspore mother
cells (mime) differentiate. The tetrads and microspores develop by the end
of January. The microspore nucleus divides in early February, and the
pollen is shed in the second week.
Female cones appear in the beginning of July (a month later than the
male cones). In early September the (small) branches bearing the female
cones bend downward. The growth of the cones is very slow during this
period. By the end of November, the megaspore mother cell (mgmc)
differentiates. The female cone undergoes a period of rest during December
and January. Reduction divisions in the mgmc take place in early February
and simultaneously the cone emerges and "opens" to expose the ovules for
pollination. From the second week of February for 20 days, a shiny
conspicuous pollination drop appears at the micropyle, in the early hours
of the morning, and dries up by about 10.00 h.
In early March, the pollen tubes are common on the nucellus; the female
gametophyte is at the free-nuclear stage. Wall formation begins, archegonia
develop early in April, and fertilization occurs by the middle of the month.
By the end of May, embryonal masses show extensive suspensor. Cotyledons
differentiate (mid-June), embryo matures (beginning of July), and dehiscence
of the cones begins in the first week of August. Seeds germinate within
15-20 days without undergoing any period of dormancy.
15.4 Podocarpaceae
The family Podocarpaceae includes trees and shrubs, and the living
representatives are predominantly distributed in the Southern Hemisphere.
There are seven genera and ca. 150 sp. Podocarpus, with nearly 106 spp.,
is the largest genus of present-day conifers. The other taxa are: Acmopyle,
Microcachrys, Pherospherea (Microstrobus), Phyllocladus, Dacrydium and
Saxegothea.
The Podocarpaceae is of great interest because it shows a remarkably
wide range of morphological characters. Most extant Podocarpaceae bear
leaves and other parts in a helix, some are opposite. The leaf varies from
sessile, decurrent and falcate with a single vein to large and flat, and contains
many parallel veins.
The male cones may be simple (characteristic of conifers) or form a part
of a compound structure. There is considerable variation in the development
of male gametophyte, and a wide range of pollination mechanism.
In some spp. the ovules are borne in cones, while in others the female
cone is so reduced that it forms a swollen fleshy receptacle with a single
terminal ovule.
222 The Gymnosperms
The Podocarpaceae is well represented in fossil record, the earliest

occurrence is in the Early Triassic. The available evidence suggests that
this family diverged from the Voltziales in the Late Palaeozoic and was
already a separate line of evolution at the onset of the Mesozoic (see Miller
1977). However, Podocarpus is the only modem genus of the group with
a record during the Mesozoic, havfng evolved by the Cretaceous.
Florin (1958) regarded the Podocarpaceae (along with Araucariaceae) as
Southern Hemisphere conifers, and indicated that this had always been so.
However, there are reports of this genus from the Northern Hemisphere.
Krassilov (1974) reports remains of Podocarpous from the Late Cretaceous
of Russia and states that the "northern" and "southern" conifers grew side
by side in Mesozoic forests and at least Podocarpus persisted in North
America until the Eocene (Dilcher 1969).
Podocarpus
It is distributed throughout the Southern Hemisphere, tropical and Southern
Africa, Australasia to Japan. It is regarded as the dominant conifer in the
Southern Hemisphere, as Pinus is in the Northern Hemisphere.
Podocarpus has been studied extensively, the various species show great
diversity in structure and development. Pilger (1926) divided it into two
subgenera: (a}-Stachycarpus and (b) Protopodocarpus. The latter is further
subdivided into four sections: Nageia, Microcarpus, Dacrycarpus and
Eupodocarpus.
Morphology
The plants are mostly tall evergreen trees. P. gracilior, the African fern-
pine, is 15 m high with a trunk of 1.8-2.9 m girth. The main trunk is
crowned with innumerable branches. The leaves are ca. 8 em in length and
4 mm in width.
Anatomy
Root. There is a taproot system whiCh shows numerous mycorrhizal

tubercles on the main and lateral roots (Saxt_on 1930, Baylis et al. 1963,
Konar and Oberoi 1969 a). The primary root is diarch (Fig. 15.4.1 A). The
parenchymatous cortical tissue has large intercellular spaces. Most of the
cells are filled with starch grains, and several contain tannin. Secondary
growth and the development of bark occur at an early stage. A fungus
infests the young roots, the cortical cells proliferate, becoming large and
nodular. The root tubercles are endogenous and represent modified lateral
roots. The large parenchymatous cells of the tubercles are filled with fungal
;hyphae and spores (Fig. 15.4.1 B, C).
Stem. Shoot Apex. The lateral shoot apices are slightly smaller than the
Coniferales 223
Fig. 15.4.1 A-C. Podocarpus gracilior. A Transection root shows secondary growth
and attached tubercle. B Transection root tubercle with abundant fungal hyphae in cortical
region. C Inset C from B. (After Konar and Oberoi 1969a)
terminal ones and slower in development in P. gracilior (Konar and Oberoi

1969a). Four cytohistological zones can be distinguished in the shoot apex
(Pillai 1963, Konar and Oberoi 1969a): (a) apical initials, (b) subapical
initials, (c) flanking zone, and (d) _pith mother cells and pith. The shoot
224 The Gymnosperms
apex shows variations in activity, size, topography and histological

organization.
A transection of young stem of P. gracilior displays a fluted outline due
to the presence of broad leaf bases (Fig. 15.4.2 A). The epidermis is distinct
and coverd by a thick cuticle. A young stem shows a narrow cortex of
parenchymatous cells. In an older stem, three or four layers of the outer
cortex and some isolated cells in the inner cortex contain tannin. Scattered
sclereids occur in the inner layers of the cortex. In the cortex just outside
the vascular bundles, there is a ring of large resin canals with a distinct
epithelial layer, one canal opposite each phloem patch. Two or three leaf
traces are also present in the cortex. A parenchymatous pith contains numerous
sclereids an-d resin cells (Fig. 15.4.2 A, B). A cambial ring appears in the
outer cortex, cutting off cork cells, which enclose many lenticels.
The vascular bundles remain separate from each other, just below the
shoot apex (Fig. 15.4.2 A). Secondary growth occurs at a very early stage.
The secondary phloem forms a narrow ring and consists of sieve cells,
parenchyma, fibres and rays. The sieve cells (in transection) are rectangular
and arranged in radial rows. Phloem rays are uniseriate and the tannin-
filled cells of phloem parenchyma are grouped in tangential bands. The
phloem fibres occur in a single ring.
A transection of wood shows distinct growth rings of variable width
(Fig. 15.4.2 C). The xylem consists of tracheids, parenchyma and rays. The
tracheids are rectangular (Fig. 15.4.2 D) and have uniseriate bordered pits
(Fig. 15.4.2 E, H), on both their radial and tangential walls. The rays are
homogeneous and consist of parenchymatous cells with thin, smooth
horizontal walls. They are uniseritate (Fig. 15.4.2 F, G), and two to ten
cells high, commonly three or four. One pit is present per cross field. The
xylem paenchyma is diffuse, abundant, and becomes prominent in the older
wood as it becomes resinous.
Leaf. The leaves of Podocarpus are typically bifacial with a well-defined

mid-rib: isobilateral in P. gracilior (Konar and Oberoi 1969a) and bifacial
in P. brevifolia. The internal structure shows a sharp distinction into a
compact palisade tissue on the adaxial and loosely arranged spongy cells
on the abaxial side. There are three resin ducts below the vascular bundle,
the central duct being slightly longer than the lateral ones (Kausik and
Bhattacharya 1977; Fig. 15.4.3 A). The bundle sheath is not distinct. The
vascular bundle is large and flattened dorsi ventrally, with files of parenchyma
between the rows of xylem and phloem.
In P. gracilior (isobilateral) the palisade is present on both sides. It is
amphistomatic with sunken stomata. The epidermis is covered by a prominent
cuticle, thicker at the margin and thinner over the stomata. The hypodermis
consists of small groups of sclerenchymatous cells below the upper and
lower epidermis except at the edges and above and below the vascular
Coniferales 225
Fig. 15.4.2 A-H. Podocarpus gracilior. A-D transection, E, H radial longisection,

F, G Tangentiallongisection. A young stem with a ring of vascular bundles and peripheral
resin ducts. B Mature stem with secondary growth. C Secondary xylem with growth
rings. D Secondary xylem from B. E Wood with ray cells . F, G Wood with uniseriate
rays. H Tracheids with bordered pits. (After Konar and Oberoi 1969a)
226 The Gymnosperms
bundles. The palisade usually comprises two layers. Some isolated fibres
occur around the vascular bundle and are dispersed throughout the adjacent
mesophyll. A single median resin canal occurs on the abaxial side of the
vascular bundle.
The transfusion tissue is lateral with a wing-like extension on the two
sides of the vascular bundle, and extends outward into the mesophyll
(Fig. 15.4.3 A). In P. gracilior, it comprises short spiral tracheids parallel
att~
Fig. 15.4.3 A-C. Podocarpus brevifolius. A, B Vertical section leaf. A Outline diagram
to show transfusion tissue on either side of vascular bundle and three resin ducts below
the bundle. B Transfusion tissue (ttr), accessory transfusion tissue (att), and vascular
bundle. C Accessory transfusion tracheid and transfusion tracheid. (After Kausik and
Bhattacharya 1977)
to the surface of the leaf. In P. brevifolia, the tracheids and a few parenchyma
cells occur lateral to the xylem, and the albuminous cells in the phloem.
An additional histological feature of considerable importance is the
accessory transfusion tissue in the mesophyll, between the palisade and the
spongy cells extending from the mid-rib to the two margins of the leaf. In
P. brevifolia this tissue is well developed. It consists of two distinct types
of cells: (a) thin-walled parenchyma containing large quantities of tannin
and other darkly staining inclusions (Fig. 15.4.3 B), and (b) thick-walled
tracheids with groups of bordered pits (Fig. 15.4.3 B, C). These cells are
Coniferales 227
elongated at right angles to the vascular bundle and form a net-like structure
(in paradermal sections) and tiers of cells (in transections).
Reproduction
Podocarpus is dioecious. Most of the plants produce male cones and the
seed set on the female plants is very low. In P. gracilior this is probably
due to failure of pollination (Konar and Oberoi 1969b ).
Male Cone
The male cones of Podocarpus conform to the general coniferous types and
do not exhibit much diversity. In P. gracilior the male cones-widely spread
along the short lateral branches-are borne on small pedicels which occur
singly (rarely two together) in the axils of foliage leaves (Fig. 15.4.4 A, B).
There are one to three short-stalked cones borne on the pedicel
(Fig. 15.4.4 C-E). When there are three cones, the middle one is the longer
of the two. A mature cone is ca. 2 em long and 3-4 mm in diameter.
Numerous sporophylls are spirally arranged on the cone axis. Each sporophyll
has a short narrow stalk, and a large, triangular, upwardly projecting terminal
region. Two ovoid sporangia occur on the abaxial surface, one on either
side of the stalk near the cone axis (Fig. 15.4.4 F-J). The two sporangia are
laterally fused. The line of dehiscence of the sporangium is transverse
(Fig. 15.4.4 J), as in other podocarps (Stiles 1912). The cone axis elongates
at maturity, separates the sporophylls and exposes the sporangia (Fig 15.4.4 D).
Microsporangium. The microsporangia (P. gracilior) initiate as small

hump.-like swellings on the abaxial surface of the sporophyll, near the base.
With further growth, the sporangium appears as a round sac-like structure,
and finally elongates parallel to the stalk of the sporophyll (Fig. 15.4.5 A-D).
Thfee or four hypodermal archesporia! cells differentiate, and divide
anticlinally and periclinally to form a group of sporogenous cells surrounded
by two or three wall layers (Fig. 15.4.5 E-G). As the divisions continue, the
sporogenous tissue increases enormously and the wall becomes five- to
seven-layered. The cells of the outermost and some of the inher wall layers
become resinous. The innermost wall layer surrounding the sporogenous
tissue differentiates into a tapetum. The tapetum is of the secretory type
(P. macrophyllous). The cells are densely cytoplasmic and some of them
become binucleate at maturity (Fig. 15.4.5 H). The tapetum is absorbed at
the time of the reduction divisions in the mother cells. As the microspores
mature, the cells of the middle layers become flattened and crushed
(Fig. 15.4.5 I, J). At maturity, the epidermal cells enlarge, stretch, their
radial and inner tangential walls become thick and develope upwardly directed
fibrous thickening (Fig. 15.4.6 C-E). A row of wall cells, running transversely,
remains small and unthickened, and dehiscence occurs along these cells
with the epidermis alone functioning as the wall of the sporangium
(Fig. 15.4.6 A-H).
228 The Gymnosperms
Fig. 15.4.4 A·J. Podocarpus gracilior. A Twig bearing male cones (me) in the axils
of leaves. B, C Male cones . D Dehisced cone . E Three cones in a cluster.
F-J Microsporophyll (mi) and microsporangia (ms) in various views. J Ventral view of
sporangium shows line of dehiscence (ld) . (After Konar and Oberoi 1969b)
In P. macrophyllous the nuclei of the sporogenous cells are large with

scattered chromatin. The outer membrane of the nuclear envelope shows
frequent invaginations, which may reach to the plasmalemma (at some
places). Inside these evaginations, there are several small vesicles with
dark contents, their nature is not known. I.K. Vasil and Aldrich (1970)
presume that this may constitute a line of transport. During the period of
growth, large multilayered complexes of endoplasmic reticulum (ER) are
scattered throughout the cytoplasm. Direct connections between these
multilayered complexes and nuclear envelope or plasmalemma have not
been observed (I.K. Vasil and Aldrich 1970).
Coniferales 229
~:
' A
I
Fig. 15.4.5 A-J. Podocarpus gracilior. A-D Longisection male cone, progressive stages
of microsporangial development. E-1 Development of microsporangium. spt sporogenous
tissue. tap tapetum. J Portion of sporangium with rnicrospore mother cells (mime). (After
Konar and Oberoi 1969b)
There is a difference in staining pattern between the cell walls of

sporogenous and tapetal cells , and that of other wall layers, in
P. macrophyllous. The cell walls of the wall layers are thicker and stain a
brighter magenta by periodic acid-Schiff' s (PAS) reaction than the walls of
tapetum and sporogenous cells. There are numerous vesicles in the cytoplasm
230 The Gymnosperms
~
\£Y
ep
g F
. .
~.
/
...,.
1''
Fig. 15.4.6 A-H. Podocarpus gracilior. A Microsp:>rophyllloogisection (outline diagram).

B Inset from A, epidermis (ep), tapetum (tap) and microspore tetrad. C-E Progressive
development of wall and line of dehiscence (ld); note pollen grains in C and thickenings
in epidermal cells inC-E. F Microsporophyll (ventral view). G, H Cross section dehisced
sporangium. (After Konar and Oberoi 1969b)
of both sporogenous and tapetal cells, which probably carry hydrolytic

enzymes. The walls of collapsed tapetal and sporogenous cells (prior to
meiosis) show a basic similarity in their chemical nature (I.K. Vasil and
Aldrich 1970).
In P. macrophyllous the sporogenous cells are rich in RNA throughout
Coniferales 231
their development. The RNA level is maximal in the mother cells prior to
meiosis. A loss of cytoplasmic basophilia has been observed especially
during meiosis II. The young microspore is poor in RNA, but the level rises
again on the formation of the antheridial cell.
Microsporgenesis. The microspore mother cells (mime) undergo reduction

division (Fig. 15.4.7 A-C). The cytoplasm of mime contracts, the wall
dissolves leaving the cytoplasmic masses with their prominent nuclei free
in the sporangium (Fig. 15.4.5 J). In P. macrophyllous, prior to meiosis, the
mime undergo a gradual stretching, and loss of PAS stainability. With the
dissolution of their walls, a thin osmiophilic layer of callose develops between
the cell wall and the plasmamembrane. This layer fluoresces weakly by the
aniline blue ultraviolet (UV) fluroscence test. It has been named a callose-
like wall by I.K. Vasil and Aldrich (1970). ·
Meiosis is asynchronous. Plasma channels are not formed between adjacent
sporogenous cells. In a microsporangium the meiotic stages in various
mime range from prophase I to telophase II. A callose wall has not been
reported during microspore mother cell meiosis.
Cytokinesis is simultaneous, and isobilateral and tetrahedral tetrads are
formed (Fig. 15.4.7 D, E). Several abnormalities occur during
microsporogenesis. Frequently, incomplete cytokinesis leads to double pollen
grains (Fig. 15.4.7 L). Wing formation is initiated in the microspores at the
tetrad stage. Rapid development takes place as soon as the microspores
separate from each other. The microspores increase in size, the exine at the
proximal end becomes thick and cutinized. The large nucleus is centrally
placed and the cytoplasm has a few starch grains (Fig. 15.4.7 F).
The development of the pollen grain wall has been described in
P. macrophyllus by I.K. Vasil and Aldrich (1970). The outermost layer, the
sexine, is laid down while the spores are still within the tetrad and callose
covering. The Golgi bodies in the young microspore are large and very
active. Small-unit membranous vesicles (ca. 0.1J.Lm) with granular electron-
dense contents are excreted by the Golgi bodies. These vesicles coalesce
during their transport towards the plasmalemma. A space is formed between
the plasmalemma and callose wall by contraction of the former. The space
enlarges in the region where subsequently air sacs develop. The
plasmamembrane shows many undulations or protrusions. The unit membrane
of these vesicles fuses with the plasmalemma. The cellulosic contents of
these vesicles is secreted outside the plasmalemma but inside the callose
layer, and represents the initiation of the primexine. At this stage, long
stretches of distinctly trilaminar lamellae or tapes are seen in the primexine.
Lateral evaginations appear to give rise to these trilaminar structures.
A deposition of sporopollenins on the outer surface of the vesicle contents
(lying below the callose layer) is the first sign of tectum of the sexine.
Deposits of sporopollenin also continue along the side of these vesicle
232 The Gymnosperms
•
.
.
. 0 E
8 c
A
Fig 15.4.7 A-L. Podocarpus gracilior. A-C Meiosis in microspore mother cell.
D, E Isobilateral and tetrahedral tetrad. F Uninucleate pollen grain. G Division of nucleus.
H Pollen grain with two prothallial cells (prl, pr2) and central cell nucleus. I Division
of central cell nucleus (to form a tube nucleus and a centrally placed generative cell) .
J Further divisions in prothallial cells result in a tissue; due to displacement the prothallial
cells do not appear in tiers . K Pollen grain at shedding stage; sterile nuclei lie free in
the cytoplasm. be body cell. tn tube nucleus. L Double pollen grain. (After Konar and
Oberoi 1969b)
contents (sides of trilaminar tapes) to form the baculae. The fine fibrillar
material of Golgi vesicles condenses against the callose wall and its
protrusions.
The nexine I is the next layer to be laid down. It is formed by the
deposition of sporopollenin on trilaminar tapes in the inner part of the
primexine during the tetrad phase. At least five such layers of tapes are
visible. The outermost layer has a richer deposition of sporopollenin than
the inner foot layers. Lipid granules form the primary source for nexine I,
Coniferales 233
and the tapes probably originate from the plasmamembrane or endoplasmic

reticulum. The nexine surrounds the cytoplasm except at the regions where
wings develop.
Further development takes place when the microspores are liberated
from the tetrads. Small granular particles of sporopollenin present between
plasmalemma and nexine I coalesce to form nexine II. A third layer, nexine
III, is also present in P. macrophyllous. This layer is formed after the
microspore nucleus has given rise to supernumerary prothallial cells (see
below). The development of nexine III is similar to that of nexine II, and
no trilaminar membranes or tapes are thus involved. This is the most electron-
dense layer of the pollen wall.
The intine is the last layer to be laid down. Numerous Golgi vesicles
take part in its development and empty their contents across the plasmalemma.
It is homogenous and without any lamellations of fibrous character.
The unit member, which plays an important role in pollen wall formation,
develops from the plasmalemma.
The nuclear envelope in a mature pollen grain consists of two membranes
with occasional pores of 0.1 Jlm in diameter. Finger-like invaginations,
involving both layers of the nuclear envelope, appear when the microspores
are still within the tetrad, and extend into the nucleoplasm in P. macrophyllous
(Aldrich and I.K. Vasil 1970), At the proximal end of these invaginations
are structures comparable to nuclear pores. Fibrillar material interpreted as
achromatin (I.K. Vasil and Aldrich 1970) fills the invaginations, and is
discharged into the cytoplasm, the nuclear envelope then becoming regular
once again. In addition to invaginations, there are evaginations of the outer
nuclear membrane extending into the rnicrospore cytoplasm. These are filled
with vesicles ca. 1 Jlm in diameter, surrounded by unit membranes. Nuclear
pores occur at non-evaginated portions only. A continuity between the
outer nuclear envelope and the plasmalemma has also been observed,
indicating nucleocytoplasmic transfer or exchange (Aldrich and I.K. Vasil
1970). These membranes can serve as sites for the synthesis of various
substances in preparation for meiosis. They may also contribute enzymes
for the breakdown of the mother cell wall and subsequent formation of the
callose layer. The space between the two nuclear membranes may act as a
channel for transport of exine precursors. Sporopollenin-like deposition
occurs on Golgi cisternae and elements of endoplasmic reticulum within
micros pore mother cells during rnicrosporogenesis (see Moitra and Bhatnagar
1982).
Male Gametophyte. At the proximal end, a mature rnicrospore nucleus cuts

off successively two lenticular primary prothallial cells (Fig. 15.4.7 G, H),
which undergo further divisions to form three to four cells (Fig. 15.4.7 J).
Simultaneously, or prior to the second division of these cells, the nucleus
of the central cell divides to form a large tube nucleus towards the distal
234 The Gymnosperms
end and a centrally placed antheridial or generative cell towards the proximal
end (Fig. 15.4.7 I). The generative cell divides to form a stalk and a body
cell. It is difficult to distinguish the stalk cell from the prothallial tissue,
whose cells also contain prominent nuclei (Fig. 15.4.7 J). At maturity, the
wall of all the cells (except the body cell) dissolve and the nuclei lie free
in the general cytoplasm of the pollen grain (Fig. 15.4.7 K). The cytoplasm
contains abundant starch grains. At the time of shedding, a pollen grain
shows a maximum of six free nuclei, in addition to the body cell
(Fig. 15.4.7 K).
Female Cones
The female strobili in P. gracilior is axillary and occurs at the distal end
of a vegetative shoot. The fertile branch can be distinguished from a vegetative
branch only after pollination when the ovule emerges from the subtending
bracts. Each fertile branch is ca. 2.5 em long and bears five to ten leafy
bracts (Fig. 15.4.8 A). The uppermost bract (rarely two) is fertile, and bears
in its axil a single unitegmic anatropous ovule (Fig. 15.4.8 B). At the time
of pollination it is nearly ovoid. The fertile bract does not grow further, and
remains on the side away from the micropyle. The ovuliferous scale, or
epimatium, encircles the ovule completely and is free from it at the micropylar
A B c
Fig. 15.4.8 A-C. Podocarpus gracilior. A, B Twig bearing female strobili (Js).
ov ovule. C Mature seed. (After Konar and Oberoi I969b)
end. The young ovule is externally covered by a white, shiny waxy deposition.
The mature seed (Fig. 15.4.8 C) is ca. 2.5 em in length and 2 em in
diameter. It is covered over by a thick fleshy seed coat which appears
yellowish green at mautrity. The inner part of the seed coat is hard, stony,
sharply pointed at the micropylar end, and has numerous small pits.
Ovule. In P. gracilior, the earliest stage showed an undifferentiated hump

which is carried a little above the shoot apex by a short stalk, a short
proJected nucellus in the middle, and the single integument above it. At this
stage, the ovule is more or less hemianatropous. Simultaneously, the
epimatium (another integument-like structure) appears on the side of the
Coniferales 235
ovule (Fig. 15.4.9 A). The ovule grows rapidly, becomes anatropous, and
has a wide micropylar opening. The epimatium is completely fused with
the ovule except at the micropylar end, and is slightly shorter than the
integument (Fig. 15.4.9 E).
F
Fig. 15.4.9 A-G. Podocarpus gracilior. A, E Longisectim young ovule (outline diagram).
epi epimatium. i integument. n nucellus. B Nucellus from A, megaspore mother cell.
C, D, F, G Formation of linear megaspore tetrad; note the circumjacent spongy tissue
(sg). (After Konar and Oberoi 1969b)
236 The Gymnosperms
Megasporogenesis. A megaspore mother cell (mgmc) originates from a

hypodermal archesporia! cell and becomes deeply embedded due to the
periclinal divisions of the cells above it (Fig. 15.4.9 B, C). A tapetum is
present in P. andinus, P. nivalis, P. falcatus and P. gracilior. The ovule
(P. gracilior) becomes somewhat exposed, and, for about a week, a pollination
drop is secreted at its micropylar end. As the ovule grows, the part of the
integument lying above the nucellus elongates to form a long narrow
micropylar canal, which may be occasionally curved. The cells lining the
upper part of the integumentary canal elongate inwardly, develop thickenings
in the form of striations, and close the micropyle after pollination. Later,
these cells lose their contents and look like tracheidal cells.
The redQction division in the mgmc (Fig. 15.4.9 D) results in a row of
three cells (uppermost dyad and two megaspores) in P. andinus (Looby and
Doyle 1944a), P. nivalis (Boyle and Doyle 1953), and a linear tetrad in
P. gracilior (Fig. 15.4.9 F; Konar and Oberoi 1969b), P. spicatus,
P. ferrugineus (Sinnott 1913) and P. falcatus (Osborn 1960). The chalazal
megaspore is functional (Fig. 15.4.9 G).
Female Gametophyte. The functional megaspore enlarges and becomes

vacuolate (Fig. 15.4.10 A). Free-nuclear divisions follow and the nuclei are
distributed peripherally (Fig. 15.4.10 B-D). Centripetal wall formation begins
after several hundred free nuclei are formed. In P. andinus, alveolation occurs
with an open inner face (Looby and Doyle 1944a). The gametophytic cells
are arranged in files converging towards the centre; they are thin-walled
and contain scanty cytoplasm. The two-layered megaspore membrane remains
comparatively thin in the micropylar region of the gametophyte. The latter
is without any definite shape due to its early contact with the pollen tube.
The prothallial tissue may grow around the pollen tube.
The archegonial initial is a superficial cell (Fig. 15.4.11 A). It has scanty
cytoplasm and a nucleus which grows in size. The development is very
rapid (Fig. 15.4.11 B-D, G) and a mature archegonium has a neck which
consists offour to six cells (P. gracilior, P. nivalis) or 10--15 cells (P. andinus)
arranged in one (rarely two) tier. The neck cells degenerate or become
inconspicuous soon after their formation. There is a small ventral canal
nucleus and an egg nucleus, the latter shifts to a central position and is
surrounded by a thick sheath of cytoplasm (Fig. 15.4.11 G). Gradually, the
cytoplasm of the egg becomes thick and has few vacuoles and darkly
staining masses appear, especially near the egg nucleus.
A mature archegonium is covered by a thin layer of the megaspore
membrane which sometimes breaks down due to the impact of the pollen
tube (P. gracilior).
A conspicuous jacket layer is present, its cells become multinucleate at
maturity. Occasionally, before fertilization, the nuclei may escape into the
archegonium. Groups of archegonia surrounded by a common jacket are
also frequent.
Coniferales 237
Fig. 15.4.10 A-D. Podocarpus gracilior. A Longisection central part of nucellus with
functional megaspore, and three degenerated megaspores. B, C Free-nuclear gametophyte
(whole-mount). D Portion of free-nuclear gametophyte. (After Konar and Oberoi 1969b)
There are two, rarely three, archegonia in P. andinus and P. nivalis. In

P. gracilior, the number varies from 10-12; occasionally, as many as 22
have been counted (Konar and Oberoi 1969b). The shape of the archegonium
depends on its position on the prothallus, and may be round, oval or elongate
(Fig. 15.4.11 E, F). The formation of archegonia is markedly influenced by
the presence of the pollen tube, and shows various stages of development
in the same prothallus. Immature archegonia are present along the path of
the pollen tube. The archegonia situated close to the spermatogenous (body)
cell show a more advanced stage of development than those away from it.
Pollination
At the free-nuclear stage of the gametophyte, tre ovules (P. gracilior) secrete
a pollination drop in the early hours of the day, continuously for nearly a
238 The Gymnosperms
G K
Fig. 15.4.11 A-K. Podocarpus gracilior. A-D Development of archegonium. en egg
nucleus. vcn ventral canal nucleus. E, F Outline diagrams of micropylar portion of
gametophyte at fertilization, irregular arrangement of archegonia. pt pollen tube.
G Mature archegonium shows thick cytoplasmic sheath around the egg nucleus. H Two
male gametes (mg) in archegonium. sn sterile nuclei. I Fusion of male and female nuclei.
J, K Archegonia show several nuclei in the egg cells formed by division of the unfertilized
egg nucleus. (After Konar and Oberoi 1969b)
Coniferales 239
week. The pollen grains are caught in the pollination drop and sucked into
the micropyle. Konar and Oberoi (1969b) also observed a large number of
ovules pollinated with the pollen grains of Pinus (in the micropyle), besides
the pollen of Podocarpus (Fig. 15.4.12 A, B). There was a pine forest in
the neighbourhood. The pollen grains of pine abort after some time.
Post-pollination Development of Male Gametophyte. On reaching the

nucellar surface, the pollen grain germinates immediately (Fig. 15.4.12 A, B).
The intine bursts through the exine, a small pollen tube forms at the proximal
end of the pollen grain and penetrates the nucellus but remains unbranched
(Konar and Oberoi 1969b). The tube nucleus is the largest among the
sterile nuclei and is the first to enter the pollen tube. It is followed by the
rest of the sterile nuclei. The spermatogenous (body) cell is the last to enter
the pollen tube (Fig. 15.4.12 B). In all species of Podocarpus, the
spermatogenous cell lags behind in the pollen grain.
In P. andinus the spermatogenous cell overwinters in the grain, while
the sterile nuclei·pass into the short pollen tube. Later, the spermatogenous
cell overtakes the sterile nuclei in the pollen tube and comes to lie ahead
of them. In its early stages of growth, the pollen tube remains narrow and
unbranched in the nucellus (P. andinus, P. gracilior). Later, its growth through
the nucellus is rapid, and the pollen tube comes in contact with the (enlarged)
female gametophyte, which is still at the free-nuclear stage (P. gracilior,
P. falcatus). The sterile nuclei, as well as the spermatogenos cell, increase
in size (Fig. 15.4.12 C-E).
On reaching the inner surface of the nucellar cap, the pollen tube expands
into a more or less cylindrical vesicle (bulla) and expands maximally in the
space between the nucellar cap and the female gametophyte (Konar and
Oberoi 1969b). The bulla gives off lateral branches and the narrow tubes
may themselves branch again (Fig. 15.4.13 A-E) and ramify on the inner
surface of the nucellar dome and outer surface of the female gametophyte.
The branched nature has been clearly documented from dissections by
Konar and Oberoi ( 1969b). A lateral burrowing of the branches is a normal
feature in P. gracilior and P. falcatus. The nuclear contents of the pollen
tube lie within the bulla just above the female gametophyte in P. nivalis
(Boyle and Doyle 1953), P. gracilior (Konar and Oberoi 1969b, Mehra
1991) and other members of the family Podocarpaceae.
Johri (1992) discusses the haustoria! role of pollen tubes in several
angiosperms and gymnosperms. In P. gracilior the pollen tube branches
but does not grow intercellularly in the nucellar and gametophytic tissue.
There is also no evidence of the collapse of the adjoining epidermal cells.
Therefore, a haustorial function is out of the question (see also Konar and
Oberoi 1969b). Mehra (1991), who also studied P. gracilior, concludes, on
the basis of denser cytoplasm at the tip of tubes (dissected from macerated
previously.fixed material), that the pollen tube branches have a haustorial
role. This presumption is not correct (see Johri 1992).
240 The Gymnosperms
Fig. 15.4.12 A-E. Podocarpus gracilior. A Longisection ovule (outline diagram) with
pollen grains (pg) in micropylar canal and nucellus. B Nucellar tip from A, the tip of
one of the three pollen grains has grown up to the nucellus. be body cell. Ppg Pinus
pollen grain. C Further growth of pollen tube in nucellus. sn sterile nuclei. D longisection
ovule (outline diagram). pt pollen tube. E Pollen tube from D. (After Konar and Oberoi
l969b)
Coniferales 241
Fig. 15.4.13 A-E. Podocarpus gracilior. A Cellular female gametophyte (/g) with pollen
tubes (pt) . B-E Profuse branching of pollen tubes from bulla (whole-mount) . be body
cell. mg I functional, mg2 nonfunctional male gamete. sn sterile nuclei . (A-D After Konar
and Oberoi 1969b; E after Mehra 1991)
The functional pollen tube, on reaching the female gametophyte, contains

an enlarged spermatogenous cell (Fig. 15.4.14 A). The nucleus is eccentrically
placed in the dense mass of cytoplasm (Fig. 15.4.14 B). Two or three small
sterile nuclei surround the spermatogenous cell. the remaining nuclei lie
away from it. The prothallial nuclei, tube nucleus, and the stalk nucleus
reach the female gametophyte in a healthy state. By the time cell formation
242 The Gymnosperms
begins in the free-nuclear female gametophyte, the spermatogenous nucleus

divides. In P. andinus, two unequal male gametes are formed (Looby and
Doyle 1944a). In P. nivalis, one gamete remains small and is pushed out
of the functional male cytoplasm. In P. gracilior, at first the two gametes
appear to be equal but later become unequal (Fig. 15.4.14 C-F). Gradually,
the smaller gamete is partly or wholly extruded, while the larger gamete
comes to lie in the centre of the cytoplasm. The lattter shows a thick
nuclear membrane, and a dark, densely staining homogenous nucleoplasm
with no nucleolus. Later, irregular densely staining bodies appear in the
cytoplasm of the male gametes and granular bodies appear within its nucleus
(Fig. 15.4.14 E, F). Frequently, the non-functional male gamete enlarges
slightly, shows a homogenous cytoplasm, and becomes demarcated from
the functional gamete. The functional male gamete escapes from the
cytoplasm, and enters the archegonium laterally, or through the neck cells.
Along with the functional gamete, several sterile nuclei may also enter into
the archegonium. More commonly, however, they degenerate outside the
archegonia.
Fertilization
In P. gracilior, the pollen tube displaces the neck cells laterally, demolishing
a part of the archegonial wall, and the contents of the tube are discharged
into the archegonium. The functional male gamete with the cytoplasm enters
the archegonium. The non-functional male gamete and some degenerated
sterile nuclei lie outside the fertilized archegonium. Sometimes, both the
male gametes and the sterile nuclei have been observed in the archegonium
(Fig. 15.4.11 H). The egg nucleus of an unfertilized archegonium may
divide to form a few nuclei (Fig. 15.4.11 J, K), which eventually degenerate.
The fusion of the male gamete and egg nucleus occurs in the upper part of
the archegonium (Fig. 15.4.11 1). In P. andinus, a unique phenomenon has
been described (Looby and Doyle 1944b): the archegonial cytoplasm is
discharged betwen the neck cells and pushes the male gametes backward.
This is followed by a contraction of the cytoplasm and the functional male
gamete back into the archegonium. According to Looby and Doyle (1944b),
this is a normal feature and not due to any pressure on the turgid archegonia
during collection and fixation.
Embryogeny
Podocarpus is an exceptional taxon among the conifers, as its embryogeny
conforms to several distinct types (see Roy Chowdhury 1962). The basic
plan of development is, however, uniform and consists of: (a) nuclear division
of the zygote followed by four to five free-nuclear divisions to form
16-32 nuclei, (b) arrangement of nuclei in two tiers followed by wall
formation, (c) the cells of both the tiers divide, (d) the upper tier (pU) forms
an open tier (U) and the lower a suspensor tier (S), and (e) the division in
Coniferales 243
E F
Fig. 15.4.14 A-F. Podocarpus gracilior. A Part of pollen tube with body cell (be) and
some sterile nuclei. B Body cell just before divisions. C Body cell with nearly equal
eccentric male nuclei. D-F Maturation of functional male gamete (mgl, larger) and
extrusion of the non-functional male gamete (mg2, smaller). sn sterile nucleus. (After
Konar and Oberoi 1969b)
the lower tier (p£) is not followed by a wall, so that its cells (E) become
binucleate. The binucleate condition of the cells of tier E is characteris.tic
of the family Podocarpaceae.
244 The Gymnosperms
In P. gracilior, the zygote is surrounded by dense non-vacuolate

protoplasm. The nucleus moves to the lower part of the archegonium and
divides (Fig. 15.4.15 A). These nuclei undergo four free-nuclear divisions,
forming 32 nuclei (Fig. 15.4.15 B-E). This is also characteristic of the
. .· ... . .
.•, ..· . . :.· :· ~
.. -:.:. .
.·~·~
-~ · ._
; ~ .. .
...
Fig. 15.4.15 A-1. Podocarpus gracilior. A-C 2-, 4-, and 8-nucleate proembryo. rn
relict nuclei. D, E Transection 32-nucleate proembryo through upper region. F-H Two-
tiered proembryo with nuclei dividing in one or both tiers. p£ primary embryonal tier,
pU primary upper tier. I Three-tiered proembryo, the embryonal cells (E) are binucleate.
U upper tier. S suspensor tier. (After Konar and Oberoi 1969b)
Coniferales 245
primitive species of the sections Stachycarpus, Afrocarpus, Sundacarpus

and Nageia. In the advanced sections, Eupodocarpus and Dacrycarpus, only
16 free nuclei are formed.
In P. falcatus (Afrocarpus), the proembryo shows 16 and 32 nuclei, 16
nuclei are more frequent (Osborn 1960).
During the free-nuclear phase, some of the proembryonal nuclei escape
into the upper part of the archegonium, persist till a much later stage of
embryogeny, and eventually degenerate. Looby and Doyle (1944b) consider
these as relict nuclei. They are more frequent in the sections of Podocarpus
where five free mitoses occur.
During progressive divisions, the proembryonal nuclei decrease in size.
Wall formation results in two tiers. The lower embryonal tier (pE)consists
of 9-12 cells and the upper open tier (pU) of 15-22 cells. The embryonal
cells are usually aranged in a single tier, so that each cell is in direct
contact with the cells of the open tier, or the main group of pE cells
surrounds 3-4 cells in the center. The group of inner cells are comparatively
smaller and remain in direct contact with the suspensors; some of the cells
lying below the inner cells have no direct contact with the cells of the open
tier. All the nuclei in pU and pE tier undergo an internal division
(Fig. 15.4.15 F-H). In pU, wall formation results in an upper tier (U) whose
cells remain open, and a middle suspensor tier (S). In the pE tier the divisions
are not followed by walls and the cells of this tier (E) become binucleate
(Fig. 15.4.15 I).
There is variation in the ratio of cells in the S and E tiers among different
species of Podocarpus. The total number in these tiers is determined by the
number of free mitoses and the number of relict nuclei. However, the
number of E cells in a species or a group of related species shows a narrower
range of variation than the corresponding number of S cell. In Afrocarpus,
Stachycarpus, and Nageia sections the average number of E cells is ca. ten,
in Dacrycarpus it is four, while the most advanced Eupodocarpus shows
one or two E cells.
A terminal cell in the embryonal group is reported in P. spicatus,
P. ferrugineus (Buchholz 1936) and P. andinus (Looby and Doyle 1944b).
This cell probably facilitates penetration of the embryo into the starchy
endosperm. In P. gracilior the distal end of some of the E cells develops
thick cellulose caps (Konar and Oberoi 1969b). In P. nivalis an apical
enucleate portion is cut off in each embryonal cell (Boyle and Doyle 1954).
Differentiation of Embryo. After the embryonal cells become binucleate,

the tierS elongates (Fig. 15.4.16 A, B). The nuclei of the tier U escape from
the cells, form a group and eventually degenerate. The development from
the time of fertilization to the elongation of suspensor takes 3-4 days. The
gametophytic cells around the developing proembryo become multinucleate
and filled with starch. The embryonal cells are pushed through these cells
246 The Gymnosperms
due to the elongation of the S tier. During elongation, many suspensor cells
become detached, so that only a few cells remain in contact with the embryonal
cells. The open end of such detached suspensor cells frequently proliferates
to form small groups of cells (Konar and Oberoi 1969b). Such proliferation
is common to other species (see Roy Chowdhury 1962). Occasionally, the
cells of the U tier form groups of "cells simulating embryonal masses.
The period of suspensor elongation is ca. 3 weeks. During this time the
embryonal cells are in a "resting" stage, appear collapsed, and the cytoplasm
stains deeply. Due to continued elongation, the suspensor becomes highly
coiled and lies in the cavity formed by the breaking up and degeneration
of the gametophytic tissue. After the suspensor tier ceases to elongate,
the embryonal cells become active and the nuclei and nucleoli become
prominent. The two nuclei of the embryonal cell divide simultaneoulsy
(Fig. 15.4.16 C, D), followed by wall formation to form uninucleate cells.
These cells are in groups of four, referred to as the embryonal tetrad (Looby
and Doyle 1944b), and are a characteristic feature of the family (see Doyle
1954). The divisions in different cells of the embryonal group are not
synchronous; some of the smaller inner cells may fail to divide and degenerate
(Fig. 15.4.16 C, D).
Rapid divisions in the cells of the embryonal tetrad result in distinct
lobing (Fig. 15.4.16 E), each lobe corresponds to a separate embryonal
tetrad and remains distinct during later development. Finally, the lobes
separate from each other due to the elongation of their upper cells, which
function as secondary suspensor. Each embryonal tetrad separated by cleavage
is a potential embryo (P. gracilior).
Cleavage polyembryony in Podocarpus appears to be related to the
arrangement of cells in the tier E. In P. andinus (Looby and Doyle 1944b),
P. spicatus and P. ferrugineus (Buchholz 1936), theE cells are arranged in
superimposed tiers, and there is no cleavage of the embryonal group. In
other species, such as P. amarus, P. usambarensis (Buchholz 1941) and
P. gracilior (Konar and Oberoi 1969b), and in other advanced sections
where the E cells are arranged in one horizontal tier, each embryonal cell
represents a potential embryo.
In P. gracilior, most of the embryonal groups may become suppressed
at a very early stage, so that ultimately there is only a single mass of
embyonal cells at the tip of the massive secondary suspensor. For some
time, meristematic activity and divisions in all planes in the lower half of
the embryonal mass adds to the over-all size, while the other half continues
to contribute to the massive secondary suspensors.
During early ontogeny, a dermatogen-like layer differentiates around the
proximal end of the embryonal mass, and undergoes periclinal divisions.
By the time the cotyledons are initiated, a stem tip and a root tip can also
be clearly discerned. In the central part of the embryo, between the root
and shoot tip, the cells are elongated and the procambial strand passes into
Coniferales 247
''; • •1.
<:;:;:;~q~~ft~:.:~)·~)
Fig. 15.4.16 A-E. Podocarpus gracilior. A, B Three-tiered proembryo,at the beginning

of suspensor elongation. C, D Longi-and transection of embryonal tier, some of the
embryonal cells have degemerated. E Lobing of embryonal tier after formation of
embryonal tetrad. (After Konar and Oberoi 1969b)
each cotyledon. Before any further organization, the embryo elongates

considerably. The mature embryo shows a well-marked stem and root apex.
In some mature seeds, the embryo has a well-developed plumular leaf
lateral to the shoot apex.
The fleshy seeds of P. andinus and P. ferrugineus fall to the ground before
the embryo matures (cf. Cycas , Ginkgo and Gnetum , where further
development of the embryo takes place only after the seeds are shed). In
P. andinus, the oldest embryo showed initial stages of cotyledonary
development (see Roy Chowdhury 1962).
248 The Gymnosperms
Seed
In addition to the integument, the seed coat includes a thick layer of
epimatium. The tissues of the integument and epimatium are difficult to
demarcate, as they are completely fused with each other except for a short
distance at the micropylar end.
At the time of pollination, the integument consists of several layers of
thin-walled cells, some of which, in the hypodermal region, contain tannin.
At the free-nuclear stage of the gametophyte, some cells of the integument
at the level of the nucellar beak contain tannin and resinous material. As
the ovule enlarges the tanniniferous layer extends towards the micropyle
and around the nucellus (Fig. 15.4.17 A-D). Simultaneously, the cells around
the female gametophyte and interior to the tannin layer become
sclerenchymatous (Fig. 15.4.17 A-D). This thickening extends towards
the micropyle to the tanniniferous layer. During and after the cellular
stage of the gametophyte, the seed coat differentiates into three zones
(Fig. 15.4.17 E, F): (a) an inner zone of sclerenchymatous cells around the
nucellus; (b) a middle two-/three-layered zone of tannin-filled cells, and (c)
an outer several-layered zone of parenchymatous cells. Stomata are present
on the epidermis. During the maturation of archegonia and early embryogeny,
two to three layers below the epidermis become completely resinous. A
few scattered tracheids are also present in the outer parenchymatous region.
The hard, stony sclerenchymatous zone becomes exposed after the seed is
shed and the two outer zones disintegrate.
The mature seeds of P. henkelii at shedding have a moisture content of
ca. 62%; when incubated on a moist substrate they show slow and sporadic
germination (Palmer and Pitman 1972). The removal of epicuticular wax,
the epidermis, or the entire epimatium, leads to rapid water uptake and
germination (Noel and van Staden 1975). The epimatium acts as a barrier
to germination by restricting water uptake, and also permits a rapid and
uncontrolled water loss, which can lead to loss of viability (Noel and van
Staden 1975, Dodd and van Staden 1981). The seeds are at a double
disadvantage by their inability to imbibe water readily and control dessication,
which probably contributes to the limited plant regeneration in nature (von
Breitenback 1965).
Short-term stroage of seeds (up to 18 months) is possible by holding
seeds of high moisture content at 4 oc (Dodd and van Staden 1981). Over
a 16-week period of storage, the seeds remain metabolically active and
further embryonic growth occurs. Dodd et al. (1989) investigated the
biochemical changes and fine structure of the female gametophyte and
embryo of such stored and scarified seeds, during the course of germination.
After a 3-day incubation at 25 °C, the cells of the root tip (at the electron
microscope level) show increased vacuolation, numerous amyloplasts and
lipid mobilization. By day 6, there is measurable embryonic growth and
ultrastructural evidence of synthetic activity by the presence of abundant
Coniferales 249
c E F
Fig. 15.4.17 A-F. Prodocarpus gracilior. A-D Longisection ovule (outline diagrams).
epi epimatium. fg female gametophyte. i integument. n nucellus. of outer fleshy layer.
stl stony layer. tan tanniniferous zone. A-C Free-nuclear gametophyte. D Cellular
gametophyte at fertilization. E Longisection of mature seed with persistent female
gametophyte, and dicotyledonous embryo. F Transection mature seed through
hypocotyledonary region of embryo. (After Konar and Oberoi 1969b)
endoplasmic reticulum (ER), ribosomes, and dictyosomes. Fine-structural

changes occur in the female gametophyte and suggest mobilization of reserve
food. By 9 days of incubation, biochemical studies indicate a gradual decline
in lipid and protein in the female gametophyte and embryo. At day 6, a
decline in the embryonic starch levels contrasts with the increase in the
female gametophyte. Between days 3 and 6, minor changes occur in the
sugar level of the female gametophyte in contrast to the increase in embryonic
tissues. These changes coincide with the first phase of germination. The
retention of high moisture and the evidence of metabolic activity suggest
that, following scarification, the transition between maturation and germination
250 The Gymnosperms
is characterized by a continuation of earlier synthetic activity and reserve

inter-conversions.
Germination. There is no dormancy period, and young seedlings develop

within 3 months (P. gracilior). The germination is epigeal (Fig. 15.4.18 A-E).
The radicle emerges after the stony layer cracks laterally. The stony layer
is cast off at an early stage and the hypocotyl elongates carrying the cotyledons
covered with remnants of the female gametophyte (endosperm). There are
two, rarely three, cotyledons, which persist for a long time. The juvenile
leaves are short and arranged in loose whorls (Fig. 15.4.18 F-H).
~
~
A
~ B
E
Fig. 15.4.18 A-H. Podocarpus gracilior. A Mature seed. B-E Germination of seed.
F -H Later stages of seedling with cluster of leaves. (After Konar and Oberoi !969b)
Chromosome Number
The haploid number, n = 17, has been determined from endosperm squashes
in P. neriifolius (Mehra 1988). Three chromosomes are isobrachial with
median primary constriction (marked M) and six chromosomes with terminal
or near-terminal primary constriction (Fig. 15.4.19 A).
In P. macrophyllus the haploid number is n = 19. Three chromosomes
are isobrachial, ten heterobrachial, and six have terminal or near-terminal
primary constriction (Fig. 15.4.19 B).
Coniferales 251
A B
Fig. 15.4.19 A, B. A Podocarpus neriifolius (n = 17), three chromosomes isobrachial
with median primary constriction (marked M), eight chromosomes whith subterminal
primary constriction, and six chromosomes with near-terminal primary constriction.
B P. macrophyllus (n = 19), three chromosomes isobrachial, ten heterobrachial, six with
terminal primary constriction. (After Mehra 1988)
Two types of reproductive cycles have been recognized in Podocarpus
(Doyle 1954: a 2-year type, as in P. ardinus (Looby and Doyle 1944b),
P. falcatus (Osborn 1960) and P. gracilior (Konar and Oberoi 1969b), while
in the other type, the cycle is completed in 1 year, as in P. nivalis (Boyle
and Doyle 1954).
15.5 ARAUCARIACEAE
The family Araucariaceae comprises two extant genera, Agathis and

Araucaria. These are tall evergreen trees reaching a height of 60 m or
more. Agathis has 13 spp. (Whitmore 1980a) and Araucaria 18 spp.
(de Laubenfels 1972). They are confined to the Southern Hemisphere, with
a very restricted distribution (see Chap. 1).
The Araucariaceae has a long and extensive fossil record. The remains
of plants from the Southern Hemisphere are also recorded from the Northern
Hemisphere. The fossil record indicates that this family was considerably
larger, more diverse and widespread during the Mesozoic. Unquestionable
araucarian remains extend into the Jurassic; there were numerous taxa in
the Northern Hemisphere at that time. Petrified seed cones from the Middle
Jurassic of Argentina indicate thatAraucaria had evolved by that time (Calder
1953). While there are several reports of araucarian remains in the Triassic
(Bock 1954, 1969), these are mostly impression fossils with dubious affinities
to the family (Stockey 1982). The members of Araucariaceae first appear
with certainity in the Late Triassic, seem to reach a peak of diversity during
the Jurassic and decline towards the end' of the Cretaceous (Miller 1977).
252 The Gymnosperms
Araucaria
The name is adopted after Arauco, a province of Chile, the native habitat
of A. araucana (monkey puzzle tree). According to Endlicher (1842),
Araucaria comprised two sections: Eutacta and Columbea. Wilde and Eames
(1952) divided Araucaria into four sections based on extant species.
a) Section Eutacta Endlicher. Leaves reduced, thick, often keeled,

imbricate and erect. Male cones solitary, terminal, female cones on long
peduncles. Ovulate cone scales thinly winged, indehiscent, the seed retained
on the scale at shedding. Vascular supply of bract-scale unit single at
source. Germination epigeal. Cotyledons subsessile, freed from seed at
germination. Seedling not fleshy; 15 spp., including: A. biramulata Buchholz,
A. columnaris (Forster) Hooker, A. cunninghamii Aiton ex Lambert, A.
heterophylla (Salisbury) Franco, A. memorosa de Laubenfels.
b) Section Columbea Endlicher Emend.Wilde and Eames (1952). Leaves

large, generally thin, flat. Male cones axillary, usually two to several on
foliaceous branches. Ovulate cone scales nut-like, wings absent, indehiscent,
the seed retained on the scale at shedding; vascular supply of bract-scale
unit single at source. Germination hypogeal. Cotyledons long-stalked at
germination, retained in seed coats, stalks not fused. Seedling fleshy, without
a long subterranean dormant period; two spp. A. araucana (Molina) K. Koch,
and A. angustifolia (Bertoloni) 0. Kuntze.
c) Section lntermedia. White (1947). Leaves large, generally thin, flat,

spreading, sometimes slightly imbricate; juvenile leaves needle-like, flat,
small. Male and female cones axillary; ovulate cone scales samara-like
with broad thin wings, indehiscent; seed retained on the scale at shedding.
Germination epigeal. Cotyledons subsessile, freed from seed coats at
germination. Seedling not fleshy. Three spp: A. hunsteinii K. Schumann, A.
klinkii Lauterbach, and A. schumanniana Warburg.
d) Section Bunya.Wilde and Eames (1952). ·Leaves large, flat, spreading

or slightly imbricate. Male cones axillary, female cones subsessile or on
short (2 em long) peduncles. Ovulate cone scales large, heavy with woody
rings; dehiscent, large "seed" shed from the scale at maturity. The vascular
supply of bract-scale unit double at source. Germination hypogeal. Cotyledons
long-stalked at germination, retained within seed coats, the stalks fused
into a hollow cylinder. Seedling fleshy with a long subterranean dormant
period. One sp: A. bidwillii Hooker.
Morphology
The young evergreen trees are symmetrical, clothed with branches from
base to apex, the old trees have the trunk clean of branches for the greater
Coniferales 253
part of their height; branches are horizontal and in whorls, leaves persist
for many years, and are extremely variable in morphology. They may be
sharply pointed, needle-like and imbricate (A. memorasa; Fig. 15.5.1 A, B)
or broad and imbricate (A. angustifolia, A. bidwillii, A.ldinkii; Fig. 15.5.1 C, D).
Fig. 15.5.1 A-D. A, B Araucaria memorosa. A Juvenile foliage. B Mature foliage and
young female cones (jc) at pollination. CA. klinkii and D A. bidwillii, mature foliage .
(After Stockey 1982)
254 The Gymnosperms
The leaves are spirally arranged, clasp the stem and overlap, or may occur
in two or more ranks by means of a basal twist. They are leathery, lance-
shaped and sharply pointed, or awl-shaped and four-angled , or triangular.
The size and shape of the leaf varies on different parts of the same tree.
This taxon is dioecious, occasionally the male and female cones are
borne on different branches of the. same tree.
Anatomy
Stem. The wood of Araucaria is mostly whitish or very light coloured.

In A. brasiliana, irregular red streaks are present. Leaf traces occur and are
often numerous.
The wood of extant araucarias can be distinguished readily from other
conifers by the absence of resin canals or resin cells (Fig. 15.5.2 B). Resin
occurs in the ray cells and axial tracheids as resin plugs (Jane 1970).
Multiseriate circular bordered pits are present in two to five vertical rows,
which are characteristically alternate and appear hexagonal due to crowding
(Greguss 1955; Fig. 15.5.2 A). Pitting on the ray cells is cupressoid and
rays are 1- 20 cells high. The family is also peculiar in lacking crassulae.
-.. ....
... ....," £1. I
"''
II .r
~:.:.
u P"'
TXI.l
·..W
~
l
..
r
'Kilt
I
..J ..
_:J.,
.J_j
I.
~
.I
n
,...
I
I
... :J' ]I l. l,l
~""'.~~.lil ,,~
-
.,. .r"l' I. T T
T¥"J:
Fig.15.5.2 A, B. Araucaria heterophylla, wood anatomy. A Radiallongisection shows

characteristic alternate pitting. B Transection, note absence of resin canals. (After Stockey
1982)
Leaf. The cuticle of Araucaria leaf is thick and the micromorphology of

juvenile, adult, herbarium and preserved leaves has been studied with scanning
electron microscopy (SEM). The micromorphological features reveal much
Coniferales 255
more variability than was presumed previously (Stockey and Ko 1986).

The external and internal characters of abaxial and adaxial cuticle have
been characterized for the four sections of the genus.
The outer surface of Araucaria leaf is relatively smooth (Fig. 15.5.3 A).
There are no stomatal papillae, "Florin rings" (Buchholz and Gray 1948),
or other surface ornamentation. However, irregularly shaped platelets of
unknown origin are scattered over the cuticular surface frequently on the
small needle-like leaves of Section Eutacta. All the species have stomatal
plugs (Fig. 15.5.3 B) sunken into the stoma on both adaxial and abaxial
surfaces. These plugs (perhaps wax) are commonly composed of fused rods
(Fig. 15.5.3 C), or are nearly solid (Fig. 15.5.3 D).
The stomata are sunken to the level of the hypodermis in discontinuous
rows. They are arranged regularly on broad leaves regardless of the Section
affinities. The stomata are also present in groups or clusters (A barnieri).
They occur on both adaxial and abaxial surfaces in widely spaced rows in
species with needlelike imbricate leaves (A. cunninghamii; Stockey and Taylor
1978a, b).
The stomatal orientation varies. It is parallel (vertical) in the section
Buniya (Fig. 15.5.3 E, F), Columbea and Intermedia. In the Section Eutacta
it is oblique and perpendicular (horizontal; Fig. 15.5.3 G).
Subsidiary cells vary from three to seven (Stockey and Ko 1986). Four
subsidiary cells (two polar and two lateral; Fig. 15.5.3 E, G) are common
for the genus; five subsidiary cells are also common in smaller or needle-
like leaves.
The cuticle on the subsidiary cells is slightly pitted. Depending on the
species, regardless of sectional affinity, on the stomatal surface the cuticle
may be oval, elliptical, central or polygonal. On the guard cell surface the
cuticle is mostly reticulate over the entire or part of the surface, especially
near the stoma. It is a diagnostic character for distinguishing the araucarian
species.
The cuticular flange between the subsidiary cell and guard cells is usually
serrated with a few species showing a smooth edge. Prominent polar
extensions occur, more often in smaller leaves of the section Eutacta.
The epidermal cells are usually elongate and rectangutar with pitted
cuticular surface (Fig. 15.5.3 H).
According to Stockey and Ko ( 1986), study of the cuticular morphology
indicates a need for further taxonomic work on Araucaria.
The leaves of Araucaria have a complete layer of sclerified hypodermal
cells interrutped only by stomata (Vasiliyeva 1969). In Agathis, the sclerified
hypodermal layer is incomplete. The mesophyll contains palisade and spongy
parenchyma, sclerids and unusually enlarged cells. Bamber et al. (1978)
examined these unusual cells (which are present only in Araucaria). The
lack of cytoplasm and the presence of numerous partitions in the lumen
characterize these cells. The partitions divide the cell lumen into
256 The Gymnosperms
Fig. 15.5.3 A·H. Cuticle. A, D, G, H Araucaria huntsteinii. A Adaxial cuticle shows

smooth surface and stomata. B, C, E, FA. bidwillii. B Stomatal plugs. C Stomatal plugs
of fused rods. D Nearly solid stomatal plug . E, F Adaxial cuticle. E Stomatal apparatus
with two polar and two lateral subsidiary cells. F Orientation of stomata parallel to the
long axis of the leaf. G Horizontally oriented stomatal apparatus shows thick cuticle
especially on subsidiary cell surfaces. H Adaxial cuticle. inner surface shows pitted
epidermal flanges. (After Stockey and Ko 1986)
Coniferales 251
compartments. Bamber et al. (1978) suggest that these cells can be referred
to as compartmented cells. Histochemical tests indicate that the partitions
are pectinaceous.
The function of the compartmented cells has not been determined. The
high prop0rtion of the mesophyll zone (which they occupy) suggests some
function in relation to the stroage of carbohydrates. This is supported by
the strong positive PAS reaction for carbohydrates (by the compartmented
cell partitions). There was no effect on cell contents after 7 days' complete
shading from light. The abundance of these cells in the mesophyll zone of
all Araucaria leaves examined indicates that they have (or have had in the
past) an important physiological role. Compartmented cells appear to be of
taxonomic significance as they are the only definite anatomical character
by which Araucaria and Agathis can be distinguished.
Reproduction
A survey of the literature reveals that Araucariaceae, as compared to other
families of the Coniferales, have received little attention in the study of
reproductive biology. There are a few brief reports on the life cycle of
Agathis australis (Eames 1913), Araucaria brasiliensis(= A. angustifolia-
Burlingame 1913, 1914, 1915) and A. araucana (Favre-Duchartre 1960a,
Hodcent 1971 ), and male cone development in Agathis robusta (Kaur et al.
1981). This is possibly due to the restricted distribution of the two taxa
(Kaur et al. 1981 ).
Male cone
The male cones are large, ca. 20 em in length (Araucaria rulei). There is
a central axis with numerous microsporophylls arranged spirally
(Fig. 15.5.4 A). Each microsporophyll bears 8-15 microsporangia on its
abaxial surface (Fig. 15.5.4 B). The sporangia are elongated. The pollen
_putput of each cone may be as high as 10 000 000 (see Spome 1965). The
pollen grains are wingless.
Microsporogenesis. Due to the amoeboid nature of only a few tapetal

protoplasts in Araucaria, Hodcent (1965) has reported the formation of
limited periplasmodium. Towards the end of meiosis in the microspore
mother cell, the tapetal cells begin to degenerate and are consumed by the
time pollen grains are formed.
Male Gametophyte. The microspore nucleus divides periclinally, and two

prothallial cells are formed which divide repeatedly and produce 13-40
cells (cf. family Podocarpaceae, where three or four cells are formed). The
walls between these cells break down, so that the free nuclei remain irregularly
distributed in the cytoplasm. The antheridial cell divides into a tube cell
and a generative cell. The latter divides anticlinally into a stalk cell and
258 The Gymnosperms
Fig. 15.5.4 A, B. Araucaria columnaris. A Shoot with microsporangiate cones.

B Transection microsporophyll with several microsporangia on abaxial surface. sta
stalk. (After Bierhorst 1971)
Coniferales 259
body (spermatogenous) cell (Fig. 15.5.5 A-E). The spermatogenous cell

divides and produces two equal (Burlingame 1915, Ghose 1924) or unequal
(Burlingame 1913, Eames 1913) male gametes.
B
tn
be be
Fig. 15.5.5 A-E. Araucaria cunninghamii, male gametophyte. A Uninucleate pollen

grain. B Pollen grain with two prothallial cells. C Lower prothcil.lial cell divided anticlinally.
D Further multiplication of prothallial cells; also note tube (tn), body (be) and stalk (sc)
cells. E The enlarged stalk cell, body cell, tube cell, and free nuclei of prothallial cells;
the walls have collapsed. (After Chamberlain 1935)
Female Cone
The female cones are spherical (Fig. 15.5.6 A, B), ca. 30 em in diameter,
and have deciduous scales. The bract and the ovuliferous scale are only
partially fused, the tip of the ovuliferous scale is free and« constitutes the
so-called ligule. There is normally only one or two (less common) reflexed
ovule(s) on each cone scale. At a relatively early stage in development, the
ovule fuses (to a large extent) to the subtending bract. A flap of cone scale
tissue covers the ovule and its micropyle is exposed at a basal notch. This
flap is free of the ovule and can be easily peeled off (Fig. 15.5.7 A-D). The
seeds are deeply embedded in the scale tissue. The cones shed the scales
at maturity; the entire cone shatters. In A. bidwilli, the scales are shed;
however, the entire cone abscisses, scattering the scales on impact. The
seeds can then be removed from the cone .scale.
Megasporogenesis. A deep-seated megaspore mother cell differentiates

260 The Gymnosperms
Fig. 15.5.6 A, B. A Araucaria angustifolia, immature ovulate cones and mature foliage.
B A. bidwillii, female cone at pollination. (After Stockey 1982)
in A. brasiliensis (Burlingame 1914). It is assumed that the reduction divisions

occur, tetrads are formed, and the chalaza! megaspore functions
(P. Maheshwari and H. Singh 1967).
Female Gametophyte. Over 2000 free nuclei are produced (Burlingame

1914) before wall formation. According toP. Maheshwari and H. Singh
(1967), alveolation in the Araucariaceae needs to be reinvestigated.
The number of archegonia varies from 3 to 25 , remain confined to the
upper half or one third of the gametophyte, and form a ring around the apex
of the prothallus (P. Maheshwari and Sanwal 1963). Although apparently
sunken, the archegonia probably arise from the superficial initials which
later come to lie at a lower level, due to upward growth of the gametophytic
cells. Not all the archegonia mature or are fertilized (P. Maheshwari and
Sanwal 1963).
A single-layered archegonial jacket differentiates, and may rarely be
absent between two archegonia. A binucleate condition of the jacket has
been recorded by Burlingame (1914) in A. brasiliensis. Thick and thin areas
Coniferales 261
Fig. 15.5.7 A·E. A Araucaria klinkii, cone-scale complx. B·D A. columnaris. B Cone-
scale complex, adaxial view, arrow indicates opening of the ovular pouch. bs bract scale.
ovs ovuliferus scale. C Ovule (from B) after removing cone-scale flap. D opening of
the ovular pouch (from B). EA. bidwilli, excised embryo shows a small embryo (at
left), dark area corresponds to calyptro-periblem, the cotyledons are folded. (A, E After
Stockey 1982, B-D after Bierhorst 1971)
occur in the inner wall of the jacket cells. In exceptional cases, the jacket
may be double-layered at places (see Konar and Moitra 1980).
The neck is dome-shaped, comprise about 12 wedge-shaped cells in a
single tier, and a narrow passage is left in the centre. Frequently, the entire
neck is shed.
A ventral canal nucleus is present in the archegonium of Araucaria
(P. Maheshwari and H. Singh 1967), but Burlingame (1914) failed to
observe it.
262 The Gymnosperms
Pollination
Probably there is no pollination drop mechanism. The pollen grains fail on
the ovuliferous scale, germinate in situ, the pollen tubes grow Oike fungal
hyphae) towards the micropyle and are the longest (along with Agathis,
Saxegothea, Tsuga dumosa and .T. heterophylla) in gymnosperms (see
H. Singh 1978). The micropyle is symmetrical and non-stigmatic and the
nucellus has a beak which projects beyond the micropyle.
Embryogeny
The zygote divides and both the nuclei lie in an irregularly shaped mass of
densely staining cytoplsm (Fig. 15.5.8 A), slightly below the centre of the
archegonium (A. cunninghamii-Haines and Prakash 1980). The central
location of the proembryo of Araucaria is a specialized feature in contrast
to the basally situated proembryo in other conifers, cycads and Ginkgo. Four
synchronous mitoses follow, and there is no evidence of any irregularity in
...:. .
.· . .
·.~·:·.: : ·:·. '
.:.r .... :
~>'<,..c'. . ...
~
····~
· ·.
~.· I
··: ,_~~·'.
. ·'?£~· I , . ..
~-··
H
Fig. 15.5.8 A-1. Araucaria cunninghamii, proembryogeny. A Binucleate proembryo.
B proembryo after second mitosis. C Metaphase of third division. D 16-nuclcate (only
5 nuclei visible) proembryo. E Fifth mitosis, nuclei at metaphase. F 32-nucleate (only
7 nuclei are seen) proembryo, nuclei spherical and irregularly arranged. G Nuclei (only
11 are seen) angular and regularly arranged. H Sixth mitosis . I 64-nucleate (only 15
nuclei are seen) proembryo. (After Haines and Prakash 1980)
Coniferales 263
these divisions. The cytoplasm of the proembryo, containing the randomly

scattered nuclei, is clearly demarcated from the degenerating archegonial
cytoplasm, particularly at the lower end. The proembryo becomes spherical
(Fig. 15.5.8 B-I), unlike other conifers. At the 32-nucleate stage (which
lasts considerably longer than 2-, 4-, 8-, and 16-nucleate stages) the nuclei
become angular and regularly arranged (Fig. 15.5.8 G). After the sixth
mitosis (Fig 15.5.8 H), there are 64 free nuclei in the proembryo
(Fig. 15.5.8 I) followed by wall formation in A. cunninghamii, A. bidwilli,
A. heterophylla and A. araucana. Although the details are not known, the
64-nucleate proembryo is either free-nuclear or cellular.
The two-tiered proembryo consists of a primary upper tier (pU) and a
primary embryonal group (pE) with a cap. The cells of the pU tier and the
cap are in a single tier, each arranged in a hemi-spherical plane. They
jointly enclose the embryonal group (Fig. 15.5.9 A, B; A. bidwilli). The cells
of the pU tier remain open towards the distal end and subsequently elongate,
as do also the cap cells, though not as markedly.
The cells of both the pU and pE tiers divide. In the former, the division
leads to the formation of the upper tier (U) and the suspensor tier (S). In
the pE tier, the cap cells divide anticlinally and the daughter cells become
arranged in a single tier. The cells of the embryonal group divide irregularly
and in A. cunninghamii the proembryo consists of a U tier of 31-28 cells,
S of 31-38 cells and E of 18-26 cells+ cap 32-44 cells. In A. bidwilli the
U tier is 21-32 cells, S 21-32 cells and E 18-22 cells+ cap 40-62 cells.
In A. heterophylla the U tier comprises 32-35 cells, S 32-35 cells and
E 18-24 cells + cap 36-44 cells.
In a cellular proembryo of Araucaria, the U and S tiers are curved
(Fig. 15.5.9 A, B) while they are more or less horizontal in other conifers.
Other features of interest in the proembryogeny are: (a) six free mitoses
(the highest in conifers), (b) cell wall formation, (c) elongation of S cells
independently of the final mitosis and (d) the formation of cap. According
to Haines and Prakash (1980), Araucaria is the most primitive of the conifers,
with affinity closest to the primitive podocarps.
The embryogeny of Araucaria resembles other conifers (Haines 1983).
Closest to the axis of the proembryo, the laterally situated cap cells elongate
(Fig. 15.5.9 A-D). The cell walls are not markedly thick, and intercellular
spaces do not occur (A. bidwilli; Fig. 15.5.9 F, G). During this phase, the
suspensor cells also elongate, begin to separate at their distal end, and
accumulate abundant starch grains (Fig. 15.5.9 C). The elongation is
subsequently evident in two regions of the suspensor:
a) The separated distal part which curls and exhibits thickening of cell
walls to form an anchorage (Fig. 15.5.9 C).
b) Elongation in the unseparated region pushes the proembryo out of the
archegoinum through the nutritive tissue of the corrosion region. The latter
is formed below the archegonia in the central region of the gametophyte
concurrently with the development of proembryo.
264 The Gymnosperms
A c
cp
G
Fig. 15.5.9 A-G. Araucaria bidwillii, embryogeny. A-D Longisection proembryo.

A Two-tiered proembryo shows primary upper tier (plf) and primary embryonal tier
(pE). B Three-tiered proembryo; upper tier (U), suspensor (S) and embryonal (E) tier;
the latter comprises embryonal cells and cap cells (cp). C, D Proembryo with elongated
cap and suspensor cells. C Suspensor anchorage. E Coiled suspensor with embryonal
mass at tip. F, G Transection embryonal cells surrounded by cap cells. (After Haines.
and Prakash 1980)
Coniferales 265
The suspensor cells do not divide. In a fully elongated suspensor, the

cells rem~in attached to one another as a column, although there are spaces
between them. The region next to the tip of the proembryo and the
comparatively longer region towards the archegonium remain more or less
linear, while the intervening area is markedly coiled (Fig. 15.5.9 E). The
individual suspensor cells increase in diameter from the archegonium towards
the tip, ultimately reach four to five times their original diameter. Only four
or five suspensor cells remain in contact with the tip of the proembryo. The
remaining cells cease to elongate during the development of the suspensor.
As the proembryo grows downward to the gametophytic tissue, the
corrosion region is digested and a cavity is formed. Although there is some
degeneration of cell contents, ahead of the embryo the major activity occurs
in the vicinity of the suspensor. The corrosion region and corrosion cavity
develop in both pollinated and unpollinated ovules. The cap cell contents
often show signs of degeneration during later stages of suspensor elongation.
Simple polyembryony is a regular feature; cleavage polyembryony has
not been reported in the family Araucariaceae (Haines and Prakash 1980).
In this respect, according to Doyle and Brennan (1971, 1972), the
Araucariaceae must be considered as the most primitive conifer family.
A mature excised embryo of A. bidwillii (Fig. 15.5.7 E) shows a small
embryo, a dark calyptro-periblem and folded cotyledons (Stockey 1982). In
th'e species of Araucaria, in the mature embryo, there are variations in size,
number, length of the cotyledons, hypocotyl and root cap, differentiation of
stomates, and the extent and arrangement of the vascular and secretory
tissues. According to Haines (1983), all the above features lend support to
the generally recognized division of the taxon into four sections.
Chromosome Number
The karyotypes are similar in A. angustifolia, A. bidwillii and A. columnaris,
studied from the endosperm and root tip squashes (Mehra 1988). In A. bidwillii
and A. angustifolia the endosperm squashes (n = 13) showed four
chromosomes heterobrachial and nine isobrachial (Fig. 15.5.10 A). The
root tip squashes in A. columnaris (2n =26) showed 8 heterobrachials and
18 isobrachials (Fig. 15.5.10 B).
15.6 CEPHALOTAXACEAE
The meagre fossil record of the family Cephalotaxaceae extends from the
Jurassic. It is speculated that this family may have evolved from the
Ernestiodendron line of evolution (through Palissaya) because the sterile
part of each fertile dwarf shoot is highly reduced. According to Miller
( 1977), the present fossil record of the family does not confirm this hypothesis.
Cephalotaxus
The Cephatotaxaceae is represented by a single living genus, Cephalotaxus,
266 The Gymnosperms
Fig. 15.5.10 A, B. A Araucaria angustifolia , squash preparation of endospenn cells

shows n.= 13; four chromosomes heterobrachial (h), nine isobrachial. B A. columnaris,
squash of root-tip cell, 2n = 26; 8 chromosomes heterobrachial, 18 isobrachial. (After
Mehra 1988)
Coniferales 267
with six species (Pilger and Melchior 1954).
Morphology
The plant iss shrub or small tree up to 15 ·m high; all species are dioecious.
The branches may be opposite or in whorls. The young branchlets are
green, prominently grooved, and show stomata as minute white dots. The
buds have numerous overlapping scales. The leaves are arranged in a decussate
and distichous fashion (Fig. 15.6.1 A). They remain functional for at least
Fig. 15.6.1 A-C. Cephalotaxus drupacea. A Young twig, three branches from previous
year's shoot, leaf arrangment decussate and distichous. B Alternate arrangement of leaves
on shoots from near-soil level. C Shoots with branches after 2 or 3 year's growth.
(After H. Singh 1961)
268 The Gymnosperms
3 years. The leaves are linear and pointed at the apex (Fig. 15.6.1 B). The
dorsal surface is dark shiny green with a conspicuous mid-rib. The lower
surface shows two broad silvery bands of stomatic lines. On vertical shoots
the leaves are spreading, while on lateral shoots they are arranged in two
opposite rows. In Cephalotaxus drupacea, some branches arise from the
main stem near the ground level, grow vigorously and bear alternate leaves
(Fig. 15.6.1 B). Lateral buds on these branches lack scales and unfold in
the same year in which the shoot is produced. After 2-3 years, the branches
bear decussate and two-ranked leaves (Fig. 15.6.1 C). Winter buds are
covered by numerous scales.
Anatomy
The anatomy of root and stem has not been studied.
Leaf. The leaves are distinctly bifacial (Kausik and Bhattacharya 1977).
A vertical section of the leaf of C. harringtonia shows: (a) Epidermis covered
by thick stratified cuticle, the cells are thick-walled with a prominent lumen.
(b) Sunken stomata in longitudinal rows on the abaxial surface. (c) Mesophyll
of uniseriate palisade layer, spongy parenchyma with irregular cells and
large intercellular spaces. In C. fortunei transversely elongated, loosely
arranged parenchyma cells occur between the palisade and spongy tissue
(Fig. 15.6.2 B). Several cells of the mesophyll contain tannin. A single
median resin duct is present (Figs. 15.6.2A; 15.6.3 A-C). The duct is
surrounded by two or three (Fig. 15.6.2 C) layers of compactly arranged
cells; some of the cells of the outer layer contain tannin. (d) The single
vascular bundle does not have a bundle sheath. Both protoxylem and
protophloem become crushed. The metaxylem and metaphloem are almost
equal in amount. The transfusion tissue is associated with the vascular
bundle throughout its length. The transfusion tissue is especially prominent
in the terminal region (Fig. 15.6.3 A), and forms an arc adjacent to the
xylem. In the middle of the leaf, this tissue extends in a wing-like arrangement
from each side of the xylem (Figs. 15.6.2 A; 15.6.3 B). At the base, only
a few transfusion cells are present close to the xylem (Fig. 15.6.3 C).
Richly cytoplasmic albuminous cells are present adjacent to the phloem
(Fig. 15.6.2 D). The lignified cells of the transfusion tissue are designated
tracheids because of their wall structure (Hu and Yao 1981). They are
variable in size and shape (Fig. 15.6.3 D-G) but shorter and wider than the
tracheids of the xylem. Transfusion tracheids next to the xylem are generally
longer and narrower than those further from it. The secondary walls have
bordered pits and reticulate thickening (Fig. 15.6.3 E-G). A similar
lignification and type of wall is also common in the metaxylem tracheids
of the vein. The transfusion tracheids of mature leaves lack protoplasts
(Griffith 1971).
According to Hu and Yao (1981), the transfusion tissue in the family
Coniferales 269
Fig. 15.6.2 A-D. Cephalotaxus fortunei. A, B Vertical section of leaf. A Diagrammatic.

B Transversely elongated loosely arranged parenchyma cells between palisade and spongy
tissue. C Resin duct. D Vein and transfustion tissue (ttr). ac albuminous cell. (After
Kausik and Bhattacharya 1977)
Cephalotaxaceae is similar to that in the Taxaceae. It is of the Taxus type,

i.e. lateral in groups, wing-like and mostly with bordered pits and spiral
thickenings.
270 The Gymnosperms
::...------~- tlf
ph
- --r-----,- rd
~~ 8
~---1--X
~~----~--tfr
......___---1'----- ph
c
Fig. 15.6.3 A-G. Cephalotaxus harringtonia. A-C Diagrammatic vertical sections of
leaf at tip (A), middle (B) and base (C). pa palisade. ph phloem. rd resin canal.
ttr transfusion tissue. x xylem. D-G Transfusion tracheids. D Tracheid shows first-order
frame work. E-G Mature tracheids. (After Griffith 1971)
Reproduction
Male Cone
The short, unbranched lateral shoots, covered with scales, mostly arise in
the axils of the newly emerged shoot of the male tree. In the axils of the
upper scales of these shoots, six to eight compactly arranged cones apear
(Fig. 15.6.4 A-C). A male cone consists of a short sessile axis with 15-20
microsporophylls arranged spirally.
The microsporophylls vary in shape (Fig. 15.6.4 0-G). A sporophyll
consists of a stalk bearing three or four (rarely two) abaxial sporangia
pointing toward the cone axis, and a steriie flattened region which points
outward and upward (Fig. 15.6.4 D, G).
Microsporangium. A plate of hypodermal cells differentiates on the abaxial

surface near the base of each microsporophyll (Fig. 15.6.5 A-C). Later, due
to partial sterilization, two to four groups of archesporia! cells become
delimited (Fig. 15.6.5 D); each is the site of a microsporangium. Periclinal
Coniferales 271
8 F
at G
c B
Fig. 15.6.4 A-G. Cephalotaxus drupacea. A Male cones covered by scales borne on
short lateral shoot (sh). B Short lateral shoot, male cones exposed. C Male cone with
subtending bract. D-G Microsporophylls. (After H. Singh 1961)
divisions in archesporial cells give rise to the primary parietal layer, which
organizes a four-layered wall, and the primary sporogenous layer. The
latter divide repeatedly so that a mature microsporangium has a mass of
sporogenous cells surrounded by the wall layers and the epidermis
(Fig. 15.6.5 E). The innermost wall layer forms the tapetum (Fig. 15.6.5 K),
its cells becoming binucleate at the microspore mother cell stage
(Fig. 15.6.5 L). The tapetum is of the glandular type and collapses during
the reduction divisions in the mother cells (Fig. 15.6.5 M). At maturity, the
epidermis develops fibrous thickenings, while the remaining wall layers
become crushed (Fig. 15.6.5 N). The sporangia dehisce along a line facing
the stalk of the rnicrosporophyll. The epidermal cells do not develop thickening
in this region (Fig. 15.6.5 0-Q). The upper portion of the sporophyll
c
forms
the sterile flattened region. Its cells take a uniformly dense stain, and
occasionally show a resin duct (Fig. 15 .6.5 F, G). The microsporangia
grow toward the axis of the cone, remain united at the base, but become
free of each other at the tip (Fig. 15.6.5 G-J).
Microsporogenesis. The rnicrospore mother cells (mime) contain abundant

starch (Fig. 15.6.6 A). During meiosis, the cytoplasm contracts from the
wall and a special callose wall is secreted around it. Twelve bivalents can
be counted at diakinesis (Fig. 15.6.6 B). Cytokinesis occurs after meiosis II
(Fig. 15.6.6 C-E), and tetrahedral and isobilateral rnicrospore tetrads are
formed. Several abnormalities during micmsporogenesis have been reported
(see H. Singh 1961).
272 The Gymnosperms
~
~
~
~
Q
Fig. 15.6.5 A·Q. Cephalotaxus drupacea. A, B Cross- and longisection of young
sporophylls show hypodermal archesporium. C, F, G Longisection male cones.jb fertile
bracLms microsporangia. rd resin duct. C Young male cone. D Transection sporvphyll,
initial stage of three sporangia. E Differentiation of wall layers. F, G Male cones at
early (F) and late (G) stage. H-J Microsporophyll, sections at /, J marked in H.
K Microsporangium (cross section) shows sporogenous tissue (spt) and tapetum (tap) .
L Sporangium at resting stage; some of the tapetal cells are binucleate. M, N Wall layers
at reduction division of micros!XJre mother cell (M), and exodermis with fibrous thickenings
at shedding stage of sporangium (N). 0-Q Cross sections of mature (0) and dehisced
sporophyll (P, Q). (After H. Singh 1961)
Male Gametophyte. The microspores are released from tetrads after the
digestion of the callose wall and break down of the original wall of the
Coniferales 273
• , '"
.y
I
al --~""
Fig. 15.6.6 A-J. Cephalotaxus drupacea , microsporogenesis and male gametophyte.

A-D Meiosis I of microspore mother cell. E Cytokinesis. F, G Uninucleate microspores.
H Division of microspore nucleus. I, J Two-nucleate pollen grains. al antheridial cell.
tn tube nucleus. (After H. Singh 1961)
mother cell. The microspores are spherical and small with a centrally situated
nucleus (Fig. 15.6.6 F), and a few starch grains in the cytoplasm. The intine
is thick while the exine remains thin (Fig. 15.6.6 G). According to Gullvag
(1966), the exine consists of two layers. The inner layer is lamellated and
the outer one comprises orbicules and/or a sculptured granular layer. The
nucleus divides to form a lenticular antheridial cell and a large tube cell
(Fig. 15.6.6 H, 1). Prothallial cells are absent. The pollen grain enlarges to
about twice its original size, the exine ruptures, and the intine appears
thinner due to expansion (Fig. 15.6.6 J). Starch grains disappear and the
pollen grains are shed at the two-nucleate stage.
Female Cone
The female cones usually develop close to the shoot apex, since the resting
buds mostly arise in that position (Fig. 15.6.7 A, B). Occasionally, the
cones develop in the middle or at the base of the branch. It is somewhat
difficult to determine the point of origin of the female cones because the
bud scales from which they arise and the "leaves" which subtend them look
alike. These lateral organs are not true leaves but are termed so as they are
produced by the shoot apex during the period of foliage leaf formation
(H. Singh 1961 ). A study of the formation of the lateral organs, in different
seasons in a year, shows that the cones are borne in the axil of the lowermost
two to four leaves on some newly formed shoots.
The female cones are small with a short stalk and consist of five to
seven pairs of opposite and decussate bracts (Fig. 15.6.7 C). The fleshy
nature of the cone axis conceals the exact arrangement of bracts, and is
274 The Gymnosperms
ff
.' B ~\
~~ c
Fig. 15.6.7 A-E. Cephalotaxus drupacea, female cones. A Cones near the apex and
middle of branch. B Four cones and vegetative shoot (vs), from a resting bud. C Cone.
D Two ovules of a seed scale complex replaced by foliage leaves (ff). E Mature ovule
on a cone. (After H. Singh 1961)
revealed only by a study of the vascular anatomy. Two ovules are borne in
the axil of each bract (Fig. 15.6.7 C), except the lowest pair. There is a
small ridge-like outgrowth of the cone axis between the two ovules of each
bract. Florin ( 1938-1945) termed the two ovules and the outgrowth between
them the seed-scale complex. The ovules that enlarge in the following
spring are green, and become red and fleshy on ripening. Generally, one or
two ovules mature in a cone (Fig. 15.6.7 D, E) although as many as five
seeds may sometimes mature.
Normally, the apex of the female cone becomes inactive after producing
the bracts and the ovules. However, in a few cones, the apex remains
surrounded by numerous bud scales. According to Favre-Duchartre (1957),
the proliferation of such a bud may give rise to a female cone in the
following year.
Ovule. An outgrowth develops in the axil of every fertile bract of a

young female cone (Fig. 15.6.8 A, B). Its apex can be demarcated into four
cytohistological zones typical of a vegetative shoot apex (Fig. 15.6.8 C).
Thus, the outgrowth apparently represents a scondary axis. The two ovules
Coniferales 275
8
rm
Fig. 15.6.8 A-D. Cephalotaxus drupacea, megasporangium. A Longisection young female

cone. B Inset from A Initiation of secondary axis. C Later stage, secondary axis; tip
shows typical organization of a shoot apex. ai apical initials. fm flank meristem. rm rib
meristem. D Initiation of two ovules (ov) , from secondary axis. (After H. Singh 1961)
originate as lateral protuberances from this secondary axis (Fig. 15.6.8 D).
In younger stages, the growth occurs mainly by periclinal di!visions which
characterize a leaf primordium.
A young ovule, with a conspicuous integument, shows three to six
hypodermal archesporia! cells (Fig. 15.6.9 A). The latter divide periclinally
to form the sporogenous layer and the primary parietal layer (Fig. 15.6.9 B).
A massive nucellus is formed (Fig. 15.6.9 C, D) by repeated periclinal
divisions of the primary parietal layer. The nucellar cells in the vicinity of
the sporogenous layer, at the chalaza} end, also undergo periclinal divisions
to form a distinct tussue, which in later stages contains large compound
starch grains (Fig. 15.6.9 E, F). This tissue (designated pavement tissue by
H. Singh 1961) becomes crushed during tbe enlargement of the free-nuclear
female gametophyte.
276 The Gymnosperms
Fig. 15.6.9 A-F. Cephalotaxus drupacea, megasporangium. A-C Longisection, ovules.

A Hypodermal archesporium. B Periclinal division in archesporium forms primary parietal
and sporogenous layer. C Ovule in the first (winter) resting stage, some integumentary
cells contain tannin. D Ovule at the time of pollination. E Sporogenous tissue. F Pavement
tissue. (After H. Singh 1961)
A few days before pollination, at the micropylar region, two or three

layers of nucellar cells degenerate and form a rudimentary pollen chamber
(Fig. 15.6.10 A-C). Simultaneously, some of the inner epidermal cells of
the integument lining the micropyle become active (Fig. 15 .6.10 B). After
pollination, these cells grow inward, undergo a few transverse divisions
close the micropylar opening (Fig. 15.6.10 D-F), and effectively seal the
freshly deposited pollen. These cells eventually acquire a very thick wall.
Coniferales 277
rtr-1--- -->r-E
t-rr-----J'- pt
Fig. 15.6.10 A-H. Cephalotaxus drupacea, megasporangium. A-C Longisections upper

part of nucellus show formation of pollen chamber, and active inner epidermal cells of
integument (B). D Outline diagram of ovule. mgmc megaspore mother cell. pt pollen
tube pv pavement tissue. E Inset marked E in D to show closure of micropyle.
F Transection ovule shows closed micropyle. G Inset marked G in D shows tannin-
containing epidermal cells of nucellus and integument. H Longisection ovule in the second
winter. (After H. Singh 1961)
Some epidermal cells at the junction of the nucellus and the integument
accumulate tannin (Fig. 15.6.10 G). Figure 15.6.1 0 H represents the condition
of the ovule during the second winter.
278 The Gymnosperms
Megasporogenesis. One of the sporogenous cells enlarges, especially along

its longer axis, the nucleus also enlarges, cytoplasm becomes "frothy", and
it functions as the megaspore mother cell (Fig. 15.6.11 A). The non-functional
sporogenous cells persist in the vicinity of the mother cell. Meiosis I and II
result in a linear tetrad, and the chalaza! megaspore functions
(Fig. 15.6.11 B, C). The non-functional megaspores degenerate without any
definite sequence (Fig. 15.6.11 B, C).
Female Gametophyte. The functional chalaza! megaspore becomes

vacuolate, its nucleus divides repeatedly, and an elongated free-nuclear
female gametophyte is formed (Fig. 15.6.12 A). During this process, the
circumjacent nucellar cells are crushed. The free nuclei become arranged
in a single layer along the periphery, leaving a central vacuole. At the
32-nucleate stage, the gametophyte has a two-nucleate (rarely four-nucleate)
finger-like projection at the micropylar end (Fig. 15.6.12 B, C). This projection
remains conspicuous until about 128 nuclei have been formed (Fig. 15.6.12 D).
Later, it is no longer prominent (Fig. 15.6.12 E). A similar micropylar
projection of the free-nuclear gametophyte has also been reported in other
taxa. In Taxus it is enucleate (Dupler 1917), in Torreya (Coulter and Land
1905), Austrotaxus (Saxton 1934) and Saxegothaea (Looby and Doyle 1939)
it contains only one or two nuclei.
During later stages, the oblong gametophyte is limited by a thin membrane
(Fig. 15.6.12 E, F). Wall formation is initiated after several hundred nuclei
have been formed (4096 nuclei according to Favre-Duchartre 1957).
Cellularization occurs by centripetally advancing walls (Fig. 15.6.12 G)
and the cells show a large central vacuole with the nuclei distributed either
along the periphery of the gametophyte or the lower end (Fig. 15.6.12 H).
The earlier formed cells divide periclinally even before the central region
becomes cellularized. As the gametophyte matures, the nuclei gradually
become reduced in size. Thin-walled cells have scanty cytoplasm and are
arranged in files which converge towards the centre (Fig. 15.6.12 G, H).
Binucleate cells are common. As the embryo matures, the cells of the
gametophyte become densely cytoplasmic and accumulate starch grains
and oil globules.
After the gametophyte has become cellular, two to five cells at the
micropylar end enlarge, become densely cytoplasmic, the nuclei enlarge
and function as the archegonial initials (Fig. 15.6.13 A). The adjoining
cells undergo several periclinal divisions and differentiate as the jacket
layer. At first, these cells are isodiametric, but later elongate. The jacket
cells may contain two nuclei.
The archegonial initials develop into the archegonium (Fig. 15.6.13 B-F).
The neck of the archegonium comprises two cells (occasionally three or
four) mostly arranged in one tier; their outer tangential walls are always
thick. During the "foam" stage of the archegonium, the cells of the
Coniferales 219
c
Fig. 15.6.11 A-C. Cephalotaxus drupacea, megasporogenesis. A Megaspore mother cell.
B, C Linear tetrad; degenerated non-functioning megaspores. (After H. Singh 1961)
280 The Gymnosperms
Fig. 15.6.12 A-H. Cephalotaxus drupacea, female gametophyte. A Longisection nucellus

shows eight-nucleate gametophyte. B, D, E Whole mount of free-nuclear gametophytes.
B, C, D Micropylar projection with nuclei. F Inset marked Fin E.G Part of gametophyte
(longisection), to show wall formation. H Inset marked H in G. (After H. Singh 1961)
gametophyte at the micropylar end elongate upward and divide periclinally.

As the archegonium matures, these cells become richly cytoplasmic and
frequently show two nuclei. They are especially prominent during the
development of the embryo. A similar large cone of cells is also present in
Podocarpaceae (see H. Singh 1961). The neck cells appear sunken, so that
each archegonium has its own archegonial chamber (Fig. 15.6.13 G, H).
The central cell elongates considerably, has a pointed chalaza! end
(Fig. 15.6.13 H), and during the foam stage it contains many irregular and
darkly staining bodies. Its nucleus is densely chromatic, has a prominent
nucleolus and lies just below the neck cells. In a mature central cell, the
cytoplasm is dense and contains round protein granules. The nucleus divides
to form the egg and an ephemeral ventral canal nucleus of equal size; they
are not separated by a wall (Fig. 15.6.13 D, E). The egg nucleus migrates
downward and enlarges considerably (Fig. 15.6.13 F). Minute darkly staining
bodies appear along the inner surface of the nuclear membrane, the nucleolus
and the chromatin become indistinct. A mature archegonium is long, narrow
Coniferales 281
Fig. 15.6.13 A-H. Cephalotaxus drupacea, archegonium. A-E Development of

archegonium. cl central cell. j jacket. ni neck initial. F Mature archegonium (upper part).
G, H Micropylar region of female gametophyte shows archegonial complex (G) and
mature archegonia (H). (After H. Singh 1961)
and pointed at the chalazal end. It contains dense cytoplasm and numerous
protein globules (Fig. 15.6.13 E, F). In a mature egg nucleus, stained with
282 The Gymnosperms
Schiff's reagent, the chromatin appears as a small dot in the nucleus

(H. Singh 1961). At the time of fertilization, the large nucleus is full of
nucleoplasm and several darkly staining round bodies which are unlike
nucleoli. The archegonia are situated slightly below the tip of the gametophyte
(Fig. 15.6.13 G, H), and separated from each other by four to eight layers
of cells. Occasionally, the archegonia lie in a somewhat lateral position.
Rarely, two or three adjacent archegonia have a common archegonial cavity
(Fig. 15.6.13 G).
Pollination
The pollen grains land on the pollination drop exuded from the micropyle,
imbibe water and other substances from the fluid, become heavy and sink
down in the micropyle, settling on the degenerated tip of the nucellus (a
weakly developed pollen chamber). It is likely that the ruptured exine is
cast off when the pollen grain is caught in the pollination drop. The intine
enlarges at the (tube cell end) proximal end to form a short but broad
pollen tube which grows through the nucellus (Fig. 15.6.14 A, C). During
its growth, the pollen tube crushes the nucellar cells and reaches the female
gametophyte when the archegonia are almost mature. Rarely, a pollen tube
may reach the archegonia in the foam stage. Such an archegonium matures
earlier than the other archegonia in the same gametophyte. This behaviour
indicates a correlation between the adjacence of the pollen tube and maturation
of the archegonium (H. Singh 1961).
The tube nucleus migrates into the pollen tube. The antheridial cell
enlarges considerably, and divides at right angles to the long axis of the
pollen tube to form the stalk and body cells (Fig. 15.6.14 B). The stalk cell
lacks a elear wall, but is surrounded by dense cytoplasm. Both the cells
move down and lie free in the cytoplasm of the tube (Fig. 15.6.14 C).
When they are close to the tube nucleus, the cytoplasm around the stalk
nucleus becomes indistinguishable. The pollen tube contains a body cell,
and stalk nucleus and tube nucleus-the latter two being similar to each
other (Fig. 15.6.14 C). During the growth of the pollen tube, the stalk and
tube nuclei usually precede the body cell, which remains closely associated.
The contents of the pollen tube lie at its tip (H. Singh 1961 ), or may lie far
behind (Favre-Duchartre 1957). The pollen tube persists in this stage for
nearly a year.
Post-Pollination Development of Male Gametophyte. In the following

year, the pollen tube grows rapidly and reaches the tip of the female
gametophyte when the archegonia are almost mature (Fig. 15.6.14 D). The
pollen tube enters the archegonial cavity, comes in contact with the neck
cells of the archegonium, and swells at the tip (Fig. 15.6.14 E). By this
time, the body cell has enlarged considerably. Its prominent nucleus divides,
but no wall is laid down between the two nuclei (Fig. 15.6.14 F, G). These
Coniferales 283
E
H
Fig. 15.6.14 A-H. Cephalotaxus drupacea, post-pollination development of male
gametophyte. A Two-celled pollen tube. al antheridial cell. tn tube nucleus. B Antheridial
cell has divided. bl body cell. sc stalk cell. C Pollen tube in the nucellus; stalk and body
cells are lying close to the tube nucleus. D Longisection upper part of ovule. E Pollen
tube has grown up to the tip of archegonium. nk neck cells. F Division of body cell.
G, H Mature male gamete (mg). (After H. Singh 1961)
are small, and have dense chromatin and a prominent nucleolus. At maturity,
the male gametes enlarge, the nucleolus is no longer distinguishable and
the chromatin also becomes inconspicuous (Fig. 15.6.14 H). They may be
equal (Lawson 1907; Favre-Duchartre 1957, H. Singh 1961), or unequal
(Coker 1907, Sugihara 1947, Kaur 1958). The cytoplasm of the body cell
and the pollen tube contains small, round, densely staining bodies
(Fig. 15.6.14 G, H).
Fertilization
The neck cells begin to collapse as the pollen tube comes in contact with
284 The Gymnosperms
them, and a passage is formed for the tube to enter the egg cell. The male
gametes separate from each other due to the dissolution of the wall of the
body cell; the stalk and tube nuclei degenerate. The tip of the pollen tube
comes to lie inside the archegonium, where it bursts, and one or both the
male gametes, together with some cytoplasm of the body cell, enter the egg
cell (Fig. 15.6.15 A). As the male gamete approaches the egg nucleus, the
cytoplasm in the upper part of the archegonium becomes vacuolate
(Fig. 15.6.15 B-D). When the male gamete and egg come in contact with
each other, the male nucleus becomes lenticular and both become enveloped
in the male cytoplasm (Fig. 15.6.15 B). The nuclear membrane "dissolves"
at the point of contact, and the chromatin of the two nuclei come to lie
close to each other (Fig. 15.6.15 C, D). Numerous fibrils appear in the
fusion nucleus. The two chromatin groups integrate and the fibrils become
arranged in a bipolar spindle. The metaphase plate is slightly oblique to the
long axis of the archegonium (Fig. 15.6.15 E).
Embryogeny
The division of the zygote nucleus is intranuclear and occurs in situ. The
resulting two nuclei, enclosed in the male cytoplasm, migrate to the lower
part of the proembryo. Numerous vacuoles in the cytoplasm mark the path
of descent of these nuclei (Fig. 15.6.16 A-C). Both the proembryonal nuclei
divide simultaneously (Fig. 15.6.16 A) and the resultant four nuclei separate
from each other as the sheath of the male cytoplasm disappears. The upper
part of the proembryo becomes much vacuolated while the lower portion
has dense cytoplasm with numerous protein glubules. The two regions are
separated by a large vacuole. These four nuclei divide twice simultaneously
(Fig. 15.6.16 B, C) and wall formation is initiated at the 16-nucleate stage.
the nuclei become progressively smaller as the proembryo advances in age.
A variable number of large nuclei, which degenerate later, have been
observed in the upper region of the proembryo. They could represent:
(a) the second male gamete and its derivatives, (b) supernumerary sperms,
(c) persistent ventral canal nucleus or (d) relict and lagging nuclei (see
H. Singh 1961).
Wall formation in the proembryo results in a lower embryonal group of
10-13 small cells (pE} and an upper tier of 3-6 cells open towards the
archegonium (pU}. The lowermost one or two cells of the embryonal group
are large, have dense cytoplasm, a prominent nucleus, and constitute the
cap cells (Fig. 15.6.16 D). One more division in the embryonal group
increases the number of cells to 20-26. The upper tier divides transversely
to form the lower suspensor tier (S) and an open tier (U} which later
degenerates (Fig. 15.6.16 E, F). By this time, the cytoplasm of the
archegonium forms a dense mass or plug (Fig. 15.6.16 F). The suspensor
tier elongates and pushes the embryonal tier into a cavity in the centre of
the female gametophyte (Fig. 15.6.16 G, H). This cavity is formed by the
Coniferales 285
Fig. 15.6.15 A-E. Cephalotaxus drupacea, fertilization. A Longisection upper part of

archegonium to show pollen tube and two discharged male gametes. en egg nucleus.
mg male gamete. rn relict nuclei. B Middle portion of archegonium shows egg nucleus
and lenticular male gamete. C, D Integration of male and female chromatin. E Zygote
nucleus in metaphase. (After H. Singh 1961)
286 The Gymnosperms
..-tt·v.
·· ·11.:
~"r*'~-mg
l,i
Fig. 15.6.16 A-H. Cephalotaxus drupacea. A-C Development of proembryo. A Two

proembryonal nuclei in division; the second male gamete (mg) is also in division in the
upper part of proembryo. B, C Four-and eight-nucleate proembryo, nuclei in division.
D-H Embryogeny. D Proembryo after wall formation. cp cap. E Proembryo shows upper
tier (U), suspensor tier (S) and embryonal tier (E) with cap cells. F Degenerated remains
of upper tier and cytoplasm of archegonium (pi); the cap cells have degenerated and
rosette embryos ( re) initiated. G Later stage, elongated suspensor. H Longisection
micropylar region of female gametophyte shows embryos in the corrosion cavity. (After
H . Singh 1961)
Coniferales 287
degeneration of cells and progresses toward the chalaza! end. Some of the
suspensor cells divide transversely and give rise to numerous small rosette
embryos, which eventually degenerate. All the cells of the young embryo
contain starch.
By the time the suspensor has elongated, the cap cells degenerate and
are replaced by three or four adjacent embryonal cells which enlarge. Rarely,
a cap cell is binucleate. Ultimately, these cells also degenerate.
The basal cells of the embryonal mass elongate to form embryonal
suspensor, which pushes the embryonal mass further down into the female
gametophyte. The suspensor cells collapse by this time. Occasionally, the
cells of the embryonal suspensor proliferate to form a small embryo. As the
embryonal mass grows, the suspensors become massive. Of the one or two
embryonal masses (derived from as many fertilized archegonia), one takes
the lead and develops into a mature embryo. Cleavage of the embryonal
mass is rare.
The leading embryo remains undifferentiated. Its outermost layer undergoes
periclinal divisions, and the dermatogen, periblem and root initials become
distinct. The two cotyledonary primordia also differenttate. In a mature
embryo, the root tip initials are very distinct while the stem tip is still
undifferentiated. The various histological regions of the stem tip can be
discerned when the seed germinates.
Seed
At the time of pollination, the integument comprises several layers of thin-
walled cells and some cells contain tannin. As the megaspre mother cell
differentiates in the ovule, resin ducts appear in the chalazal region. They
are narrow and lined by an inner layer of epithelial and an outer layer of
tannin cells. The lower part of the integument becomes meristematic, and
several layers thick. These cells contribute to a major part of the seed coat.
At the free-nuclear stage of the gametophyte, the lower portion of the
integument enlarges considerably. The innermost layer of the integument
comprises narrow and vertically elongated cells which becqme crushed by
the enlarging gametophyte. There is a prominent band of tanniniferous
cells in the central region of the integument. Stomata are present in the
epidermis (Fig. 15.6.17 D). At a later stage of development, some of the
hypodermal cells acquire thick pitted walls, and seven or eight layers of
cells inner to the tanniniferous band form the sclerenchymatous stony layer
(Fig. 15.6.17 A, B, E, F). Outer to the stony layer is the red fleshy layer,
which contains scattered thick-walled and tannin cells, numerous resin ducts,
and is traversed by two vascular bundles. The epithelial lining of the resin
duct degenerates. The cells inner to the stony layer form the papery layer,
which has a mosaic surface due to the presence of patches of tannin cells
(Fig. 15.6.17 G, H).
A mature seed is large, consists of a thick coat, and the embryo extends
288 The Gymnosperms
{jj emb--r.IJ
~fg~
A B
~~
G .. , il H
Fig. 15.6.17 A·J. Cephalotaxus drupacea, seed and seed-coat. A-C Longisection of
seeds containing embryos at different stages of development. emb embryo. fg female
gametophyte. D-F Portion of integument in transection. rd resin duct. st stoma. stl stony
layer. D Free-nuclear gametophyte. E, F At the stage of ovule shown in A, B. G Seed
after removal of stony and fleshy layers. H Mosaic surface of papery layer.
I, J Longisections of female gametophyte, from seeds shed from the plant, show elongation
of embryo. (After H. Singh 1961)
Coniferales 289
to a short length of the massive female gametophyte (Fig. 15.6.17 C). On

shedding, the outer fleshy layer disintegrates and the seed is covered by the
stony layer, which is sharply pointed at the two ends. The embryo grows
and extends to almost the entire length of the gametophyte (Fig. 15.6.17 I, J).
The seeds undergo a long period of dormancy lasting 2 to 3 years.
Germination. During germination, the stony layer cracks laterally and

longitudinally and the radicle emerges. As in other conifers, the germination
is epigeal. The hypocotyl and cotyledons are fleshy, and the spirally arranged
juvenile leaves are longer but narrower than the mature leaves. Cotyledonary
leaves persist for more than 2 years after germination.
Chromosome Number
The haploid chromosome number in C. griffithii, C. mannii, C. harringtonia
var. drupacea and C. fortunei is n = 12, as determined from endosperm
squashes, and their karyotypes are identical (Mehra 1988 ). Eleven
chromosomes are isobrachial. The single heterobrachial chromosome bears
a terminal satellite on the short arm. One isobrachial chromosome possibly
bears a satellite at/ near the terminal end but it is not distinct (Fig. 15.6.18 A).
Meiosis has been studied in C. mannii. Eleven chromosome pairs have
terminal chiasmata at both ends and form 0-shaped configuration. One pair
has a single chiasma at one end (Fig. 15.6.18 B).
II
Fig.15.6.18 A, B. Cytology. A Cephalotaxus grifithii, endospenn nuclear mitosis shows
n = 12. h heterobrachial chromosome, arrow shows constriction. B C. manii, microspore
mother cell shows bivalents. (After Mehra 1988)
Based on cultivated plants in Dehra Dun (India), Cephalotaxus drupacea
has a 2 year reproductive cycle.
The male cones initiate during August, and the microsporangia over
winter in the microspore mother cell stage. Development is resumed in the
middle of March. Dehiscence and shedding of pollen occurs during spring,
pollen grains land on the ovule, and produce short pollen tubes. The antheridial
cell divides, and stalk and body cells migrate to the lower portion of the
290 The Gymnosperms
pollen tube by April. No further development takes place for about a year.
Then, development is resumed, and the male gametes are formed and liberated
into the archegonium by mid-April.
The female cones initiate during late autumn, and the ovules overwinter
in the sporogenous cell stage. Pollination occurs during spring, followed by
ovule enlargement, megasporogenesis and formation of the free-nuclear
gametophyte. The ovule undergoes the second winter rest. Development is
resumed during spring, and archegonia are formed. Fertilization takes place
by mid-April. The embryo develops quickly and the seeds mature by August.
The ovule needs 1 year and 9 months from appearance to maturation into
a seed (H. Singh 1961).
15.7 PHYLOGENETIC CONSIDERATIONS OF

CONIFERALES
The knowledge of primitive conifer groups has expanded as a result of

investigations by a large number of researchers since the pioneering work
of Rudolf Florin (see Florin 1939, 1944, 1945, 1950a, 1950b, 1951, 1954).
Florin compared the fertile dwarf shoot of Cordaianthus (Cordaitales) with
the ovuliferous scale of modem conifers and documented several intermediate
stages within the Voltziales. Steps in the evolution (of an ovulate conifer
cone from Cordaitean strobilus) are as follows: (a) a reduction in internode
length, (b) a flattening of the short shoot, (c) fusion of the sterile parts of
the short shoot, (d) fusion of the ovule axis to the scale and eventual
elemination of this axis, (e) total reduction in the number of fertile parts,
and (f) reduction in the numbers of ovules (and a change in ovular position
in some genera).
Since the work of Florin, much has been learnt about the evolution and
reproductive biology of Coniferales. Systematic investigations of fossil cone
vasculature and resin canal distribution, leaf cuticles, seed integuments and
embryo structure have increased our knowledge of conifer evolution (Stockey
1982). The living conifer families appear to have originated earlier than
was formerly presumed. All the families of modem conifers (with the
exception of Cephalotaxaceae, and perhaps Pinaceae) appear to have an
Upper Triassic to Lower Jurassic appearance (Miller 1977). Several present-
day taxa are evident in the Mesozoic and can be recognized by the Late
Triassic or Early Jurassic. Many genera described by Florin are being
reinvestigated. Miller (1977) pointed out that many of the intermediate
Voltzian conifers are present in the Triassic and overlap the earliest of the
modem conifer families. Thus, conifers have an extensive geological record,
with many of the modem groups representing very ancient lineages. According
to Miller (1977), it is doubtful that modern families evolved from the
Voltziaceae, as they are mostly Late Triassic to Jurassic in age (see Chap.
14). Precursors should be sought in Palaeozoic to Early Triassic sediments,
Coniferales 291
(early records of the families based on foliage alone are disputed). New
families have been erected for many of the newly discovered fossils. The
Cheirolepidaceae have seed cones with deciduous scales and large persistent
bracts and are unlike any other known conifer. With the discovery of pollen
cones (Tamaxellia, Classostrobus) containing Classopollis pollen and their
associated foliage types (Frenelopsis, Pseudofrenelopsis), the family is now
better understood. The Cheirolepidaceae apears to have been widespread
during the Mesozoic, and its genera may have occupied a number of diverse
habitats (see Stockey 1982). Further work is necessary, especally of the
plants within the Cordaitales and Progymnospermopsida to understand their
diversity and specialization. This approach will give a clearer picture of
possible conifer ancestors. The studies of fossil conifers has added to our
understanding of the extinct forms. With the discovery of additional fossil
conifers in Late Palaeozoic to Early Mesozoic sediments, we are coming to
a closer understanding of their origin and subsequent radiation. Additional
well-preserved Triassic conifers will further expand our knowledge of the
diversity of coniferophytes (Corditales, Voltziales, Coniferales), a group
that probably was at its zenith during the Mesozoic.
16. Taxales
Taxales is a small group of plants comprising 5 living taxa and 20 species,

included in a single family, Taxaceae (Dallimore and Jackson 1966):
Amentotaxus (4 spp.), Austrotaxus, Pseudotaxus (= Nothotaxus), both
monotypic, Taxus (8 spp.) and Torreya (6 spp.).
Taxaceae
The Taxaceae appeared during the Late Triassic or Early Jurassic (Miller
1977). The earliest remains attributed to the family are leafy twigs and
attached arillate fruits of Palaeotaxus (Florin 1948) from the Lowermost
Jurassic. Amentotaxus is present by the Late Cretaceous. Thus, the fossil
record extends from the onset of the Jurassic, and the early forms show
ovulate reproductive structures. However, the fossils are so like those of
living forms that they provide no clue about the ancestry of the family.
The plants are evergreen shrubs or small trees, profusely branched, with
small spirally arranged linear leaves. The wood is pycnoxylic and the tracheids
have tertiary spirals; resin canals are absent. The plants are mostly dioecious,
only sometimes monoecious; male cones are small and rnicrosporangiophores
have two to eight peltate pollen sacs. Solitary, arillate ovules terminate a
dwarf shoot with decussate bracts. The embryo is dicotyledonous.
Taxus
Taxus, commonly known as yew, the leaves, shoots and seeds have poisonous
properties. The active principle is taxine although other alkaloids are also
present. Both fresh and partly dried shoots contain alkaloids; the withered/
dried shoots are considered more toxic in action than the fresh foliage. The
poison content may vary in male and female trees, or in different trees. The
scarlet aril around the seed is, however, harmless. The plants have to be
fenced from cattle. From the inner bark (T. brevifolia) taxol is obtained which
has therapeutic qualities (see Chap. 23).
Morphology
Taxus is one of the common hardy evergreen shrubs or trees. T. baccata
can reach a height of 20 m, with a massive trunk 7 m or more in girth,
Taxales 293
while T. wallichiana, exceptionally, may reach 30m in height and 5 min

girth. The dorsiventralleaves are linear, 2-3 em long with recurved margin,
tapering apex, dark green above and yellowish green beneath (Fig. 16.1).
They are spirally arranged but appear distichous due to a twist in the short
leaf base (T. wallichiana). The mid-rib is prominent, with two bands of
stomata on either side on the undef surface. The scale-like leaves on the
fertile shoots are opposite and decussate.
Fig. 16.1. Taxus wallichiana, twig to show arrangement of leaves and arillate ovules.
(Photograph, courtesy Professor B.D. Sharma, Jodhpur)
Anatomy
Stem. A distinct tunica is absent. The zonation of the apical meristem is

similar to Taxodiaceae and Pinaceae, except that the frequency of periclinal
divisions (in all regions) varies considerably as the growing season advances.
The secondary wood is formed from a single persistent cambium. The
wood of Taxus shows specific xylotomical features (Greguss 1955). The
wood parenchyma is generally absent, but may be present in T. baccata
and T. brevifolia (and in the root of T. cuspidata). Distinct growth rings
develop due to the difference between the flattened and thick-walled late
wood tracheids and the thin-walled early tracheids of longer lumen
(T. brevifolia). The transition from early to late wood is usually gradual.
The most characteristic feature is the presence (in longitudinal tracheids)
of spiral thickenings, which originate from the tertiary layer and lie side by
294 The Gymnosperms
side with bordered pits. The horizontal ray cells have an uneven thickness
with pit-like depressions. Tangential ray-cell walls are always smooth and
identures are fairly frequent. Pits (cupressoid) are relatively small and three,
four or six pits are present in the cross-field. The pit apertures are slit-like,
oblique/vertical but never horizontal.
Leaf. A vertical section of leaf shows mesophyll differentiated into palisade

and spongy tissues; Resin ducts absent (which distinguishes it from the
conifers; Fig. 16.2 A). The distinct bundle sheath is a ring of regularly
arranged prominent cells (Fig. 16.2 B). The vascular bundle is large and
dorsiventrally flattened, parenchyma develops between the rows of xylem
and phloem; transfusion tissue occurs around the vascular bundle, and
tracheids and some included parenchyma are confined to wing-like extensions
lateral to xylem. However, most of the parenchyma also extends adaxially
and abaxially to the bundle. The albuminous cells are especially prominent
in this region, those lying within the bundle sheath being very large and
also vesicular (Fig. 16.2 B).
c B
Fig. 16.2 A-C. Taxus baccata, leaf, vertical sections. A Outline diagram. B Transfusion
tissue and vascular bundle. ac albuminous cells. tp transfusion parenchyma. ttr transfusion
tracheid. C Stomata. (After Kausik and Bhattacharya 1977)
The stomata are haplocheilic; subsidiary cells develop papillate extensions

(Fig. 16.2 C) which form a deep epistomal chamber so that the guard cells
appear to lie deep in it.
Taxales 295
Reproduction
Male Cone
The male cones are stalked globose heads in the axil of leaves. Each male
cone axis bears ca. 10 decussate sterile scales and 6-14 symmetrical and
perisporangiate rnicrosporophylls, each bearing 6-8 reflexed sporangia. They
are terminally arranged in small cones on short shoots (Fig. 16.3 A).
Fig. 16.3 A, B. Taxus sp. A Longisection male cone. B Longisection, fertile shoot with
terminal ovule. (After Bracegirdle and Miles 1973)
Microsporangium. An ultrastructural and cytochemical investigation of

the development of the microsporangium of T. baccata reveals many
distinctive features (Pennell and Bell 1985). The early development of the
archesporia! cells is not synchronized (cf. other gymnosperms investigated).
Groups of degenerated cells are present in the archesporium which disappear,
presumably by resorption. The tapetum differentiates from the outer layer
of the archesporium. The sporogenous cells can be recognized about 3
weeks before meiosis. There is a tendency towards synchrony and diad
formation in a sporangium, almost simultaneously.
Microsporogenesis. After meiosis I is completed, vesicles accumulate in

the equatorial plane of the spindle, coalesce and give rise to a sinuous wall.
This wall fluoresces brilliantly when stained with Calcofluor White and
with aniline blue, while the wall surrounding the diad fluoresced only with
aniline blue. This wall (and not the dividing wall) contains a fibrillar layer.
After meiosis II is completed, the four microspores lie in one plane, so that
the symmetry of the tetrad is regularly isobilateral (Pennell and Bell 1986).
The partitioning walls formed at telophase of meiosis II fluoresce brilliantly
with aniline blue. In the electron microscope these walls appear similar to
those giving rise to the diads, but there is a thin layer of fibrillar material
296 The Gymnosperms
evident at the site of the middle lamellae of all the partitioning walls. The
successive wall formation in the mime of Taxus distinguishes it from many
conifers. The cytoplasm of the four spores is often different, although no
differences are detectable in the parent cells, those derived from a diad
being less dense than those derived from the other. A small proportion
(ca. 5%) of the tetrads in a sporangiw'n degenerate. Although there is inequality
in the frequency of ribosomes between the spores of a tetrad, partial
degeneration within a tetrad has not been observed (Pennell and Bell1986).
The microspores are released into the loculus by rapid dissolution of the
callosed wall of the pollen mother cell; the formation of the sporoderm
begins. Osmiophilic droplets emerge from the spore protoplast and enter
the wall. The droplets coalesce to form an outer layer on which up to six
sporopollenin lamellae, probably of tapetal origin, are deposited (the fibrillar
layer can no longer be recognized). The accretion of a single layer of
sporopollenin droplets, in no recognizable pattern, gives rise to the outer
verrucose part of the exine. The sporoderm becomes resistant to acetolysis
and fluoresces when stained with Auramine 0.
The cytoplasm of the mature spores is vacuolate. Plastids dedifferentiate,
nucleoli reappear but synaptonemal-like complexes and nuclear vacuoles
are no longer present. Large vacuoles persist in the cytoplasm and many
others develop from the dictyosomes. A single cisterna of endoplasmic
reticulum often encircles the nucleus. The microspores are shed in this
condition (Pennell and Bell 1986).
During meiosis, the tapetal cells elongate tangentially, later they become
round. Many cells are binucleate and later become trinucleate. The sinuous
radial walls contain a single fibrillar layer. Elongated amyloplasts, dictyosomes
and isolated cisternae of endoplasmic reticulum are frequent. Large vesicles
and vacuoles give the tapetum a foamy appearance (under the light
microscope). About 3 weeks before anthesis, plastids are no longer visible
in the tapetum. Orbicules appear simultaneously in the VI ails of the tapetal
cells and in the loculi of the sporangium. With the rupture of the inner
membrane, the tapetum becomes partly amoeboid. A peritapetal membrane
(often overlain with orbicules) can be identified at this time. Cytochemical
tests indicate that, from the beginning of meiosis, the tapetum is rich in
acid phosphatase (Pennell and Bell 1986).
A mature microsporangium splits, curls and liberates pollen grains. The
pollen grains are wingless and at the uninucleate stage.
Ovule
The ovules are solitary, arillate and terminate a modified dwarf shoot with
decussate bracts (Fig. 16.3 B).
The initiation of the ovule (in Taxus) is by a transformation of the shoot
meristem (Loze 1965). The apical initials give rise to the nucellus, the
integument arises from the flank meristem and the pith meristem. The
Taxales 297
subapical initials and the flank meristem contribute to the chalaza! portion
of the ovule (Fig. 16.4 A, B). The epidermal cells of the dome-shaped
nucellus undergo periclinal divisions to form a nucellar cap. A discreet
epidermis is not distinguishable in the young nucellus. The cells of the
outermost exposed layer divide periclinally to form the inner archesporia!
cells and the outer layer (Fagerlind 1961 ). The cells of the outer layer again
divide periclinally and the derivatives divide repeatedly to form the nucellar
tissue. The dual origin (parietal tissue and nucellar cap) of the nucellus is
well established in Taxus (Dupler 1917, Sterling 1948a, Pankow 1962).
Fig. 16.4 A-F. A-C, F Taxus baccata, D, E Taxus sp. A, B Longisection, tip of fertile
shoot. A After initiation of last pair of bracts (br). B Nucellus (n) and integument (i).
C, D Development of aril. E Longisection ovule with a short aril (a) at the base. F Inset
F marked in E. (A, B, E, F After Loze 1965, C, D after Pankow 1962)
The integument is fused to the nucellus, except at the apex, and becomes
stony. Usually, two feebly developed vascular bundles (represented by phloem
strands) traverse upward inner to the stony layer. The xylem of these strands
extends up to the chalaza. They alternate with the uppermost pair of scale
leaves, and correspond to the two primordia from which the integument
arises. The aril also has a stub of vasculature at the base. There is a plate
meristem across the axis, at the base of the ovule. The meristem grows
upward at the periphery and around the ovule, and forms the aril
(Fig. 16.4 C-F). The latter persists in a primordial state until after fertilization,
298 The Gymnosperms
when it develops and ultimately extends beyond the seed. The solitary erect
seed borne in a fleshy cup-like aril ripens in one season. The aril becomes
scarlet and is attractive to birds (Fig. 16.1).
Megasporogenesis. At the time of differentiation of the megaspore mother

cell (mgmc ), the ce11 lineages of nucellus (from above) converge towards
the centre. The cells in the upper half of the nucellus are larger, isodiametric,
and have large vacuoles and thick walls. In the lower half the meristematic
cells are smaller, stain densely, and divide transversely. Approximately at
the junction of these two zones, there is a small central area with ca. 20
cells (as seen in longisection). These cells have dense cytoplasm, large
nuclei, take deeper stain than the circumjacent cells and differentiate as
sporogenous tissue. Simultaneously, the adjoining cells divide parallel to
the periphery and produce rows of cells which do not enlarge and appear
to radiate from the central area (Fig. 16.5 A-E).
The megaspore mother cell lies almost in the centre of the sporogenous
tissue. In T. canadensis, usually one cell, occasionally two, functions (Dupler
1917). InT. baccata the mgmc can be distinguished from the nucellar cells
by their larger size (up to 50 Jlm in diameter) and the composite nature of
their boundary walls. The combined wall is fibrillar, 300-500 nm wide,
and the middle lamella cannot be readily distinguished. Adjacent to the
protoplast of the mother cell, there is an additional layer (up to 120 nm
thick) of denser amorphous material. Plasmodesmata have not been observed
in the wall of the mother cell, although they are frequent in the nucellar
cells. Within the protoplast of the mother cell, the mitochondria and plastids
lie close to the chalaza! pole and form a distinct layer beneath the nucleus.
The envelopes of these organelles are associated with lipid droplets (Pennell
and Bell 1987).
The development of ovules on the same tree is synchronized (T. baccata).
The prophase in the mgmc lasts ca. 1 week and corresponds with the
germination of the pollen on the nucellus (Pennell and Bell 1986). Dividing
nuclei have rarely been seen. The cytoplasmic organelles in the diads are
distributed unequally, the mitochondria and plastids are confined to the
chalaza! cell. Subsequent division in the micropylar and chalaza! dyad
leads to aT-shaped tetrad (T. baccata-Pennell and Bell 1987). Most of
the mitochondria and plastids are included in the chalaza! megaspore, which
alone develops further, while lipid droplets are present in all the megaspores.
In T. canadensis after meiosis a linear tetrad is formed and any one or
all the four megaspores may develop (Fig. 16.5 A-E) so that supernumerary
gametophytes are common. Sometimes, the upper two megaspores enlarge
and divide while the lower two abort. Normally, the larger chalaza! megaspore
functions (Sterling 1948a). Although as many as three megaspores may
germinate (Fig. 16.5 C), only one forms the gametophyte (T. cuspidata); the
adjoining sporogenous tissue breaks down. The localized growth of the
Taxales 299
\
\
\
\
I
I
I
G J
I
~~~
~b
~(>;.
.~
_.:;:::::::::--- c
.
-~·
'
•
d
Fig. 16.5 A-K. Ta.xus cuspidata,longisection ovule, megaspore and female gametophyte.
A·E Linear megaspores, the position of functional megaspore varies. F -H Free-nuclear
female gametophyte. H Two gametophytes with opposite tentpole. I Cellular gametophyte,
indentation on the right is due to pressure of pollen tube (pt). J Uppn portion of mature
gametophyte with tent pole, archegonia and pollen tube. K a-d Neck cells in surface
view. (After Sterling 1948a)
300 The Gymnosperms
gametophyte gives it a flask-shaped appearance with a tent-pole-like tip.

The micropylar tip of the gametophyte does not enlarge and the-adjoining
sporogenous tissue persists as a deeply staining cap. When twin gametophytes
develop, the persistent sporogenous tissue is crushed by the second
gametophyte. Such gametophytes are usually superposed, the upper one
enlarges upward so that its tent pole faces the chalaza and the base the
micropyle (Fig. 16.5 A).
The megaspore membrane of Taxus resembles that of gymnosperms
generally (Pettitt 1966). This membrane is formed later and is not a direct
continuation of the wall of the megaspore mother cell. It may be formed
from the interactions of proteins characteristic of the two phases. It probably
continues to isolate the gametophyte from potentially informational molecules_
from the sporophyte (Pennell and Bell 1987).
The tapetum is poorly developed or absent. Following the absorption of
the sporogenous tissue, the developing gametophyte comes into contact
with the nucellar cells.
A central vacuole appears before the division of the megaspore nucleus
(Fig. 16.5 D,E). The division continues until (T. baccata) 250 after ca. 8
mitoses (Pennell and bell 1987), 256 (Jager 1899) or more than 280
(T. cuspidata, Sterling 1948a) nuclei are formed. The nuclei form a single
layer in the peripheral cytoplasm (Fig. 16.5 F-H). During the free-nuclear
stage, the gametophyte is enveloped in a thin, tenuous, pliable megaspore
membrane, often intimately associated with the circumjacent nucellar tissue.
In the living material, it can be dissected with much difficulty.
At the time of cellularization the gametophyte is pear-shaped/spherical
with an apical tent pole. Frequently, during development, the gametophyte
becomes distorted due to the growth of the pollen tubes (Fig. 16.5 I). The
gametophyte may be at various stages of development, four-nucleate to
mature archegonia. The pressure by the pollen tubes indents the gametophyte
and its outline changes; even the tent pole may become indented. However,
there is no "erosive" effect of the tube on the gametophyte (Sterling 1948a).
Frequently, while the tent pole is present at the younger stage, it may not
remain· distinguishable in a mature gametophyte.
Initiation of walls follows alveoli formation (Sokolowa 1890, Sterling
1948a). The alveolar cells form a honeycomb-like structure containing nucleus
in each hexagon (Fig. 16.5 I). According to Pennell and Bell (1987), in
T. baccata, wall formation begins shortly after completion of nuclear divisions
and the gametophyte becomes cellular in about 1 month. New walls arise
between the nuclei in positions marked out by the accumulation of cisternae
of the endoplasmic reticulum. It is not clear how the cisternae, which
coalesce to give rise to the walls, come to lie symmetrically between the
nuclei. Secondary spindles, or any other form of microtubular participation
in the siting of these walls, have not been detected. Jager (1899) suggested
that the nuclei and associated cytoplasm of the gametophyte of Taxus become
Taxales 301
enclosed directly by growing walls. According to Pennell and Bell (1987),

in the cellular development of the female gametophyte, Taxus therefore
resembles Gnetum (Sanwal 1962) and Welwitschia (Martens 1963), where
the nuclei and the associated cytoplasm of the gametophyte become enclosed
directly by growing walls.
The first formed cells of the gametophyte usually have large vacuoles
and lightly staining cytoplasm. Subsequently, divisions occur at the micropylar
and, later, at the chalazal end. The latter cells are small and stain deeply.
The micropylar zone, with archegoniaL initials, is generally three to five
cells deep. The enlargement of the gametophyte crushes the basal and
lateral nucellar tissue. At the same time, the enlargement of the archegonia
results in the widening of the apical part of the gametophyte. At the time
of fertilization, the gametophyte is often cordate.
The superficial cells (in the micropylar region), which function as
archegonial initials, have a basal vacuole. All the initials develop
simultaneously. A periclinal division cuts off a neck initial and a central
cell. The neck initial divides to form a single tier of two to four neck cells
(Fig. 16.5 Ka-d). The central cell has a conspicuous basal vacuole. As it
elongates, the nucleus situated near the neck in dense cytoplasm moves to
the centre of the archegonium; the cytoplasm becomes frothy-the foam
stage (Fig. 16.5 J). With the maturation of the archegonium, the dense
granular cytoplasm forms a sheath around the egg nucleus. Numerous
archegonia develop simultaneously and some of them are in contact with
each other. There are 4-8 archegonia in T. canadensis, 5-11 in T. baccata
and 6-25 (usually 8-14) in T. cuspidata. InT. cuspidata, the archegonia may
form a sheath around the invaginated gametophyte. Occasionally, when
there are two or more gametophytes in the same ovule, the archegonia may
develop towards either the chalazal or the micropylar end (Dupler 1917).
In the lower gametophyte, the archegonia abut on the pollen tube, which
grows between two gametophytes. In T. cuspidata some archegonia become
superposed and are presumed to be derived from periclinal division of a
single initial; the internal archegonia lack neck cells.
In T. canadensis, the nucleus of the central cell has not been observed
to divide, and probably functions directly as the egg (Dupler 1917). In
T. cuspidata the ventral canal nucleus is rarely formed. The archegonial
jacket is single-layered around an archegonium, or a group of archegonia
(T. cuspidata - Sterling 1948a). The jacket cells are smaller and denser
than the adjoining cells.
According to Pennell and Bell (1987),. earlier stages in the development
of the archegonium in Taxus is as follows. Two or four archegonial initials
appear. The cells are large (ca. twice the diameter of the nucellar cells),
occupy peripheral position, and are in contact with the megaspore membrane.
Each cell divides to give rise to a large cell. It is taO Jlm in diameter,
surmounted by a single tier of neck cells, and surrounded by tangentially
302 The Gymnosperms
elongated jacket cells. The primary_ cell divides and gives rise to a well-
defined neck canal cell (this is an unusual observation by the authors since
neck canal cell is absent in gymnosperm. It is likely to be ventral canal
cell) of 140 by 25 J..lm, and a central cell with highly organized cytoplasm.
The transverse wall which separates the two cells in the archegonial canal
is generally uniformly thick. Further nuclear divisions do not occur. A
mature archegonium is round, the egg cell is ca. 250 by 175 J..lm. According
to Pennell and Bell (1987), it remains to be resolved whether the cell
equivalent to the central cell becomes the egg cell directly (P. Maheshwari
and H. Singh 1967), or the nucleus divides and the resultant upper nucleus
degenerates quickly (Konar and Moitra 1980).
The cytoplasm of the egg cell is zoned. The outer cytoplasm has prominent
granular bodies, 2-5 J..lm in diameter (cf. "grandes inclusions" in the egg
cells of conifers, see Chap. 2). They are derived from hypertrophied plastids
which have encapsulated part of the cytoplasm (there is a complete absence
of recognizable plastids in the mature cell). A conspicuous feature is the
presence of bundles of microtubules which radiate from the nuclear envelope
and possibly stabilize the zonation of the cytoplasm.
Pollination
Pollination occurs around the time the megaspore mother cell is undergoing
meiosis. The collections made soon after pollination show pollen tubes on
the irregular surface of the nucellus, the megaspore mother cell undergoes
meiosis, or a four nucleate gametophyte (T. cuspidata).
A pollination drop is secreted by the apical cells of the nucellus, and the
secretion continues even when the micropyle is amputated (Ziegler 1959).
The chemical composition of the pollination drop is comparable to that of
the extract of nucellar cells. The high concentration of sucrose in the
pollination drop is attributed to the occurrence of acid phosphatases in the
nucellus. The enzymes release sucrose from sucrose phosphate. Several
amino acids, peptides, malic and citric acids are also present in the pollination
drop.
The wingless pollen grains caught in the pollination drop imbibe the
fluid, shift from the micropyle and come. to lie against the lowermost surface
of the drop. The fluid (along with the pollen) is eventually reabsorbed by
the ovule and pollen reaches the nucellus.
Post-Pollination Development of Male Gametophyte. The development

of male gametophyte following pollination to fertilization was investigated
by Dupler (1917) and Sterling (1948a). The pollen is shed at the uninucleate
stage, a prothallial cell is not cut off, and the microspore nucleus functions
directly as the antheridial initial. The nucleus divides ca. 2 months before
fertilization, when the pollen grain has germinated and the tubes have
grown in the nucellus. The antheridial and tube cell are formed, and the
Taxales 303
antheridial cell divides to form the stalk and body cell. In T. cuspidata, the
stalk nucleus remains closely adpressed to the body cell on its entry into
the pollen tube. Eventually, the stalk nucleus moves close to the tube nucleus
located ahead of the enlarging body cell, and just behind the tip of the tube.
The two sterile nuclei, embedded in abundant cytoplasm, are identical in
size and structure. In the beginning, the body cell is more or less conical
with its enlarged nucleus situated at the base. It enlarges, becomes spherical,
and has dense cytoplasm with a distinct membrane. The body cell nucleus
is eccentric, located diametrically opposite to the two sterile nuclei. Before
marked expansion of the pollen tube, the sterile nuclei usually lie below the
body cell, but their relative position changes during later stages. Thus, the
entire complex may revolve as much as 180° along its horizontal axis so
that the sterile nuclei come to lie above the body cell. The much-enlarged
body cell (60 J.Lm in diameter) forms two unequal male cells. According to
Favre-Duchartre (1960b), a wall is absent between the two gametes which
should be considered as nuclei. In T. baccata, under in vitro conditions,
Rohr (1973a) observed two equal male cells unlike those produced in vivo.
The pollen tubes expand in a characteristic way and the sac-like structure
may cover the entire apex of the female prothallus. The dense cytoplasm
at the tip of the tube becomes vacuolate. The sperm cells mature and the
sterile nuclei can still be distinguished.
As the pollen tube grows through the nucellus, it becomes wider,
occasionally twists and branches. Just before it reaches the gametophyte,
the tube often branches once, and one branch grows on each side of the tent
pole. The tip of each branch expands a good deal, either on the shoulders
of the gametophyte, or over its apex, and often becomes as broad as the
prothallus itself. When several tubes grow simultaneously, they "pile up"
on each other so tha'f the sac-like expansions of successive tubes form a
heap above the prothallial apex. Occasionally, the tubes grow up to the
gametophyte and meet below the prothallus (Saxton 1936, Sterling 1948a).
The wall of the pollen tube is quite thick.
Fertilization
According to the variability in the development of the gametophyte
(T. cuspidata), fertilization may take place in 4 weeks or more. At fertilization,
the prothallus becomes relatively smaller as compared to its size at maturity.
After fertilization in Taxus sp., conspicuous growth occurs in the gametophyte,
which is in contrast to the condition in the conifers (particularly Pinaceae),
where the gametophyte is fully grown, or nearly so, at fertilization.
The male gamete contains dense granular material and stains more deeply
than the much longer egg nucleus. The functional male gamete comes to
lie close to the egg nucleus in the centre of the egg. The cytoplasm above
the mating nuclei is highly vacuolar and looks frothy. The male gamete
"sinks" into the egg nucleus, and they move to the base of the archegonium.
304 The Gymnosperms
The mating gametes, as well as the early proembryonic nuclei, are embedded
in a highly granular, deeply staining cytoplasm at the base of the egg. The
dense cytoplasm appears different in consistency and staining capacity from
the cytoplasm around the unfertilized egg nucleus, and may even include
some cytoplasm of the male gamete. Whether the male and female nuclear
material integrates completely or· not requires further study. In two
preparations, Sterling (1948a) observed only a single resting nucleus at the
base of the egg. All other preparations. of this stage showed either the two
mating nuclei or the telophase of the first mitotic division.
Embryogeny
During division of the zygote, the spindle lies transversely to the long axis
of the archegonium. The resultant two free nuclei lie close together in the
dense cytoplasm at the base of the egg cell. Occasionally, the division may
take place in the centre of the archegonium, and the two nuclei migrate to
the base. The next division is also in the transverse plane, but at right
angles to the earlier division, and produces a tier of four nuclei perpendicular
to the long axis of the archegonium. Following further division, the nuclei
become progressively smaller and irregularly distributed in the proembryonic
cytoplasm.
The walls are laid down at the 16- or 32-nucleate stage on spindle fibers
which extend between all the nuclei. With wall formation, the nuclei become
evenly distributed in the dense cytoplasm, and the cells lie approximately
in two tiers: a primary upper tier (pu) and a basal primary embryonal tier
(pE). The pu tier divides transversely and produces an upper tier (U) of "open"
cells, and a lower tier of closed cells which develop into the suspensor (S).
Simultaneously, or just before the suspensor elongates, the cells of the
basal group divide irregularly. Thus, from the base upward, a proembryo
(T. cuspidata) comprises a group of (6-14) deeply staining embryonic cells
(E), followed by another group of (9-13) suspensor cells (S), topped by a
tier of (9-3) cells (U) "open" to the archegonial cytoplasm (Fig. 16.6 A).
The proembryo occupies the lower half of the archegonium (Fig. 16.6 A, B).
Towards the neck of the archegonium, several supernumerary nuclei (from
the pollen tube) may persist in the archegonial ·cytoplasm; eventually they
disintegrate.
Soon after the organization of the proembryo, the suspensor cells elongate
and thrust the embryonal cells through the base of the archegonium
(Fig. 16.6 B). There is dissolution of the cells in the central region of the
gametophyte to form a corrosion cavity (cf. conifers). Meanwhile, at its
chalazal end, the gametophyte enlarges considerably. In the lower region,
the cells divide repeatedly and the numerous cells appear small. With further
development, the gametophyte becomes spherical and later pear-shaped
and remains so until seed ripening.
The densely cytoplasmic embryonic cells at the tip of the suspensor
Taxales 305
D
E
Fig. 16.6 A-G. Taxus cuspidata, embryogeny. A Longisection three-tiered proembryo

shows embryonal (E), suspensor (S) and upper tier ( U) . B Suspensor. C Early embryo
shows embryonal mass (e) at the tip of elongated suspensor, and embryonal tubes (et).
D-G Development of embryo, longisections. D, E Embryonal mass shows anticlinal and
periclinal divisions. F Cells marked X represent the focal zone (jz). G Cell walls oriented
around the focal zone. (After Sterling 1948b, 1949a)
remain quiescent during their passage into the endosperm. Divisions in the
embryonic group result in the formation of successive secondary suspensors,
the embryonal tubes (et; Fig. 16.6 C) which elongate autonomously. The
latter may elongate, while the original suspensor cells remain relatively
short. Most of the suspensor cells elongate synchronously and present a
tiered configuration. The suspensor cells enlarge continuously and their
diameter increases. The nucleus and most of the cytoplasm is located at the
tip. After elongation, a variable number of suspensor cells may separate
from one another with or without embryonic cells attached to their tip.
These embryonic cells form a meristematic group simulating the original
embryonic group. This cleavage or splitting results in the production of two
to four embryos, each consisting of embryonic cells borne on several suspensor
cells. More often, the embryo develops without such cleavage. Separated
single suspensor cells may also show some proliferation.
306 The Gymnosperms
The open cells generally degenerate, but some of the completely walled
cells may undergo a few divisions.
The embryonal primordium gradually becomes massive by periclinal
and anticlinal divisions at the free tip; the rib-meristem activity (behind the
apex) contributes to its mass as well as to that of the suspensor (Sterling
1948b).
Sterling (1949a) has studied the meristematic development, and tissue
differentiation of the embryonic group of cells in T. cuspidata.
After elongation of the suspensor tier, the embryonic cells become active
and, along the surface of the young embryo, a group of densely cytoplasmic
cells divides periclinally and anticlinally (Fig. 16.6 D, E). As the tip enlarges,
oblique di_visions also occur in the superficial cells. With continued
enlargement, anticlinal divisions predominate on the forward flanks of the
apex, while periclinal divisions continue at the summit. Growth activity is
limited to a small group of initials along the surface of the massive embryonal
apex. From these initials, all the other cells of the embryo (appear to)
diverge in periclinal rows. The anticlinal cells appear as concentric arcs
radiating from the initial region, indicating successive growth increments
in the apical development. After apical growth has occurred in the young
embryo for some time, a focal area of lighter-staining cells differentiates
just behind the apex (Fig. 16.6 F). The focal area enlarges and adjacent
cells divide with walls concentric to this area (Fig. 16.6 G). The concentric
cells divide along the lateral limits of the focal area, are arranged in a cup-
shaped group, somewhat flared below the apex of the embryo. These cells
constitute the procambial cylinder; pith is absent. There is a group of distinct
cells with relatively large spherical nuclei at the base of the procambial
cylinder-the root generative meristem.
As the focal group becomes less active, the cells produced by the lateral
concentric divisions build up buttresses of tissue on the flanks of the free
apex of the embryo (Fig. 16.7 A, B). Deeply staining cells in these buttresses
have a histological continuity with the cells of the elongating procambial
core of the hypocotyl. Cotyledonary primordia develop from the buttresses
by predominantly subepidermal activity; later growth of the cotyledon occurs
by the activity of the superficial initials. The procambial strand in each
cotyledon, continuous with the procamb1al core, develops acropetally behind
the apex of the cotyledon (Fig. 16.7 C-F).
The root generative meristem produces a tissue simulating the "column"
of the pinaceous root cap, only after marked elongation of the hypocotyl
and enlargement of the cotyledon. This tissue is very indistinct, there is no
juncture zone on the outer surface of the embryo, and the epidermal and
several hypodermal layers continue from the cotyledons into the suspensor
(Fig. 16.7 C). There are usually two, occasionally three, short cotyledons.
The fleshy aril (which covers the seed) comprises thin-walled cells rich
in cell sap. The epidermis has numerous stomata. The cells situated below
Taxales 307
Fig.16.7A-F. Taxus cuspidata, longisections of early and late embryos. A Early embryo
shows concentric divisions around focal zone (jz). B Root, generative meristem. C Part
of hypocotyl and root cap (rc) region, the epidermal and two hypodermal layers are
continuous from the hypocotyl to the root cap. D-F Initiation and development of
cotyledons (cot). (After Sterling 1949a)
the outer epidermis are isodiametric, those next to the inner epidermis are
flattened. The cells in the middle zone are large and vacuolate. The base
308 The Gymnosperms
has a stub of vasculature. During earlier stages the cells are chlorophyllous,
later several of them contain tannin.
On the basis of morphological and ontogenetical evidence, Andre ( 1956)
and Loze (1965) interpret the aril as the~econd integument, which originates
as a foliar organ. This interpretation is erroneous, since an integument
cannot have a foliar origin.
Seed
Both chalaza and integument contribute to the seed coat. There is an increase
in the number of cell layers, the sarcotesta is absent (in Taxales), while
sclerotesta and endotesta differentiate. A plate of thick-walled cells develops
at the chalaza, is one to several layers of palisade-like simple pitted cells,
and is contiguous with the sclerotesta. Several traces enter the chalaza, pass
through the basal plate, and traverse the length of seed coat through endotesta
(see Schnarf 1937).
Germination. The germination of seed is hypogeal and the first two linear
cotyledonary leaves pierce the soil. The primary root persists.
Chromosome Number
In Taxus wallichiana, the haploid chromosome number (as determined
from endosperm squashes) is n=12 (Fig. 16.8). Mehra (1988) states that
the karyotype could not be analyzed precisely but there are definitely
three isobrachial chromosomes (Fig. 16.8 I, II, VI), three hetero-
brachial chromosomes (Fig. 16.8 III, V, VII), and the rest apparently have
..
XI
VI IV
Ill
II
Fig. 16.8 Taxus wallichiana, endosperm cell with 12 chroma;omes (squash preparation).
(After Mehra 1988)
Taxales 309
terminal or near-terminal primary constriction (Fig. 16.8 IV, VIII, IX, X,

XI, XII).
Taxus shows a 1-year life cycle. According to Sterling (1948a), the entire
development from megaspore formation to seed maturity takes place within
the space of a single season.
The position of the Taxaceae in the Coniferales has long been disputed.
Sahni (1920) first suggested that Taxus, Torreya and Cephalotaxus should
be separated and placed in a phylum of their own. Florin (1948, 1951)
agreed, but considered Cephalotaxus a true conifer. He (Florin 1954, 1955,
1958) considered the taxads to be sufficiently distinct to form a separate
class, the Taxopsida, and segregated the Taxaceae from the other conifer
families. The main basis for such a treatment is the distinctive type of
reproductive organs in the taxads. The female reproductive structures are
not organized into cones, in either living or fossil genera. The taxads appear
to deviate from the conifers in the single terminal seed (enclosed in an aril)
borne on fertile shoots, and the perisporangiate microsporophylls. In the
Coniferopsida, the ovule is a lateral organ and its position is consistent
with the general interpretation of a conifer cone as a conduplicate strobilus
(see Miller 1977). Similar axial and terminal ovules are associated with the
extinct taxa Paleotaxus (Triassic) and Taxus jurassica (Jurassic), a feature
of great antiquity. However, the epidermis of Paleotaxus is quite different
from that of Taxus. According to Stewart (1983), this is surprising, because
several characters are in common with Taxus.
Because of lack of clear fossil evidence, the origin of the perisporangiate
microsporophyll, notably in Taxus and later in Pseudotaxus, has long been
a subject of speculation. Although the taxads have been recognized as
advanced in several morphological characters, the perisporangiate
microsporophyll is considered primitive. Wilde (1975) interpreted the male
cones in Taxus and Pseudotaxus as evolved from compound structures (similar
to those of Cephalotaxus), presenting evidence to show that the axis of
these vestigial cones is a reduced fertile branch on which each fertile branch
unit (cone) is reduced to a single terminal perisporangiate microsporophyll,
attached directly on the fertile branch. Such sporophylls resulted from the
phylogenetic fusion of two or more apical, hyposporangiate microsporophylls.
The male structures in Taxus are, therefore, interpreted as extremely reduced
and not primitive (Wilde 1975).
According to H. Singh (1978), the only novel feature in taxads may be
their ovules, and this character alone cannot be a sufficient justification to
treat them as a separate order or class. Embryologically, the taxads are like
the conifers in several respects: (a) unwinged pollen and absence of prothallial
310 The Gymnosperms
cells in the male gametophyte (as in Cupressaceae, Taxodiaceae and

Cephalotaxaceae), (b) archegonia occur singly (as in Pinaceae,
Podocarpaceae), and (c) a general pattern of embryogeny, as in conifers.
While various studies (Keng 1969, Harris 1976 and others) have not
solved the problem, they indicate a disagreement with the present separation
of the taxads and conifers, and emphasize a reconsideration of the
relationships. The subject therefore deserves prime attention in future research
(Miller 1977).
17. Gnetopsida
The extant taxa Ephedra, Gnetum and Welwitschia were earlier placed in
a single order, Gnetales, and a single family, Gnetaceae. These three taxa
(considered to be the highest evolved among the gymnosperms) lack fossil
record except for the remains of pollen of Ephedra from the Eocene
(54 my B.P.), and pollen-like remains of Ephedra and Welwitschia from
the Permian (280 my B.P.) (Delevoryas 1962). Certain common features of
these taxa are comparable to angiosperms: (a) vessels are present in the
wood, (b) the microsporangia and ovules are borne on fertile shoots which
form compound strobili, (c) the ovules are enclosed within one or two
envelopes, in addition to the integument and (d) the upper part of the inner
integument extends into a long tubular structure, the micropylar tube.
In spite of these common features, the heterogeneous nature of the order
has been apparent, and resulted in its constantly changing taxonomic treatment.
By the end of the 19th century, the order Genetales comprised three families
with one genus each. Further knowledge, especially about reproduction,
revealed significant differences between the three genera. Florin (1931,
1933b) and Eames (1952) supported the establishment of three independent
orders: Ephedrales, Welwitschiales and Gnetales, each comprising a
monogeneric family. The phylogenetic consideration of these taxa are
discussed in Chap. 21.
18. Ephedrales
The plants are herbs, woody shrubs or Hanas, leaves free, scale-like, and
the stem jointed and green. Secondary wood has vessels. The plants are
dioecious with compound male and female strobili. The ovule is surrounded
by two envelopes, the inner projects as a long tube and archegonia are
present. The embryo is dicotyledonous.
Ephedraceae
Ephedra
Morphology
The plants are highly branched herbs, shrubs or Hanas. Ephedra campylopoda
is a cultivated ornamental with pendulous branches, and is commonly grown
in hanging baskets. It grows naturally between the crevices of the Wailing
Wall in Jerusalem (Fig 18.1 A). E. triandra grows into a small tree, and
E. gerardiana is a perennial herb. E. foliata is a scrambling shrub that may
reach a height of about 6m, may climb up the trees, or walls, or, in the
absence of a support, may spread along the ground.
The stem is long, jointed, fluted and green. It has distinct nodes and
internodes (Fig 18.1 B). The internodes in most species are longitudinally
ridged and the ridges of the successive internodes stand on alternate radii.
The shoot comprises indeterminate as well as determinate branches, although
the distinction between the two is not as well marked, as in Pinus. On each
node, the indeterminate shoot bears three, occasionally four, leaves in a
whorl. The axil of some of the leaves bears determinate shoots with leaves
in opposite and decussate pairs (E. foliata, Tiagi 1966). On a node, the
leaves stand on a radii .of the ridges of the internode below. Since the
ridges of the successive internodes alternate, so do the leaves of the successive
nodes. Branching is axillary (Fig. 18.1 D), and additional accessory buds
arise below and at the base of axillary buds. An intercalary meristem occurs
above each node (Fig. 18.1 E, im, ima), and it produces most of the internodal
tissue. Later, it forms either abscission layers, or may mature as transverse
bands of sclerified parenchyma.
Ephedrales 313
Fig. 18.1 A-E. A Ephedra campylopoda, rooted plants between the stones of the Wailing
Wall in Jerusalem. B-D E. gerardiana . B Vegetative twig. C, D Scale-like leaves fused
at the base to form a sheath. E E. antisyphilitica, longisection of node with axillary bud
(ab), intercalary meristem (im), and intercalary meristem of axillary shoot (ima). (After
Bierhorst 1971)
The scaly leaves are deciduous. They may be opposite or whorled

(Fig. 18.1 C, D). Each leaf receives two traces which have distinct origin
(see below). The leaves are ultimately shed after the formation of an abscission
layer.
Anatomy
Root. The roots of E. foliata have a long cap. The apical meristem can
be distinguished into: (a) a discrete layer of stelar initials, (b) the columella
and its initials, and (c) a common region for the initiation of cortex and
peripheral region of the cap (Pillai 1966).
In cross section, the root shows epiblema, cortex, endodermis, pericycle
and a diarch vascular region (Fig. 18.2 A).
314 The Gymnosperms
n
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bf------;>~
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Fig. 18.2 A-C. Root and stem anatomy. A, B Ephedra sp. A Transection young root.
m.x metaxylem. px protoxylem. B Transection of old stem. C. E. californica, tracheary
elements from secondary xylem; a, b tra:heids (tr) with bordered pits, c-e vessel members
(ve) with foraminate plates (jpp) on end walls . (C after Esau 1953)
Stem. In longisection the shoot apex in E. altissima (Strasburger 1872,

Gifford 1943) and E. fragilis var. campylopoda (Seeliger 1954) shows a
well-defined tunica and a corpus (Fig. 18.3). A tunica layer is also reported
in Gnetum, Araucaria brasiliana, (Strasburger 1872), and permanent shoots
of Taxodium distichum (Cross 1939). In Cryptomeria japonica (Cross 1941 ),
Agathis lanceolata (Sterling 1958) and a few Araucaria spp. the tunica layer
is rather restricted (Griffith 1952). The presence of tunica and corpus is a
regular feature in angiosperms. The stem of Ephedra is also capable of
elongation by an intercalary meristem present at the base of each node
(Fig. 18.1 E).
Transection of the stem exhibits ridges and furrows (Fig. 18.2 B). A
single-layered epidermis is covered with a thick cuticle. Sunken stomata
occur in rows in the furrows only. The stomata are of the haplocheilic type,
i.e the two guard cells originate from a common mother cell, and the
Ephedrales 315
Fig. 18.3. Ephedra altissima. Longisection of shoot apex, the tunica layer is well
defined. (After Foster and Gifford 1959)
neighbouring epidermal cells become modified as subsidiary cells. Below

the epidermis, in the region of the ridge, patches of sclerenchymatous tissue
are very conspicuous. The cortex comprises palisade-like chlorenchyma,
which actively photosynthesizes and compensates for the scale-like leaves.
Besides the photosynthetic zone, chloroplasts also occur in thin-walled
parenchymatous cells of the cortex. The endodermis is distinct, while the
pericycle is ill-defined.
In Ephedra, the leaves are opposite and decussate, or in whorls of three
or four. Accordingly, there is variation in the number of vascular bundles
from species to species (see Sporne 1965). In E. foliata (Tiagi 1966), the
vascular anatomy of the indeterminate shoots which bear three or four
leaves in whorls is fundamentally alike. A transection through the middle
region of an internode of a stem, which bears leaves in whorls of three,
shows three ridges and three alternating furrows (Fig. 18.4 A). There are
15 vascular bundles arranged in three groups of five each, below the three
ridges (Fig. 18.4 A). In the subnodal region all the five bundles (under a
ridge) become laterally united to form a single vascular arc (Fig. 18.4 B, C).
Slightly higher up, each of these three arcs splits radially into two bundles
(Fig. 18.4 D). Two traces each, one from either side of the gap thus formed,
are first given off to the leaf, subsequently another pair to its axillary
branch (Fig. 18.4 E, F). The two branch traces quickly divide twice in a
316 The Gymnosperms
Fig. 18.4 A-M. Ephedrafoliata, serial transections through successive nodes and two
internodes of an indeterminate shoot with leaves in whorls of three. bt branch trace.
It leaf trace. (After Tiagi 1966)
radial manner to form the stelar ring of eight bundles of the axillary shoot
(Fig. 18.4 F, G). Simultaneously, two vascular strands between any of the
three gaps become laterally united to form a single arc (Fig. 18.4 H, I). As
the leaf sheath and the axillary shoots separate from the stem at the node,
each of the arcs radially splits into a group of five separate vascular strands
below each of the three ridges of the next higher internode (Fig. 18.4 J-M).
Ephedrales 317
This process of fusion, branching and regrouping is repeated below every

node. Consequently, the three sets of five bundles below each ridge in an
internode alternate in position with their counterparts in the internode above
but, again, become superposed in the alternate internodes. The longitudinal
course of the vascular bundles in the stem through three successive internodes
and two nodes in shown in Fig. 18.5.
In the determinate shoots with leaves in opposite and decussate pairs,
the vascular anatomy is similar to that of the indeterminate shoots except
that under each ridge there are only four bundles (the middle bundle is
absent).
According to Tiagi (1966), the primary vascular system in Ephedra
(based on his work on E. foliata) is best interpreted as a perforated, ectophloic
type of dictyostele where the breaks in the vascular cylinder internal to the
furrows and the ridges are leaf-cum-branch gaps and perforations, respectively.
Marsden and Steeves (1955) made a detailed study of the primary vascular
system in a number of species of Ephedra bearing two or three leaves at
a node. They interpreted the primary vascular system as an eustele in which
the internal bundles are continuous with the leaf traces.
The xylem consists of tracheids, vessels and xylem parenchyma. The
vessels originate from the pitted tracheids. Transitional forms between typical
tracheids and foraminate vessel members, characteristic of Ephedra, have
been observed (Fig. 18.2 Ca-e). The plates show a uniseriate arrangement
of pores in early-formed elements, while in the later-formed plates most
pores are two-ranked, or irregularly grouped. There is a persistent primary
cambium but not much secondary wood formation occurs, so that the vascular
cylinder does not become thick.
A transection of stern of Ephedra with secondary growth appears more
like an angiosperm than a gymnosperm, because of the deep origin of the
periderm, slight increment of phloem, multiseriate and dilated phloem ray
and ring-porous xylem (Fig. 18.2 B).
A peculiar anatomical feature is the presence of a diaphragm-like plate
of cells at the base of each internode. This makes the stem easily separable
at the nodes.
The phloem in Ephedra combines characteristics of both angiosperms
and gymnosperms, and has some unique features of its own.
Alosi and Alfieri (1972) investigated the structure of phloem in
E. californica and E. viridis. The conducting phloem lies outside the cambial
zone. It consists of parenchymatous rays, sieve cells, and axial parenchyma
which includes albuminous cells. In the non-conducting phloem the sieve
elements cease to function. This region is comparatively small due to the
formation of annual rings of periderm which originate deep within the
previous season's phloem (Fig. 18.6 A). The annual periderm arises in
spring at the same time as the vascular cambium is reactivated. Fusiform
prenchyma cells which lie internal to the periderm develop into fibre sclereids.
318 The Gymnosperms
CABACCABAC CABAC
Fig.18.5. Ephedrafoliata, vascular skeleton of three successive nodes and two internodes
of an indeterminate shoot with leaves in whorls of three. The skeleton has been cut
longitudinally on one side and spread open. Approximate levels of transections A-L are
shown in Fig. 18.4. bt branch trace. It leaf trace. (After Tiagi 1966)
All sclerified cells in both the axial and ray systems generally contain
crystal sand in the region of the middle lamella.
The immediate phloem derivatives resemble the cambial initials. They
are long, have primary pit fields in their primary cell walls, small radial
Ephedrales 319
Fig. 18.6 A, B. Ephedra californica. A Radial sections of bark to show (left to right)
old periderm (pd), collapsed phloem, non-conducting phloem (np), conducting phloem
(ph) with fusiform parenchyma (jp), cambial zone (cz), xylem (x) and a vessel (ve) .
B Tangential section to show differentiated phloem with oblique anticlinal divisions,
sieve areas and sieve elements. (After Alosi and Alfieri 1972)
diameter, and oblique end walls. The cellular contents take a light stain, the
nucleus is granular with several nucleoli. An ovoid body appears near the
nucleus in the differentiated cell. It resembles the largest nucleolus in shape
and staining. Because of its positive staining reaction with Ponceau S, this
body has been referred to as the slime body (as in the angiospermous sieve
elements). However, ultrastructural studies by Behnke and Paliwal (1973)
failed to confirm this.
The nucleus flattens and in some cells appears to curl around the perphery
of the cell in a bracelet-like fashion. It may be irregular in outline, but
maintains its granular character and chromaticity. Degeneration of the nuclear
membrane has not been observed. Necrotic nuclei (as in Pinaceae) are
frequent in the mature sieve cells.
Mature sieve elements vary in shape from extremely tapered (Fig. 18.6 B)
to blunt-ended. They remain thin-walled with no apparent secondary
thickening. The secondary sieve elements are of two categories based on
their length: ca. 400 and 220 jlm long. The former are direct derivatives of
the fusiform initials, whereas the latter arise after a transverse division in
their precursors. This shortening of sieve elements has been considered an
advanced character (see Paliwal 1992).
In a radial view, the end walls comprise a row of closely spaced sieve
areas (Fig. 18.7 A) similar in size and shape to lateral sieve areas
320 The Gymnosperms
Fig. 18.7 A, B. Ephedra californica. A, B Radial sections of phloem. A Sieve areas

with sieve fields on overlapping radial end walls of sieve elements. B Lateral sieve areas
on radial walls. (After Alosi and Alfieri 1972)
(Fig. 18.7 B). Within the sieve areas, the pores are generally ~ggregated
into groups (Fig. 18.7 A), termed sieve fields. Each pore measures
ca. 0.8 J1 m. Sieve ~reas do not occur on transverse walls between sister
cells (two sieve cells or a sieve cell and albuminous cell).
The differentiation of sieve areas begins very early, and progresses very
rapidly in developing sieve elements. Presumably, full perforation of the
sieve-area pores occurs just before the element becomes functional.
Numerous conspicuous albuminous cells are present in the axial system
of Ephedra. They may occur in radial files, or scattered in the conducting
phloem. The albuminous cells have dense protoplasm, rich in elongated
mitochondria, plastids, granular ER, free ribosomes and a prominent nucleus.
They are connected to the sieve cells by branched plasmodesmata on their
side, fusing with a sieve pore on the sieve cell side (see Paliwal 1992). The
two daughter cells from a transverse division may not differentiate into a
similar cell type; one may develop into a sieve cell, the other into an
albuminous cell or phloem parenchyma cell.
There is a remarkable intergradation between the elements of phloem:
sieve cells ~ albuminous cells ~ parenchyma. It is difficult to distinguish
between certain sieve cells and albuminous cells, and also between some
albuminous cells and other phloem parenchyma cells. Although most sieve
Ephedrales 321
elements and some parenchyma cells may occur as long fusiform cells, any
of the three cell types may arise from a daughter cell following transverse
division of a fusiform phloem mother cell. That an albuminous cell of
Ephedra forms directly from a single division of a phloem mother cell
distinguishes it from albuminous cells of most other gymnosperms, and is
comparable to the specialized parenchyma cells, including companion cells,
of higher plants.
Reproduction
Terms like strobili, inflorescence, flower, bract (commonly used for describing
angiosperms) are often used for Ephedra, Gnetum and Welwitschia for
comparison only; there are no phylogenetic implications.
Ephedra is typically dioecious, although bisporangiate cones
(Pearson 1929) have been recorded on the same plant.
Male Strobilus
The male strobili are in clusters at the nodes of branches (Fig. 18.8 A),
each strobilus arising in the axil of a scale leaf so that their number depends
on the number of scale leaves. The microsporangiate cone is regarded as
a compound structure. It consists of a number of pairs of decussately arranged,
broad, cupped bracts, the lowest pair of which is sterile (Fig. 18.8 B). Each
remaining pair of bracts develops a solitary microsporangiate shoot in its
axil. This shoot continues into a short axis. The microsporangiophore
(antherophore) bears a pair of bracteoles or perianth fused at the base. In
their upper free region, the posterior bracteole overlaps the anterior one,
and completely encloses the antherophore. The latter outgrows the sheath
and protrudes at maturity, bearing a variable number of terminal anthers
(Fig. 18.8 C).
Microsporangium. Both E. foliata and E. gerardiana show a similar

development of the antherophore and microsporangium (H. Singh and
K. Maheshwari 1962). In the axil of each bract of a male cone, an antherophore
arises as a protuberance; two perianth primordia differentiate laterally
(Fig. 18.9 A, B). The protuberance has a tunica-corpus organization which
is disturbed due to periclinal divisions in the tunica soon after the
differentiation of perianth (Fig. 18.9 B). A group of hypodermal archesporia!
cells appear in the protuberance after the demarcation of the perianth
(Fig. 18.9 C, D). After some time, the protuberance becomes lobed. Each
lobe is the primordium of a sporangium (Fig. 18.9 E, F), and contains a
plate of hypodermal archesporia! cells. Later, in each sporangium, a band
of cells becomes sterile, forming two chambers (Fig. 18.9 G). The outermost
cells of the archesporia! tissue divide periclinally and give rise to an inner
sporogenous layer and the outer wall layer. The latter divides again to form
a middle layer and the tapetum (Fig. 18.9 H, I). Anticlinal divisions occur
322 The Gymnosperms
Fig. 18.8 A-C. Microsporangiate strobilus. A Ephedra chilensis, twig with

microsporangiate strobili. B, C Ephedra sp. B Male strobilus . C Mature male flower
(from B) after removal of bracts, the antherophore bears sporangia at its apex and
protrudes by displacing the bracteoles. (A After Foster and Gifford 1959, B, C after
Bierhorst 1971)
in the wall layers to keep pace with the growth of the anther. With further
growth of the sporangium, the middle layer becomes flattened and crushed.
The tapetal cells enlarge, become densely cytoplasmic, and show two to
four nuclei when the microspore mother cells enter prophase.! (Fig. 18.9 J).
The tapetum begins to degenerate soon after the reduction divisions are
over. Only the epidermis persists in the mature microsporangium; its walls
Ephedrales 323
Fig. 18.9 A-K. Ephedra gerardiana, development of microsporangium. A, C, E

Longisection of male flowers at progressive stages of development. br bract. col column.
pe perianth. B Column from A. D Later stage to show perianth and a plate of hypodermal
archesporia! cells (arc). F Sporangium from E. G Two groups of archesporia! cells
formed by sterilization of sporogenous (spt) tissue. H-J Tra11section of sporangium.
ep epidermis. mid middle layer. tap tapetum. K Epidermis from mature sporangium.
(After H. Singh and K. Maheshwari 1962)
thicken and show a wavy outline (Fig. 18.9 K). The sporogenous tissue
increases due to repeated mitotic divisions.
In a young microsporangium of Ephedrafoliata (see Moitra and Bhatnagar
1982), there is a difference in the pattern of staining of the walls of
sporogenous and tapetal cells, and the other wall layers. The cell walls of
wall layers are thick, and take a brighter magenta stain with periodic Schiff's
(PAS) reaction than the walls of tapetum and sporogenous cells (Fig. 18.10 C).
324 The Gymnosperms
The cell walls of the epidermis and middle layer are mostly of cellulose,
and of tapetal and sporogenous cells mostly pectinaceous. There is no
starch in the microsporangium until the microspore mother cell stage
(Fig 18.10 A); it then appears in the tapetum and wall layers when the
microspores are formed. In the wall layers, starch is consumed during and
after meiosis. The polysaccharides become depleted in the wall layers but,
simultaneously, increase in the locular fluid.
Fig. 18.10 A-D. Ephedra foliata, microsporogenesis. A Longisection of microsporangium

to show starch (sr) in wall layers and tapetum (tap). spt sporogenous tissue. B Mature
pollen grains. C Longisection of microsporangium stained with spirit blue, tapetal orbicules
and pollen grains (pg) take up similar stain. D Association of pollen grains with tapetal
orbicules (to). (After Moitra and Bhatnagar 1982)
In E. foliata, an increase in Feulgen stainability occurs at three stages:

(a) just before meiosis, (b) at the interphase between the mother cell meiosis
and microspore mitosis, and (c) in the antheridial cell.
Throughout their development, the sporongenous cells are rich in RNA.
The maximal level in mother cells reaches prior to meiosis. A loss of
cytoplasmic basophilia has been observed, especially during meiosis II.
The young microspore is poor in RNA but the level rises again at the time
of antheridial cell formation.
The archesporia!, later sporogenous, cells are rich in total proteins. The
level is high in the microspore mother cells, but low in microspores. The
protein content in the rnicrospores increases sharply before microspore mitosis
(Fig. 18.10 B). The antheridial nucleus in the pollen grain has less protein
than the tube nucleus
Ephedrales 325
Microsporogenesis. Before meiosis, the microspore mother cells undergo

a gradual stretching, thinning and loss of PAS stainability. Their walls
dissolve and a thin layer of callose develops between the cell wall and the
plasma membrane. With the advance of meiosis, callose increases in thickness.
The wall formation is simultaneous (as in most gymnosperms), the callose
grows centripetally, and separates the four nuclei into a tetrad (Fig. 18.11
A-C). Numerous vesicles of Golgi origin are active at anaphase II and
telophase II. They become arranged in the region of the future wall. Later,
a cellulosic wall separates the resultant cells. The four microspores are
usually arranged in a tetrahedral or decussate fashion. Meiosis is asynchronous,
so that meiotic stages range from prophase I to telephase II, in various
microspore mother cells in a microsporangium.
A peritapetal membrane completely covers the tapetum and developing
pollen grains, in Ephedra foliata. It is acetolysis-resistant and can be separated
as a bag containing pollen grains.
Alongwith the development of tapetal membrane, tapetal orbicules are
formed in the tapetal cytoplasm which completely mask the developing
pollen grains (Fig. 18.10 C, D). The staining intensity of these orbicules
increases, accompanied by an increase in the intensity of the pollen exine.
Young microspores have bright autofluorescent nuclei, which suggests that
the haploid microspore may have the capacity to synthesize sporpollenin
(see Moitra and Bhatnagar 1982).
Changes in DNA, RNA and protein level in the tapetal cells have been
studied. The tapetal nuclei show a sharp increase in chromatin stainability
before meiosis in mirospore mother cells. The nuclei in a number of tapetal
cells undergo regular mitotic divisions followed by inhibited cytokinesis.
This results in the formation of binucleate, rarely multinucleate cells. (It is
somewhat diffic~lt to distinguish the changes in tapetal cells with respect
to mother cells because of asynchronous meiosis). The tapetal nuclei show
a higher Feulgen stainability in younger sporangia than in older sporangia.
The role of tapetal cells as a source of nucleic acid is not fully understood.
In E. foliata, Feulgen-positive material extruding from tapetal cells has not
been observed. There is, however, a reduction in staining intensity of nuclei
at later stages of meiosis and thereafter.
Mehra (1949) studied the effect of sulphanilamide on the division
mechanism of the body nucleus in pollen grains of Ephedra foliata (n = 7),
E. sinica (n = 14), E. saxatilis (n = 14) and E. intermedia (n = 14). The
grains were germinated in artificial culture in the natural secretion which
oozes out during pollination. In cultures containing germinating grains at
metaphase, anaphase or telophase, the addition of 0.5% sulphanilamide
caused immediate collapse of the spindle mechanism followed by the splitting
and scattering of chromosomes. With 0.2% sulphanilamide, there was no
effect on the mitotic phenomena; while 0.4% was at the critical level and
had a paralyzing effect on the spindle mechanism. The effects of the chemical
326 The Gymnosperms
Fig. 18.11 A-1. Ephedra gerardiana, microsporogenesis and male gametophyte.

A-C Formation of microspore tetrad. D Uninucleate microspore. E-H Development of
male gametophyte. prl first prothallial cell. pr2 second prothallial cell. tn tube nucleus .
I Mature pollen grain at shedding stage. be body cell. sc stalk cell. (After H. Singh and
K. Maheshwari 1962)
on chromosomes are: the kinetochore region becomes prominent; occasionally

a high degree of contraction occurs, and the chromosomes become straightened
due to the absence of a spindle mechanism. Sulphanilamide (in effective
dose) produces effects similar to colchicine which is, however, required in
relatively diluted strength (Mehra 1949).
The pollen grain wall shows an outer exine and an inner intine. In
Ephedrale,s 327
Ephedra monosperma (Gullvag 1966) the exine comprises two layers, the
outer of orbicules and/or a sculptured granular layer, and a lamellated inner
layer. In E. foliata the exine is ornamented with 16 meridional ridges with
crests which show prominent undulations (Tiagi 1966). The intine is the
last layer to be laid down and consists of cellulose only (E. foliata). Ephedra
pollen is polycolpate (see Khoshoo and Ahuja 1963).
Male Gametophyte. After separation from the tetrad, the microspores

become ovoid in E. gerardiana (H. Singh and K. Maheshwari 1962). The
nucleus moves to the lower end and divides to form the first lenticular
prothallial cell (Fig. 18.11 D, E). The next division cuts off the second
prothallial cell followed by cleavage of the cytoplasm (Fig. 18.11 F, G).
Sometimes, the second prothallial cell may lack the upper wall; both prothallial
cells are ephemeral. The central large nucleus divides and produces the
tube cell and antheridial nucleus. The latter has a distinct cytoplasmic
sheath and gives rise to the stalk and body cell (Fig. 18.11 H, 1). The
mature pollen grain is five-celled at shedding (Fig. 18.11 I); it is spindle-
shaped and yellow.
Dehiscence of microsporangia is poricidal, the pores .are located at the
tip of sporangia.
Female Strobilus
The female strobili are borne in the axil of leaves at the nodes (Fig. 18.12 A).
Rarely, there may be a single strobilus which terminates the main axis. Tbe
number of strobili varies with species: Ephedra distachya L. usually produces
three strobili from each axil of foliage leaves, while E. equisetina bears only
one (Takaso 1984). Each strobilus (cone) consists of three or four pairs of
decussately arranged bracts (Fig. 18.12 A) followed by one or two (or
three) ovules (Fig. 18.13 A-1) per strobilus. The number of ovules varies
from species to species. Each ovule has an outer (oe) and an inner envelope
(ie) which enclose thenucellus (Fig. 18.13 C-G). At maturity the ie extends
to form a long micropylar tube, the longest known is gymnosperm
(Fig. 18.13 I).
Ovule. The ovule is initiated by a transformation of the apical meristem

(Fagerlind 1971). This is marked by periclinal divisions in the outermost
layer of the lateral shoot apical meristem. Takaso (1984) studied the
histological changes in the apex of the female strobilus, and the initiation
of the ovule in E. distachya and E. equisetina. There is a uniseriate dermal
layer in the shoot apex, and residual shoot apex. Two relatively large ovular
primordia, semi globose in outline, originate from the axil of the uppermost
bract (Fig. 18.13 A, B). In contrast to the vegetative shoot apex, periclinal
divisions occur frequently in the dermal layer of apex of the female strobilus
and of the ovule. These divisions begin at the apex of the female strobilus
328 The Gymnosperms
Fig. 18.11 A, B. A Ephedra chilensis, twig bearing megasporangiate strobili (str).

B Ephedra sp., note pollination drop at the tip of ovule. (A After Foster and Gifford
1959, B after Bierhorst 1971)
before the initiation of the ovule, or simultaneously at the apex as well as

in the peripheral region. The shoot apex increases slightly in width towards
the uppermost pair of bracts and the two ovular primordia and the
residual shoot apex (between the primordia) become clearly discernible
(Fig. 18.13 A, B).
The early ontogeny of the two ovular envelopes (ie and oe) was studied
by Takaso (1985). The oe arises from all round the base of the ovular
Ephedrales 329
Fig.18.13 A-I. Scanning electron micrographs of ovules at various stages of development.

A-G Ephedra distachya . A, B Apical and lateral views of (swollen) ovular primordia.
br baret. C-G Initiation of outer ( oe) and inner (ie) envelopes. H, I E. equisetina.
H Later stage of development of ovule, the inner envelope has one-sided growth. I Nearly
mature ovule. The inner envelope forms a long micropylar tube with an elliptic opening
which extends beyond the outer envelope. (After Takaso 1985)
330 The Gymnosperms
primordium, except at the dorsal side, so that it is horseshoe-shaped

(Fig. 18.13 E, F). Later, it becomes continuous but grows more rapidly
laterally so that oe appears as two opposite projections (Fig. 18.13 F). The
mature envelope is a stout structure; most of its cells are tanniferous. It is
laterally vascularized.
The ie appears as a small annular swelling after the initiation of oe. It
grows faster on the ventral side than on the dorsal side and become asymmetric
(Fig. 18.13 G). At maturity, the ie forms a long, asymmetric micropylar
tube which grows beyond the nucellus and has an elliptic orifice
(Fig. 18.13 H, I). The greater part of ie is uniformly thin, except the basal
part which is thick and massive.
In a young ovule the two envelopes are inserted at nearly the same
levels, while in later stages the ie is set at a much higher level.
The morphology of the oe has long been disputed and, at one time, it
was regarded as the outer integument. According to Mehra ( 1950) and
Eames (1952), the oe is the result of fusion of two perianth leaves or
bracteloles. Eames regards the inner envelope alone as the true integument.
According to Takaso (1985), histologically, the oe in Ephedra distachya
and E. equisetina is formed by derivatives of both the dermal and subdermal
cells of the ovular primordium, i.e. it is of dual origin. The inner envelope
is formed exclusively by derivatives of the dermal cells (except at the basal
region), i.e. it is of dermal origin. Thus, morphology and anatomy do not
support the homology between t:he two envelopes. Rather, the outer envelope
resembles vegetative leaves more than the inner one.
The young nucellus shows one to several hypodermal archesporia! cells
which divide periclinally, and form the inner pnmary sporogenous cells
and the outer primary parietal layer. The latter undergoes further divisions
and a massive parietal tissue is formed. The epidermal cells of the nucellus
also divide perclinally and form a nucellar cap (H. Singh and K. Maheshwari
1962) which adds to the already massive nucellus. In Ephedra americana
(Pankow 1962) and E. gerardiana (H. Singh and K. Maheshwari 1962), the
formation of parietal tissue (with some exceptions) has been confirmed. In
E. foliata the hypodermal archesporia! cell divides to produce a primary
parietal cell and a megaspore mother cell (P. Maheshwari 1935), while in
E. helvetica and E. intermedia (Mehra 1950) the archesporia! cell functions
directly as megaspore mother cell. Occasionally, two hypodermal cells and
two tetrads have been observed in E. foliata. The megaspore mother cell
(Fig. 18.14 B) undergoes reduction division and gives rise to linear
(Fig. 18.14 C) or T-shaped tetrad.
A well-defined thin-walled hypostase is present deep in the chalaza,
where vasculature for central portion of the ovule ends (Fig. 18.14 A, E).
Female Gametophyte
The chalaza! megaspore of the tetrad usually functions. Following the first
Ephedrales 331
D E
Fig. 18.14 A-E. Ephedra gerardiana, megasporogenesis. A Longisection female cone.

br bract. fg female gametophyte. hy hypostase. ie inner envelope. oe outer envelope.
B-D Longisection nucellus to show sporogenous cell, linear megaspore tetrad, and four-
nucleate gametophyte. E Longisection lower portion of ovule to show hypostase and
vascular trace (vt) . (After H. Singh and K. Maheshwari 1962)
division, the two nuclei move towards opposite poles and a vacuole is
formed between them. Further nuclear divisions result in a number of free
nuclei which lie on the periphery of a large central vacuole (Figs 18.14 D;
18.15 A-D). The free-nuclear divisions are not simultaneous. Fig. 18.15 D
shows a gametophyte with most of its nuclei at metaphase, the remaining
nuclei at the micropylar end being at anaphase or early telophase.
Fig. 18.15 E is a wholemount 'o f a top-shaped free-nuclear gametophyte of
E. foliata. The maximum number of free nuclei in the gametophyte varies
with species: 256 in E. trifurca (Land 1904), 500 in E.foliata (P. Maheshwari
1935), and nearly I 000 in E. distachya (Berridge and Sandy 1907).
Ephedra lacks a spongy tissue. The mechanics of enlargement of megaspore
332 The Gymnosperms
c 0 F
Fig. 18.15 A-G. Female gametophyte. A-D, F, G Ephedra gerardiana. A-D Longisection
of free-nuclear female gametophyte at progressive stages of development. E E. foliata,
wholmount of free-nuclear female gametophyte. F Longisection of ovule, outline
diagramme for G. fg. female gametophyte. G Cellular female gametophyte from F.
(After H. Singh and K. Maheshwari 1962)
membrane during the coenocytic phase of female gametophyte has been

described by B. and C. Moussel (1973)! in E. distachya. During the mitotic
cycle, the Golgi apparatus gives out numerous vescicles (of variable
morphology) (Fig. 18.16) which store polysaccharides. These vescicles may
fuse and form concentration vescicles with fibrillar contents, and an
eccentrically placed blob of electron-dense material. The simple or
concentration vescicles pour their contents outside the plasma membrane
and increase the surface area of the megaspore membrane (Fig. 18.16).
Cell wall formation in Ephedra seems to follow the usual plan of
alveolation (P. Maheshwari and H. Singh 1967). The gametophyte
differentiates into two distinct regions: (a) the micropylar part composed of
radially-elongated thin-walled hyaline cells with scanty cytoplasm, and (b)
Ephedrales 333
• .,__vc1
•
Fig. 18.16. Ephedra distachya, formation of megaspore membrane. Diagrammatic

representation of golgi (go) activity in female gametophyte during mitotic cycle.
Production of two types of vescicles (vel and vc2), formation of concentration vescicles
(cv), deposition of contents of vel or concentration vescicles on the megaspore membrane
(mm). pm plasma membrane. (After B. Mousse! and C. Mousse! 1973)
the chalazal part of compact polygonal cells of smaller size but with dense
cytoplasm (Fig. 18.15 F, G). By the time archegonia are mature, the lower
half of gametophyte differentiates further into an upper region of actively
dividing polygonal cells, and a lower region of relatively larger cells with
dense cytoplasm and large nuclei. The lower part of the gametophyte is
narrow and tubular and grows deep into the chalazal region. It consumes
the protoplasmic contents of the circumjacent cells, which ultimately collapse.
Thus, the mature gametophyte can be differentiated into three zones: (a)
the upper fertile, (b) the middle storage, and (c) the lower haustoria! zone.
In E. trifurca (Land 1907) and E. distachya (Lehman-Baerts 1967), during
embryo formation the apical cells of the gametophyte divide actively and
form a plug, which closes the pollen chamber. In E. foliata (P. Maheshwari
1935), this plug is formed even before the archegonia are mature, and
simulates the tent pole.
Archegonial initials appear (Fig. 18.17 A) at the micropylar end of the
cellular gametophyte. One to six archegonia develop singly. P. Maheshwari
( 1935) reported two archegonia within a common jacket in E. foliata.
A mature archegonium has a Jong and massive neck (longest in any
334 The Gymnosperms
vcn
Fig. 18.17 A-G. Ephedra gerardiana, development of archegonium. A, B Longisection

of micropylar part of female gametophyte. A Archegonial initial (ari). B Archegonial
initial divides and forms a central cell (d) and a neck initial (ni). C-E Development of
archegonium. E Central cell nucleus at metaphase. F Portion of longisection of nucellus
(n) and female gametophyte; note the pollen chamber (pc). G Longisection of mature
archegonium, note ventral canal nucleus (vcn) and jacket (j). (After H. Singh and
K. Maheshwari 1962)
Ephedrales 335
gymnosperm) of 30-40 cells. They merge into the adjoining cells of the
gametophyte and it is not possible to make an exact count (Fig. 18.17 D).
The newly formed neck cells have a thin wall and show vacuolate cytoplasm
rich in ribosomes and mitochondria (B. Moussel1972). During the maturation
of the neck, Golgi vescicles deposit a thick wall across the plasma membrane.
Eventually, the cytoplasm and its organelles degenerate and the cell wall
becomes prominent. The neck cells facilitate the passage of the pollen tube.
Each archegonium is surrounded by a two-layered jacket of thin-walled
elongated cells (Fig. 18.17 G). The jacket cells originate by repeated transverse
divisions of the cells adjacent to the archegonial initials. The cells are
mostly multinucleate. Frequently, the nuclei of the neck and jacket cells
migrate into the archegonium, where they divide and produce many
micronuclei, or may enlarge and simulate embryonal nuclei. H. Singh and
K. Maheshwari (1962) observed (in one preparation of E. gerardiana) one
of the jacket nuclei so close to the egg that it could easily be mistaken for
a male nucleus.
The central cell enlarges (Fig. 18.17 B), its cytoplasm becomes thinly
distributed, and is highly vacuolate in initial stages (Fig. 18.17 B-F). This
is the foam stage. The vacuolated archegonium gradually matures. Its large
nucleus at the tip (Fig. 18.17 C, F) divides mitotically to give rise to the
egg and ventral canal nuclei (Fig. 18.17 G). A wall is not formed between
the nuclei. The egg nucleus moves to the center of the archegonium. It is
surrounded by a dense sheath of cytoplasm which sometimes extends
downward to a considerable extent in E. gerardiana (Fig. 18.17 G).
Pollination.
Ephedra is believed to be wind-pollinated, but there are also reports of
insect pollination (see Bino et ·al. 1984). The stickiness of pollen also
suggests entomophily. In E. aphylla and E. campylopoda, pollination is
partially entomophilous. The adaptation to insect pollination is shown by
nectarial secretion which occurs in both male and female flowers. In
E. aphylla, the male flowers produce one to ten nectar droplets, before
anthesis, mainly on the outer cover (perianth) in the functional reproductive
unit (Bino et al. 1984). The droplets persist for 2-4 days. In the female
flowers, the single droplet of nectar is secreted on the outer cover before
anthesis and continues till anthesis. Amino acid is present in the exudate
in appreciable quantity and remains constant, irrespective of the stage of the
anthers. In addition to nectar, there is a single pollination droplet, at anthesis,
at the tip of the micropylar tube (Fig. 18.12 B). It persists for 2-4 days
when accessible to insects or 10-16 days when there are no insect visitors.
Usually, the amino acid is not detectable (Bino et al. 1984). There is no
appreciable difference between the volume and sugar concentration of nectar
drops of male and female flowers and pollination drop. The sugar content
is independent of temperature, time and location on the plant, but there is
336 The Gymnosperms
relation with relative humidity (RH). During the day, in the area studied,
RH is usually below 60%, the sugar content is over 80%, while during the
night, the RH is occasionally 90% or higher and the sugar content is lower.
A chemical analysis of the pollination drop also shows the presence of
peptides, malic and citric acid (Ziegler 1959), inorganic phosphates and
sugars (especially high-25% ).
Among the frequently encountered insect visitors are Lucilia caeson
(Calliphoridae), Metasyrphus corollae, Episyrphus balteatus and Scaeva
albomaculata (Syrphidae). Only nectar is consumed while pollen consumption
is negligible (as shown by an analysis of gut content).
The pollen grains are caught in the glistening pollination drop, and are
sucked with the fluid column (present in the micropylar tube) which becomes
thicker due to evaporation (Strasburger 1871).
During archesporia! development, the nucellar tissue, directly above the
female gametophyte, disorganizes and becomes obliterated. Thus , a deep
and conspicuous pollen chamber is formed. The upper portion of the
gametophyte is exposed at the base of this chamber (Fig. 18.18 A), a
unique feature in Ephedra. The pollen grains come into direct contact with
the female gemetophyte.
The pollen swells when caught in the pollination drop. The exine ruptures
irregularly and is cast off before the pollen germinates. The intine is
thin.
Pollination in Ephedra occurs when the ovule has fully formed archegonia.
After pollination, the outer envelope of the ovule shows small papillate
outgrowths directed towards the inner envelope. As the ovule matures,
these papillae elongate, become thick-walled (Fig. 18.18 B, C), and help to
close the space between the two envelopes. It may also seal off the micropyle
by pressing inward on the micropylar tube (H. Singh and K. Maheshwari
1962).
Fertilization
The pollen grains germinate within a few hours of entering the pollen
chamber. The body cell divides and forms two sperm nuclei. The latter
may be of the same size (E. foliata, E. sinica), or markedly unequal
(E. altissima, E. saxatilis). The pollen tube is narrow and short, and pushes
its way between the neck cells of the archegonium. The tip ruptures and
discharges four nuclei (tube, stalk cell nucleus, and two male gametes) into
the archegonium. One of the gametes fuses with the egg and the other may
fuse with the ventral canal nucleus near the upper part of archegonium
(Fig. 18.19 A, B 1, B 2). According to Khan (1943); " ... The type of double
fertilization seen in Ephedra may have no phylogenetic significance at all
and may simply be the natural outcome of a tendency towards fusion between
any two nuclei of opposite sexual potencies that happen to lie in a common
chamber". Moussel (1978), Friedman (1990a, b, 1991) have also reported
Ephedrales 337
A
c
Fig. 18.18 A-C. A Ephedra foliata, longisection of ovule at pollination. ie inner envelope.
n nucellus. oe outer envelope. pc pollen chamber. B, C E. gerardiana, longisection
of micropylar canal at two stages of development of papillate projections from
outer envelope . (A After P. Maheshwari 1935, B, C after H. Singh and K. Maheshwari
1962)
double fertilization in E. distachya, E. nevadensis and E. trifurca. Double

fertilization in Ephedra (and in angiosperms) is a significant occurrence in
reproduction (for details see Friedman 1992a, b, 1994).
According to Land (1907), the interval between pollination and fertilization
in E. trifurca may be as short as 10 h.
Embryogeny
The embryogeny of only a few species of Ephedra has been studied. The
338 The Gymnosperms
~
~
Fig. 18.19. A-1. Ephedrafoliata, fertilization and proembryogeny. A Longisection of

ovule at double fertilization. BH B2 Nuclei from A. B1 Male nucleus in contact with egg
nucleus. B2 Second male nucleus in contact with ventral canal nucleus. C. Longisection
of two-nucleate proembryo, note radiating cytoplasm. At the micropylar end persists a
large ventral canal nucleus, and a few small nuclei of the neck or adjacent cells, at the
lower end a jacket nucleus prior to entry into proembryo. D-F Development of eight-
nucleate proembryo. G, H Longisection egg cell shows five (G) and six (H) proembryos.
I Mature proembryo. (After Khan 1943)
Ephedrales 339
zygote nucleus divides in situ; the two nuclei move apart (Fig. 18.19 C),
occasionally to the two poles of the zygote. Two more mitoses follow
which result in eight nuclei (Fig. 18.19 D). Each nucleus is surrounded by
a cytoplasmic sheath which radiates strands (Fig. 18.19 C-F) and takes a
dense stain. At the eight-nucleate stage, a wall develops around each nucleus
(Fig. 18.19 G, H) followed by cleavage, which results in eight units. According
to Foster and Gifford (1959), this is precocious type of cleavage
polyembryony. The proembryo is spherical. The cytoplasm next to the
nucleus takes a denser stain and does not show vacuoles; the outer part is
lighter and highly vacuolated. The nucleus contains numerous nucleoli and
granular, irregularly scattered chromatin (Fig. 18.19 1).
Each cell of the proeinbryo puts out a tubular outgrowth (Fig. 18.19 H).
The nucleus may divide before the outgrowth is formed, or it may move
into the tube and divide. A transverse wall is laid down, which gives rise
to an embryonal cell and a suspensor cell (morphologically embryonal
suspensor), which elongates (Fig. 18.20 A). A periclinal division in the
embryonal cell is followed by a longitudinal division in the lower of the
two cells to form three cells (Fig. 18.20 B). Further divisions give rise to
the embryo proper; a multicellular secondary suspensor differentiates adjoining
the suspensor cell (Fig. 18.20 C). The lower end of the embryo produces
two cotyledons and the shoot apex. A root cap of column (horizontally
oriented cells in the central region) and pericolumn (peripheral cells which
are steeply inclined or almost vertical) differentiates (Deshpande and
Bhatnagar 1961).
Several embryos begin to develop within a single ovule, but only one
reaches maturity (Fig. 18.20 D, H). The large number of embryos is due
to a combination of simple and cleavage polyembryony. A well-defined
tunica layer originates in the embryo even before the development of
cotyledons.
Seed
Following fertilization, the ovule undergoes marked changes, in the zygote
(development of embryo), female gametophyte (development of endosperm)
and integument (development of the seed coat).
During post-fertilization stages, in restricted areas the female gametophyte
goes through meristematic activity. The micropylar part grows upward into
the deep pollen chamber and may plug it partially. Starch accumulates in
the cells of the female gametophyte in the micropylar region before
fertilization, and in the axial cells after fertilization (B. Mousse! 1974).
Many of the gametophytic cells become binucleate. During the development
of embryo, storage products accumulate only in the lower half of the
endosperm. In a mature seed the endosperm is derived mainly from the
lower part of the gametophyte. It can be distinguished into four zones from
periphety inwards:
340 The Gymnosperms
Fig. 18.20. A-H. Ephedra foliata, development of seed. A, B Two- and three-celled
embryonal unit with elongated suspensor. C Later stage in embryo development.
D Longisection of upper part of ovule shows six embryos . E Transection (part of) outer
envelope. ep outer epidermis . iep inner epidermis. F Transection inner envelope passing
through the micropylar region. G, H Trans- and longisection of mature seed . There are
three vascular bundles in the outer envelope in G. (After Khan 1943)
Ephedrales 341
a) The haustorial zone is a single layer of especially differentiated outer

tangential cells which are richly cytoplasmic. The outer walls have a thick
and uneven deposition which projects like stalacites into the cell cavity.
The cytoplasm has several small vacuoles, mitochondria and different profiles
of ER, but has only a few plastids. The nucellar cells in the vicinity of
haustoria! zone degenerate and become compressed.
b) The proteolipid and starch zone differentiates as the apical embryonal
cell divides to initiate the formation of embryonal mass.
c) The starchy zone
d) The central zone is very narrow and devoid of storage products.
The accumulated food reserves in tPe endosperm are utilized at the time
of seed germination.
The nucellar cells, in the micropylar region, become multinucleate (Khan
1943). Those around the pollen chamber persist in the seed and some of
them become thick-walled. The remaining nucellus becomes crushed and
appears as a band lying near the endosperm.
The inner envelope (integument) has two layers of cells. The cells of the
outer layer are slightly thick while those of the inner layer are thick-walled
(Fig. 18.20 F). The cells of the outer envelope (oe) are thick-walled
(Fig. 18.20 E) and form the main protective layer in the seed.
The bracts surrounding the seeds become thick and fleshy. The lower
pair is small while the upper pair of bracts is large and covers the seed
(Fig. 18.20 H). In Ephedra, only the outer envelope is vascularized
(Fig. 18.20 G). Three bundles enter the ovule and remain unbranched; only
a few tracheids differentiate at the base of the nucellus (H. Singh and K.
Maheshwari 1962).
Chromosome Number
The haploid chromosome number of 7 and 14 is characteristic of Ephedra:
E. foliata n = 7; E. sinica, E. saxitilis and E. intermedia n = 14 (Mehra
1949). The basikaryo type is composed of five median-submedian and two
subterminal chromosomes (see Khoshoo and Ahuja 1963).
E. saxatilis is an allo-polyploid (Mehra 1947). Besides normal haploid
pollen grains, a small percentage of diploid pollen grains is also formed.
The latter condition arises due to incomplete cytokinesis of the microspore
mother cell after meiosis II. The tetrad may contain two diploid microspores
or one diploid and two haploid microspores.
Ephedra shows a non-synchronous reproductive behaviour. In a plot of
E. foliata in the Delhi University Botanical Garden, initiation of cones,
anthesis, pollination and ripening of seeds occurred (in different cones)
during February-May (see H. Singh 197&). Thus, at the same time, young
342 The Gymnosperms
ovules and mature seeds can be collected from any one plant. In E. gerardiana,
the cones appear in April and the seeds ripen towards the end of September.
The bracts of the female cone become fleshy and red on ripening, which
makes the plant very attractive (H. Singh and K. Maheshwari 1962). In
E. campylopoda, the bracts, perianth leaves and other parts of female flowers
are bright yellow and later turn fiery red.
.19. Welwitschiales
Welwitschiaceae
Welwitschia
Morphology
The adult sporophyte of Welwitschia has no parallel in the plant kingdom.
It resembles a gigantic turnip, and reaches a diameter of more than 1 m.
The stem consists of a tough axis shaped like an inverted cone, and has a
deep apical depression. Most of the stem is buried in the sandy soil, its
lower part tapering abruptly to an extremely long carrot-like tap root which
penetrates the soil between 1.0 and 1.5 m deep before it splits into numerous
thin roots. How far downward the roots extend is still uncertain. The exposed
portion of the stem consists of a massive, woody, concave disc which bears
two opposite and decussate strap-shaped leaves (Fig. 19.1 A, B). These
leaves grow continuously throughout the life of the plant but very slowly,
ca. 10-15 em in a year (Von Willarl1985), from a basal meristem, and the
tip dies back continuously. A mature leaf is ca. 3 m long and 1-1.5 mm
thick. The distal end of the leaves splits and becomes frayed and extends
in a twisted or contorted manner along the surface of ground. Old plants
may be more than 1000 years (Bierhorst 1971). A specimen ca. 2000 years
old is shown in Fig. 19.1 A.
The plant produces only three pairs .of foliar organs (Martens 1977a,
Martens and Waterkeyn 1963a, b, 1964a, b):
a) The first is a pair of cotyledons which have a limited growth, and are
not shed.
b) The second is the pair of large, persistent leaves which appear at right
angles to the cotyledons.
c) Lastly, two more leaf primordia, the scaly bodies, appear which has
a long life, and increase in height, breadth and thickness, due to an intercalary
basal meristem comparable to that of the persistent leaves. The initi~l shoot
apex also produces two short epicotyledonary internodes which increase in
diameter in a manner similar to the hyp9cotyl. The apical meristem then
aborts (cause unknown); its loss does not lead to the development of axillary
344 The Gymnosperms
Fig. 19.1 A, B. Welwitschia mirabilis. A Ca. 2 000-year-old plant, note opposite strap-
shaped leaves. B Central part shows woody concave disc. (A After Von Willart 1985,
B after Rodin 1953a)
buds and branches (as in most plants) in Welwitschia. Rather, an intercalary

and permanent meristem, which widens indefinitely, arises at the leaf base.
The vegetative activity is thus restricted to the basal leaf meristem, and
Welwitschiales 345
extends to neighbouring tissues so that the hypocotyl increases in diameter

due to intercalary growth. After a gap of 12-50 years, buds are formed on
the centrifugal and axillary crests of the crown. They produce branches
with scale-like leaves (nodal bracts according to Rodin 1963), internodes
and axillary buds. These leaves are different from the two persistent leaves.
The buds and leafy branches are vegetative in the beginning. Later, cones
are formed due to hormonal determination of the branches and further
vegetative activity is completely lost (Martens 1977a). The adult and fertile
plant, therefore, produces an indefinite number of leaves, in addition to the
three whorls on the primary axis. The succession of these two phases (primary
axial growth and development of fertile branches) is separated by a long
time-interval. Thus, Welwitschia can produce new roots, buds, branches
(vegetative production) even after 100 or 1000 years (Martens 1977a).
The leaves grow slowly and the tip dies back continuously. Consequently,
the leaves have a continuous developmental gradient spanning many years,
which makes it possible to study the behaviour of every tissue age, unlike
other plants (Von Willart 1985). The length of the leaf is determined by
environmental factors. It may dry completely and resume growth when
conditions improve. During the period of drought, antelopes and zebras
feed on the leaves, and occasionally pull the ribbons out of the groove. The
leaf will still grow if the meristem is not damaged.
Welwitschia is classified as a hydrostable plant because even a prolonged
drought does not affect its water content significantly (see Von Willart
1985). There is not much diurnal change in the water content of the leaf.
The stomata remain open during the day, and there is tremendous loss of
water. However, the water conduit present functions, and water loss by
transpiration is balanced without much delay. The sponge-like roots may
act as a water store, balancing temporary water deficits, but the demand for
water is undoubtedly supplied from a water source to which it must be
connected. The accompanying flora cannot exploit this source and suffers
much more from prolonged drought. It is not conclusively proved yet whether
Welwitschia is able to make use of dew and fog to balance the daily water
loss. Wherever the water conduit is disturbed, water loss appears to be
regulated by reduction of leaf area.
The photosynthetic capacity and the carbon balance, over a period of
24 h, show that leaves of all ages are able to take up C02 during (parts of)
the day. The C02 uptake decreases with increase in leaf age. Only the first
half of the leaf has a carbon gain while the older parts show carbon loss.
Nevertheless, the carbon balance of the entire leaf is positive, otherwise the
leaf length is reduced. With improved water supply, even old parts contribute
to a carbon gain. A plant with such a slow growth rate must keep as much
of the leaf blade alive as possible.
The Welwitschia leaf has a high reflectivity. Only 55% of the incident
global radiation is absorbed, nearly 40% is reflected. The temperature of
346 The Gymnosperms
the soil shaded by the leaf is cooler than the temperature of the lower
surface, which results in a net loss of long wave radiation. Transpiration
also plays an important part in energy dissipation. High transpiration is
necessary to prevent lethal temperature of the leaf. When it cannot be
maintained (due to shortage of water) the tissue at the tip dies (probably
due to lethal temperature). Besides carbon balance, lethal temperature at
the leaf tip may regulate leaf length.
According to Von Willart (1985), the key to understanding the biology
of this peculiar plant lies in the fact that Welwitschia has a connection to
water sources in the soil (which cannot be exploited by other plants) and,
therefore, does not norinally suffer from water shortage. With a guaranteed
supply of sufficient water, a high transpiration rate is possible, which prevents
lethal leaf temperature, and a large leaf can be maintained.
The surface of the stem is covered with a ridged, corky layer nearly
2 em thick in the upper region. The portion of stem above the point of
insertion of leaves is called the crown, while the lower portion is called the
stock. The crown is marked by a series of ridges, each of which extends
to nearly half the circumference of the stem. They bear scars of old, fallen
inflorescences. The ridges on the ~tock are less prominent.
The taproot is long. It forms lateral roots, at various depths, depending
on the soil.
Anatomy
Root. Primary Structure. A transection of root of a young seedling shows

(Fig. 19.2 A) a piliferous layer which bears root hairs followed by one- or
two-layered parenchymatous cortex, and an endodermis with Casparian
bands. The pericycle remains single-layered above the protoxylem (px) points,
while in the remaining area it undergoes a precocious periclinal division.
The cells of the outer layer are large, their inner cell wall is thin and
weakly suberized. The inner layer consists of smaller cells with cell contents.
The stele is diarch. The two xylem groups join in the middle or remain
separate. The metaxylem (mx) consists of large and the px of narrow tracheids.
The two phloem groups are separated from the xylem by one or two small
cells with dense contents which later form the cambium. Thick cellulosic
fibres are present between the phloem and the pericycle (Fig. 19.2 A, B).
Two groups of secondary xylem tracheids alternate with large primary
mx. The internal layer of the two-layered pericycle becomes the initial of
the phellogen. This protective tissue is weak and is compensated for by
several layers of cellulosic fibres which completely encircle the stele cylinder
(except above the px points). An extrafascicular cambium is absent, which
results in reduced radial growth above the two vascular plates. This double
depression of the radicle is manifested externally in its bilateral symmetry
(Martens 1977b).
Welwitschiales 347
sp
sx
Fig. 19.2 A, B. Welwitschia mirabilis. A Transection of main root from 5-month-old

seedling. c cortex .ffiber. sp secondary phloem. sx secondary xylem. B Vascular region
from A. (After Martens 1977b)
Stem. In a young plant the main axis, below the crown, has a ring of
collateral vascular bundles (Fig. 19.3 A). In the old plant there is a
348 The Gymnosperms
Fig. 19.3 A-D. Welwitschia mirabilis . A Transection through smooth and cylindrical
part of axis below the crown, from a 15-year-old plant. B Transection of vascular system
of crown below the level leaf of meristem; there are two transverse series of bundles.
C A bundle from B. xp xylem pole. D Coiled vessel isolated from mature plant. (After
Bierhorst 1971)
saucer-shaped mass from which vascular traces extend to the leaves, the
inflorescence and the single taproot. The cortical parenchyma becomes
meristematic, and a phellogen produces an outer corky tissue (Rodin 1963).
Within the flanges of the crown are two parallel series of bundles with the
xylem on the inside (Fig. 19.3 B, C). Traces from these bundles extend
towards the surface, presumably to old reproductive axes. The stem is
heavily sclerified.
In early-formed tracheary elements, circular bordered pits are present
between the gyres of the secondary wall helix. Vessel elements have single
apical or lateral circular pores, oriented transversely or obliquely. In occasional
Welwitschiales 349
vessel members, the pores are in pairs or, rarely, a foraminate plate has
three pores (Bierhorst 1971). The later-formed thick-walled elements have
a complex rarniform bordered pit system. A peculiar feature is the occurrence
of isolated, unconnected vessels of about 12 cells, which are coiled into
tight worm-like masses (Fig. 19.3 D).
Electron microscopic observations indicate that most of the primary and
secondary xylem and phloem elements are arranged in radial rows. In the
phloem, two types of cells, namely, sieve cells (structure similar to Ephedra
and Gnetum) and parenchyma cells, have been described. The plastids in
sieve cells undergo changes such as the formation of starch grains, proliferation
of internal membranes, and decrease in the density of the matrix. Osmiophilic
globules persist in the plastids (but disappear from the cytoplasm). These
plastids, unlike those of Pinus, lack crystalline inclusions and fibrillar material
at all stages of differentiation. The starch grains present are club-shaped (as
in Gnetum).
A mature plasmalemma-lined sieve cell contains nucleus, mitochondria,
ER and plastids with starch granules. The plastids lack internal membranes
and their matrices are hyaline (very clear) unlike the plastids from immature
cells. The mitochondria undergo no apparent structural modifications. The
sieve cells lack ribosomes, dictyosomes, and microtubules. The vacuoles of
young sieve cells and the lumen of mature ones occasionally contain a
coarse fibrous substance, similar in appearance to that observed in the
vacuoles of parenchymatous cells in the leaf. It should not be confused
with slime or P-protein, which is totally absent (see Paliwal 1992). During
maturation, the tonoplast, which delimits the vacuolar contents from the
cytoplasm in young cells, ceases to be identifiable and the sieve cell appears
to contain a single large central cavity (Evert et al. 1973a, b).
An unusual feature of Welwitschia is the presence of "spicular" cells
(Hooker 1863) throughout the plant body: in leaves, stem, reproductive
branches and cone scales. The large, mostly unbranched, sclereids contain
numerous crystals embedded in the cell wall. They are closely packed and
interlaced, which makes the tissues extremely hard and tough.
The wall of a mature sclereid consists of a thin cellulose primary wall.
After the sclereid attains maximal size, calcium oxalate crystals are deposited
on its inner wall. The secondary wall layers are formed by further deposition
of wall materials, the first layer covering the primary wall and crystals.
A mature sclereid becomes slightly lignified.This is followed centripetally
by the highly lignified second wall, which is the thickest layer of the
secondary wall. The third and innermost layer is cellulosic, the lignified
and inner cellulose layers laminated. The lumen is very small. Numerous
simple pits are present on the sclereids, the outer and inner extremities of
the pits are conically flared (Rodin 1958). At the tips of the sclereid is a
branched aperture which perhaps functions like a pit, but is never conically
flared.
350 The Gymnosperms
Leaf. A mature leaf of Welwitschia is isolateral, amphistomatic with sunken

stomata. It has large longitudinal and small oblique vascular bundles which
anastomose (Fig. 19.4 A, B). The mature leaves have numerous sclereids
(Fig. 19.4 D).
A vertical section of a leaf (Fig. 19.5 A) shows:
a) The epidermis is a single layer of cells on the upper and lower surface
of leaf. The outer wall of epidermal cells is heavily thickened and consists
of a layer of cuticle. The median layer comprises powdery crystals of
calcium oxalate, and extends laterally to the subsidiary cells. The innermost
cellulosic layer is very thick, specially in a mature leaf. A thin discontinuous
layer between the crystal and the inner cellulosic layer contains cutin (Martens
1977b).
The sunken stoma have a stomatal chamber and the development is
syndetocheilic.
b) The mesophyll consists of three or four layers of distinct palisade
cells on both sides, and a compact central zone of spongy parenchyma
(Fig. 19.5 A). The palisade parenchyma is the chief photosynthetic tissue.
The chloroplasts in the central mesophyll decrease toward the middle region
of leaf. Calcium oxalate granules are present in its cell walls. Hypodermal
cellulosic fibres occur in the massive bundles, which may extend to the
inner margin of palisade parenchyma. The bundles on the upper surface of
the leaf are smaller than those on the lower surface. Secondary walls develop
first in the fibres near the epidermis, and maturation proceeds inward.
A paradermal section of leaf shows (Fig. 19.4 C) stomata between the
bundles of fibres. Occasionally, gum canals occur in mature leaves adjacent
to the phloem between vascular bundles. The spindle-shaped unbranched
canals are formed by enlargement of irregular chains of mesophyll cells
(their walls break down).
c) The vascular system consists of collateral bundles, the xylem faces
the upper and the phloem the lower side of the leaf. Fibres surround the
vascular bundles, followed by transfusion tissue (Fig. 19.5 B). The latter
ensheaths the vascular bundle and strengthens it. Its cells are shorter than
the xylem elements, have reticulate or scalariform thickenings, and small
simple or bordered pits in the thin-walled areas. In anastomosing bundles,
fibres are absent.
Protoxylem tracheids develop from the first procambial strand of the
leaf primordium (Rodin 1958). The tracheids have annular or spiral
thickenings. The protoxylem is eventually crushed. The metaxylem replaces
it, and is the functional xylem. The earlier-formed metaxylem intergrades
with the protoxylem and has spiral or annular thickenings with large uniseriate
bordered pits. Later metaxylem has vessels and tracheids with spiral, reticulate
or scalariform wall thickenings. Large uni- or biseriate bordered pits are
common. There is a single perforation on the slanting end wall between
two adjacent vessel elements. As many as four bordered pits may also
Welwitschiales 351
Fig. 19.4 A-D. Welwitschia mirabilis. A Paradermal section of mature leaf shows parallel
longitudinal and smaller anastomosed vascular bundles. B Longisection of leaf, the bundles
are oblique. ttr transfusion tissue. x xylem. C Oblique paradermal section of mature
leaf, note the position of stomata in relation to hypodermal fibre (hj) bundles.
D Longisection of a large, brancred crystalliferous sclereid (sci) in leaf. (After Rodin 1958)
352 The Gymnosperms
Fig. 19.5 A, B. Welwitschia mirabilis. A Transection of mature leaf. hfhypodermal

fibres. B Vascular bundle from A./fibre. sci sclereid. ttr transfusion tissue. (After Rodin
1958)
occur on the same end wall. The vessel elements closely resemble the
metaxylem tracheids, except for the larger diameter and presence of
perforations.
Although tissues resembling secondary xylem and phloem occur in the
collateral bundles of the leaf, they are only primary tissues, since a fasicular
cambium has not been demonstrated decisively (Rodin 1958).
At the apex of the leaf primordia, toward the tip of the procambial or
tracheidal strand, several hypodermal cells become vacuolate, enlarge and
become tubular. Their walls thicken and the cells develop into cellulosic
fibres, and lignified and crystalliferous sclereids. They form a protective
cushion between the epidermis and the apical net of vascular traces (Martens
1977b).
Reproduction
Welwitschia is a dioecious plant. The inflorescences, borne in clusters,
arise adventitiously from within the cortical tissues adjacent to a groove
from which the leaf grows. They branch in a dichasial manner, each branch
terminating in a cone with opposite and decussate scales (Figs. 19.6 A;
19.7 A, B). According to Martens (1959), ontogenetically, both male and
female flowers have the same opposite and decussate plan as seen in their
floral diagrams (Fig. 19.8 A, B). The cone bracts in both sexes are broad
and ovate with tapering thin lateral wings.
Cone Bracts. The male cone bract is thickest at the base and gradually
tapers toward the apex, while the female cone bract is quite thin near the
base and thick toward the apex. The flattened seed lies in a pocket formed by
the thin area, (Fig. 19.7 E), and the mature cone forms a compact structure.
Welwitschiales 353
Fig. 19.6 A-E. Welwitschia mirabilis. A Three male cones. B Abaxial view of male
flower. C, D Longi- (C) and transection (D) of mature male cone. E Cleared male cone
bract. (A, B, DAfter Bierhorst 1971, C, E after Rodin 1963)
There is a similarity in the venation pattern in the cone bracts of male

and female cones (cleared bracts facilitate study). The two vascular bundles
are widely separated when they enter the cone bracts. The veins branch
dichotomously, with a large number of short dichotomies in the distal
354 The Gymnosperms
Fig. 19.7 A-E. Welwitschia mirabilis. A Female inflorescence. B Two female cones.
C Female flower. br bract. D Successive stages to show initiation of ovule (whole
mount). E Mature seed. (A After Rodin 1963, B, C, E after Bierhorst 1971, D after
Martens 1959).
region (Fig. 19.6 E) where the bract lies exposed. A major part of the
female cone bract is winged and devoid of vascular tissue. A large number
of fibres and sclereids are present in the distal portion of a mature female
cone bract, and nearly obscure the vascular system in cleared material. The
cone scales become scarlet red at maturity.
Male Strobilus
A mature microsporangiate strobilus is four-angled, 20-30 mm long and
8 mm wide. It bears broad decussate bracts in four ranks (Fig. 19.6 D) and
each bract has in its axil a flower (Fig. 19.6 A). Each flower consists of
Welwitschiales 355
Fig. 19.8 A, B. Welwitschia mirabilis. A, B Male and female flower, floral diagram.
(A After Martens 1961, B after Martens 1959)
(from outside inward) (Fig. 19.8 A): (a) two pairs of a~ymmetrically disposed
bracts, (b) six microsporangiophores, which are fused basally into a sheath
(Fig. 19.6 B); (c) a pair of minute bracts at the base on either side of the
centrally located non-functional ovule. The latter has a single integument,
which extends upward and flares out at the apex. Each microsporangiophore
bears three fused sporangia with their lines of dehiscence radiating from
near the point where the septa converge (Fig. 19.6 B). In each flower all
the stamens mature at the same time. Flowers at the base of the cone mature
first (Fig. 19.6 C). Each stamen in a flower has a single vascular bundle.
Microsporangium
Each microsporangium has an outer layer of cells followed by two tapetal
layers enclosing the central fertile tissue.
Male Gametophyte. The precise sequence of development of the male

gametophyte has not yet been worked out (Martens and Waterkeyn 1974).
A pollen grain at shedding is three-celled; a tube cell, a generative cell;
there is a controversy regarding the third cell, interpreted as a prothallial
cell or a stalk cell, which aborts even before pollination. The pollen grains
are shed through a vertical slit.
356 The Gymnosperms
Female Strobilus
The ovulate strobilus is comparatively thicker and comprises broad decussate
bracts (Figs. 19.7 A, B; 19.8 B). A mature cone is ca. 60 to 80 mm long
and 25 mm wide. Each female flower is subtended by a cone scale, and
consists of a nucellus enclosed by two envelopes and two small lateral
bract-like structures (Figs. 19.7 C, 19.8 B). The inner envelope extends into
a long tubular micropyle (Fig. 19.7 C) and is the true integument. The outer
envelope is called a perianth.
The ovules arise in the axil of bracts, but the exact morphological nature
of meristem which forms the ovule is not known. The outer envelope arises
first as the annular hump on the ovular primordium followed by the
integument, which is also annular (Fig. 19.7 D). The integument does not
cover the nucellus. During later stages of development, the perianth forms
a conspicuous laterally extending fibrous wing in the seed (Fig. 19.7 E).
The wing has a complex structure which varies according to age, level and
the zone (median or lateral) of the seed (Martens 1975). The integument
extends apically as a thin micropylar tube.
A discrete epidermis has not been observed in the young nucellus (Fagerlind
1961). The cells of the outermost exposed layer divide periclinally to form
the inner archesporia! cells and the outer layer. The parietal tissue develops
from the (non-functional) archesporia! cells.
Megasporogenesis. A single megaspore mother cell can be distinguished

in the third or fourth layer of the nucellus, and during further development
its walls become thick (Fig. 19.9 A). Reduction divisions do take place, but
the walls are not formed, so that the development of the gametophyte is
tetrasporic (Fig. 19.9 A-D). Extrafloral embryo sacs have been observed in
the cone axis (Fig. 19.9 E). However, they degenerate at an early stage.
Female Gametophyte. The spongy tissue is absent in Welwitschia (Pearson

1929, Martens and Waterkeyn 1974). The coenomegaspore (Martens 1971)
undergoes eight to ten free-nuclear divisions which are synchronous. A
central vacuole is absent. The free-nuclear gametophyte shows gradual
vacuolation from the chalazal to the micropylar pole (Fig. 19.9 F, G).
Walls are laid down by free cell formation (Fig. 19.9 H), and the cytoplasm
becomes divided into irregular multinucleate c~mpartments with cell walls.
Two zones differentiate; a smaller, fertile micropylar and a larger, sterile
chalazal zone (Fig. 19.9 I, J). The micropylar cells are vacuolate with
usually two to eight nuclei, whereas the chalazal cells are dense and have
6-12 or more nuclei (Fig. 19.9 J). Although the partitioning ofthe gametophyte
is simultaneous, the walls and vacuoles differentiate more rapidly in the
micropylar region (Martens and Waterkeyn 1974). The micropylar cells,
along with the contents, form a cluster of embryo sac tubes or prothallial
tubes (Fig. 19.10 A, B). These tubes pierce the megaspore membrane and
Welwitschiales 357
Fig. 19.9 A-J. Welwitschia mirabilis, megasporogenesis rutd development of female

gametophyte. A Megaspore mother cell. B, C Telophase I and II; cell plate is not formed.
D Four-nucleate coenomegaspore. E Longisection of floral axis shows extrafloral
megaspore mother cell (above), and coenomegaspore (below). F Longisection of free-
nuclear gametophyte, the nuclei are scattered all over the homogenous dense cytoplasm.
G Longisection of upper portion of gametophyte, later stage, cytoplasm vacuolate.
H Portion of gametophyte shows laying down of walls. I Outline diagram for J; micropylar
(miz) and chalaza! zone (caz) of endosperm. J Cellular details of I. (A-E After Martens
1963, F-J after Martens and Waterkeyn 1974)
grow in all directions in the nucellus. Each tube grows independently of the
rest and normally ends in an enlarged swelling, called the fertilization bulb
358 The Gymnosperms
Fig. 19.10 A-F. Welwitschia mirabilis. A, B Longisection of upper portion of nucelli

(n) with female gametophyte. caz chalaza! zone of endosperm. est endospermic tube.
miz micropylar zone of endosperm. pt pollen tube. C-E Elongated endosperm tube. F
Tip of endosperm tube shows (swollen) fertilization bulb. (After Martens and Waterkeyn
1974)
(Fig. 19. 10 F), at about half the height of the nucellar cone (Fig. 19.10 A).
This level is reached independently of and often before pollination, and
eventually exceeds it. Occasionally, the tubes may over-reach the epidermis.
The growth zone of the prothallial tube depends on the accumulation of
starch in a limited part of the nucellar cone. Branching in the tubes is rare.
The nuclei in a tube lie close together during early stages, but later they
become arranged in a row (Fig. 19.10 C-F). All the nuclei in the embryo
sac tube remain small and are potentially female nuclei. The prothallial
tubes obtain their nourishment from the nucellar cells, and thin degenerated
remains lie in contact with the tubes.
The haploid nuclei in the polynucleated compartments of the chalaza!
region undergo mitosis and the daughter nuclei fuse . Finally, a tissue with
polyploid nuclei is formed.
The lipid megaspore wall increases in thickness during the growth of
endosperm. Later, it becomes thin due to stretching.
Welwitschiales 359
Pollination
In the ovule, a honey-like fluid, the pollination drop, is secreted at the tip
of the micropylar tube, in the morning hours (around 09.00 h). It persists
during the day and disappears before sunset (Pearson 1929). It is probable
that the pollination drop may attract insects. No secretion has been observed
on the male flowers. The anthers bend over the disc, suggesting the possibility
of insect visits. It has not yet been established whether pollination is effected
by insects or by wind.
A pollen chamber is formed in the apical papilla of the nucellus by loss
of nuclear material and an axial stretching of the hypodermal layers followed
by the breakdown of the walls. Finally, a concave chamber is formed
(Fig. 19.11 A, B). The pollen germinates in the pollen chamber (Fig. 19.11 B),
or at the base of the micropylar tube (Martens and Waterkeyn 1974).
Fig. 19.11 A-E. Welwitschia mirabilis. A, B Formation of pollen chamber. A Nucellar

tip with empty cells and ruptured walls; papillate cells at the base of future chamber.
B Broken cell walls of nucellar tip form a veil (ve) on the papillate cells. nc nucellar
cuticle. pg germinated pollen grain. C-E Pollen tube. g generative cell. mg male gamete
(nuclei). tn tube nucleus. (After Martens and Waterkeyn 1974)
360 The Gymnosperms
Post-Pollination Development of Male Gametophyte. The exine splits

(into valves). The pollen has a tube nucleus and a generative cell
(Fig. 19.11 C). The growth of the pollen tube through the nucellus is inter-
and intracellular. The tube nucleus enters first (Fig. 19.11 D). While two
male gametes have been observed (Fig. 19.11 E), the division of the generative
cell has not been observed.
Fertilization
The pollen tubes grow downward to the fertilization bulb (Fig. 19.12 A, B),
syngamy occurs and a zygote is formed within the fertilization bulb. It is
surrounded by a distinct cellulose wall. Thus, zygote formation and its
further development occurs in the maternal cytoplasm (Martens and Waterkeyn
1974).
:~·"" ·-
•
Fig. 19.12 A-D. Welwitschia mirabilis. A, B Zygote in the fertilization bulb with
binucleolate nucleus, dense cytoplasm, and cellulose wall; two haploid nuclei (arrowed)
in the residual cytoplasm of the bulb in A. fbl fertilization bulb. n nucellus . pt pollen
tube . z zygote. C, D Division in zygote to form proembryo initial (pre) and primary
suspensor (S). (After Martens and Waterkeyn 1974)
Embryogeny
The zygote has a prominent cell wall (Fig. 19.12 A, B). It elongates, divides
inside the fertilization bulb: and gives rise to a long suspensor cell and a
small proembryo initial (Fig. 19.12 C, D). the intercalary growth of the
suspensor pushes the proembryo towards the endosperm through the broad,
free cavity of the endospermic tube. The successive divisions of the proembryo
initial produce a terminal group of two (Fig. 19.13 A) and later four cells.
Welwitschiales 361
Fig.19.13 A-G. Welwitschia mirabilis. A Two-celled apical cell. est endosperm tube.
pt pollen tube. B Transverse division of the apical quartet. C, D Mitosis (C) and oblique
division (D) in the apical quartet. E, F Proembryo in the endosperm; initiation of cortical
tier (ci) of secondary suspensor. cp cap cells. epl embryonic plate. miz micropylar zone.
S primary suspensor. F Periclinal division in embryonic plate; wall of spearhead has
thickened (partially due to an outer incrustation). G Hemispherical homogeneous mass
of meristematic tissue formed by anticlinal and periclinal divisions in cap cells; two
cells form embryonic plate. (After Martens and Waterkeyn 1974)
The terminal cell group gives rise to a horizontal plate of inner cells
(Fig. 19.13 B). The latter grows (after the proembryo penetrates the
endosperm) peripherally and forms the first cortical tier or ring which
covers and strengthens the primary suspensor. A second cortical tier similarly
covers the first ring, and the two together form the secondary suspensor.
362 The Gymnosperms
Further mitoses and oblique divisions (Fig. 19.13 C, D) in the apical

quartet eventually form a group of inner and outer cells which divide
anticlinally. The components of the outer layer elongate to form the cap
cells, and the inner layer forms the embryonic plate (Fig. 19.13 E). The
proembryo acquires the shape of a spearhead (Fig. 19.13 E, F). The cap
cells are large, distinct, thickened by an external incrustation and better
adapted for endosperm penetration than absorption (Martens and Waterkeyn
1974). Further development in the proembryo stops until it penetrates the
endosperm and reaches the level beyond the insertion of the integument.
Development is resumed, and anticlinal and periclinal divisions in the cap
cells produce a meristematic, homogeneous and hemispherical or conical
tissue (Fig. 19.13 G). The embryo is still undifferentiated although orientation
of cellular rows shows a root centre opposite to the shoot apex, and a
coleorhiza. Meanwhile, the peripheral cells of the embryonal mass contribute
to the formation of a massive secondary suspensor. In a mature ovule the
primary and secondary suspensors are folded in a suspensor sack above the
endosperm.
Late embryogeny has been investigated in Welwitschia. A mature embryo
shows vertical orientation of the two cotyledons, prominent shoot apex, a
root centre, voluminous coleorhiza with curved cell rows and the axial
rows of the columella. The cotyledons overreach the shoot apex and elongate;
a hypocotyl is present. A non-vascular outgrowth of embryo remains
embedded in the gametophyte, represented at the mature embryo stap,e by
an annular bulge (collar) in the embryonal axis. As the seed germinates, the
collar gives rise to the "feeder" (Fig. 19.14 A; Bower 1881).
The endosperm (gametophyte) develops from a part of the chalazal zone
of the gametophyte and undergoes a long period of growth. The outer
lateral cell layers and the entire deep zone maintain their meristematic
activity during post-fertilization stages. In the micropylar and the central
region of the endosperm, there is gradual vacuolation and the cells stretch
from the centre to the upper pole. The suspensor sack differentiates in this
zone.
After the nuclear fusions in the multinucleate chalaza! compartment, the
mitoses in the polyploid endosperm cells are followed by wall formation.
As the endosperm development advances, the size of cells, nuclei, number
of nucleoli and chromosomes in a nucleus, becomes reduced in some areas.
This has been interpreted as depolyploidization (Martens and Waterkeyn
1974). It results in the formation of a heterogenous tissue made of large
and small cells and nuclei.
The main reserve tissue is relatively poor in starch, but rich in proteinaceous
granules; their accumulation compresses and deforms the nuclei. The
dermatogen differentiates late and is devoid of inclusions.
Welwitschiales 363
Fig. 19.14 A-C. Welwitschia mirabilis. A Longiscction of germinated seed.

cot cotyledons. fr feeder. nt nutritive tissue. B Seedling, cotyledons above ground.
C Eighteen-month-old seedling, cotyledons still healthy. (A After Bierhorst 1971,
B after Rodin 1953b, C after Von Willart 1985)
364 The Gymnosperms
Seed
The seeds are winged. The wing and the seed-coat develop from the perianth
(see Female Strobilus). The integument is papery.
There is a double system of vascular bundles in the seed-coat (Martens
1971, 1974). The outer system has two unbranched traces, while the inner
system has eight traces which traverse the inner part of the seed and extend
up to the base of the integument.
In the chalazal region there is a cupule of spirally thickened tracheids
intermixed with thin-walled cells. This extends basally and is not connected
with ovular vascular supply. According to Martens (1974), this tracheidal
cupule is the relict of nucellar vasculature.
Germination. When the seed germinates, the radicle breaks through the
testa (and not through the micropyle), and it is always directed downward
(Rodin 1953b). The two cotyledons break through the testa opposite the
point where the roots emerge. Figure 19.14 A shows a germinated seed in
longisection. The cotyledons are lanceolate and dark red, gradually changing
to green within a week. The cotyiedons are about 5 mm long and 2 mm
broad when they first appear above ground (Fig. 19.14 B), and reach
approximately 3 em in length and 5 mm in width when the seedlings are
about 6 months old. They remain healthy for several years. A pair of
decussately an·anged plumular leaves appear (Fig. 19.14 C), and a meristem
becomes active at the base of the leaf at a very early stage.
The venation pattern is similar in both cotyledons and foliage leaves, i.e.
longitudinal veins and oblique anastomosing veinlets. Sclereids and vessels
are absent in the cotyledons.
Many changes take place at the ultrastructural and biochemical levels
when the seed imbibes water for germination (Bornman et al. 1979 a, b, c,
Butler et al. 1979a, b, c). The radicle and the compressed hypocotyl are
separated by a uniform bulge (collar) which is the future feeder. The
embryonic collar cells are activated first after hydration, followed by the
gametophytic cells. The latter activates from the embryo outwards, and the
rate and sequence suggest the presence of stimulatory factors (in the embryo)
which diffuse into the gametophyte and induce and/or enhance metabolic
activity therein. Within 36-48 h of hydration, starch, protein and lipid
reserves in the collar and developing feeder are degraded, and the rapidly
developing seedling becomes dependent on nutritive material in the
gametophyte until the plumule emerges. Feeder cells in contact with the
gametophytic tissue develop numerous small wall projections invested with
plasmalemma and a large number of mitochondria, and act as transfer cells.
The collar gives rise to the feeder, which is ca. 5 mm long, at about 36 h
following imbibition. The cell walls are mucilaginous and the feeder and
gametophyte adhere firmly to each other, which facilitates translocation of
nutrients. Butler et al. (1979c) confirms the hypothesis of Bower (1881)
Welwitschiales 365
that the feeder functions as an absorptive organ. The feeder may have a
secondary function to provide the emerging plumule with a firm base.
The reserve material within protein bodies of the embryo and gametophyte
is a protein-carbohydrate complex. The immediate digestion of protein body
reserves in the embryo and gametophyte interface zone indicates the presence
of preexisting hydrolytic enzymes within the protein bodies. However, the
enzymes responsible for the breakdown of reserve food in the outer
gametophytic tissue are synthesized de novo. Protein hydrolysis precedes
lipid digestion, which indicates that some of the resulting free amino acids
are used in de novo synthesis of lipases. Lipid bodies, microbodies,
mitochondria and amyloplasts are characteristically encircled with ER.
The mean dry mass of the gametophyte decreases by approximately 47%
by day 5, and the amount of lipid decrease is 75% and protein 25%. Some
of the hydrolyzed fatty acids and amino acids are utilized in the gametophyte,
while the major part thereof is probably converted to sugars, which, together
with free amino acids, are transported via the feeder to the embryo to be
utilized in seedling growth.
Chromosome Number
The somatic chromosome number is 42 (K.hoshoo and Ahuja 1963). The
karyotype is peculiar and all the chromosomes appear to be rod-shaped
with probably terminal centromeres. The chromosomes are asymmetrical
with pronounced size difference. The longest pair is 3.25 times longer than
the shortest pair, wi~ a gradual transition in size. Two of the chromosomes
in the complement are satellited, which coincides with two nucleoli in the
metabolic nucleus.
20. Gnetales
Gnetaceae
Gnetum
Gnetum, with more than 30 spp., is confined to the humid, moist regions
of the world (see Chap. 1).
Morphology
Gnetum plants are shrubs, trees or climbers with twining stems, and in
general habit resemble a dicotyledonous plant (Fig. 20.1 ). The main stem
bears two types of branches: a short shoot of limited growth (internodes
present), and a long shoot of unlimited growth (internodes absent). The
difference between the types of shoots is not very pronounced. In the
climber G. ula, the short shoots bear leaves higher up near the branches of
the supporting tree. The leaves lie crowded in one plane, in decussate pairs
so that the branch looks like a pinnate leaf. The lamina is large, oval, entire
with reticulate venation and gives a typically dicotyledonous appearance.
Several Gnetum spp. have articl!lated stems. The joints consist of two
parts-one immediately above and the other immediately below the node,
the two being separated by an annular groove. The internodes are shed at
the joint, and lie in large numbers below the plant, resembling bones.
Anatomy
Root. The root is diarch. The cortex comprises several layers of large
polygonal cells filled with starch grains and numerous thick-walled fibres.
An endodermis and a five- or six-layered pericycle are present. The secondary
growth is normal; the radial walls of tracheids have uniseriate bordered pits
with conspicuous bars of Sanio. The vessel elements have smaller multiseriate
pits and the bars of Sanio are inconspicuous or absent. The xylem elements
in the root are larger than those in the stem, and some of them contain
starch grains in later stages. The phloem consists of thin-walled broad rays
packed with starch grains, which makes the wood of the root comparatively
soft as against that of the stem, where the phloem ray is thick-walled and
pitted (P. Maheshwari and V. Vasil 196la).
Gnetales 367
Fig. 20.1 Gnetum gnemon, a branch with female cones (jc). (After P. Maheshwari and
V. Vasil 1961a)
Stem. There is a distinct tunica-corpus organization. The single layer of

tunica extends over the shoot apex from the axil of the first pair of leaf
primordia. The cells at the apex are larger and more vacuolate than those
368 The Gymnosperms
in the flanks. Periclinal divisions occur in the tunica during initiation of the
foliar buttress. The corpus comprises two or three layers of subapical initials,
central mother cell zone, flanking layers and pith rib meristem.
Immediately following elevation of a pair of leaf primordia, the apex is
at its minimum volume. It is a shallow cap of subapical initials and a few
central mother cells. Also, a rib meristem may be absent above the first
node. It is, however, reconstituted as the apex increases in size.
Transection of a young stem shows a thick cuticle followed by a single-
layered epidermis with a thick outer wall, and papillate and rectangular
cells. The stomata are sunken. The parenchymatous cortex is 12- to
16-layered. The outer four to six layers are chlorophyllous, followed by a
large number of scattered fibres. In older stems there is a conspicuous
sclerenchymatous zone (Fig. 20.2 A-C) in the inner part of the cortex. It is
formed from large parenchymatous cells which become lignified. These
cells have branched and unbranched pit canals. The primary vascular bundles
(20-24) are collateral and endarch (Fig. 20.2 A-C). They are arranged in
a ring separated by broad medullary rays of considerable height. The wood
consists of tracheids and a few vessels, and the phloem comprises sieve
cells and phloem parenchyma. The endodermis and pericycle are not
discernible. The pith is parenchymatous when young; later, the cells towards
the vascular ring become thick-walled, lignified and have numerous pits.
Several laticifers occur scattered in the cortex and pith.
Laticifers of Gnetum are cellular. They originate as single cells, enlarge
and ramify (Martens 1971, Carlquist 1996). Intercellular spaces are associated
with their pectic-rich walls.
In the arborescent type (Gnetum gnemon ), the secondary growth is normal.
In climbing species (G. ula and C. africanum ), a new cambium differentiates
at various points in the inner part of cortex and gradually become a continuous
cylinder. It produces a normally oriented ring of xylem and phloem wedge-
shaped bundles separated by medullary rays. The first ring stops growing
at the commencement of the outer ring. Successive rings continue to form;
some may remain incomplete, which results in an eccentric arrangement of
the rings (Fig. 20.2 B), or an eccentric position of the pith. In G. ula, the
anomalous rings of vascular bundles appear to be extrastelar.
The phellogen arises irregularly in the epidermis and initially spreads
unequally. Later, in older internodes, the ring of cork cells becomes complete.
The periderm is thin and has lenticels.
The xylem consists of highly tapering tracheids, vessels and xylem
parenchyma. The latter are round or elongated with simple pits.
Vessel structure was studied by Muhammad and Sattler (1982) in
G. gnemon and G. montanum by light microscopy (bright field, phase contrast
and Nomarski interference optics), SEM and TEM. The tracheary elements
earliest to mature are narrow with annular thickenings followed (in the
ontogenetic sequence) by longer and wider elements with annular-helical
Gnetales 369
Fig. 20.2 A-C. Gnetum uta. A, B Transection of young and old stem to show accessory
(A) and eccentric (B) rings of vascular bundles. C Vascular bundles from A.
sci sclerenchyma. (After P. Maheshwari and V. Vasil 1961a)
and later only helical thickenings with compound gyres. In annular and
helical elements, the gyres are made up of three elementary strands
(Fig. 20.3 A). Pits range from slit-shaped to circular, and are formed either
between the elementary strands of one gyre or between adjacent gyres. In
both species, scalariform pits (Fig. 20.3 B) occur in addition to the circular
pits. Larger pits occur in contact with other tracheary elements, while vessel
elements in contact with parenchyma have smaller pits. In the nodal region,
comparatively large pits with prominent borders occur, especially on the
inclined walls of tracheids (Fig. 20.3 C). The pit membrane is thin and does
not have a torus (cf. angiosperms), unlike conifers (Fig. 20.4 A). Intervessel
370 The Gymnosperms
Fig. 20.3 A-C. A, C Gnetum montanum. A SEM tracheary element from stem shows
compound gyre. B G. gnemon, vessel from petiole with scalariform pits seen through
perforation plate. C Bordered pits on tracheids from protoxylem. (After Muhammad
and Sattler - 19~2)
pits have well-developed vestures (Fig. 20.4 B), which are absent in tracheids.
Both primary and secondary xylem have vessels. Each vessel has two,
usually similar, perforation plates at the two ends. The perforation may be
scalaroid (vertical row of circular perforations; Fig. 20.5 A), scalariform
(row of transversely elongated perforations; Fig. 20.5 B), foraminate (round
perforations in alternate or horizontal rows-commonly in the nodal region
of stem; Fig. 20.5 C), and simple perforation plate (Fig. 20.5 D). The
simple perforation plate is more common in the late metaxylem, especially
in the secondary xylem. The breakdown of the wall between the perforations
may lead to their fusion horizontally, vertically or haphazardly so that a
simple perforation plate is formed. There is no evidence that scalaroid,
scalariform, foraminate and intermediate perforation plates are formed in a
sequence (Muhammad and Sattler 1982).
Carlquist (1994, 1996) and Carlquist and Robinson (1995) do not agree
that perforation plates in Gnetum are sealariform or sealariform-like.
Both species of Gnetum demonstrate a wide range of variation in the
shape of pits and perforations.
The primary and secondary rays are broad, and comprise 10-20 thick-
walled pitted and radially elongated cells, which occasionally contain calcium
oxalate crystals. In a tangential section the rays appear boat-shaped, and
Gnetales 371
Fig. 20.4 A, B. Gnetum montanum, SEM tracheary element from stem. A Two pits with
intact membranes (arrow). B Vessel with vestured pits. (After Muhammad and Sattler
1982)
372 The Gymnosperms
Fig. 20.5 A-D. Gnetum montanum. A-D Vessels show scalaroid (A), scalariform (B),
foraminate (C) and simple perforation plate (D). (After Muhammad and Sattler 1982)
Gnetales 373
consist of polygonal cells. In addition, uniseriate or biseriate rays have

been observed and may be two, three or several cells high.
Paliwal and Behnke (1973) studied the primary and secondary phloem
in G. gnemon. It comprises only two cell types-the sieve cells and phloem
parenchyma (this term is preferred to "albuminous cells" or "companion
cells" by the above authors)-which are ontogenetically unrelated and
alternate with each other. A regular filing of both the cell types is lost
during secondary growth. A sieve cell is larger than a phloem parenchyma
cell, and has a parietal layer of cytoplasm, which occasionally clumps
together on one side of the cell. The phloem parenchyma cell has dense
plasmatic contents.
The secondary growth is initiated in a normal manner. The transection
of a young stem shows three to five layers of cambial cells within the
vascular bundles. Ray initials are present in the region of medullary rays,
but are absent in the phloem tissue itself. The latter is delimited from the
cortex by a two-layered parenchymatous sheath followed by sclerieds, and
broad rays are present laterally.
Transection of an older stem shows several layers of obliterated crushed
and dark phloem cells (op, Fig. 20.6 A, B). In none of their preparation
could Paliwal and Behnke (1973) recognize the deposition of thickening
material in older cells of phloem tissue.
In the active phloem, the parenchyma cells are filled with starch. In the
older phloem, they are empty and crushed like the sieve elements. Parenchyma
cell differentiation (of all types including those associated with the sieve
cells) reveals features comparable to those of the companion cells in
angiosperms. The cambial initial-destined to become a parenchyma cell-
becomes rich in cellular organelles. The ribosomes are abundantly distributed
in the cytoplasm and are deposited on the ER. Microtubules, dictyosomes
and mitochondria with well-defined cisternae are also present. Plastids may
have ovoid, hemispherical or elongated profiles. These contain thylakoids,
a dense matrix, and accumulate varying quantities of starch. The nucleus
persists for the entire life of these cells. Cell sap is retained in several small
vacuoles, which generally fuse to form a central cavity. They are devoid of
proteinaceous contents (see Paliwal 1992).
A differentiating sieve element in G. gnemon can be discerned from its
neighbouring parenchymatous cell by its very long nucleus. The changes in
ER during sieve cell differentiation are significant. The granular ER in the
young sieve cells tends to be modified by a swelling of interacisternal
spaces. After the loss of ribosomes, the ER assumes a more tubular structure;
tubules spread throughout the cell lumen but tend to form aggregates. The
ER tubules lje parallel to each other within the aggregates, or may become
twisted. Frequently, the ER complexes are associated with the degenerating
nuclei or with the plastids. Dictyosomes and microtubules also disappear.
Finally, the ER, plastids and mitochondria are the only organelles present
374 The Gymnosperms
Fig. 20.6 A-D. Gnetum gnemon. A, 8 Transection (part of stem) at advanced stage of
secondary growth, shows crushed primary phloem (op) and thick callose deposits on
sieve plates (spl) . C Radial longisection of phloem, the sieve areas with end-plate
(arrows). D Tangentiallongisection of phloem between two medullary rays (mr); close
grouping of sieve areas on the sieve plate, indicated by callose deposition. (After Paliwal
and Behnke 1973)
Gnetales 375
in the "living" protoplast (characteristic ~ondition in gymnosperms). The

central cell sap cavity occasionally contains fibrillar material that mixes
with and disappears within the protoplast after degeneration of tonoplast.
P-protein has not been detected either by light or electron microscopy in
young or mature sieve elements (Paliwal and Behnke 1973).
In tangential section of phloem, the sieve plates are arranged on oblique
end walls (Figs. 20.6 D; 20.7 A-D). In a radial section each end wall
(Fig. 20.6 C) appears to be made up of 7-19 sieve areas. The sieve areas
on the lateral walls between two contiguous sieve cells are more widely
spaced than those on the end walls.
At maturity, the tapering sieve cells lack cellular contents (Fig. 20.7 A)
and plastids occur throughout their length. One to four phloem parenchyma
cells remain connected to a sieve cell. They have transverse, oblique or
tapering ends. Their lateral association with the sieve elements is concluded
from callose plugs which occur on the lateral sieve areas, and take a deep-
blue stain with resorcin blue. A well-organized nucleus and other cytoplasmic
contents, including plastids, are present even at maturity.
Sieve cells and phloem parenchyma cells are ontogenetically unrelated,
but at any time of phloem differentiation, these cells are laterally connected
by specialized plasmatic strands.
At no stage was a positive staining reaction observed for proteinaceous
substances with mercuric bromophenol blue. This indicates that P-protein
(slime) is not produced during the differentiation of sieve cells or phloem
parenchyma cells.
Leaf. A vertical section of a leaf shows a well-marked cuticle on epidermal

cells with undulate walls on both surfaces, and the mesophyll of a single
layer of short palisade cells and a well-developed spongy tissue. Stellately
branched sclereids, fibres and latex tubes occur near the lower epidermis,
especially in the mid-rib region. The vascular bundles are arranged in a
curve. The xylem consists of vessels, tracheids and parenchyma. Below the
xylem phloem cells are arranged in regular rows. In Gnetum ula, pitted and
thick-walled cells form a narrow patch just outside the phloem and towards
the protoxylem.
Stomata, in addition to leaves (lower surface), occur on young stems,
axes and collars of male and female cones, and on the two outer envelopes
of the ovule.
Nautiyal et al. (1976) studied the aerial parts for epidermal structure and
ontogeny of stomata in G. gnemon, G. montanum and G. ula.
The aerial parts in the three species show irregularly dispersed stomata
and cork warts. The stomatal development is mesoperigenous. The mature
stomata are either tetracytic with two mesogene lateral subsidiaries and two
perigene polar neighbouring cells, or multicytic with three to seven
subsidiaries. The latter may be partly or wholly amphicyclic. Developmental
376 The Gymnosperms
Fig. 20.7 A-D. Gnetum gnemon, tangentiallongisection of phloem. A Phloem shows

sieve cells (sic) with oblique sieve plates and phloem-parenchyma (pp) cells. B Single
sieve plate; association of sieve cells and phloem parenchyma cells is obvious by lateral
sieve areas (arrows). C Oblique sieve plate between two sieve cells, sieve area dark.
D Sieve plate. (After Paliwal and Behnke 1973)
Gnetales 377
studies show that the multicytic stomata are formed by subsequent radial
or tangential divisions in the lateral subsidiaries and polar neighbouring
cells.
Takeda ( 1913) reported syndetocheilic development of stomata in the
leaves of Gnetum. However, P. Maheshwari and V. Vasil (1961b) report
haplocheilic development (Fig. 20.8 A-E) in G. ula and G. gnemon .. A few
. stomata on the leaves show parallel subsidiary cells, which arise from
adjoining epidermal cells, and do not seem to have a common origin with
the guard cells. Twin stomata (Fig. 20.8 F) also occur, originating from two
adjacent stoma mother cells.
Fig. 20.8 A-F. Gnetum gnemiJn, development of stomata on lower epidermis of leaf.
A-C Stomatal iritials. D Guard cells.E Older stoma. F Twin stomata. (After P.Maheshwari
and V. Vasil 1961b)
A wide variety of sclereids develop in the collars of the cones, the

perianth of male flowers, and the outer two envelopes of female flowers in
G. ula
378 The Gymnosperms
Laticiferous elements are scattered all over the plant in G. ula and
G. gnemon. These are long and, occasionally, branched tubes filled with a
latex-like substance. Those present in the pith are initiated from rows of
parenchymatous cells during the growth of the embryo.
Reproduction
Gnetum is dioecious. The male and female strobili consist of a stout axis
which bears a basal pair of opposite and connate bracts, and usually six to
eight superposed cupules or collars. The cupules arise as annular protuberances
in acropetal succession (Takaso and Bouman 1986), and an annular rim is
formed at the axillary position of each cupule. The cupules are tightly
packed in a young strobilus (Figs. 20.9 A; 20.10 A), and the latter do not
elongate appreciably during initiation of the annular rims. Later, the internode
elongates so that the annular rims separate from the next upper cupule and
the axillary position becomes pronounced. According to V. Vasil (1959),
however, the annular rim develops as a hump from a few cells lying below
each collar. Then this annular meristem comes to lie between the collar
which bears it and the one below it, and gives the impression of axillary
origin (Fig. 20.10 B-H).
Male Strobilus
Three to six rings of male flowers develop basipetally above each collar
(Fig. 20.9 B). A .single ring of abortive ovules (Figs. 20.9 B; 20.10 F-H)
may occur above the flowers. The flowers in different rings are arranged
alternately.
A male flower consists of a stalk bearing two unilocular anthers enclosed
in a perianth (Fig. 20.9 D). On maturity, the stalk elongates and pushes the
anthers (through an opening in the perianth) beyond the collars of the cone
(Fig. 20.9 C, E).
Microsporangium. In a young anther, the hypodermal archesporia! cells

divide and give rise to a multicelled archesporium (Fig. 20.11 A-C), the
outermost layer divides periclinally and differentiates into the parietal and
sporogenous cells. The former divide again to produce an outer wall layer
and the tapetum (Fig. 20.11 C). The narrow tangentially elongated wall
layer cells become compressed during meiosis of the microspore mother
cells. The tapetal cells increase in size, have dense cytoplasm and are
binucleate (Fig. 20.11 C, G). In G. gnemon, the outer tapetal layer is sculptured
by the deposition of globules (Fig. 20.11 G), which are 8-110 J.Lm3 in volume
(Carniel 1966). In the beginning, the sculpturing appears coarse, followed
by finer globules (Fig. 20.11 H, I), which give the reaction for sporopollenin.
In G. gnemon, they appear on the outer 'tangential walls of the tapetum,
while in G. africanum they appear even on the inner side of the tapetal
cells (Waterkeyn 1959). Eventually, the wall layer and tapetum are absorbed
and only the epidermis persists in the mature anther (Fig. 20.11 D, F).
Gnetales 379
Fig. 20.9 A-E. A, C-E Gnetum ula, B G. gnemon. A Panicle of male cones. B Rings
of male flowers and an ovular ring above each collar. C Portion of male cone at dehiscence.
D Longisection of male flowers, two anthers are enclosed within the perianth. E Dehisced
male flower. (After P. Maheshwari and V. Vasil 1961a)
380 The Gymnosperms
Fig. 20.10 A-H. Gnetum ula. A Longisection young male cone. B-H Portions of
longisection to show development of upper ovular and four rings of male flowers.
(After P. Maheshwari and V. Vasil 196Ia)
Gnetales 381
-
.
'
Fig. 20.11 A-1. A, D, G-1 Gnetum gnemon, B, C, E, F G. uta. A-C Longisection of

antherophores. A, B Hypodermal archesporium. C Differentiation of wall layers, tapetum
and sporogenous tissue. D Portion of wall with site of dehiscence (ld) and two-celled
pollen grains. E Longisection dehisced male flower. F Inset from E marked F to show
epidermal thickenings. G Wall layers at tetrad stage, orbicules deposited on the outer
tangential tapetal wall. H, I Outer tangential wall in surfac:e view to show fine and
coarse orbicules. (A-F After P. Maheshwari and V. Vasill96la, G-1 after Carnie! 1966)
During dehiscence, the epidermal cells become radially enlarged, and

the outer wall becomes slightly thickened and cutinized. From the inner
tangential walls, fibrous bands of thickenings run upward and outward to
the outer wall of each cell (Fig. 20.11 F). The anthers open along a double
row of smaller cells, which extend vertically on either side from the tip to
some distance below the anther sac (Fig. 20.11 D, E).
Microsporogenesis
The sporogenous cells undergo further divisions, and the microspore mother
cells contain dense cytoplasm and prominent nuclei (Fig. 20.12 A). Prior
to meiosis, the protoplasts recede and a special mucilaginous wall is secreted
between the protoplast and the mother wall. The meiosis is not sychronous
even in the same loculus (P. Maheshwari and V. Vasil 1961 a).
The reduction divisions are simultaneous (Fig. 20.12 A-E), and cytokinesis
takes place by centripetal furrows. The tetrads are tetrahedral (Fig. 20.12 F),
isobilateral and decussate. As the microspores enlarge, the callose wall is
gradually absorbed, the original wall breaks down, and the young microspores
382 The Gymnosperms
F
G
Fig. 20.12 A-G. Gnetum uta. A-E Microspore mother cell (A), meiosis I (B), and
II (C-E). F Tetrahedral tetrads. G Tetrad before liberation of microspores, callose wall
more or less consumed. (After P. Maheshwari and V. Vasil 196la)
are released. Even before separation, the microspore wall shows minute
spiny protuberances. Gradually, the wall differentiates into a thick, spiny
exine and a thin intine(Fig. 20.12 G). According to Gull vag ( 1966), the
pollen grains of G. ula, G. gnemon and G. montanum have a single layer
of lamellated exine. This needs confirmation.
Gnetales 383
Male Gametophyte. The mature pollen grain is shed at the three-celled

stage (P. Maheshwari and V. Vasil l961a, Martens 1971). The precise
sequence by which the three nuclei/cells arise has been investigated by
Negi and Madhulata (1957), Waterkeyn (1959), Martens (1971), Swamy
(1974) and others. The division of the microspore nucleus results in a small
lenticular and a large cell (Fig. 20.13 A-C). The lenticular cell rounds up,
does not divide further nor take part in the development of the pollen
tube, and degenerates. The nucleus of the larger cell divides again
(Fig. 20.13 D, E) and, of the two nuclei, one hyaline with a large nucleolus
and is the first to enter the pollen tube. The second nucleus is rich in
be
Fig. 20.13 A·L. A·F, J·L Gnetum afrieanum. A·F Development of three-celled male
gametophyte. ani antheridial initial. be body cell. pr prothallial cell. tn tube nucleus.
G-1 G. ula. G Germinated (early stage) pollen grain in pollen chamber. H, I Pollen tube
with body cell (be) and tube nucleus (tn). J·L Pollen tube with tube nucleus and division
of body cell inK and two male gametes (mg) in L. (A·F, J·L After Waterkeyn 1959,
G-1 after V. Vasil 1959)
384 The Gymnosperms
chromatin, has a cytoplasmic sheath of its own (Fig. 20.13 F), and divides
in the pollen tube to give rise to two male gametes.
These three nuclei have been variously interpreted. According to Pearson
(1912, 1914), they represent the prothallial, tube and generative nuclei.
Thompson (1916) presumes that they represent the tube nucleus, and stalk
and body cells. Negi and Madhulata (1957) and Waterkeyn (1959) interpret
them as the prothallial cell, generative (body) cell, and tube nucleus
(Fig. 20.13 F).
Swamy (1974) proposed a new type of gametophyte development in
Gnetum ula. The microspore divides and forms a small generative (body)
cell on one side, and a large cell. The generative cell (which has its own
sheath of cytoplasm) eventually lies free in the cytoplasm of the large cell,
and then moves into the pollen tube and divides to form two male gametes.
The nucleus of the larger cell divides to become a binucleate cell.
Female Strobilus
A female strobilus usually has six to eight collars (Figs. 20.14 A; 20.15 A)
and a single ring of ovules above each collar (Fig. 20.14 B, C). Two to
eight (usually five or six) ovular primordia differentiate from the annular
rim. The latter becomes conspicuous before initiation of the ovules
(Fig. 20.15 B). Gradually, the ovules become visible on the upper edge of
each collar (Fig. 20.15 C). Generally, the upper few collars have no ovules.
Ovule. The cells in the epidermal and subepidermal layers of the ovular
primordium divide actively. The dermal cells undergo both periclinal and
anticlinal divisions. Simultaneously, each primordium can be distinguished
into an upper region, which bears the ovule, and a lower, cushion-like area.
Hairs differentiate from the surface of this cushion and from sterile cells
between the ovules (Fig. 20.15 C-E).
Three envelopes arise acropetally on ovular primordia and enclose the
nucellus (Figs. 20.15 C-E; 20.16 A-C). The outer envelope (oe) is the first
to differentiate. It arises laterally to the ovular primordium by peri- and
anticlinal divisions in the dermal and subdermal cells (Fig. 20.15 C, D) and
is of dual origin. At pollination, the oe increases in thickness. About 30
vascular strands are present at the level just above the junction between the
inner envelope (ie) and the nucellus, and a few of them extend almost to
the apex. Laticifer canals (do not form lateral connections) are scattered
throughout the envelope, and numerous stomata occur in the outer layer.
After the oe is initiated, the apex of the ovule shows a clear uniseriate
dermal layer. The middle envelope (me) and inner envelope (ie) have a
subdermal origin (Takaso and Bouman 1986).
The me arises next (Fig. 20.15 E). Around pollination, it grows in thickness,
only in the subapical part of the envelope, by periclinal divisions ot the
subdermal cells which elongate radially. The outer dermal cells remain
Gnetales 385
c
8
Fig. 20.14 A-E. A, B Gnetum ula, young female cone (A) with ovules (B), C-E
G. gnemon. C Cone bearing two seeds. D, E Mature seed after shedding (picked up
from ground). (After P. Maheshwari and V. Vasil 1961a)
small and bear stomata. Provascular strands extend to subapical region.

Laticifers are also present. The ie arises last. It develops as a tubular wall
around the newly outlined nucellus, grows rapidly and forms several lobes
at the apex (Fig. 20.15 A-C). It forms a micropylar tube which extends
beyond the rim of the two outer envelopes (Fig. 20.16 C). Soon after
pollination, the apical part breaks off. Stomata, sclereids and vascular bundles
are absent. However, very few ovules develop into mature seeds
(Fig. 20.14 D, E).
In G. gnemon, at pollination, the outer epidermal cells of the micropylar
tube divide and produce a massive circular structure, called a flange, around
the tip of the middle envelope (Figs. 20.16 D; 20.17 A). At a later stage,
the outer surface of the flange becomes closely appressed against the outer
envelope. After the pollen grains have entered the ovule, the dermal and
386 The Gymnosperms
Fig. 20.15 A-E. Gnetum gnemon. A Young strobilus with one bract (br) removed.
B Axillary region of cupule with annular rim (ar). C Ovular primordia on the rim, the
outer envelope is initiated laterally. Sterile hairs occur on the rim in between the ovules.
D Ovular apex surrounded by outer envelope (oe). E Ovule with middle envelope (me).
(After Takaso and Bouman 1986)
subdermal cells lining the micropylar tube divide to produce a tissue which
fills the canal and completely blocks it (Fig. 20.17 B, C). In older seeds of
G. gnemon, these cells develop thick walls with simple pits (Sanwal 1962).
Gnetales 387
Fig. 20.16 A-D. Gnetum gnemon. A Ovule with middle (me) and inner envelope (ie).
B Inner envelope shows several apical lobes. C ·ovule with protruding inner envelope
(ie inner integument). D A flange (jl) covers the apical part of middle envelope. (After
Takaso and Bouman 1986)
The cells of the upwardly directed flange , the closing tissue and the
intermediate tissue of the inner envelope contain a suberin-like substance
(Takaso and Bouman 1986). The micropylar tube above the suberized tissue
degenerates.
A few cells of the hypodermal layer of the nucellus divide periclinally
to form the parietal tissue and the sporogenous cells. Occasionally, the
cells of the second and third layer differentiate directly into sporocytes
(Takaso and Bouman 1986). The sporogenous cells divide further and produce
nearly 12 megaspore mother cells (mgmc). A nucellar cap is formed by
388 The Gymnosperms
Fig. 20.17 A-C. A Gnetum gnemon, longisection upper part of ovule to show the
formation of flange from inner envelope. B, C G. uta, longisection upper portion of
micropylar tube. B Cells of inner epidermis. C Interlocked cells, the central inner epidermal
cells extend into the micropyle. (After P. Maheshwari and V. Vasil l96la)
Gnetales 389
periclinal division of the apical dermal layer. During the development of

the nucellus, a few mgmc degenerate. While these cells are preparing for
meiosis I, a fan-shaped tissue, the pavement tissue, of radiating rows of
nucellar cells appears at the base of female gametophyte. It has dense
cytoplasm, and the developing embryo sac gradually absorbs this tissue.
Megasporogenesis. Meiosis is non~synchronous in the developing

megaspore mother cells. Walls are not laid down after meiosis I and
II, which results in the formation of a tetranucleate coenomegaspore
(Fig. 20.18 A-D). The four nuclei later move to the peripheral cytoplasm
surrounding a central vacuole.
Fig. 20.18 A-D. Gnetum africanum, coenomegaspore. A Three megaspore mother cells,
one in telophase I. B, C Meiosis II. D Four-nucleate coenomegaspore. (After Waterkeyn
1954)
Female Gametophyte. A number of coenomegaspores develop

simultaneously, but only two or three reach the 16-nucleate stage. The
nuclei undergo repeated divisions which are synchronous in the beginning.
Mitoses proceed in a wave from the base upward, and different divisional
stages can be seen in one and the same female gametophyte. In G. uta and
G. gnemon, the divisions are intranuclear, the nuclear membrane disappears
at the anaphase. Free-nuclear divisions continue till a definite number
of nuclei have been produced, which varies in different species. In
G. leptostachyum (Thompson 1916) and G. africanum (Waterkeyn 1954),
there are 512 nuclei, in G. gnemon ca. 1000 (Sanwal 1962), and in G. uta over
1500 nuclei (V. Vasil 1959). As divisions continue, the gametophyte takes
the form of an inverted flask (Fig. 20.19 A, B). The upper part is wide and
contains the vacuole, and in the narrow lower part the nuclei aggregate
densely. The female gametophyte grows towards the chalaza) end.
390 The Gymnosperms
(
'\
~
-1
:w;·l
D
I •
I
I
I
I_- _...J
B
Fig. 20.19 A-E. Gnetum uta. A, B Longisection female gametophyte. A Free-nuclear

gametophyte. B Cell formation in the lower region of gametophyte, upper portion shows
cells and free nuclei. C-E Lower region of gametophyte. C, D Formation of multinucleate
cells. D Inset marked D in B. E Uninucleate cells resulting from fusion of nuclei.
(After V. Vasil 1959)
Walls are laid down by free cell formation, which begins at the lower
end and proceeds upward (Fig. 20.19 B, C). In G. gnemon, it is initiated
Gnetales 391
simulataneously at fertilization or immediately after. In G. ula and

G. africanum, the cytoplasm in the upper swollen portion is partitioned
before fertilization. It is not regular, and results in groups of either uninucleate
or multinucleate cells and packets of free nuclei. In the lower portion, the
cells are multinucleate in the beginning, later the nuclei fuse and become
polyploid (Fig. 20.19 C-E). These cells divide, the tissue extends upward
and fills the micropylar part of the gametophyte, which contains the zygote.
In Gnetum, the archegonia are absent. In G. ula (V. Vasil 1959), when
a pollen tube enters the free-nuclear micropylar region of the gametophyte,
one or more group/s of adjoining cells becomes densely cytoplasmic. Only
one (rarely two) cells from each group function as the egg. The lower end
of gametophyte becomes partly cellular. According to Swamy (1973), the
egg in G. ula is a uninucleate or coenocytic cell (Fig. 20.20 C). The fuoctional
female nuclei are large, and in prophase. In G. africanum also certain large
and densely cytoplasmic cells in the micropylar region are potential egg
cells (Fig. 20.20 A; Waterkeyn 1954).
In G. gnemon, one or more nuclei in the vicinity of the pollen tube
become conspicuous by their larger size (Fig. 20.20 B). They show striated
cytoplasm around them, which later becomes delimited from the rest of the
cytoplasm (Sanwal 1962).
In the male cones, the female gametophyte (in the ovules) does not
develop beyond the megaspore mother cell, or when the first meiotic division
is completed (G. ula) or has 64 nuclei (G. gnemon). Rarely, only some of
the ovules are functional, develop normally and ripen into seeds (G. gnemon,
G. africanum).
Pollination
In G. ula and G. gnemon, a drop of sugary fluid exudes from the tip of the
micropylar tube and catches pollen grains. The slimy pollination drop has
appreciable quantities of reducing sugars in G. gnemon (Pijl1953). Surface
evaporation causes the contraction of the column of fluid, which brings
about the withdrawal of pollen grains down to the nucellus (Lotsy 1899).
Anemophily is probably concerned in the transfer of pollen to the female
cone, though entomophily may also be involved. The male inflorescences
emit a sweetish odour in the morning. The anthers open between 0.700 and
11.00 h. At about the same time, or earlier, nectar drops are present on
fertile and sterile (in the male inflorescences) female flowers. All these
characters are suggestive of insect pollination (Pijl 1953).
Kato et al. (1995) studied pollination biology in two Gnetum spp. in a
lowland mixed dipterocarp forest in Sarawak, Malaysia. A dioecious (shrub
species), G. gnemon var. tenerum, flowered in the evening. Both male and
female strobili emitted a putrid odour, that from male strobilus being stronger
than that from the female. Pollination droplets are secreted in the evening
from ovules on female strobili and from sterile ovules on male strobili.
392 The Gymnosperms
Fig. 20.20 A-C. A, B Longisection upper portion of female gametophyte. A Gnetum

africanum, group of cells at the upper end are (potential) eggs (ec). B G. gnemon, two
large egg nuclei (en) with radiating cytoplasm around them near the pollen tube (pt).
C G. ula, multinucleate cell from upper part of gametophyte, the egg nucleus (en) is
larger than the others. (A After Waterkeyn 1954, B after Sanwal 1962, C after Swamy
1973)
Withdrawal of droplets after pollination has not been observed. The droplets
contain sugar at a concentration of 3 to 13%. The sugar concentration is
affected by the relative humidity (usually 95% in the evening in the forest
floor habitat) of the surrounding air. A very small amount of nectar is also
secreted on the outer covers of the microsporangiophores on male strobili.
These strobili are visited by nectar-seeking moths of the Pyralidae and
Geometridae. The pollen has been observed sticking on the proboscides
and antennae of these moths.
G. cuspidatum, a dioecious climber, has cauline strobili on woody stem
near the forest floor. Its male strobili (which lack sterile ovules) emit a
fungus-like odour (different from that of G. gnemon var. tenerum) in the
evening. Nectar is secreted on collars that subtend flower rings early in the
Gnetales 393
evening (between 17.00 and 18.20 h) with sugar concentration 6 to 10%.

The strobili are visited by small flies, Lauxaniidae (Diptera), the pollen
sticking to their antenna and legs. In female strobili, the pollination droplet
is secreted on the tip of the ovules between 17.30 and 18.00 h and ceases
by 21.00 h. The mean sugar concentration of the pollination droplet is
14.7%. During flowering, the female strobili also emit an odour similar to
the male strobili.
Evaporation of the exposed droplet/nectar of these Gnetum "flowers"
seemed to be minimized by nocturnal flowering near the tropical rain forest
floor [which, according to Kato et al. (1995), could be a reason why Gnetum
survived mainly in the tropical rain forests]. The lack of showy "petals" (in
gymnosperm) is compensated for by odour.
The two Gnetum spp. flower in the understorey of rain forests where the
humid habitat is not suitable for anemophily. Therefore the odou·r and
secretion of droplets and/or nectar on exposed parts attract nocturnal
pollinators. The sugar concentration is higher than that of anemophilous
conifers (Pinus 1.2%; Mcwilliam 1958) but is lower than in entomophilous
Ephedra (72-80%; see Chap. 18). These micropylar secretions also serve to
draw pollen into the inner region of the ovule, where pollen germination
takes place. According to Kato et al. (1995), the nocturnal pollination system
of these Gnetum spp. has been derived from unspecialized entomophily,
which is presumed to be an original pollination system of the early Gnetales
and early angiosperms (Lloyd and Wells 1992). A study of the composition
of carbohydrates in pollination droplets and nectars of these Gnetum spp.
will help in the understanding of evolution of reproductive systems in
gymnosperms (Chesnoy 1993).
The ovules are pollinated when the female gametophyte is still free-
nuclear.
The pollen grains germinate in the shallow pollen chamber on the nucellar
tip (Fig. 20.13 G), and occasionally in the micropylar tube (seeP. Maheshwari
and V. Vasil 196la). Germ pores are absent on the exine. During germination,
the exine is thrown off, and the intine, nearest to the tube nucleus, grows
into a tube (Fig. 20.13 H). The pollen grains contain starch grains. In
G. ula and G. gnemon the pollen tube grows into the nucellus through
intercellular spaces. The nucellar cells are full of starch, which nourishes
the pollen tubes. The tube nucleus is the first to enter the pollen tube, the
body cell moves in after the pollen tube has traversed nearly half the length
of the nucellus. The prothallial cell remains in the grain and degenerates in
situ (Fig. 20.13 I). The body cell divides to form two male gametes
(Fig. 20.13 J-L).
Fertilization
All the pollen tubes do not reach the female gametophyte at the same time,
nor do the eggs differentiate simultaneously. As soon as one of the eggs is
394 The Gymnosperms
fertilized, wall formation begins in the upper part of the gametophyte. The
latter becomes nearly cellular by the time the second or third egg differentiates.
There are very few observations on fertilization. In G. ula the pollen tube
lies close to one of the groups of densely cytoplasmic cells present in the
upper part of the gametophyte. Only the male nucleus enters the egg; its
sheath is cast off outside the egg cell. Both male cells from a pollen tube
can function if two egg cells are present close to the pollen tube.
The observations of Swamy (1973) in G. ula are somewhat different.
The two small male nuclei are discharged into the cell containing the egg
nucleus. The functional male nucleus enlarges slightly, comes close to the
egg nucleus and enters into prophase (Fig. 20.21 A, B). The second male
nucleus stays in the peripheral zone of cytoplasm, and disintegrates. As the
male and female nuclei come in to contact, their chromatin becomes more
pt
c 0
A
Fig. 20.21 A-D. Gnetum ula . A Longisection upper part of nucellus with a cellular
female gametophyte and pollen tubes (pt). B Egg cell from A (arrow), a functional male
nucleus (c5') is in contact with the egg nucleus while the second male nucleus has
degenerated. C, D Integration of chromosomes of male and female nuclei. (After Swamy
1973)
Gnetales 395
intense. In the male nucleus the chromatin assumes the form of a short,
somewhat curved and stumpy rod-like structure (Fig. 20.21 C). The nucleoli
disappear from both the nuclei, and the two groups of chromosomes eventually
become indistinguishable (Fig. 20.21 D).
In G. gnemon, the two male nuclei from a pollen tube may fertilize two
adjacent eggs, as the zygotes often appear in pairs at the tip of the pollen
tube. The zygote nucleus is large and hyaline, and has its own cytoplasm
and limiting membrane. It is surrounded by many nuclei embedded in
dense cytoplasm (Sanwal 1962).
The nature and behaviour of the gametes during fertilization in G. gnemon
has been studied by Carmichael and Friedman ( 1995). The pollen tube
discharges two male gametes which fuse with nearby undifferentiated female
nuclei, within the coenocytic female gametophyte, and form two zygotes.
According to Carmichael and Friedman (1995), this is an expression of a
rudimentary pattern of double fertilization. The process of fertilization has
been studied with light and fluorescence microscopy and the DNA content
of various nuclei involved quantified with 4', 6-diamidino-2-phenylindole
microspectrofluorometry. It has been observed that both male and female
nuclei pass through the S-phase (DNA synthesis phase) of the cell cycle,
and double their DNA content from 1C to 2C prior to fertilization. Each of
the two zygotes (in association with a pollen tube) contains 4C DNA.
Fertilization is therefore dependent on attaining a precise stage within the
cell cycle. This reproductive cell cycle pattern is defined as G2 karyogamy
(Carmichael and Friedman 1995).
Double fertilization has been reported in Ephedra spp. [Two male gametes
are released into the archegonium, one fuses with the egg to form the
zygote, and the other fuses with the ventral canal nucleus (see Chap. 18)].
In angiosperms the zygote and the primary endosperm nucleus are formed
as a result of double fertilization. It has been proposed that double fertilization
evolved before the origin of angiosperms. The original manifestation of
double fertilization in seed plants may have led to the formation of two
embryos (Friedman 1992b, 1994, 1995).
Some angiosperms have diploid endosperm, e.g. Butomopsis lanceolata
(a primitive angiosperm) (Johri 1936). Of the two male gametes from a
pollen tube, one gamete fuses with the egg and the other with the upper
polar nucleus (lower polar nucleus is not formed). Thus, the zygote and the
primary endosperm nucleus are genetically identical but their products are
quite different. Zygote gives rise to the embryo and the primary endosperm
nucleus to endosperm-both diploid. There is no such parallel in the
gymnosperms.
The term "double fertilization" should be restricted to the fusion of one
male gamete with the egg and the other with the secondary nucleus (fused
polar nuclei) and not to their products.
396 The Gymnosperms
Embryogeny
Various investigators report both free-nuclear and nuclear division followed
by a wall in the zygote. The details of embryonal development are not very
clear. According toP. Maheshwari and V. Vasil (1961a), this confusion is
partly due to variation in different species of the genus. Several zygotes are
usually formed in the female gametophyte.
In G. africanum, successive division in a zygote and its derivatives results
in a row of cells (Fig. 20.22 A). Each of these cells gives rise to a suspensor
tube devoid of any cross-walls (Waterkeyn 1954).
In G. gnemon, the number of zygotes varies from two to four, rarely six.
According to Sanwal (1962), the zygote may directly give out a small
tubular projection on one side (as also reported by Lotsy 1899), or may
divide (Fig. 20.22 B, C) into two cells (as reported by Thompson 1916).
Each of these cells may give out a tube or only one of them may "germinate".
These tubes have been variously termed proembryonal tubes, embryonal
tubes, embryonal suspensor tubes and primary suspensor tubes. Generally,
a single tube is given out, rarely two or three (Fig. 20.22 D, E). The zygote
nucleus passes into one of the tubes, the other tubes without the nucleus
degenerate. Sometimes, two zygotes may "germinate" while they are still
in contact (Fig. 20.22 F).
In G. ula, the division of the zygote (Fig. 20.22 G) is followed by a wall
(V. Vasil 1959). The two daughter cells elongate to form suspensor tubes
(Fig. 20.22 H). According to Swamy (1973), the zygote in G. ula enlarges
and the early divisions are free-nuclear. After the four-nucleate stage, two
short processes appear at one pole of the proembryo and grow towards the
chalaza. The tubular processes branch, and at the 8- and 16-nucleate stages
the nuclei migrate into the tubes. this phenomenon continues until a large
number of uninucleate primary suspensor tubes are formed, and all the
tubes penetrate the core of the endosperm (female gametophyte) in the
chalazal direction.
In G. gnemon, the thin-walled embryonal suspensor tube grows downward,
while the developing endosperm tissue extends upward. The tubes grow
through the intercellular spaces of the endosperm, and become septate to
form long uninucleate cells. Above or below a septum, a small protuberance
appears, which later elongates into a tubular structure containing a nucleus.
Rarely, a branch is given out without any such disposition. The suspensor
tubes grow downward into the endosperm and tend to aggregate in the
centre, where the cells break down to form a conspicuous cavity. The
development of embryo is initiated in only a few suspensor tubes, and a
small cell is cut off at the tip. This cell divides transversely and then
longitudinally and a quartet is produced. Further divisions result in an
embryonal mass, and a long secondary suspensor pushes it into the endosperm.
The mature embryo shows two cotyledons and a prominent feeder.
In G. ula the suspensor tube becomes coiled in 6 to 7 month-old
Gnetales 397
Fig. 20.22 A-H. A Gnetum africanum. B-F G. gnemon. G, H G. ula. A Filament of

proembryonal cells (pee). B, D, E Zygote with one, two and three proembryonal tubes,
respectively. C Two-celled proembryo. F Two proembryonal tubes. G Division of zygote.
H Two-celled proembryos, the upper cell elongates and forms suspensor (S) tube .
(A After Waterkeyn 1954, B-F after Sanwal 1962, G, H after V. Vasil 1959)
proembryos, and ceases to branch. The nucleus divides to form a larger

nucleus with poor chromatin, and a smaller one embedded in the dense
cytoplasm at the tip (Fig. 20.23 A-C). The latter is cut off from the rest of the
tube by a thin wall and forms a pyriform cell, the so-called 'peculiar' cell
(Fig. 20.23 C, D). In some tubes it may elongate parallel to the tube and
398 The Gymnosperms
Fig. 20.23 A-J. Gnetum ula. A-D Formation of peculiar cell (pel). E-1 Development
of embryonal tissue and suspensor. H, I Formation of secondary suspensors. J Portion
of suspensor (S), multicelled secondary suspensor (ss), and embryonal cells (e) at the
tip. (After V. Vasil 1959)
remain straight, slightly curved or twisted, and is suspected of degenerating.

According to Swamy (1973), due to lack of precise information about its
formation and variable shape, this cell has been termed peculiar although
there is nothing peculiar in its ontogeny or structure. The pyriform cell
Gnetales 399
ultimately gives rise to mature embryo (V. Vasil 1959, Swamy 1973).
Morphologically, it is the embryonal cell. The seed is shed at this stage,
and further development takes place on the ground.
The pyriform cell rounds up and divides to produce eight cells
(Fig. 20.23 D-F). Further divisions are irregular, and a mass of cells is
formed . The cells towards the suspensor tube elongate, divide further and
form secondary suspensors (Fig. 20.23 G-1). The cells at the lower end
remain compact and densely cytoplasmic, and contribute to the embryo
proper (Fig. 20.23 I, J). Gradually, the massive secondary suspensor tubes
cease to function and degenerate.
The embryonal mass grows in size, and two cotyledons differentiate
which enclose the stem tip. The root apex is initiated at the same time at
the opposite end. From the side of the hypocotyl a lateral hump or feeder
arises and outgrows the hypocotyl (Fig. 20.24 A-C). The bulk of the feeder
results from the cortical cells of the hypocotyl (Sanwal 1962).-lt has a well-
developed vascular supply, is the most prominent part of the embryo, and
is longer than the hypocotyl at the mature embryo stage (Fig. 20.24 C).
Laticiferous ducts and stomata occur in a mature embryo.
Fig. 20.24 A-C. Gnetum ula. A-C Longiscction of embryo shows initiation, elongation
and vascularization of feeder (j). (After V. Vasil 1959)
Polyembryony is a regular feature. Many embryos arise by cleavage of

the embryonal mass and /or budding of the primary and secondary suspensors.
However, only one embryo matures in a seed.
400 The Gymnosperms
The bulk of the endosperm develops after fertilization, though cell

formation may commence before fertilization. In G. gnemon, the female
gametophyte is free-nuclear at the time of differentiation of the eggs. The
lower part becomes cellular when the zygotes appear in the micropylar part
(Sanwal 1962). Wall formation is initiated either simultaneously with
fertilization, or about that time. In G. uta, polyploid cells are produced at
the chalaza! end of the gametophyte before fertilization, but only uninucleate
cells are produced after fertilization (Swamy 1973). With further growth,
the endosperm broadens at the expense of adjoining nucellar cells which
are consumed laterally as well as at the base. In G. gnemon (Sanwal 1962)
and G. africanum (Waterkeyn 1954) the meristematic activity in the
gametophyte is limited to the axial region and a few outer layers, especially
at the two poles. Thus, the main mass of endosperm is formed and starch
appears first in the outer cells at the lower end, later at the micropylar end,
lastly in the inner cells. In older seeds, the cells become gorged with starch
grains and may mask the nuclei. Oil droplets have also been detected; in
later stages the cell walls show many simple pits (Fig. 20.25 A-D).
Seed
In most species the mature seed is oval and green to red. The endosperm
forms the bulk of the seed, and is surrounded by three envelopes. The
nucellus is consumed except for a thin strip at the apex.
During the development of (ovule and) seed, the level of insertion of the
inner envelope shifts (appreciably) distally. This is due to the development
of an ovular tissue between the inner and middle envelopes, accompanied
by an intercalary growth in the entire developing seed. The middle and
outer envelopes show no such shift in their levels of insertion. A special
term, endochalazal, was introduced (Bouman 1984) for such an ovular
structure.
Rodin and Kapil (1969) studied the comparative anatomy of the seed
coat of G. gnemon, G. ula, G. montanum f. parvifolium and G. neglectum.
The seed coat consists of three layers; (a) outer sarcotesta, (b) middle
sclerotesta and (c) inner endotesta (Fig. 20.26). The sarcotesta is free from
the base up to apex, and is green and succulent. It is composed of a heavily
cutinized epidermis and homogeneous parenchymatous cells. Diverse types
of sclereids with lignified walls, and numerous branched fibers and laticifers
also occur. In G. ula, G. montanum and G. neglectum, small astrosclerieds
or brachysclerids occur just below the outer epidermis. Additional
astrosclereids are scattered in the deeper tissues. Haplocheilic stomata, as
in the leaves, occasionally develop on the outer epidermis.
The sclerotesta forms the protective layer of the seed. It has numeruus
sclereids of varying shape and may sometimes extend as a basal plate.
Depending on the species, at maturity the middle layer may be nearly free
from the outer layer, or may be partially or completely fused with it.
Gnetales 401
Fig. 20.25 A-D. Gnetum gnemon. A Endosperm from lower end shows cells in division.
B Older endosperm shows proteinaceous globules in addition to starch . C Endosperm
axial region with cells in division. D Mature endosperm cells have pitted walls and
store starch. (After P. Maheshwari and V. Vasil 196la)
The endotesta is parenchymatous and membraneous. In a few Gnetum

spp. it has elongated sclereids (Rodin and Kapil 1969).
All the layers of the seed coat are vascularized (Fig. 20.26). A single
ring of vascular bundles enters the base of the seed; each bundle bifurcates,
giving rise to an outer series which supplies the outer envelope. The inner
series bifurcates again to supply the middle envelope and the peripheral
ovular/chalaza! tissue which surrounds the endosperm, and extend to the
inner envelope. In G. gnemon, on an average, 21 bundles are present in the
basal ring, while the outer, middle and inner rings contain approximately
62, 18 and 18 bundles, respectively (Ro~din and Kapil 1969).
402 The Gymnosperms
Fig. 20.26. Gnetum gnemon, seed coat. Transection of seed shows endosperm (esp),
endotesta (ent), sclerotesta (sci), sarcote-sta (sa), and vascular bundle (vb). (After Rodin
and Kapil 1969)
Germination. In several Gnetum spp., the seed does not germinate

immediately on falling to the ground as it is immature and a considerable
part of its development takes place on the ground (cf. Cycas, Ginkgo). The
root with the root cap emerges form the seed first, bends downward, enters
the soil, and gives rise to the taproot system (Fig. 20.27 A-C). Next, the
upper part of the hypocotyl elongates, pulls the cotyledons out of the seed
(Fig. 20.27 D, E) and straightens to bring up the two (occasionally three,
Fig. 20.27 K) cotyledons and stem tip in an upright position (Fig. 20.27 F).
The cotyledons expand, turn green and resemble a foliage leaf in shape and
venation (Fig. 20.27 G-J). The stem tip expands and produces the first pair
of plumular leaves, which are opposite and decussate to the cotyledonary
leaves (Fig. 20.27 L). The feeder remains inside the seed and occupies the
entire space previously filled by the hypocotyl. The endosperm tissue adjacent
to the feeder has a lighter colour and appears to be corroded by it. The
feeder continues to function as an absorptive organ till a very late stage,
and can be seen even after the second and third pair of leaves have developed
on the seedling (P. Maheshwari and V. Vasil 1961a).
Chromosome Number
The haploid chromosome number in G. ula is n = 22. The bivalents
show slight intergradational differences. At diakinesis the configurations
observed are in the form of X, L, Y, 0 and a figure 8; the number of
chiasmata in a bivalent varies from one to three. Meiosis is normal. Occasional
Gnetales 403
e •
fr
8
A
~ E
F G H
Fig. 20.27 A-L. A-J, L Gnetum gnemon, seed germination. A-E Germination of seed.
A Half-endosperm removed after emergence of root. fr feeder. F-K Development of
seedling. K G. uta, seedling with three cotyledonary leaves. L seedling shows vegetative
leaves. (After P. Maheshwari and V. Vasil 196la)
diploid grains (inferred from their larger size and form) are observed
(Mehra 1988).
=
In G. gnemon, Fagerlind (1941) noticed n 22 chromosomes at metaphase
of a single sporocyte (Mehra 1988). Of these, 11 chromosomes are longer
than the rest. During metaphase, the smaller chromosomes are mostly grouped
in the centre of the plate, while the longer chromosomes are arranged
around in a radiating fashion (P. Maheshwari and V. Vasil 196la). In
G. africanum, the haploid number is between 20 and 25 (Waterkeyn 1959).
Temporal considerations
The life cycle of G. ula (growing wild in Peninsular India) takes nearly 18
404 The Gymnosperms
months from the time of initiation of the flowers to the germination of

seeds. The seeds germinate ca. 11-12 months after they are shed from the
plant (V. Vasil 1959). In G. gnemon the seeds are shed in June/July (in
Assam, Eastern India), a few cells have been observed at the tip of a
primary suspensor. It takes 2 months for the maturation of the embryo. The
seeds germinate in September (Sanwal 1962).
21. Phylogenetic Considerations:
Ephedra, Welwitschia and Gnetum
The Gnetopsida has been studied for over a century. The scanty fossil
record, restricted to the Tertiary, does not Indicate the origin and relationship
of this enigmatic group. The discovery of Ephedripites pollen (related to
modem Gnetopsida pollen, specially Welwitschia) in the lower Cretace_ous
(Trevisan 1980) is significant, as it suggests that at least some member of
the group was present at the time of the origin of angiosperms. According
to Stewart (1983), based on fossil record, a polyphyletic origin of angiosperms
from gymnosperms appears more plausible than previously presumed. As
in progymnospermopsida (gymnospermous wood and pteridophytic
reproduction), the preangiospermopsida (proangiosperms) could have any
one,of a several combinations. Those gymnosperms, that combine a few/
some angiospermic charact.ers and form a complex from which evolutionary
lines lead to the flowering plants, should be investigated for the origin of
angiosperms.
The Gnetopsida ovule has multiple nucellar envelopes, unlike other
gymnosperms (which have a single integument only). These have been
interpreted as integumentary structures homologous with the outer integument
of an angiosperm (Stewart 1983).
Entomophilly in several spp. of Ephedra, Gnetum and in Welwitschia is
rather an evolutionary progress from the primitive gymnospermous mode
of anemophilly present in the majority of extinct and extant gymnosperms
(Cycadeoidea is an exception, where beetles were partially responsible for
pollination, Crepet 1972). It is a more economical and probably more effective
and reliable form of pollination (Bino et al. 1984). The incidence of an
apparently effective insect pollination in gymnosperms is of great interest.
Eames (1952) discusses the affinities of Gnetopsida in considerable detail,
and concludes that Ephedra is nearer to cordaites and conifers than to
Welwitschia and Gnetum. The ovule and microsporangia of Ephedra are
appendicular (i.e. terminal on the lateral appendage of a fertile shoot), and
not cauline (terminal on the shoot, as in Welwitschia and Gnetum). According
to Eames (1952), this is an important morphological difference, indicating
a wide phyletic gap between the three taxa.
406 The Gymnosperms
Embryologically also, Ephedra differs from Welwitschia and Gnetum

(see H. Singh 1978). The male gametophyte has a stalk cell. The female
gametophyte is monosporic, and alveoli are formed during wall formation.
The archegonia are well developed, there is a distinct and specialized type
of proembryogeny and a feeder is absent in the mature embryo. Welwitschia
and Gnetum, on the other hand, lack a stalk cell in the male gametophyte;
development of the female gametophyte is tetrasporic, and alveoli are not
formed during wall formation, archegonia are absent, the proembryogeny
is very different and a feeder is formed in a mature embryo. Thus, Ephedra
occupies a position isolated from the other two taxa, and may have had a
completely different origin (Sporne 1965). Welwitschia is also completely
isolated with respect to hereditary relationships (Von Willart 1985).
Phylogenetically, it must be very old, as there are no recent forms known
that relate to other taxonomic groups.
Gnetum has long held a key position in any discussion relating to the
origin of angiosperms (seeP. Maheshwari and V. Vasil1961a). Besides the
similarities it shares with other members of the group, it has features which
show a proximity to the angiosperms.
Leaf. The external appearance of the broad leaf with netted venation is
strikingly angiospermous. The development of the stomata is of the
mesoparacytic (Rubiaceous) type (Kausik 1974) which, according to Takhtajan
(1969), appears to be the basic type in the evolution of flowering plants.
Nautiyal et al. (1976) confirm that the stomata of Gnetum differ from those
of other gymnosperms (possible exception Bennettitales) and resemble
angiosperms such as some MagnoHales and Casuarina.
All the six main types of sclereids which are known in angiosperms are
also reported in Gnetum (Sharma 1975). Outside the flowering plants, the
occurrence of laticifers in G. gnemon is a rare example (Behnke and Herrmann
1978).
Shoot Apex. The shoot apex of Gnetum is similar to that of angiosperms

in the tunica and corpus organization. However, the central mother cell
zone is a reminder of gymnosperms such as cycads and Ginkgo (Johnson
1950).
There are physiological similarities between Gnetum and angiosperms in
the chlorophyll synthesis in dark-germinated seedlings (Laudi and Bonatti
1973).
Phloem. The phloem in Gnetum has sieve cells, associated with

ontogenetically unrelated parenchymatous cells (in angiosperms sieve tube
and companion cells are derived from the same cambial initial and are
successive cells in a given row). In Austrobaileya scandens (a recognized
primitive angiosperm), a tier-by-tier study of the origin of sieve elements
Phylogenetic Considerations: Ephedra, Welwitschia and Gnetum 407
and parenchyma (companion) cells led Srivastava (1970) to conclude that

true companion cells are absent. Instead, parenchyma cells and sieve elements
arise independently from different initials, and are connected to each other
by lateral sieve areas. Thus, the phloem of Gnetum also closely approaches
the angiospermous taxa (see Paliwal and Behnke 1973).
Xylem. Studies on the vessel of G. gnemon and G. montanum (Muhammad

and Sattler 1982) demonstrate a greater range of variation in the shape of
pits and perforations than known so far. It consists of typical circular pits
(of progymnosperms, pteridosperms and coniferopsids) and non-scalariform
perforation plates to scalariform pitting (as in Magnolioideae). Bliss (1921)
and MacDuffie (1921) demonstrated striking similarities between perforation
plates of Gnetum and angiosperms such as Paeonia, Cydonia and Vitis.
Most of the patterns characteristic of and comparable to angiosperms are
present, such as: absence of torus in the pit, presence of vestured pits and
compound gyres.
Gnetum thus combines both gymnospermous and angiospermous
characters. A preliminary survey of 58 characters showed that Gnetum
shares more than 60% characters with the angiosperms, and ca. 30% with
gymnosperms (see Muhammad and Sattler 1982). This hypothesis, however,
does not necessarily demonstrate an ancestral relationship to angiosperms,
but the possibility that Gnetum may have been close to the ancestry of all,
or at least some, of the taxa of angiosperms (Muhammad and Sattler 1982).
According toP. Maheshwari (1950), it appears best to conclude that, while
the angiosperms have probably passed through some such stages as are
shown by Gnetum, there is no decisive evidence in favour of any close
relationship.
22. In Vitro Experimental Studies
Experimental procedures provide excellent opportunity to study growth,

development and differentiation of vegetative and reproductive tissues. The
gymnosperms-a group of plants between the pteridophytes- and
angiosperms-have not yet been investigated as thoroughly as the
angiosperms.
There are isolated reports of experimental embryology in gymnosperms
dating back more than a century (see Norstog 1982). Such studies can be
traced to the pioneering work of Professor Carl D. LaRue, University of
Michigan, Ann Arbor (USA). He extended his concept of cellular totipotency
and developmental potentials in plants to include all aspects of reproduction
(LaRue 1954). Despite the easy availability of material, only a few
gymnosperms have been investigated, possibly because they are extremely
slow-growing plants and less amenable to cultural conditions.
Vegetative Tissues and Organs

Ball (1950) cultured tissues from young adventive shoots growing on the
burls of Sequoia sempervirens, on diluted Knop's solution with 3% sucrose
and 1 mg/1 IAA. For the first time, he successfully maintained a continuous
culture of coniferous tissue in vitro. Marginal meristems, and cambium-like
meristems around groups of tracheids and mature parenchyma cells could
be distinguished in the callus mass. The parenchymatous cells occasionally
contained tannin. Ball (1955) reported the utilization of various sugars by
the callus, the normal chlorophyllous tissue containing glucose, fructose
and sucrose. When the callus is grown on any one of these three sugars, the
tissue can produce the other two. Maximal growth is achieved on a medium
containing sucrose. When either raffinose or galactose is supplied as the
source of carbohydrates, sucrose, fructose, glucose, galactose and raffinose
are detected in the callus. According to Ball (1955), the enzyme that yields
galactose-first from the hydrolysis of excess raffinose-forms raffinose
later by condensation of excess galactose and sucrose.
Reinert and White (1956) cultured normal and tumerous (characteristic
of the species) tissues of Picea glauca, to understand the degree of malignancy
of the cells and the biochemical characters of the tumours. The cambial
In Vitro Experimental Studies 409
region is excised from the tumorous and non-tumorous portions of tumour-

bearing trees and also from normal trees. A complex nutrient medium
consisting of White's minerals, 16 amino acids and amides, 8 vitamins and
auxins was developed to raise the cultures. The tissues, grown on filter
paper moistened with the above medium, turned brown and ceased to grow.
This is due to a copper-containing enzyme (phenoloxidase) which appears
to be especially active in traumatized tissue. Browning could be prevented
by the addition of tyrosine at 40 mg/1, or phenoloxidase inhibitors like
diethyldithiocarbamate or sodium ethylxanthate. The tissues continued to
grow for some time; with tyrosine the growth was continuous. Additives
like 2, 4-D, glutamine and IAA also enhanced growth. The behaviour of
normal and tumorous tissues differed; the growth of the tumorous tissues
was faster, but they were difficult to establish. The tumorous tissues require
vitamin B 12 while the normal cells do not.
White and Risser (1964) studied conditions for maximal growth of callus
derived from tumour tissues of Picea glauca. Maximal growth was achieved
when 20 mg of callus was inoculated on an agarized substrate (0.5%) at pH
5.5, in square bottles with screw caps. The cultures were maintained at
23°C either in darkness or in diffuse light, and transferred to fresh media
every 2 weeks. Under these conditions, the mean increment per passage
was about ninefold.
Risser and White (1964) formulated a less complex medium than suggested
earlier by Reinert and White (1956). Sucrose at 5% is the most effective
carbohydrate source; it can be replaced satisfactorily only by dextrose,
while fructose, mannose and raffinose are only half as effective as sucrose.
All the 18 amino acids in the medium can be replaced by !-glutamine
(250 mg or more/1). Amongst the auxins tested, the effective growth promoters
are 10-7M 2, 4-D, 10-5M IAA and 10-3M .8-napthoxyacetic acid; 2, 4-D
gives the best result. Adenine, KN, ,8-alanine, gibberellin and folic acid (in
the presence of 2, 4-D) have no promotive effect.
The cells of Picea glauca (White 1967) grown in vitro exhibit patterned
deposits in the form of strands, furrows or channels, in the cytoplasm,
which may act as templates for cellulose and lignin deposition. The cytoplasm
is extruded into characteristic papillae (in some cells), which condense to
form distinctive extracellular patterns.
Borchert (1967) studied the physiological differences in tissues derived
from various explants, or p!ants of different ages. As an initial step, he
determined conditions necessary for in vitro culture of secondary phloem
and cambium of Cupressus lusitanica. The explants proliferated on a medium
Abbreviations: ABA abscisic acid, BA benzyladenine, BAP benzylaminopurine, BM

basal medium, CH casein hydrolysate, CM coconut milk, CW coconut water, 2, 4-D
2, 4-dichlorophenoxyacetic acid, GA gibberellic acid, fAA indoleacetic acid, IBA
indolebutyric acid, KN kinetin, MS Murashige and Skoog ( 1962), NAA napthaleneacetic
acid, TIBA tri-iodo-benzoic acid, WB White's medium, YE yeast extract.
410 The Gymnosperms
containing Heller's minerals and sucrose. When vitamins or growth substances

are added to this medium, there is little improvement in growth. However,
when they are added to Heller's medium containing higher concentrations
of mineral salts, growth is considerably stimulated.
Suspension culture technique is a very useful approach to many problems
in cellular biology. Konar (1963b) studied the cell behaviour of callus of
Pinus gerardiana, grown on Reinert's modified liquid medium. In suspension
cultures, the tissues dissociate into single cells, which later divide to form
cell aggregates.
Hasegawa et al. (1960) demonstrated the formation of lignin in the cultures
of cambial tissue of Pinus strobus, using radioactive compounds. Tissues
fed with 14 C glucose, acetic acid and shikimic acid, and the four fractions
obtained (amino acid, acid fraction, phenol and sugar) were analyzed for
radioactivity. When the tissues are fed with glucose-1- 14C, maximal activity
occurs in the acid fraction which contains shikimic acid. The extent of
conversion of glucose to shikimic acid is ca. three to five time more than
that of acetic acid; the incorporation of acetic acid into shikimic acid is
considerably lower. The authors conclude that shikimic acid and lignin are
synthesized from glucose in the isolated cambium tissue. They assume that
the aromatic nucleus of lignin is formed from sugar via shikimic acid.
Constabel (1965) reports that the extent of tannin production in the
cultures of Juniperus communis depends on growth, the presence of precursors,
and light. With increase in growth rate, there is a decrease in tannin content.
When cinnamic, s.inapic and ferulic acids are added to the medium to
determine whether these compounds function as precursors of tannin, the
level of tannin rises. However, these compounds stimulate growth at low
concentrations, but have an inhibitory effect at high concentration. Cinnamic
aid is definitely a precursor of tannin, whereas ferulic and sinapic acid also
act as precursors. There is a significant difference in tannin content of the
tissues kept under 14-h light against those kept in dark. In the dark, tissues
show higher tannin production.
Al-Talib and Torrey (1959) cultured dormant terminal buds of Pseudotsuga
taxifolia, and pointed out that isolated dormant terminal buds required oxygen,
sucrose, glucose or fructose, and light for growth. They also studied the
effect of auxins, gibberellins, kinetin and adenine sulphate on the growth
and development of buds. Auxin 10-6M had no stimulatory effect on bud
development. At a higher concentration (10-4M), the development of the
leaf is generally retarded. The addition of GA (0.1-1.0 mVl) in the medium
restricts leaf expansion and elongation of the main axis and the buds collapse
during later stages. Adenine sulphate reduces bud development and leaf
expansion. KN (0.001-1.0 mg/1) has a stimulatory effect on callus growth.
Excellent bud development and leaf expansion occur when a Seitz-filtered
solution of urea is added to the medium at 10-1M. Supplements like CM,
YE, CH, and watermelon juice are inhibitory to the development of the
bud.
A comparative study of in vitro growth and developmental pattern of·

shoot apices of different gymnosperms would be rewarding (Biswas 1994).
Microspore and Male Gametophyte

In vitro pollen culture of gymnosperms· has been successful in obtaining
normal or near-normal development. The objectives are to overcome problems
of dormancy, viability, storage and longevity. The first success in pollen
culture of a gymnosperm resulted by using the hanging drop technique.
According to Kuhlwein (1937), 2-20% (lower concentrations give better
results) sugar solution is well suited for (in vitro) pollen culture of several
gymnosperms. Branscheidt (1930) with the hanging drop meth<>4 also cultured
the pollen of Taxus and Cupressus in a medium supplemented ',with sucro~e ..
The sperm nuclei formed in about 20 days. LaRue (1954)-cultured pollen
grains of Zamia on nutrient agar. The development proceeded through several
stages until the penultimate division of the generative cell. Blepharoplasts
were formed in the sperm mother cells, but not spermatozoids. The tubes
remained active (in this stage much longer than under in vivo conditions)
and branched.
De Luca and Sabato (1979) reported, the first in vitro spermatogenesis
in Encephalartos altensteinii. The microsporangia (containing three-celled
microspores) were cultured for 8 months. Within 1 week after inoculation,
masses of pollen tubes emerged from the sporangia. ln 6 months, sperm
mother cell formed at the tip of pollen tube. On division, two flagellate
sperms were produced, which, however, did not attain maturity.
Pollen of Ephedra germinates in a medium containing ovular extracts
(Mehra 1938), and forms fully developed pollen tubes with (apparently)
normal sperm mother cells. In vivo, the pollen tubes develop rapidly in ca.
10 h (in other gymnosperms ca. "10 months), and are non-haustoria!. It may
be presu.med that specific nutritional requirements in vitro are not as
demanding as in vivo.
Male cones of Pinus roxburghii, at the microspore mother cell stage,
were cultured by Konar (1963c), in White's medium supplemented with
nucleic acids. The pollen mother cells show normal development and give
rise to mature pollen grains which germinate. The antheridial cell divides
to form the stalk and body cell.
The first subculturable callus derived from pollen is t\lat of Ginkgo ('fulecke
1953). The mature pollen is cultured at the shedding stage (Tulecke 1953,
1957, 1960). The gametophyte develops normally till the formation of
flagellated spermatozoids, but motility has not been observed. The pollen
tube forms the usual haustoria! structure (see Chap. 12). Several abnormalities
have been observed: formation of septate pollen tubes; divisions in the tube
nucleus (forms large coenocytic vesicles, each containing nearly 50 nuclei)
and, less frequently, divisions in the prothallial, stalk, body and (occasionally)
even in the sperm cells to form a cluster of cells (Fig. 22.1). Such clusters
of;J
412 The Gymnosperms
A B C 0
~--:rmal ~
~ 1 ------- - ;. 1 .
0
- -
'-
c0 - -
~
~
F
o-· Coenocytes
~
G
Fig. 22.1. Ginkgo biloba. Diagrammatic representation of normal and abnormal

development of male gametophyte under in vitro conditions ; note embryo-like structures
in E, G. (After Tulecke 1957)
from several pollen tubes give rise to a tissue of meristematic parenchymatous

cells, which is white and friable, and separates readily into groups of adherent
cells. This tissue is basically haploid and forms a subculturable callus.
Tulecke and Nickell (1960) maintained this tissue (in vitro) continuously
for a number of years, without cell differentiation. Tulecke et al. (1965)
devised a setup, the Phytostat, for obtaining continuous suspension cultures
of Ginkgo biloba pollen tissue.
Biochemical and physiological studies have been conducted to determine
the differences in the free and protein amino acids, sugars, and non-volatile
organic acids in pollen and in tissue derived from it. A strain of pollen
tissue was isolated which requires arginine (1000 mg/1) for its growth (Tulecke
1960). Canavanine, an antimetabolite of arginine, inhibits the growth of the
arginine-requiring strain. Ammonia could, to a certain extent, replace arginine.
Similar subculturable callus has been obtained from the pollen of Taxus
(Tulecke 1959), Torreya (Tulecke and Sehgall963), Ephedrafoliata (Konar
1963d) and Thuja (Rao and Mehta 1969). This tissue is haploid and remains
undifferentiated.
Razmologov (1973) observed that the generative cell of the pollen grain
of Cupressus produces a callus-like tissue in vitro. Bonga and Mcinnis
(1975) cultured rnicro-sporophylls of Pinus resinosa. The first division of
the immature pollen produces equal-sized cells. Cold treatment results in
more pollen with such cells, and an increase in callus initiation. When cold
treatment is followed by centrifugation, callusing increases significantly.
However, none of the pollen (or rnicrospore) cultures, or the resultant tissue,
regenerates buds, roots or embryos. This may be due to a higher capacity
of gymnospermous tissue for proliferation or that the correct stage of culture
for induction of embryos has not been ascertained. According to Norstog
(1982 ), the culture of rnicrospores and microgametophytes, unlike
angiosperms, do not produce the hoped-for clones of haploid tissue and
embryos. However, the production of subculturable callus indicates that
improved techniques may result in cloning and diploidization, as is now
possible in some angiosperms.
The in vitro production of mature rnicrogametophytes of cycads and
Ginkgo provides much additional information about spermatogenesis. There
is scanty information, especially at the ultrastructural level, concerning the
development of microgametophyte and sperm cells of other gymnosperms.
Rohr (1973b) made an ultrastructural study of sperm cells formed within
the cultured pollen tubes of Taxus baccata. In some of the electron
micrographs, ultrastructural aspects of well-defined sperm cells have been
observed.
Female Gametophyte
The production of plants through vegetative growth of prothallial tissues
(apogamy) is of considerable scientific interest. It demonstrates the ability
of cells to manifest almost the entire range of morphogenetic potential of
the species (totipotency). It is valuable to geneticists, horticulturists and
foresters as a means of rapid vegetative propagation. The haploid female
gametophyte can be used as an alternate source (po!len grains are readily
available but with numerous genotypes) for the production of haploids and,
being massive, is more amenable than the male gametophyte. This may be
due to its larger size, sterile location, and a distinct meristematic phase.
Du Chartre (1888) described root formation by the female gametophyte
of Cycas, which is the earliest record of regeneration by megametophyte of
a gymnosperm. Coulter and Chrysler (1904) made a similar observation in
Zamia.
LaRue (1948, 1954) successfully maintained the female gametophyte
(excised at about the time of fertilization) of Zamiafloridana and later Cycas
revoluta on simple media, and obtained regeneration. Both root and shoots
in Zamia (Fig. 22.2) and Cycas, as well as adventive embryos (in Zamia,
see below), have been reported. The leaves on the shoots are small and
closely resemble those of the seedling.
Norstog (1965b, 1967b) and Norstog and Rhamstein (1967) made a
detailed study of the initiation of callus, adventive embryos, roots and
leaves from the gametophytic tissue of Zamia integrifolia and Cycas circinalis.
The megagametophyte of Zamia responded variously to additions, deletions
and changes in the concentration of au~ins, cytokinins and amino acids
(Figs. 22.3; 22.4). Without these additives, only callusing occurred. The
414 The Gymnosperms
Fig. 22.2. Zamiafloridana. Regeneration of roots and shoots in cultured megagametophyte.

(After LaRue 1948).
addition of 2, 4-D and KN induces callusing followed by rooting. The

addition of glutamine and asparagine to the auxin-cytokinin medium results
in a con·siderable increase (from 48 to 92%) in the number of
megagametophytes which exhibit morphogenetic response. In addition, there
is regeneration of shoots as well as roots (Fig. 22.3). The organization of
the gametophyte influences the orientation of regenerated roots and shoots
(Norstog 1965b). These organs tend to form at the archegonial end of
gametophytes on media without an auxin. When auxin and KN are both
present in the media, this polarizing effect is largely offset and roots and
shoots form in other regions as well. The presence of an organized mass
of cells (megagametophyte) appears to be essential in the direct regeneration
92%
Q)
1/l
c:
0
c.
1/l
~
"E
e
Q)
Q)
a..
Q.___ __
Fig. 22.3. Zamia integrifolia, megagametophyte cultured on three different media.

A Minimal basal mineral medium. B Basal medium+ 2, 4-D + KN. C Basal medium
+ 2, 4-D + KN + glutamine + asparagine. Black callus. Horizontal lines callus + root.
Vertical lines callus +bud. Cross-hatched callus +root+ bud. (After Norstog 1967b)
of roots and shoots. The roots originate from peripheral areas of meristematic
cells, and retain the haploid chromosome number.
There is a different response on a medium with different concentrations
of auxin and cytokinin (Norstog and Rhamsteine 1967). The mega-
gametophytes are cultured on a high-auxin medium (10 mg/1 2, 4-D) in
shake cultures. When the resulting callus is transferred to a medium with
lower levels of auxin and cytokinin (1 mg/1 each of 2,4-D and KN), it
could be maintained and subcultured without any organ regeneration. When
the cells are transferred to a medium without auxin or cytokinin, embryoids
(embryo-like structures; see H. Singh 1978) differentiate. This sequence
repeatedly gives the same result. Similarly, the megagametophytic tissue of
Cycas circinalis produces subculturable callus which does not form roots,
shoots or embryoids.
The megagametophytes from mature seeds of Ceratozamia mexicana,
Cycas revoluta and Encephalartos umbeluziensis were cultured on modified
White's media (according to DeLuca et al. 1979) with 2% sucrose-with
416 The Gymnosperms
Ml megagametophyte
.·F G.: egg
n n
E ·.
2n
embryo
Jemb•yotd
D
.- L . _ _ _ l_ _ _ -----ir
H :.·
nor 2n n or 2n
Fig. 22.4. Zamia integrifolia. Megagametophytes and embryos grown on eight different
media: A Basal mineral medium without 2, 4-D and KN; B Basal mineral medium+
2, 4-D + KN (1.0 mg/1 each); C Basal mineral medium+ 2, 4-D + KN (1.0 mg/1 each)
+asparagine+ glutamine (400 mg/1 each); D Medium containing alanine (100 mg/1) +
glutamine+ asparagine (400 mg/1 each)+ adenine (10 mg/1) + NH4-malate (100 mg/1); E
Medium same as in D + 2, 4-D + KN (1.0 mg/1 each); F Modified Murashige and Skoog
medium+ alanine (100 mg/1) +glutamine (400 mg/1) + 2, 4-D + KN (1.0 mg/1 each);
G Medium same as E +2, 4-D (10 mg/1); H Basal medium+ sucrose, vitamins not added.
All media from A-G contain 2% sucrose and vitamins: (After Norstog 1967b)
or without 2, 4-D and KN. Callusing occurred in all the taxa but regeneration
only in Ceratozamia and Cycas. The callus of Ceratozamia (on BM) showed
differentiation of only adventive embryos, but later circinate leaves developed.
On a medium enriched with amin~ acids, embryos and haploid roots were
also formed. The Cycas megagametophytes, when callused on an enriched
medium, produce a large number of pseudobulbils (interpreted as root
primordia by the authors). Coralloid roots are formed when the
megagametophytes are cultured on a medium containing 2, 4-D and KN.
These roots lacked the endophytic blue-green alga but are anatomically
comparable to in vivo roots (DeLuca and Sabato 1980).
Tulecke (1964, 1967) reported that Ginkgo megagametophyte produces
a subculturable callus but, unlike Zamia, there is no regeneration of roots,
shoots and embryoids. Rohr (1977) cultured the female gametophyte of

Ginkgo, 4-6 months after fertilization. The superficial tissues turned deep
green and produced a variety of outgrowths. A tissue isolated from the
gametophyte and subcultured continues to grow even after several transfers
on the same medium (see Norstog 1982).
Borchert (1968) reports callus formation from the megagametophytes of
Pinus lambertiana, P. resinosa, P. nigra and P. mugo. Only non-subculturable
proliferations of callus is obtained from the female gametophytes of
P. resinosa (Bonga and Fowler 1970). After 5-7 weeks of culture, 10%
megagametophytes of P. nigra and about 14% of P. mugo produce a bright
green callus capable of subculture (Bonga 1974). In P. nigra, the callus
comprises haploid cells, and in P. mugo, predominantly haploid with a few
diploid cells.
Konar and M.N. Singh (1979) cultured the female gametophyte excised
from mature seeds of Ephedra foliata. Initiation of callus (Fig. 22.5 A, B)
occurs on BM (MS + 2% sucrose + 10% CM) + 2, 4-D (2 mg/1); stock
cultures were maintained on a lower concentration of 2,4-D (1 mg/1). The
subcultures on BM + 2 mg/1 KN (auxin-free medium) result in the formation
of multiple shoots and roots (Fig. 22.5 C). Transfer of callus with shoot
buds and roots on KN-free medium produced better growth of shoots only
(Fig. 22.5 D). According to Konar and M.N. Singh (1979), 2, 4-D stimulates
callus formation. However, extensive proliferation and formation of
subculturable callus is obtained only when coconut milk is added. In the
absence of CM, the tissue neither regenerates nor yields a subculturable
callus.
M.N. Singh et al. (1981) excised female gametophytes from ovules at
the archegonial stage and cultured them on BM (MS + 2% sucrose +
10% CM). Growth and morphogenetic responses of the explants to auxins
(at different concentrations) and their interactions with cytokinins has been
studied. BM + 2, 4-D (2 mg/1) induces profuse callusing \Vhich subsequently
produces roots. NAA (4 mg/1) is optimal for callus growth and rooting.
Combinations of 2, 4-D and KN are more effective in inducing roots and
shoot buds, than a combination of 2, 4-D and BAP. The addition of BAP
(0.05 mg/1) to a medium containing optimal concentration of NAA (4 mg/1)
results in the formation of a large number of roots. A high concentration
of BAP (8 mg/1) stimulates shoot bud formation. The root tips and callus
have haploid number of chromosomes (n = 7).
Bhatnagar and M.N. Singh (1984) cultured female gametophyte of Ephedra
foliata for inducing haploids. The age of the explant is important to determine
patterns of regeneration. Explants cultured before archegonia formation do
not respond.
Longitudinal halves of the female gametophyte have been used as explants.
Transverse halves (to ascertain the role of tile archegonium on morphogenetic
responses) when inoculated on the BM (MS+2% sucrose+ 10% CM
418 The Gymnosperms
Fig. 22.5 A-D. Ephedrafoliata, female gametophyte. A, B 2- and 4- week-old culture

on BM + 2 mg/1 2, 4-D . C Callus with shoot and root on BM + 2 mg/1 KN, 4 weeks
after subculture from BM + I mg/1 2, 4-D. D Shoots differentiate from the callus . (After
Konar and M.N. Singh 1979)
+2 parts 10-6 NAA showed no significant difference (between the micropylar

and chalazal halves) in callus growth and roots differentiated in both halves.
As compared to longitudinal halves, in transverse halves there is much less
callus growth and fewer roots.
In Ephedra, 2, 4-D causes callus formation, regeneration does not occur.
With the addition of CM (10%) the female gametophyte produces callus
and roots differentiate. However, CM-in the absence of sucrose, even
though 2, 4-D at 2 parts 10-6 is present in the medium-supports scanty
callus growth. CM has a promotory effect on growth only when the medium
has been supplemented with essential growth substances (Bhatnagar and
M.N. Singh 1984).
The regeneration of roots depends upon the addition of NAA (substitute
for 2, 4-D). NAA at 4 parts 10-6 is optimal for the growth of callus and
differentiation of roots per culture of the female gametophyte at both
archegonial and mature embryo stages.
Callus growth and morphogenetic development are strongly influenced
by the ratio of NAA and BAP, and the developmental stage of the
gametophyte. Root regeneration depends on (the presence of) NAA, while
BAP has a modifying effect. When lower concentrations of BAP (0.05--0.5
parts 10-6) is incorporated in the medium containing (0.05-4 parts 10-6) NAA
(explants at the archegonial stage), BAP promotes the rhizogenic effect of
NAA. The roots differentiate in 91% cultures (Fig. 22.6 A), and the average
number of roots in the explant is higher than in the explants at the mature
embryo (stage), which show roots in only 33% cultures. At higher
concentration of BAP (6.0 parts 10-6 and 8.0 parts 10-6), the explants at the
archegonial stage differentiate shoot buds (Fig. 22.6 B), while those at the
mature embryo stage do not respond at all. Callus on explants at the
archegonial stage produces the maximal number of roots at 1 part 1o-6 BAP,
and the mature embryo stage at 0.5 part 10-6
Callus initiation, growth and regeneration are optimal at 4 parts 10-6
NAA. The addition of 0.05 part 10-6 BAP in the medium (containing optimal
concentration of NAA) increases the rate of callus growth, percentage of
cultures forming roots, and the number of roots. BAP (above 6 parts 1o-6 )
does not stimulate growth of callus, and does not effect root differentiation.
However, a higher (2-4 parts 10-6) concentration of BAP (along with NAA
0.5 part 10-6) improves the differentiation of shoot buds (but the roots are
fewer) at the archegonial stage (Fig. 22.6 C, D).
In interaction studies of 2, 4-D and KN, the cultured female gametophyte
showed significant differences in growth and regeneration of root and shoot
buds. Shoot buds regenerated at lower concentration of 2, 4-D (0.05-2
parts 10-6 ) and different concentrations of KN (6 parts w-6 ), and formed
normal shoots (Fig. 22.7 A). Further growth is inhibited at 0.5 part 10-6 and
2 parts 10-6 2, 4-D. When grown on BM without 2, 4-D and KN, growth
is resumed, and new roots are also formed. This response indicates that
420 The Gymnosperms
Fig. 22.6 A-D. Ephedra foliata, female gametophyte at archegonial stage. A 4-week-
old culture on BM + 0.05 part 10-6 NAA + 0.05 part 10-6 BAP shows callus with roots.
B, C, D 6-week-old culture . B On BM + 0.05 part 10-6 NAA + 6.0 parts 10-6 BAP
shows formation of root and shoot buds. COn BM + 0.5 part 10-6 NAA + 2.0 parts
10-6 BAP; callus mass with roots and shoots. DOn BM + 0.5 part 10-6 NAA + 4.0
parts 10-6 BAP shows differentiation of roots and shoots. (After Bhatnagar and M.N.
Singh 1984).
2, 4-D at relatively high concentrations, together with KN, has an inductive

effect on shoot bud initiation; its continued presence is not necessary for
further growth. 2, 4-D (0.05-2 parts 10-6), when combined with lower
concentrations of KN (0.05 part 10-6 and 0.5 part 10-6 ), generally favours
root formation. A combination of 2, 4-D (0.05 part 10-6 ) and KN
(4 parts 10-6) is optimal for the induction of root and shoot buds. Higher
concentrations of KN (6 and 8 parts 10-6 ) inhibit rooting (Fig. 22.7 B).
The callus cells and root tips have haploid chromosomes (Fig. 22.7 C);
rarely, a few cells are diploid (Fig. 22.7 D).
According to Gauthert (1966), a differentiated cell can undergo
dedifferentiation and provide new cells that are likely to exhibit various
other types of specializations under in vitro conditions. The formation of
an organized structure begins with meristemoids (Torrey 1966). The
meristemoids are small, and comprise densely cytoplasmic compact groups
of cells, which have prominent nuclei (Fig. 22.8 A). They respond to
directional stimuli (within the culture) to form a primordium (Fig. 22.8
B, C), which can give rise to root, shoot bud or an embryo-like structure.
The meristemoids in the female gametophyte cultures of Ephedra are located
on the surface, or embedded in the callus tissue (Bhatnagar and M.N. Singh
1984). The deep-seated meristemoids organize only into root primordia,
but the peripheral ones give rise to root as well as shoot bud primordia.
Initially, there is no vascular connection between the shoot bud and the
callus, but it is established later (Fig. 22.8 D, E).
Nagmani and Bonga (1985) report embryogenesis in subcultured callus
of Larix decidua. The megagametophytes are excised from immature ovules.
The entire (coenocytic or prearchegoinal cellular) gametophyte is sown on
the modified nutrient medium (MS 1962 or Litvay et al. 1981, either at full
strength or with all macro- and microelements-except FeSo 4 and
Na 2 EDT A-reduced to half-strength). Gametophytes at an advanced stage
of development are cut longitudinally (archegonia, proembryos/embryos
and suspensors are removed) and each half is placed with its cut surface in
contact with the medium. After a small gametophytic callus has been formed,
it is transferred to hormone-free medium. Thereafter, the callus is subdivided
and subcultured (on the same medium) every 2 weeks. After 2 to 3 weeks,
a large number of embryoids with suspensor differentiated. The embryoic!s
comprised small isodiametric cells with dense cytoplasm, large nuclei and
several large nucleoli. Most embryoids remain small, but some develop
into plantlets (the ploidy could not be determined). After six to eight
subcultures, the number of embryoids gradually declined.
In gymnosperms, haploid cultures have been established more often from
megagametophytes than from microsporophylls, microsporangium or pollen.
The cultures of female gametophytes may prove to be a useful means of
achieving haploid and doubled-haploid trees of gymnosperms incorporating
desirable genetic characters. In cycads, megagametophyte culture demonstrates
a considerable capacity for regeneration. Further investigations are necessary.
422 The Gymnosperms
Fig. 22.7 A-D. Ephedrafoliata, female gametophyte. A, B 6-week-old culture. A On

BM + 0.05 part l0-6 2, 4-D + 1.0 part I 0-6 KN shows shoot and root formed from
ca llus . BOn BM + 0.05 part l0-6 2, 4-D + 6.0 parts l0-6 KN ; large number of shoot
buds. C, D Acetocarmine squash of p-dichlorobenzene-treated root tip of in vitro-
differentiated root from callus shows n = 7 haploid (C) and 2n = 14 diploid (D)
chromosomes. (After Bhatnagar and M.N. Singh 1984)
Fig. 22.8 A-E. Ephedrafoliata, transections of callus . A Meristematic zones. B Shoot

bud primordia along the periphery. C Shoot bud with apical meristem and two leaf
primordia. D, E Vasculate nodules and root. E Inset from D shows vascular connection
between root and callus . (After Bhatnagar and M.N. Singh 1984)
Embryo
Studies on embryo culture are of great importance in determining the factors
influencing the development of embryos for overcoming dormancy, and for
rearing hybrid embryos.
424 The Gymnosperms
The earliest report of embryo culture in gymnosperms was by A. Schmidt

in 1924 (see Durzan and Campbell 1974). He cultured embryos of Pinus
sylvestris in organic nutrient solution. Li (1934) and Li and Shen (1934)
cultured embryos of Ginkgo and demonstrated that the excised embryos of
gymnosperms can germinate and produce plantlets, on relatively simple
nutrient media, which replace the female gametophyte. LaRue (1936) cultured
the excised embryos of a number of plants-Pinus resinosa, Picea canadensis,
Tsuga canadensis and Pseudotsuga menziesii (taxifolia)- and pointed out
that the immature embryos (2-4 mm in length) grow and develop into
plantlets. Subsequent investigations have shown that the excised mature
embryo is capable of giving rise to a seedling independently of the
megagametophyte.
Radforth and his associates (Radforth 1936, Radforth and Bonga 1960,
Radforth et al. 1958) cultured proembryos of Ginkgo and Pinus nigra. One
of their objectives was to assess the developmental potentialities of the
embryo itself. It was observed that the embryo develops radially and forms
callus-like masses (suspensor is not formed). In Ginkgo (Radforth et al. 1958)
the archegonia-containing free-nuclear proembryos (without suspensor
formation) are excised along with the minimum amount of circumjacent
gametophytic tissue, and cultured on nutrient agar. Cell divisions occur in
the cultured proembryos but there is no suspensor formation. The absence
of polarity in the embryo is perhaps due to its uniform accessibility to
nutrients (Fig. 22.9 A, B).
Sterling ( 1949b) cultured immature embryos of larch (Larix sp. ). In some
cultures, the proembryonic mass cleavaged, but none of the units completed
normal development. Similar results were reported by Loo and Wang (1943),
Radforth and Pegoraro (1955), Berlyn (1962) and Konar and Oberoi (1965).
This is partly due to suboptimal culture media. Thomas (1970) cultured the
undifferentiated proembryos of Pinus on Heller, Halperin, and Murashige
and Skoog (1962) media. The proembryo survived up to 48 h on all three
media, but up to 8 days only on Halperin medium. However, there was
neither cell division nor cellular elongation.
Norstog and Rhamstine (1967) observed that in Zamia integrifolia the
embryo cultures with suspensors formed normal dicotyledonous (occasionally
polycotyledonous) embryos. Normal or near-normal differentiation occurs
on media without auxin and KN. In a relatively low concentration of
2, 4-D (0.01-0.1 mg/1), and KN (0.05-0.5 mg/1), a subculturable callus is
formed. The embryos differentiate on transfer to an auxin-free medium.
Cells and tissues of Zamia (apparently) are much more plastic in their
morphogenetic responses, than other gymnosperms. This plasticity was
elucidated after a trial of 78 different media, of these 8 showed marked
selective effects on growth and development (Norstog 1967b, Norstog and
Rhamsteine 1967).
V. Vasil (1963) obtained callus from the embryos of Gnetum ula cultured
A B
Fig. 22.9 A, B Ginkgo biloba . Embryos of same age in vitro (A) and in vivo (B) . Note
absence of suspensor (formation) and polarity in embryo in A; the embryo has a distinct
polarity in B. (After Radforth et a!. 1958)
at the embryonal cell (the so-called peculiar cell) stage, on White's basal
medium supplemented with (500 mg/1) CH and YE: The callus comprised
thin-walled cells of various size and shape. In a subculture, after 4 weeks
the callus formed uniseriate filaments, or small buds, or proembryos. Attempts
to induce differentiation of cotyledons in these proembryos were unsuccessful.
In gymnosperms, the culture of very young embryos (proembryos) has
not been successful. Normal patterns of embryogenesis do not occur in
vitro even with partial or complete growth. The cultured embryos usually
exhibit radial symmetry/asymmetry rather than axial organization.
Using partially mature Ginkgo embryos, Ball (1956a) observed that the
primary root originates endogenously between the base of the hypocotyl
and the hypocotyl cap. He concluded that the hypocotyl meristem is not
synonymous with the root meristem. Further studies (Balr 1956b)
demonstrated the developmental potential of the root initials, and a longitudinal
split generates two new complete apices (Fig. 22.10 A, B).
The root and shoot of Ginkgo embryos have different requirements with
respect to sugars (sucrose, glucose, raffinose, galactose, levulose), IAA,
glutamine, and CM. The uptake of these substances occurs only through
the cotyledons [Ball (1959); Fig. 22.11)], which indicates that the cotyledons
may not only take up and distribute growth regulators but also mediate the
morphogenesis of root and shoot.
426 The Gymnosperms
Fig. 22.10 A, B. Ginkgo biloba. Effect of a longitudinal incision at the tip of the
hypocotyl of embryo on root development in vitro A Longisection embryo immediately
after longitudinal incision at the tip of hypocotyl. B Same, 9-day-old culture, each half
of the hypocotyl has grown into a normal root. (After Ball 1956b)
Ginkgo embryos could be grown in vitro, with or without cotyledoas.

However, the growth of cotyledons, hypocotyl and root ceased after about
15 days (Li 1934). According to Bulard (1952), normal growth of embryos
occurred only when the cotyledons were in contact with the nutrient medium.
Brown and Gifford (1958) demonstrated the role of cotyledons in affecting
root growth in Pinus lambertiana embryos grown in vitro. They devised a
double-medium technique in which the embryos were suspended with the
cotyledons embedded in agar, and the hypocotyl tip remained immersed in
a liquid medium contained in an inner vial. Excellent root growth occurs
if the roots remain immersed in a sugar-free medium. However, root
elongation occurs only in the presence of sugar.
In P. lambertiana Greenwood and Berlyn (1965) reported physiological
polarity in sections of embryos. The hypocotyl of the dormant embryos
were cut into 2-3 mm segments and cultured in inverted tubes on Knop's
medium containing agar (0.8%) and sucrose (4%). When the segments
were inoculated with their morphological basal end downward in air, roots
emerged at this end. On reversing the orientation, root did not differentiate.
Fig. 22.11. Ginkgo biloba, embryo cultured with only the cotyledons embedded in
medium. There is a well-developed root and shoot. (After Ball 1959)
Berlyn and Miksche (1965) studied the in vitro growth behaviour of

mature embryos of P. lambertiana. The embryos show good growth even
428 The Gymnosperms
without cotyledons. They point out that the removal of shoot meristem (of
the embryos) suppresses root development. The orientation of the excised
embryo in relation to gravity and light alters embryo growth. On placing
the culture tubes (containing the embryos) in a vertical position in light, the
growth was much better than in the tubes placed horizontally. In the dark,
the cultures in a horizontal posi~ion showed a very high growth rate. However,
this differential effect of light or orientation occurred only in excised embryos;
the embryos with gametophytes showed no such response. The authors
conclude that the gametophyte inhibits tropic responses of the embryo.
Normally, the germinating embryos of gymnosperms develop chlorophyll
in the dark, while excised embryos on nutrient media do not. However, a
germinating Pinus embryo (with the female gametophyte removed) may
maintain chlorophyll synthesis for a short period in the dark (Bogorad
1950, Sacher 1956). In cultures, when the embryos are suspended with the
cotyledons embedded in agar media containing yeast, malt and female
gametophyte extracts, Engvild (1964) observed that apparently a specific
chlorophyll synthesis-stimulating molecule is not involved; chlorophyll
synthesis could be promoted by sucrose, B-vitamin, urea and a mixture of
amino acids.
Le Page-Degivry (1968, 1970, 1973a, b, c) and Le Page Degivry and
Garello (1973) investigated the dormancy of the embryo of Taxus baccata.
Immature embryos cultured on agar media grew very slightly. When cultured
in a liquid medium first and transferred to agar media (after 8 days),
germination occurred. According to these investigators, the inhibitors for
germination leach out of the embryos in liquid culture. An analysis showed
that the excised uncultured embryos, as well as the liquid medium (in
which the embryos had been cultured for 15-20 days) contained substances
with properties of ABA; the extracts of the cultured embryos lacked ABA.
Following leaching, treatment of embryos with gibberellic acid, or chilling
(at 4°C), broke the dormancy. In Ginkgo also, gibberellic acid promotes
in vitro germination of excised embryos (Bulard and Le Page-Degivry
1968).
Excised embryos of Pinus strobus were cultured on MS medium
supplemented with different growth hormones (Minocha 1980). On BM
containing 3-6% sucrose, the embryos developed into plantlets; with 1-2%
sucrose, the root primordia did not grow satisfactorily. The addition of
GA3 (0.01-20 mg/1) suppressed the growth of root primordia; while auxin
(NAN2, 4-D/IBA-each 0.01-2 mg/1) led to callus formation. It showed
no organogenesis. IBA (1-5 mg/1) induced adventitious shoots from the
hypocotyl of some embryos. Similar shoots were formed at the tip of
cotyledons when TIBA (0.5-1 mg/1) was added to the medium (Minocha
1980).
The excised embryos of Peseudotsuga menziesii reared on MS medium,
with or without the female gametophyte or its extract, exhibited striking
differences in size. Obviously, the female gametophyte contains some

substances which have a growth-promoting effect on the embryo. It is
thermostable and diffusible in agar. The extract also has a synergistic effect
with CM in promoting morphogenesis in suspension cultures of Pseudotsuga
(Mapes and Zaerr 1981).
Sommer et al. (1975) cultured the embryos of Pinus palustris on Gresshoff
and Doy (1972) medium (modified) for the first 4-6 weeks. Numerous
adventitious buds along elongating cotyledons were formed. The buds, when
excised and grown on modified Risser and White (1964) spruce medium,
formed roots and vigorous plantlets.
The excised embryos of Pinus ponderosa formed callus and initiated a
few buds from the tip of cotyledons when placed on medium devised by
Sommer et al. (1975) and Cheng (1975). When they were placed on Reilly
and Washer's (1977) medium (full or half-strength dilution); or on Cheng
(1975) medium (half-strength dilution), buds differentiated along the entire
length of those cotyledons in contact with the medium. For bud initiation,
BA must be present in the medium; the addition of auxin increases the
yield of excisable cotyledonary buds (Ellis and Bilderback 1984).
Konar and Oberoi (1965) cultured the embryos of Biota orienta/is, at
different stages of development, on White's medium with various supplements
such as 2, 4-D (1-10 mg/1), IAA (0.1-5.0 mg/1), CM (10-20%), CH (1-5
mg/1) and YE (500-1000 mg/1). Other media were also tested: Crone (see
Radforth and Pegaro 1955), Butenko and Yakovleva (1962) and Tepfer
et al. (1963). Sucrose (2%) was used as the carbon source and the cultures
were maintained at 25 ± 2 °C with ca. 8 h of light:
Embryos cultured before the initiation of cotyledons showed occasional
proliferation; those with cotyledons and shoot apex developed rapidly. Within
3-4 days, the cotyledons mature and tum green; the hypocotyl swells and
a short root develops at the basal end. On WB + CM (10 mg/1) + 2, 4-D
(1 mg/1) + IAA (0.1 mg/1), the embryos develop into small seedlings in 15
days. On WB + IAA (0.1 mg/1), the hypocotyl proliferates and forms a
massive friable green callus. In these embryos, the root and the shoot are
initiated but their further growth is arrested. In the absence of initiation of
root and shoot, the hypocotyl fails to callus.
In Crone's medium, the root develops and reaches 15 mm in 20 days;
several well-developed secondary roots are also formed. The development
of shoot is, however, suppressed. In Butenko's medium the cotyledons in
90% of embryos mature within 1 week. Concurrently, a short taproot is
formed at the radicular end, followed by slight callusing of the hypocotyl
and appreciable swelling of the cotyledons (Fig 22.12 A). In these cultures,
the shoot and root failed to grow further. The 8-week-old cultures had five
to seven small swellings on the inner surface of the cotyledons and become
prominent within another week (Fig. 22.12 B, C). These swellings gradually
differentiated into embryo-like structures (embryoids) with two prominent
430 The Gymnosperms
Fig. 22.12 A-D. Biota orienta/is. A Young seedling with swollen hypocotyl (hyp) and
cotyledons (cot) bearing embryoids (e). ca callus. r root. B, C Embryoids. D Young shoot
from cotyledonary embryoid. (After Konar and Oberoi 1965)
cotyledons. Finally, they germinated and formed shoots with normal leaves.
When the embryoids are subcultured with a small amount of cotyledonary
tissue, normal shoots are produced with spirally arranged leaves (Fig. 22.12 D).
In none of the media did the embryoids develop a root. However, the two
cotyledonary leaves and the dome-shaped apex compare well with a normal
plantlet (seedling). This is probably the first report of induction of embryoids
on the cotyledons of a gymnosperm.
Konar and M.N. Singh (1980) reported continuous callus cultures and
regeneration of shoots from hypocotyl, cotyledons and callus obtained from
mature embryos of Pinus wallichiana.
The excised segments of hypocotyl, when cultured on modified MS
containing a very low concentration of salts supplemented with 0.1 mg/1
NAA, give rise to a continuously growing callus. Higher concentration of
NAA (l , 3 mg/1) inhibits callus growth. Ex plants grown on BM + different
concentrations of BAP (0.1, 0.5, 1, 3 mg/1) show a general increase in size

within 12 days. At 0.1 mg/1 callusing occurs within 24 days-the callus is
green and its growth slow. Better growth results with increase in BAP
(0.5 mg/1). At 1 mg/1, regeneration of shoot buds with interphase of callus
has been observed but the percentage of regeneration is low. At a higher
concentration of BAP (3 mg/1), callus growth is prolific.
In mature embryos excised and cultured on BM + 0.1 mg/1 NAA, the
cotyledons tum light green and elongate five times in 7 days. Callusing
occurs after 12 days and complete dedifferentiation of embryos after 3
months. This medium was supplemented with different concentrations of
ammonium nitrate (200 mg/1 to 1400 mg/1) to determine the optimal
concentration to support maximal callus growth. The lowest concentration
(200 mg/1) brings about faster growth. The increased concentration brings
about a slow decline in callus growth. Callus grown on BM + 0.1 mg/1
NAA, when subcultured on BM + 1 mg/1 BAP, shoot buds differentiated
after about 30 days (Fig. 22.13 A). Callus with shoot buds when transferred
to MS + 2% sucrose+ 10% CM, forms shoots (Fig. 22.13 B).
The embryos cultured on BM + 1 mg/1 BAP show a general increase in
size, and numerous shoot buds differentiate along the surface of the hypocotyl
(Fig. 22.13 C) after 24 days. On transfer toMS+ 2% sucrose+ 10% CM,
shoots develop. On BM + 1 mg/1 BAP + 2 mg/1 NAA, the cotyledons
ceased to elongate, and appeared "succulent". The cotyledons and hypocotyl
form callus when they are in contact with the medium. The green and
compact callus can be subcultured continuously on the same medium. Within
3 weeks, primary leaves and shoot buds become visible along the surface
of the cotyledon and hypocotyl. Excised shoot buds, cultured on MS +
2% sucrose+ 10% CM, differentiated into shoots, and the tissue in contact
with the nutrient medium dedifferentiated and formed callus (Fig. 22.13 D).
Bhatnagar et al. (1983) made preliminary investigations on organ
differentiation in tissue cultures of Cedrus deodara and Pinus roxburghii.
In C. deodara, callus could be induced from mature female gametophyte
and mature embryo. Adventitious shoot buds formed on the hypocotyl of
embryos cultured on MS + KN 2 mg/1. The embryos of P. roxburghii, cultured
on a medium with BAP 1 mg/1 and NAA 2 mg/1, differentiated primary
needles and shoot buds on the cotyledons.
In Welwitschia mirabilis the growing point of the stem is extremely
short-lived; further growth is from a meristem at the base of each leaf
which persists throughout the life of the plant (see Chap. 19). To obtain
differentiation of shoot apices, embryo, somatic tissue and single-cell cultures
give best results (Button et al. 1971)
The seeds of W. mirabilis contain inhibitors for germination (see Button
et al. 1971 ). However, the excised embryos from surface-sterilized seeds
grow very well on an agarized substrate containing MS +organic additives
(mg/1 glycine 5.0, inositol 50, calcium pantothenate 0.03, thiamine
432 The Gymnosperms
Fig. 22.13 A-D. Pinus wallichiana, subculture of callus from exctsed embryos.
A 40-day-old culture on BM + 1 mg/1 BAP shows shoot bud. B 4-month-old culture on
MS + 2% sucrose+ 10% CM transferred from A shows shoot formation . C 30-day-old
culture on BM + I mg/1 BAP, shows shoot bud (formation) along the surface of hypocotyl.
D 4-month-old culture on MS + 2% sucrose + 10% CM - transferred from BM
+ I mg/1 BAP + 2 mg/1 NAA,- shows shoot differentiation; tissue in contact with the
medium has dedifferentiated from callus. (After Konar and M.N. Singh 1980)
hydrochloride 0.3, niacin 1.0, pyridoxine hydrochloride 0.05) + CH 400 +

sucrose 4 x 104 + NAA 1.0 or 5.0 mg/l. There is prolific callus development
from the hypocotyl-root axis. It is friable and consists of abnormally large
(75-200 J..lm in diameter) and variously shaped cells. Several cells showed
conspicuous nuclei, nucleoli and well-developed vacuoles traversed by
numerous thick cytoplasmic strands which showed active cyclosis. These

cells are (potentially) totipotent.
Small segments of this callus are transferred to 500-ml culture flasks
containing 10 mlliquid medium (as above) but containing CM 10% (v/v)
instead of NAA. The flasks (attached to a culture wheel) are rotated at
1 rpm. The alternate flooding and aeration of the callus (in the flasks)
promoted disintegration of the callus and cell multiplication. The cell
suspension (mounted on microslides in buffered methylene blue) was
examined under the phase-contrast microscope. The cells were spherical,
cylindrical or like fungal hypha. The majority of the cells were live, did not
take the stain, and showed cyclosis. Other cells were irregular (in shape)
and took the stain. The cells proliferated by equational division, and by
budding from papilla-like outgrowths on the callus.
Conifer Biotechnology
The practice of silviculture and tree improvement is important in forestry.
Renewal of forests is obligatory to ensure future crops. The yield depends
on the quality and vigour of the trees. However, there may be qualities/
traits which are not easy to acquire or resolve. In such areas, biotechnology
has significant impact on tree improvement. The object of conifer
biotechnology is plant propagation and improvement (as in conventional
methods).
Somatic Embryogenesis
Embryogenesis that does not begin with the zygote is asexual or somatic.
It is the process by which somatic cells develop into differentiated plants
through characteristic zygotic stages.
There are two types of somatic embryogenesis: (a) Direct somatic
embryogenesis-the embryo develops directly from the explant tissue. (b)
Indirect somatic embryogenesis-the embryo develops from callus or cell
suspensions.
Cells which can develop into somatic embryos have embryogenic
competence. The selection of the correct developmental stage of an explant,
appropriate media and environmental conditions, and repeated transfers are
generally necessary for successful embryogenesis. Immature zygotic embryos
are the best explants. Embryos from mature stored seeds have also been
used (such seeds are available throughout the year). However, the recovery
of plantlets from somatic embryos is poor. Somatic embryos occur in a
repressed state of development in an induction media and require a
differentiation media for maturation.
Hakrnan et al. (1985) studied the development of somatic embryos initiated
from immature (at various developmental stages) embryos of Picea abies,
on basal medium gelled with 0.5% agar and supplemented with 2, 4-D
(10-5M) and BA (5 x 10-6M) for tissue cultures, and with activated charcoal
434 The Gymnosperms
(1% w/v) for embryo development. The immature embryos produced a

highly friable and embryogenic callus which could be maintained in
subcultures. Polarized and organized somatic embryos, with long highly
vacuolate cells (suspensor) at one end, and a group of meristematic cells
(embryonal) at the other end are formed. These structures are comparable
to the early stages of normal zygotic embryogeny.
In subculture, the embryos form a bipolar shoot-root axis with an
independent and closed vascular system. Frequently, either the shoot or the
root meristem fail to differentiate. Embryogenic tissue obtained on agarized
media could be transferred to liquid media and maintained in subcultures
for at least 6 months. Differentiation of somatic embryos has also been
observed in liquid cultures. This investigation shows that cultures with a
high potential for initiation of somatic embryos have been obtained from
immature embryos of P. abies. Similar results were also obtained from
immature embryos isolated from seeds; the cones had been stored at 4°C
for up to 6 months. This procedure significantly extends the period of
availability of proembryos and immature embryos.
Gupta and Durzan (1986a, b) obtained callus in Pinus lambertiana from
the radicle of mature zygotic embryos. However, immature embryos showed
a greater degree of competence. About 1-2% of the embryos developed
from the callus, and produced plantlets. Lelu et al. (1987) reported production
of somatic embryos from seedling cotyledons of Picea abies. NAA with
BA gave t.he best results. Becwar et al. (1987) developed a plating-counting
method for Picea abies. Nagmani et al. (1987) compared somatic
embryogenesis in Picea glauca and P. abies. Hakman et al. (1987) studied
it in Picea glauca using light and electron microscopy. Gupta and Durzan
(1987a) used the term polyembryogenesis to describe the somatic embryo
cultures of Pinus taeda. A four-nucleate free-nuclear stage occurs, followed
by segregation of the nuclei and delineation of a proembryonal cell. According
to these authors, the pine somatic embryogenesis system truly represents
zygotic polyembryony.
It is possible to cryopreserve somatic embryo-competent cells of Picea
alba and Pinus taeda in liquid N2 ( -196 oq and recover viable cultures
(Gupta et al. 1987). This is also true for Picea glauca (see Dunstan 1988).
For encapsulation the somatic embryos (Picea abies, Pinus taeda and
P lambertiana) are mixed with 1.5% sodium alginate and dropped singly
into a 50-100 rnM solution of Ca(N03) 2• A coating of calcium alginate
formed around the embryos and hardened to form capsules during a
20-30-min incubation. Encapsulated somatic embryos have been stored for
6 months at 4 °C without loss of viability. The plants from encapsulated
embryos develop on vermiculite, perlite, sand or peat, but at very low
frequencies (Gupta 1988). Somatic embryos of Piceaglauca can be maintained
for extended periods with repeated subcultures in liquid suspensions. With
the use of bioreactors, it would eventually be possible to produce large
quantities of elite genotypes on a regular schedule.
The advantages of somatic embryogenesis are (a) A large number of

plantlets can be produced. (b) Cell suspensions which are amenable to
bioreactor technology can cut down labour, time and cost. (c) Encapsulated
somatic embryos with controlled dormancy could form an efficient storage
and delivery system, as in natural seeds. (d) Ce.ll-suspension somatic
embryogenesis system is better suited for genetic engineering techniques
(Gupta 1988).
The development of a method of embryogenesis in vitro for conifers is
an important breakthrough and a rapidly growing culture system is available
(Dunstan 1988).
A culture system comparable to somatic embryogenesis is gametophytic
embryogenesis. It originates from haploid gametophytes, gynogenesis from
female gametophyte (see Nagmani and Bonga 1985, described on p 421)
and androgenesis from male gametophyte (microspore).
The culture of gametophytic tissue is of interest to tree breeders for use
as homozygous diploid (doubled haploid) plants to study inheritance.
The first report of conifer protoplast (described below) regeneration to
somatic embryos is in Pinus taeda and Picea glauca (Gupta and Durzan
1987b). Somatic embryo-competent cell cultures were used as the starting
material; 95% protoplast viability was obtained by using 4-day-old suspension
cultures of somatic embryos of Pinus teada.
The regeneration of somatic embryos from protoplasts of individual cells
of conifers is useful for propagation and provides a method for genetically
improving species by somatic hybrization.
Somatic Embryogenesis in Some Conifers
Species Explants Investigator/s
Abies alba Immature zygotic embryos Schuller et al. (1989)

Larix decidua Megagametophytes Nagmani and Bonga (1985),
von Aderkas et al. (1987),
von Aderkas and Bonga (1988)
L. eurolepis Immature zygotic embryos Klimaszewska (1989),
von Aderkas et al. (1990)
Picea abies Immature zygotic embryos Hakman et al. (1985), Hakman and
von Arnold (1985)
Mature zygotic embryos Jain et al. (1988) Verhagen and
Wann (1989), von Arnold and
Hakman (1986), von Arnold
(1987)
Mature zygotic embryos Krogstrup (1986)
and 3-7-day-old seedlings Lelu et al. (1987)
Megagametophytes Simola and Santanen (1990)
P. glauca Immature zygotic embryos Attree et al. (1989), Hakman and
Fowke (1987), Lu and Thorpe
(1987)
Contd
436 The Gymnosperms
Species Explants Investigator/s

P. glauca Mature zygotic embryos Tremblay (1990)
12-30-day-old seedlings Attree et al. (1990)
P. sitchensis Immature zygotic embryos Krogstrup et al. (1988)
Mature zygotic embryos von Arnold and Woodward
(1988)
Picea mariana Immature zygotic embryos Hakman and Fowke (1987),
Tautorus et al. (1990)
Mature zygotic embryos Tautorus et al. (1990)
12-21-day-old seedlings Attree et al. (1990)
Pinus caribaea Immature zygotic embryos Laine and A. David (1990)
P. elliottii Immature zygotic embryos Jain et al. (1989)
P. lambertiana Mature zygotic embryos Gupta and Durzan (1986a)
P. radiata Immature zygotic ell).bryos Chandler et al. (1989)
P. strobus Immature zygotic embryos Finer et al. (1989)
P. taeda Immature zygotic embryos Gupta and Durzan (1987a),
Becwar et al. (1990, 1991)
Pseudotsuga Immature zygotic embryos Durzan and Gupta (1987)
menziesii
Sequoia 7-day-old seedlings Bourgkard and Favre (1988)
sempervirens
Genetic Transformation
Genetic transformation is the incorporation of native or modified gene in
a recipient organism. The "foreign" genes can be inserted into the conifer
genome in vitro by two methods: (a) Direct gene transfer. (b) Agrobacterium-
mediated transfer. In the first method, the transfer is by microinjection (see
Mild et al. 1987). It requires a micromanipulator and microsyringe to deliver
small volumes of plasmid directly into the recipient nucleus. Microinjection
reduces the regenerative capacity of protoplasts but transformation frequency
is high .
.Agrobacterium is a naturally occurring bacterium which is capable of
transforming a wide variety of plants by introducing DNA from its tumour-
inducing plasmid. It has a wide host range and has been reported to be
infective in gymnosperms (De Cleene and De Ley 1976). Conifer cultures
that lend themselves to Agrobacterium transformation are somatic embryos
and protoplast-derived cells. Scanty work has been reported in conifers (see
Dunstan 1988).
Protoplast Culture
Inbreeding for tree improvement requires a long time to produce successive
generations; many trees can not be easily inbred due to problems of self-
incompatibility and in-breeding depression. Isolation and fusion of protoplasts
have stimulated considerable interest, specially in the development of new
conifer hybrids. The protoplast is plasmalemma-bound without a wall. It is
useful for propagation, for plant breeding by somatic hybridizatiop or genetic
transformation, and is also of use in resolving questions of basic plant

biology. The protoplast is obtained by removing the cell wall constituents
enzymatically in an osmotic solution, which slightly plasmolyzes the cell.
However, improved techniques need to be developed for isolating protoplasts
in high frequencies; for separating viable protoplasts from intact cells, cellular
debris, and the cell wall-degrading enzymes used to isolate the protoplasts;
for fusing the protolplasts; to distinguish between hybrids and non-hybrids
among the fusion products, and for regenerating whole plants from the
protoplasts.
The most frequently studied coniferous material is plantlet cotyledon-
derived protoplasts: Pseudotsuga menziesii (Kirby and Cheng 1979), Pinus
pinaster (A. David and H. David 1979), P. coulteri (Patel et al. 1984). Other
sources of protoplasts are (haploid) pollen (Duhoux 1980), root (Faye and
A. David 1983), and primary needles of plantlet buds of Pinus pinaster
(H. David et al. 1986).
Micropropagation
Micropropagation is the foremost area of plant biotechnology in terms of
commercial application. Micropropagation through tissue cultures is a means
of producing a new plant and a large number of plants can be obtained
from a very small amount of tissue from an elite species. The multiplication
potential of this technique is immense. It offers a new dimension and
unparalled opportunity for forest tree improvement. (see Kamosky 1981).
The utilization of tissue culture in the propagation. and genetic improvement
of conifers promises several benefits, such as faster development and
multiplication of selected and improved genotypes, and the transfer of genes
between non-fertile parents. However, improved techiques have still to be
developed. Also, genetic differentiation of cells in culture creates uncertainty
as to the fidelity of reproduction by micropropagation (Berlyn et al. 1986).
Horgan and Holland (1989) developed a reliable method of rooting
micropropagated shoots from mature Pinus radiata. There was difficulty in
achieving consistent results in the beginning, but finally a high percentage
(78%) of rooting was achieved. Shoots that had been cold-stored prior to
rooting did not survive. Maximal rooting occurs in a free-draining peat-
pumice-perlite medium in propagation trays maintained under controlled
environmental conditions. A well-managed watering regime and gradual
conditioning of the shoots from the day of setting enhanced survival and
rooting. Removal of callus from the base of the shoots prior to an auxin
treatment significantly improved survival (30-70%) and rooting (16-56%).
It was further improved by a 5-week prerooting treatment in nutrient medium
containing 6% sucrose. The shoots formed vigorous plantlets, which could
be successfully acclimatized to full sunlight within 4-6 weeks of transfer
to root trainers. Plantlet survival ranged from 90 to 100%.
The number of roots and their percentage in tissue-cultured Pseudotsuga
438 The Gymnosperms
menzieii is influenced by the rooting substrate, sucrose concentration and

boron in the rooting medium, shoot height and shoot generation. Peat-
perlite is a better substrate (produces 70% rooted shoots) than agar
(0% rooted shoots). In the former, organized cell divisions occur and are
associated with tracheidal nests, while in agar the proliferation is unorganized
and not restricted to the tracheidal nests. Optimal sucrose concentration is
4% to produce nodular or rooted shoots. With the addition of 3 mg/1 boric
acid, 100% of the shoots rooted and the mean root number was 11. The
number and percentage of roots is higher with shoots 3 em long; shoot
responses are more (80%) in third and fourth generation than in second
generation (36%) shoots (Mohammad et al. 1989).
A number of factors have been identified which affect the rooting and
acclimatization of conifers. The quality of shoot, age of the donor, clone,
temperature and substrate influence the production of root. Biochemically,
root initiation is a high-energy process which requires a continuous supply
of free sugar from the medium. One of the most important factors is auxin,
its type, concentration, mode of applications and duration of exposure. When
used alone, it is usually one of the synthetic auxins NAA (0.05-0.5 mg/1)
or IBA (1-10 mg/1). Combinations of NAA, IBA or IAA (10-20 mg/1)
have been more effective with Pinus radiata, P. rigida, Picea abies, and
Pseudotsuga menziesii (see Mohammad and Vidaver 1988). The type of
auxin used affects the formation of callus at the shoot base, and the number
of roots per rooted shoot.
During acclimatization, it is important to minimize water stress; new
growth is necessary for the plantlets to become fully autotrophic. The quality
of the root system is critical, a well-.branched fibrous system is essential for
good performance in the field. It is influenced by the rooting substrate, root
pruning and the container. Further work is needed; the immediate areas are:
role of sucrose in rooting and subsequent plantlet survival, the potential
role of ectomycorrhizal associations prior to or during transfer stages, and
the cause(s) of early maturation following transfer to the greenhouse
(Mohammad and Vidaver 1988).
Some gymnospermous forest tree species from which entire plants have
been regenerated through tissue cultures (based on Karnosky 1981).
Species Explant InvestigatorIs
Araucaria Shoot tip Haines and de Fossard (1977)

cunninghamii
Cryptomeria Hypocotyl, stem Isikawa (1974)
japonica
Picea abies Megagametophyte Bonga (1977)
P. glauca Hypocotyl Campbell and Durzan (1976)
P. sitchensis Embryo, needles, Webb and Street (1977)
shoot tips
Contd
Species Explant Investigator/s

Pinus elliottii Cotyledon Sommer and Brown (1974)
P. palustris Cotyledon, embryo Sommer et al. (1975)
P. pinaster Cotyledon, hypoco- H. David et al. (1978)
cotyl
P. radiata Cotyledon, hypo- Reilly and Washer (1977),
cotyl Smith et al. (1982)
P. rigida
P. sabiniana ]-Cotyledon Brown and Sommer (1977)
P. virginiana
P. taeda Cotyledon Sommer and Brown (1974)
P. strobus Embryo Minocha (1980)
Pseudotsuga Cotyledon Cheng and Voqui (1977)
menziesii
Sequoia Juvenile shoot Ball (1978)
sempervirens
Thuja plicata Cotyledon, Coleman and Thorpe (1977)
shoot tip
Tsuga heterophylla Cotyledon Cheng (1976)
To meet the ever-increasing demand on the forests, therefore, traditional

improvement and silvicultural practices along with developing biotechnologies
are required.
23. Economic Importance
The gymnosperms are predominantly woody plants. They are frequently

used in landscaping of parks and gardens because of their evergreen habit
and symmetrical appearance.
General Aspects
Various Cycas species are cultivated in the garden as palms.
Ginkgo biloba is grown in groups or as avenue trees. The male trees are
preferred, as the ripened ovules (on female plants) have a disagreeable
odour, like that of rancid butter. However, the male and female plants
cannot be distinguished in the vegetative state. The plant is exceptionally
resistant to attacks of insects and fungi and can be grown successfully in
modern cities.
The conifers are much valued, and several taxa are grown wherever the
climate permits; Thuja, Biota, Araucaria, Juniperus and Pinus are quite
popular. Cupressus funebris is commonly planted in China and India in
cemeteries. Cryptomeria japonica lends beauty to the groves and gardens
of Japan. It forms the famous avenue which leads to the temples and tomb
of Icyasu (founder of Tokugawa dynasty) at Nikko. The avenue, laid out
in the 17th century, has been maintained by replacing dead or damaged
trees. Much attention was paid to forest administration by the Tokugawa
Government. Tree felling was restricted. Certain trees, such as Chamaecyparis
obtusa, C. pisifera, Sciadopitys verticillata, Thuja standishii and Thujopsis
dolabrata, the celebrated "five trees of_Kisco" have been strictly preserved.
Agathis alba is grown as an ornamental tree in Java. Juniperus horizontalis-
the creeping juniper-spreads over the ground, and is used for covering
exposed banks. Several horticultural shrub varieties of J. virginiana have
been developed for ornamental plantings (Hemmerley 1970). They vary
widely in foliage colour which may be "white", gold-tipped or blue-green,
and turn purple in winter. The shape of the plant may be pyramidal, spreading
or globose. They need abundant sun, but can grow in sandy or dry soil
where other shrubs will not grow. Chamaecyparis obtusa var. compacta is
a dense-growing dwarf with very short branches and branchlets. C. obtusa
var. nana is a spreading green-leaved bush. Both the varieties are suitable
Economic Importance 441
for rock gardens. Cephalotaxus is a spreading bush which makes excellent

screens in the garden; it withstands pruning well and is used as a hedge
plant. Taxus spp. are widely planted in gardens. It has several erect branches
from the base, stands clipping well, and is one of the best hedge plants,
used extensively in topiary work.
Various species of Abies, Picea, Pinus, Pseudotsuga and Juniperus are
popular as Christmas trees. In Europe Abies alba, and in the USA A. balsamea
are preferred (they hold the needles longer than other species). Before
World War II, Juniperus virginiana was the traditional Christmas tree
(Hemmerley 1970). Bonsai is an ancient Japanse art form (planting of trees
in shallow trays or pots). The trees or shrubs are grown indefinitely in
proportionately small containers and treated with certain miniaturization/
dwarfing techniques to impart in them an appearance of age. Cycas revoluta,
various species of Pinus and Juniperus are very popular for growing bonsai.
In Japan, Chamaecyparis obtusa, Pinus densiflora, P. parviflora, P. thunbergii
and Podocarpus nagi are largely used for dwarfing. However, almost any
conifer will make a dwarf plant.
Fresh or dried leaves of various cycads are utilized as decorative material.
The foliage of Cycas revoluta is used for making wreaths, floral decorations
and/or artificial palm trees for window display. The leaves are boiled in
water, soaked in a preservative solution for several days (Thieret 1958), or
plunged in boiling seawater. The leaves become leathery and do not break
on drying.
The gymnosperms are economically important fm: their timber, in the
manufacture of paper and board, resins, tannins, essential and fatty oils, as
food supplements and pharmaceuticals.
Woods
Most of the commercially important gymnospermous woods are obtained
from conifers and taxads. Anatmoically, these woods comprise tracheids,
xylem parenchyma and xylem rays. They lack xylem fibres for the most
part, and the high cellulose content imparts a softer texture than the wood
of angiosperms (seeP. Maheshwari and H. Singh 1960). In Larix decidua,
Agathis and Taxus, the wood is very hard. In conifers, the heartwood and
sapwood are not always well defined. The distinction between spring and
autumn wood is usually better marked in trees from temperate regions than
from warmer regions. A few coniferous woods have a definite taste, for
example bitter (Dacrydium colensoi) or astringent (Agathis australis), while
others are greasy tq the touch (D. colensoi, Taxodium distichum).
Most coniferous woods are straight-grained, light-coloured, light-weight
and strong in comparison to their weight. They can be easily worked, have
good nail-taking properties, take a fine finish with sharp tools, and polish
and paint well. These qualities make them suitable for a wide range of
work where strength and durability are not essential: furniture, cabinet
442 The Gymnosperms
making, flooring, interior decoration, for general carpentery, building purposes,

joinery, for boats and shipbuilding, poles, posts, railway sleepers, cooperage,
veneers, etc. Some of the taxa have scented wood or are beautifully figured
and are, therefore, specially valued for making chests, cabinets, and for
panelling. All species in a genus usually have the same odour, except
Chamaecyparis nootkatensis which is different from other species.
Araucariaceae. The wood of Agathis is strong, durable, pale yellow to

yellow-brown with a silky, even texture and lustrous surface. A. australis
is the principal timber tree of New Zealand. The wood has several uses,
including making piano parts, artificial limbs, and reproduction of antique
furniture, especially in France (Whitmore 1980 a, b). Occasionally, the
wood of Araucaria araucana has dark brown fixed knots which contrast
strikingly with the rest of the wood. _Logs of A. angustifolia and
A. cunninghamia are used for plywood manufacture.
Cupressaceae. The wood of Callitris is fragrant and shows distinct

heartwood and sapwood. It is sometimes beautifully figured. Due to the
presence of phenol and other chemical compounds, the wood is resistant to
white ants. It is valuable as timber, and mostly the wood of C. glauca is
used. In Chamaecyparis, the wood is durable, finishes with a glossy surface,
and has a pleasant lasting spicy odour which repels moths and insects.
C. lawsoniana is one of the most valuable timbers of North America. The
fragrant essential oil present in the wood is a powerful diuretic, which
makes it necessary for workers in the saw plills to occasionally change to
other woods (Dallimore and Jackson 1966). The wood is also used for
aeroplanes, canoe paddle, and organ pipe making. C. nootkatensis is
considered to be one of the best woods for battery separators. C. obtusa has
white-, straw-coloured or pink wood and is popular in Japan. It is used as
a base for superior lacquer ware. Due to its straight and even-grained
nature, the wood can be cut up into very thin shavings and plated into hats.
The bark is used for roofing, as it is decay-resistant. C. pisifera is used for
bentwood articles. Cupressus wood is very durable, and has a spicy-resinous
odour which is insect-repellent. C. sempervirens (gopherwood) wood is
reputed to be one of the four woods used in the construction of the cross
upon which Christ was crucified (Baumann 1960). The wood lasts indefinitely
under water. It has been a favourite wood for coffins ever since the Egyptians
introduced it. It is considered an excellent furniture wood in France and
Italy. The doors of St. Peter's in Rome were made from this wood, and
lasted for nearly 1000 years (P. Maheshwari and H. Singh 1960). The
timber of C. torulosa is often used in India in temples. When burnt as
incense, it forms very little ash. The wood is suitable for second-grade
pencils, battery separators, and in aircraft manufacture.
Juniperus wood is fragrant, reddish or reddish brown, and rarely damaged
by insects. It is very durable and especially resistant to weather conditions.

J. virginiana has a cream-coloured sapwood with projections into the dark
red heartwood which gives it a distinct appearance. It is durable even when
in contact with the soil, and is resistant to microbial decay. It was the
standard wood for pencils until it became scarce. J. macropoda wood is
suitable for pencils in India; it is also burnt as incense. J. recurva and
J. wallichiana wood are also burnt in Buddhist temples. The wood of
Libocedrus is reddish brown, -resinous, fragrant, durable and easily worked.
The heartwood of L. decurrens contains a volatile oil and small quantities
of carvacrol and hydrothymoquinone-the last two highly fungicidal
(Anderson 1967). ·
All the high-grade woods are used in the .manufacture of lead pencils,
and venetian blinds. The wood of Thuja is fragrant, non-resinous, very durable,
and yellowish or reddish brown. The heartwood of J. plicata is very durable,
and contains tropolones (thujaplicins), which are extremely toxic to wood-
destroying fungi (Anderson 1955). The wood is used for glasshouse
construction, beehives, and outhouses. In the USA, native Americans used
it for canoes by hollowing out the wood, and splitting the trunks for their
totem poles.
Pinaceae. The colour of wood in Abies varies from white-yellowish to

reddish brown. It is light and used for packing cases, matchwood, wood
wool, aircraft work, light camp furniture, and plywood. The odourless wood
is much in demand for butter, lard and grocery boxes. Cedrus wood is very
durable, oily, fragrant, insect-repellant, and rot-resistant. The wood of
C. libani was extensively used for coffins and mummy cases by the ancient
Egyptians (seeP. Maheshwari and Biswas 1970). In India, C. deodara is
a general utility wood and is the strongest of the Indian coniferous woods.
Due to the presence of oil, the seasoned heartwood is very durable and
rarely attacked by white ants or fungi. It is, however, difficult to glue or
polish, and unsuitable for painting and varnishing, as resin oozes out from
the knots even after seasoning.
The wood of Larix is one of the heaviest, strongest and toughest of the
softwoods. It is durable even in contact with the ground. The wood is
extensively used for garden furniture, and outdoor feeding racks and troughs
for cattle. The plant has a naturally curving lower part which is ideal for
boat- and shipbuilding. Some of the important species are Larix decidua,
L. laricina, L. leptolepis and L occidentalis.
Picea wood is soft, odourless, long-fibered, white or pinkish, black or
dark-brown, works well, and finishes with a satiny surface. It is resonant
and is prized for making sounding boards of pianos and bodies of violin
and organ pipes. The timber of P. abies has a natural lustre, and is extensively
used for toys, carving, wood chips for hats, and fruit baskets. The wood
from Romania is excellent for making sounding boards for musical
444 The Gymnosperms
instruments. It is popular as dairy and kitchen table-top and dressers as the

wood can be washed and scrubbed to give a clean appearance. P. sitchensis
wood is the most valuable of all spruce woods because of its combined
qualities of strength and lightness. It is used for plywood for special laminates
in aeroplane and glider construction.
Pinus timber is commercially important and valuable. The soft pines
have a straight-grained, uniform, easy-to-work wood, comparatively free of
resin. The hard pines have resinous, heavy, hard, strong and durable wood;
heartwood and sapwood are well defined. The group is heterogenous, each
species has distinctive wood. It is highly inflammable and can be used as
a torch. P. lambertiana yields large quantities of unblemished, high-quality
timber. The wood of P. palustris has a world-wide reputation. It is very
hard, strong, durable, resinous, and occasionally has decorative markings.
It is esteemed for heavy construction work such as bridge and naval
architecture. P. roxburghii timber is moderately hard, resinous, light and
non-durable. It is extensively used for packing cases, cheap furniture and
charcoal. P. sylvestris wood is superior, and has excellent lasting qualities.
The treated best-grade wood, when laid on a good concrete foundation,
withstands very heavy wear, as shown by blocks which remained in use for
over 13 years on Westminster Bridge (London) where a continuous and
heavy traffic flows (Dallimore and Jackson 1966). Some other notable
timber-yielding species are: P. ponderosa, P. resinosa, P. taeda, P. strobus
and P. wallichiana.
Pseudotsuga wood is variable in character. The heartwood is reddish- or
brownish yellow and sapwood creamy yellow with a pronounced difference
in spring and summer wood. Resin ducts are usually present in groups. The
tracheids have spiral thickenings, otherwise superficially the wood resembles
Larix. The tall tree gives a clear timber which can be used for heavy
construction work. The wood is especially suited for bending. In the USA,
most veneer and plywood is made from this plant (P. Maheshwari and
H. Singh 1960). The brownish wood of Tsuga normally lacks resin ducts;
the heartwood is darker than sapwood. The wood is tenacious and holds
nails well, hence its value as a box wood. The important species are
T. heterophylla and T. canadensis.
Podocarpaceae. Podocarpus is commonly called yellow wood because

of its colour; occasionally it may be brown or reddish. The wood is not
economical for cheaper manufactures as it takes nails only when it is bored
and is more expensive to work. Several species are important timber trees
in their native countries. P. ferrugineus and P. totara wood resists teredo
(shipworm moluscs) and is used for marine work, such as piles for bridges
and dock. The yellow or reddish wood of Dacrydium is resinous, durable,
and occasionally beautifully figured. D. biforme wood is durable even on
contact with the ground. D. colensoi wood is of low specific gravity, very
strong, elastic and does not shrink much on seasoning. D. intermedium wood
is strong, durable, resinous, and highly inflammable.
Taxodiaceae. The wood of Cryptomeria japonica is fragrant, strong,

durable, coarse and variably grained, and is not readily attacked by insects.
It is a widely used timber in Japan. The heartwood of Sequoia sempervirens
is a rich dull red, and sapwood yellowish white. It is soft, fine-grained,
easy to work, and durable when exposed to weather or soil. It does not
warp or shrink readily. The wood has many uses, since it is available in
large dimensions free from defects. The plywood can also be used in exposed
situations. Sequoiadendron gigantea wood has a limited use. Both the trees
are preserved for their scarcity. Taxodium distichum wood is soft, straight-
grained, non-shrinking, and has a characteristic sour odour. The heartwood
varies from red to almost black, and the sapwood is white or yellowish
white. It is very durable in wet places, resistant to insects, and inimune to
white ants. It is used for construction of water tanks, water pipes, and is the
best all round timber for chemical tanks and vats, since it does not impart
any colour or taste to the products. It is particularly suitable for greenhouse
construction, cooling towers and roof planks of dye houses.
Taxaceae. The wood of Taxus is strong, oily, elastic, close-grained arid

very durable (tertiary spiral thickenings are present in the tracheids). It is
the heaviest of the softwoods, with good mechanical properties; it works
well, and finishes with a smooth glossy surface. The heartwood of T. baccata
is reddish brown, toning with age and exposure, the sapwood is pale yellow-
white. It has irregular growth rings, which give it a decorative value.
Transections of the base of old trees may show several distinct "hearts",
which are due to erect shoots growing from near the base, thickening and
being overgrown by the main trunk. Such sections are prized as tabletops.
In ancient times, the wood was popular for bows and it is still used in
archery. The wood of Torreya nucifera is lustrous, and durable under water.
It is used for water pails and Japanese chess men.
Ginkgoaceae. Ginkgo biloba wood is light and brittle. In China and Japan
it is used for chess boards and chess men.
Paper and Board

Paper can be made from any natural fibrous material depending on the
amount, nature, softness and pliability of the cellulose present in the cell
wall. At present, nearly 95% of the paper produced in the world comes
from wood, and in the USA and Canada (the largest paper producers), 85%
of the wood pulp is made from coniferous wood. Conifers are important
because of the greater average length of the fibres and larger percentage of
long fibres per given volume of wood.
446 The Gymnosperms
The wood is converted into pulp or fibrous mass by various processes:

(a) Ground wood or mechanical process, (b) Chemical processes.
In the mechanical process, the pulp is produced by pressing the logs
against revolving pulpstone in the presence of water. The resins, lignin and
other undesirable materials are not removed, and they resist bleaching agents,
which causes the paper to turn yellow. The fibres do not felt readily and
the pulp is poor in quality. Usuillly, the mechanical pulp is blended with
a varying percentage of chemical pulp to give the paper the necessary
strength. Ground-wood pulp is mostly used for newsprint (blended with
20% sulphite pulp).
In chemical processes, the wood chips are subjected to chemical treatment/
cooking which removes most of the lignin and other materials, and nearly
pure cellulose fibres remain. For pulping softwoods, the chemical processes
such as sulphite and sulphate or kraft are used. The sulphite process has an
acidic reaction. Bleached sulphite pulp is used in the manufacture of higher
grades of paper. The sulphate or kraft process operates under alkaline
conditions. This method is adopted for coniferous wood, especially pines,
which have a high resin content. The digesting solution dissolves resins,
wax and fat from the wood. The pulp is mostly used in the manufacture of
strong grades of paper such as kraft paper, used for wrapping, paper bags·
and paperboard. The important conifers used extensively for the manufacture
of paper and board are: Pinus sp. for kraft paper; Picea, Abies and Tsugd
for higher grades of writing and printing paper; Pinus, Picea and Pseudotsuga
for thermal and sound insulation boards. From Picea and Tsuga a number
of derived products are also obtained, such as transparent films, photographic
films, artificial sponges, sausage casings and lacquers. The bast fibre of
Sequoia sempervirens is formed into sheets for battery separators (Isenberg
1956). Other conifers used in the industry are: Thuja plicata, Agathis
palmerstoni and A. microstachya; Araucaria angustifoila, A. bidwilli and
A. cunninghami; Larix laricina, L. occidentalis and L. decidua.
Resins
Resins are complex plant exudates which vary in their chemical composition,
and are related to the terpenes or essential oils. They are insoluble in water
but are soluble in vegetable oils and organic solvents such as alcohol, ether
and carbon bisulphide. When heated, resins melt with progressive distillation
of volatile oils as the temperature increases. The conifers are one of the
major resin-yielders in the world. Resins harden gradually as their oil
evaporates, which makes them invaluable in industries like paints and varnish,
lacquers, paper-sizing and medicine. They are dissolved in solvents and
painted on the given surface; after the oil and solvent evaporate a thin
waterproof coating remains on the surface. Resinous substances have long
been used as waterproof coatings. The Egyptians varnished their mummy
cases with them. In most conifers, the resin remains mixed with either
abundant essential oil (oleo-resin) or very little of it (hard resins). In others,

the resin may be mixed with gum (gum resin) as in Araucaria and it is not
possible to separate them.
Hard Resins
The hard resins are usually solid, more or less transparent, and brittle. They
are an excellent source of varnish as they have a low oil content, and
dissolve readily in alcohol. Some of the uses are: (a) Printing inks. Resins
are ingredients of speciality inks which include gloss, non-rub, non-scratch
food carton, candy wrapper, cellophane, soap wrapper, gravure inks, etc.
These inks are applied to surfaces other than paper. (b) Adhesives,
(c) Pyrotechnics (fireworks industry), and (d) Linoleum.
Copal. The term copal is of Mexican origin, and is applied to a large

group of resins obtained from a variety of plants. It is of varying hardness,
and has relatively high melting point. The softer copals are almost completely
soluble in cold alcohol, and used as such for spirit varnishes. Harder copals
need heat treatment or "running" to render them soluble in drying oils.
There are three principal copals, two are from conifers.
a) Kauri Copal. This is obtained from Agathis australis, the most important
tree of New Zealand. The copal is chiefly fossil in nature and is collected
from sites of present or former kauri forests. It is dug up from ridges and
swamps which furnish the bulk of the supply, and the pieces may weigh as
much as 45 kg. The range gum is the best grade of kauri. An inferior bush
gum is obtained by tappping living trees. A small amount of resin exudes
naturally and forms green gum. The resin is extremely valuable for varnish
and especially suited for marine and outdoor work. It has a high depth of
gloss combined with elasticity and durability. For a long time, this resin
held a premier position in varnish trade. An inferior grade of kauri copal
is also used for making linoleum.
b) Manila Copal. This is obtained from Agathis alba distributed throughout

Malaysia. The first shipments of this copal were from Manila, and the
name has persisted. In Malaya, the trees occur only at higher elevations.
The tapped resin does not harden there for some unknown reason (Mantell
1950), and is known as syrup copal. The resin exudes naturally; during
high winds and storms, cracks and fissures are formed in the tree. The resin
flows out from them and accumulates as large lumps up to 18 kg in weight,
in the forks of trees from where it is collected. Resin is also obtained by
systematic tapping. The resin is present in the bark; pieces of it are removed
and the exposed surface is cleaned. After every collection, bark is removed
from upper edge of the wound for a new opening. Resin also occurs in a
fossilized state and is collected from the earth. It consists of large, irregular
448 The Gymnosperms
milky pieces with a yellowish interior. In Borneo, a semi-fossil grade

known as Pontianac is obtained and is the hardest variety of copal. The
resin is used in oil and spirit varnish and paints, lacquers and in making
linoleum. It is used as a sizing material and has numerous other applications
such as preparation of plastics, driers, adhesives, oil cloth, printing inks,
and waterproofing compositions.
Amber. This is a fossil, water-insoluble tree resin which has attained a

stable state after various changes such as loss of volatile constituents, oxidation
and polymerization processes and a prolonged burial period. Amber is a
general word for resins which are heterogeneous and differ in chemical and
physical properties. It occurs throughout the world in widely separated
localities such as Burma, Japan, Alaska, North, Central and South America,
and many European countries as well as near-Eastern countries. When one
speaks of amber, it is primarily the fossil resin which is found chiefly on
the shores of the Baltic sea, and it is the most important commercial amber.
Chemically, it is a resin from a number of extinct conifers of which Pinus
succinifera is the principal source. These plants flourished during the Eocine
on the shores of a former sea.
Amber is exceedingly hard and brittle, the color varies from yellow to
brown or even black. It occurs in several forms, the most important is
succinite. When rubbed, it takes a high polish, becomes negatively electrified,
and emits a characteristic aromatic odour.
Amber was highly prized by the ancient peoples; the Greeks and Romans
for jewellery, beads and other ornamental purposes. Certain magical properties
were attributed to it. At present, it is used for the mouthpiece of pipes, and
holders for cigars and cigarettes. It is also used in medicine and X-ray
therapy. Human blood does not coagulate if kept in amber containers. The
darker grades yield a valuable varnish, but it is too expensive to be of much
use. Scientifically, amber is of interest because remains of insect or plant
material are occasionally embedded in it. These existed when the tree was
alive and the resin was fluid. This helps in the study of evolution of life
on this earth.
Sandarac. Sandarac is a pale yellow resin obtained from Tetraclinis

articulata (Vahl) Mast. (African Sandarc), or various species of Callitris
(Australian Sandarc). The resin is formed between inner and outer layers
of the bark and exudes as small "tears". The hard white, rather brittle, spirit
varnish is particularly adapted to the finishing of metal objects, to which
it gives a good lustre. One of its largest applications is as a primer for metal
surfaces, where it provides a high degree of adhesion. It is used as paper
and leather varnish, for glass and procelain cements, spirit varnishes and
lacquers for photographic work, and as a constituent of incense. Many of
its uses are in combination with mastic (resinous exudation of trees of
Pistacia, Anacardiaceae) and elemi (oleo-resins of different origin belong

to members of the Burseraceae). Mastic sandarc has been used for the
preservation of old paintings (Mantell 1950). An alcoholic solution of sandarc
on cotton wool is used as a temporary filling for teeth.
Oleoresins
The oleoresins have a substantial amount of essential oil which m~es it
almost liquid in nature, and they have a characteristic aroma. The two
components can be separated by distillation. Turpentine is an oleoresin
obtained exclusively from conifers.
Turpentine. The pine oleoresin is called pine gum, pine pitch or turpentine.
It is a viscous, honey-like liquid which is obtained, for commercial use, by
tapping living trees. On distillation the turpentine yields the essential oil
(spirits of turpentine) and rosin (also called colophony). Bqth of them are
immensely useful, and important industries have grown around them. The
turpentine or Naval Stores industry is one of the oldest forest industries. In
- _I
the 17th century, the wooden sailing vessels used large quantities of oleoresin
and gave the name·naval stores to the industry. The United States leads in
production, followed by France, whose products are of the highest quality.
Spain is third, followed by the European countries, India and East Asia.
In the USA, the major turpentine pines are Pinus palustris and P. caribaea;
some of the other species of importance are P. taeda, P. ponderosa,
P. lambertiana and P. contorta. In Europe, P. pinaster, P. halepensis,
P. nigra, P. pinea and P. sylvestries are the common yielders. In India,
P. rOitburghii, in the East Indies, P. merkusii, and in the Phillipines, P. khasya
are turpentined.
The trees are 'tapped for oleoresin. The method of tapping varies in
different countries although the basic principal is the same. In the USA, in
early days, tapping methods were very destructive and injurious to the tree.
Cavities known as "boxes" were cut near the base of the tree and the
oleoresin collected in them. Improved methods of tapping, called the Henry
system, which involves shallow chipping and the use of several types of
cup-and-gutter system followed. In a tree for turpentining (exceeding
23 em in diameter), basal incisions are made in the trunk and metal gutters
are slipped into it. These guide the oleoresin into a metal cup suspended
below the gutters. A strip. of bark and wood (approximately 1.75 em deep)
is removed just above the gutter for a distance of one-third of the
circumference of the tree. This stimulates the flow of resin and induces the
formation of J;Iew ducts. At present, the recommended practice is bark
chipping and acid treatment. The surface is prepared by removing a strip
of bark only by using especially designed scrapers, and no wood is removed.
The fresh streak is then sprayed with 50% H2S04 • The acid stimulates the
flow of resin by not allowing the wound to heal quickly (Andersort 1955).
4SO The Gymnosperms
A similar effect :5 shown by inoculation of Fusarium lateritum f. pini (Clapper

1954). Such streaks produce 50-100% more oleoresin during the first week,
and continue more than normal yield during the second and third week.
The interval between chipping can be increased from the usual 1 week to
2~3 weeks. This technique leaves the tree base in better condition for
subsequent utilization as plywood or lumber.
Much turpentine and rosin are also obtained from old pine stumps and
logging waste, by steam and solvent processes.
The. oleoresins collected in the cup are emptied into larger vessels and
transported to the factory, where they are washed and cleaned before
distillation. The distillate is collected in barrels, where the oil of turpentine
rises and is run off for storage. The residue (rosin), while still hot, is run
through a series of screens to remove impurities and is transferred to a
cooling vat. It is then put into slack barrels where it completely hardens
within 24 h.
The oil of turpentine has many uses. Due to its properties as a solvent,
it acts as a thinner in the paint and varnish industry. It is useful for printing
cloth, especially cotton and wool, in perfumery, pharmaceutical and allied
industries, and as a solvent for rubber and gutta-percha.
Rosin is even more important in industry. Depending on grade and quality,
it has several uses, such as in the manufacture of soap, varnishes, paint
driers, oil cloth, linoleum, sealing wax, adhesives, plastics, etc. Superior
grades are used for paper sizing, enamels and ointments. Palustric acid has
been discovered from rosin which is useful in paper sizing.
Canada Balsam. This oleoresin is obtained from Abies balsamea

(balsam fir), the most widely distributed conifer in North America and
Canada. The oleoresin is located in elongated, schizogenous ducts or blisters
on the bark. It is collected by using a pot with a spout, cut at an angle,
which is forced into the blisters, and the oleoresin drains out. Most of the
collection is made in the province of Quebec in Canada. Each tree yields
approx. 230-285 g in a year. Canada balsam is a viscid, yellowish or
greenish substance which does not granulate or crystalize on drying. It is
transparent, and has a high refractive index, approximating that of glass.
This property makes it extremely suitable as a mounting medium for
microscopic objects, and as cement for lenses in optical work. It is also
used as a fixative for soap and perfumes and as a component of collodion
and several plasters. Pseudotsuga taxifolia and Tsuga canadensis also have
oleoresin with similar properties and uses.
Venice Turpentine. This is obtained from Larix decidua. The resin ducts
are located in the heartwood of the tree. To collect the resin, a hole is bored
into the trunk; one single hole may suffice for the whole life of a tree. The
trees are tapped in spring. The oleoresin is a yellowish/greenish liquid and
has a characteristic odour and taste. It is used in varnishes, lithographic

work, veterinary medicine and histology.
Tannins
Tannins are organic compounds, glucosidal, have an acid reaction, and are
astringent. They are useful because of their ab~lity to unite with the proteins
of animal skin to form a strong, flexible, resistant insoluble substance
known as leather. Tannins also react with salts of iron to form dark blue
or greenish black compounds which form the basis of common inks. They
are useful in medicine because of their astringent nature. Tannins are used
in the petroleum industry as a dispersant to control the viscosity of mud in
oil-well drilling. The bark of Tsuga canadensis has 8-14% of tannin and
has been in use in the USA as the chief domestic source. In Europe, Larix
decidua, Picea abies, and in New Zealand Phyllocladus trichomanoides are
used. The latter is utilized for glove leather, as it contains a bright orange-
yellow dye in addition to tannin, which gives the kid gloves their particular
shade.
Conifers do not form very important tannin yielders but small industries
develop wherever large quantities of the bark are available.
Essential Oils
Almost all conifers with resin ducts can yield essential oils, but they are
not always commercially important. Steam distillation of the young branches
and adherent leaves,. wood and sawdust yields the oil, which is used
extensively in preparations of deodorants, room sprays, disinfectants,
preparations of bath salts, perfumery and medicine. Some other uses of the
oil are: Himalayan cedarwood oil (Cedrus deodara) and red 'cedarwood oil
(Juniperus virginiana) are used for clearing tissues in histological work,
and for use with the oil immersion lens of the microscope. Oil of juniper
(J. communis) is used as an essence for flavouring several European liquors,
such as gin, which owe their characteristic aroma to it. Oil of cade
(J. oxycedrus) is widely used in the treatment of chronic eczema, other
skin diseases, preparation of medical soaps, and healing of cuts and cutaneous
diseases of animals. Oil of .savin (J. sabina) was once used as an
antirheumatic, and vermifuge but has lost much of its importance due to its
disagreeable odour, irritating effect and toxicity. Huonpine wood (Dacrydium
franklinii) oil is highly germicidal, and is used as a preservative of casein
and other nitrogenous products; it exterminates powder pests and furniture
borers, and is an effective insect-repellant; also used in scenting transparent
soaps.
Fatty Oils
The seeds of several gymnosperms have fatty oils, mostly not available for
oil-milling, as the seeds are eaten either raw or roasted. The fleshy layers
452 The Gymnosperms
of seeds of Macrozamia contain a bright orange oil, which resembles palm

oil in its physical ·and chemical properties. Cephalotaxus drupacea seeds
yield a fatty oil which is used as an illuminant, and is of local importance
in Japan. The small brown seeds of Torreya nucifera yield an oil called
kaya-no-abura, which is used traditionally for cooking in Japan (Burke
1975). It is also used for paints and other technical preparations (Eckey
1954). The seed kernels of Gnetum ula yield 14.2% fixed oil. It is used in
South India for massage in rheumatism, as an illuminant, and for edible
purpose.
Tall oil is a by-product of the sulphate pulp industry. The waste liquor
from pine pulp mills, after treatment, results in the crude tall oil which is
refined by steam distillation. The composition of the crude oil is approximately
20-60% fatty acids, 10-60% resin acid and 5-24% unsaponifiable material,
mainly sterols (Hildititch 1956). It has innumerable uses. Products made
with the oil are asphalt emulsions, wetting agents, binders, cement addition
agents, waterproofing agents, boring oils, cutting oils, sulphonated oils,
mould lubricants etc. During World War II, for the quickly constructed
landing fields laid out in swampy land to be strong enough for bombers,
a small amount of tall oil was used in the paving, which induced the wet
earth to dehydrate and stick to the asphalt. Tall oil is the lowest-priced of
all organic acids and also a source of components such as abietic acid in
unpolymerized form, oleic and linoleic acids and sterols.
Food Supplements
Young succulent leaves of various Cycas spp. (C. circinalis, C. pectinata,
C. revoluta, C. rumphii, C. siamensis), and Gnetum (also the inflorescence)
are cooked and eaten as vegetables in their native countries.
The starch present in the pith and cortex of the stem and endosperm of
the seeds of cycads is extracted and used as a food. Most cycads yield a
large amount of starch (from the stem) at about 7 years of age; the best
time for extraction is prior to a flush of new leaves. The male plants have
more starch than female, and the starch content also varies from season to
season. The plant is felled and the innermost cylindrical axis of the stem
is removed. It is sliced into thin, oval or circular discs, spread upon mats
and sun-dried. When crisp, it is pounded into flour, sifted and mixed in
water and poured into a vessel, allowed to stand till the starchy substance
is deposited at the bottom. The clear liquid is drained off, and the precipitate
rolled about between boards until spherical pellets called sago are formed.
Various grades such as bullet sago and pearl sago are formed. The majority
of the manufacturers make an amorphous flour, which is stored. The starch
is, however, a poor man's food, or used in times of scarcity in several areas
in southern and southeastern Asia, New Caledonia, Indo-China, Malaya,
India, Burma, Sri Lanka and Fiji.
It is more economical to extract starch from the seeds than from the
stem; for the latter the entire plant has to be destroyed. A Cycas plant produces
annually about 550 seeds, which yield 11early the same amount of starch
(ca. 2.26 kg) as a 1.25-m-long stem (Thieret 1958).
The various parts of the plants and the starch extracted from cycad seeds
and stems often contain a toxic principle, which should be removed before
using it as food.
The pith of Encephalartos is used to make kaffir bread by the aborigines
of South Africa. The pith is scooped out and buried in the earth to rot for
2 months. It is taken out, kneaded with water, made into cakes and baked
in embers under the ashes. Dutch colonists of South Africa gave the name
kaffer brood boom, hottentot brood boom or simply brood boom (bread
tree) to several Encephalartos spp. These names are still in use.
Roasted seeds of Ginkgo are eaten at feasts in China and Japan to promote
digestion and diminish the effects of drinking wine (Dallimore and Jackson
1966).
Pine seeds are rich in fats and proteins. The seeds of several species are
large enough, have a good flavour, and are edible. Some of the species are
P. pineae (Europe), P. cembra (Europe, Siberia), P. armandi (China),
P. gerardiana (India, Afghanistan), and P. cembroides, P. edulis,
P. monophylla, P. sabiniana, P. parryana and P. counteri (North America).
P. pineae has been used as a food item in the northern Mediterranean
region for over 2000 years. The nut is referred to as pignolia (England),
pinone (Italy) and pignon (France).
In Italy, the kernels are largely used for maKing confectionery. Chocolate
manufacturers mix them with cocoa (Theobroma cacao) and the product is
a great delicacy. Traditionally, the kernels are used in soups. Raw or roasted
kernels are also eaten as dessert. In North America, the processing of nuts
has been mechanized. The kernels are dried or roasted in oil or manufactured
into nut-coffee and other sweets and candies. The seeds of Pinus edulis
were a staple diet of the Navajo Indians.
The Aborigines in Queensland depend on the ripe seeds of Araucaria
bidwilli for food. They travel long distances to the groves or forests and
feed on the seeds, which are very fattening. There is a restriction by the
government upon felling these trees in one tract of hilly country, where
several trees are reserved for the natives. They apportion the trees among
themselves, so that each tribe has its own trees, which are again divided
amongst families. The trees are thus handed down from generation to
generation (Dallimore and Jackson 1966). A. araucana seeds are similarly
used by the natives of Chile, and A. angustifolia in Brazil (Howes 1948).
The seeds of Torreya nucifera are an important article of food in Japan.
The seeds of Gnetum ula and G. gnemon are eaten roasted or cooked. The
seed kernel is mashed, moulded into cakes or biscuits, dried in the sun and
fried in boiling oil (P. Maheshwari and V. Vasil 1961a).
454 The Gymnosperms
A sugary exudation with cathartic properties is obtained from the

heartwood, particularly from charred and wounded trees, or from sawed
wood of Pinus lambertiana (s\Igar pine). It is sometimes used as a substitute
for sugar.
Spruce Beer. This is a fermented liquor made from an extract of twigs

and leaves of Picea abies. It is then mixed with treacle or other sugary
substances (Dallimore and Jackson 1966).
Pharmaceuticals
Ephedra, known as ma-huang, has been a common medicine in China for
over 5000 years (see Trease and Evans 1983).
The chief sources of the drug are E. sinica, E. equisetina, E. intermedia,
E. gerardiana and E. major. The ephedras contain ca. 0.5-2.0% of alkaloids,
which varies with species, seasons and age of the plant. The maxium is
reached when the plants are ca. 4 years old, and still flowering. The alkaloids
present are 30-90% ephedrine (and its isomers) and pseudoephidrine, which
were isolated in 1887, although their pharmacological value was discovered
only much later, and have been in extensive use during the present century.
Physiologically and chemically, ephedrine resembles epinephrine (adrenaline).!..
a hormone-like substance with a stimulating action on the sympathetic
nervous system~ The roots also contain a number of macrocyclic alkaloids
(ephedradines) which have hypotensive properties. However, the medical
properties of root and stem are in opposition to each other (Shiu-Yiilg Hu
1969). The root is prescribed for checking excessive perspiration in a weak
patient.
Green branches of Ephedra are collected in autumn when it has the
highest level of ephedrine. They are dried completely, first in shade and
later in the sun. The best material is dry, thick, light green, solid with a
bitter taste and little odour.
Ma-huang is hot, bitter and warming. It is prescribed for typhoid, bad
colds, fe_ver without sweat, pain all over the body, in joints, swelling of
ankles, short breath, etc. The anti-inflammatry action is due to a recently
discovered oxazolidone related to ephedrine (Trease and Evans 1983). Among
the main clinical applications of ephedrine are for bronchospasm, as a nasal
decongestant, and in certain allergic disorders.
Taxol. Taxol is a drug obtained fro!Il the dried inner bark (phloem-cambial
tissues) of Taxus brevifolia (Pacific yew). It has unique therapeutic qualities
against ovarian cancer, breast cancer, non-small-cell lung cancer, melanoma
and colon cancer. Taxol is a complex diterpene-a 20-carbon taxane
containing a .rare oxetane ring and an amide side-chain with antineoplastic
properties (see Edgington 1991). Wani et al in 1969, discovered and named
it 'Taxol' (Wanf et al. 1971, Edgington 1991). It is considered to be the
most important novel natural anticancer drug after 15 years search by the
National Cancer Institute (NCI) in USA. It is regarded as the prototype of
a new class of cancer chemotherapeutic agents (Cragg et al. 1993, Chen
1990). The drug has been approved for clinical treatment of ovarian and
breast cancer by the Food and Drug Adrninstration (FDA) in the USA (see
Zhong 1995).
Taxol catalyzes rapid microtubule formation, stabilizing them against
depolymerization. This gives the drug two tumour fighting mechanisms:
(a) It freezes the mitotic spindle, prevents the depolymerization that
pulls the chromosomes into the two halves of the rapidly 'dividing tumor
cells. This suspends the cells at G 2 or M phase, ultimately killing it.
(b) Taxol inhibits cell migration, which may prevent the spread of
metastatic cancer cells (Edgington 1991).
Taxus is a slow-growing plant of restricted geographical distribution.
The amount of taxol in the bark is relatively low - ca. 0.01% of the dry
weight. Ca. 7000 kg bark is required to produce 1 kg of taxol by current
bark extraction procedures (Cragg et al. 1993). To produce 1 g of the
substance, 3 or 4 trees (at least 60 years old) are required. T. brevifolia is
the only commercial source for the drug at present. However, other species
of Taxus (or Austrotaxus - see Zhong 1995) also produce taxol and are
potential sources (Fett-Neto et al. 1992).
Total synthesis of taxol has not been achieved; semi-synthetic methods
have been developed. The tissue of T. brevifolia has been successfully
cultured to produce taxol, related alkaloids and alkaloid precursors
(Fett-Neto et al. 1992). Tissue and cell cultures of other species of Taxus
have also been established (see Zhong 1995)- T. cuspidata, T. canadensis,
T. baccata, T. yunnanensis, T. chinensis, T. chinensis var mainei, T. globosa,
T. wallichiana, T. floaridana. Most of them contain taxol or taxol-like
compounds (Kang and Hou 1993). In in vitro cultures, callus can be induced
from any tissue from any part of the plant - bark, cambium, needle, stem,
seed, aril and roots. For optimum yield of taxol, the bark or cambial tissue
is preferred (Fett-Neto et al. 1992). Due to the slow growth of Taxus spp.,
tissue and cell cultures of Taxus are also recalcitrant and require continuous
effort for the production of taxol. The possibility of commercializing large-
scale 'yew' cell culture is being considered as the demand for taxol is
continually increasing (Taxus species may become extinct if its exploitation
continues).
Scientists are also exploring alternative sources for taxol. Taxomyces
andreanae, an endophytic hyphomyceteous fungus associated with the phloem
of Taxus brevifolia, also produces taxol. The fungus is cultured in semi-
synthetic liquid medium and produces taxol and related compounds.
Biotechnology may ultimately enhance the production of taxol by Taxomyces
andreanae (Babu et al. 1993).
In Cycas a mucilaginous and transparent gum exudes from the stem
456 The Gymnosperms
when it is wounded. Later, it hardens and turns light brown. The gum
swells when placed in water, becomes colourless and transparent. It produces
rapid supuration when applied to malignant ulcers. The gum is also used
as an antidote for snake and insect bites. Other gum yielders are Dioon,
Encephalartos and Macrozamia.
The gymnosperms are indeed very useful in our daily life.
24 Concluding Remarks
Seasonal Changes in Secondary Growth. A comparative comprehensive

study of the seasonal variations in anatomy would be very useful for uses
of the timber (Sharma 1994). Cambial growth research is a productive field
for future studies. The mechanism by which the vascular cambium originated
in early plants is not fully understood. Factors such as rainfall, temperature
and photoperiod directly affect the ontogeny of tissues. The influence on
the development of vascular cambium (meristem that produces secondary
vascular tissues) and its derivatives is closely related to the effects of
internal influences-physiological, structural and genetic. Because of this
interaction between external and internal influences, investigations of vascular
tissues of the plant as a whole are necessary (see Munting and Willemse
1987).
Little is known about the influence of external factors on the development
of secondary phloem. The phloem and xylem may differentiate simultaneously,
or phloem differentiation may precede or. follow the differentiation of xylem.
Functional sieve cells are present throughout the year in the secondary
phloem of various Pinaceae (Alfieri and Evert 1973). The last-formed sieve
cells overwinter, and remain functional throughout the next growing season.
According to Alfieri and Mottola (1983), the differentiation of phloem may
follow xylem differentiation. The secondary phloem affects cambium activity
and, through the cambium, the secondary xylem. Internal influences on the
secondary phloem are more pronounced than external. The external and
internal factors have a direct influence on sieve tubes and sieve cells,
during development as well as at maturity. Therefor.e further investigations
are necessary to understand the formation of secondary phloem. According
to Denne (1976), the major seasonal variation in tracheid size and thickness
of wall in seedlings of Picea sitchensis is likely to be associated rather with
changes in the production of· hormones, than with variation in substrate
availability.
Population and Wood Requirement. The world population is growing

fast and the non-renewable resources are dwindling. The demand for wood
and wood products is continuously rising, through direct burning of fuel,
458 The Gymnosperms
production of charcoal, alcohol (methane), and through the use of cellulose

by the plastic industry. Cell and tissue culture techniques can introduce
new genetic variation and reduce barriers to crossability amongst species,
and thus assume a role supporting biotechnology (Mohammad and Vidavar
1988).
Pollution. There have been several studies on the harmful effects of air
pollutants on plants. Inhibition of seed germination, retarded tree growth,
and damage to leaves are some of them (see Renzoni et al. 1990). Pinus
pinea is very sensitive to pollution, and it may be used as an environmental
indicator of air pollution. In vitro germination responses have been investigated
from two localities in Italy (San ,Rossore National Park, and Pisa Town
Centre) characterized by different ~ypes and levels of pollutants. The pollen
from San Rossore showed a higher rate of germination (80%) while those
from Pisa showed only 40%. Also, pollen from Pisa had several morphological
anomalies, such as reduced size and wrinkled grains. The data suggest that
there is higher pollution in Pisa than in San Rossore. This has an adverse/
disturbing effect on sporogenesis, microspore maturation and development
of pollen grains. The reduction in pollen fertility caused by pollutants may
lead to a decrease in seed production (Renzoni et al. 1990). Similar
investigations in other taxa would be fruitful.
Symbiosis. The study of coralloid roots in cycads is fascinating. Since its

discovery in Cycas revoluta by Reinke (1872), little work has been done
on this aspect (see Chap. 8), and knowledge about coralloid root growth
and development is inadequate. There is no information on the transition
stage between precoralloid and coralloid roots. The identification of specific
stages of development based on distinct morphological characteristics would
provide a more accurate basis for further anatomical and ultrastructural
investigations. The study of the processes involved in the transition of
apices from the precoralloid to coralloid condition, the mechanism involved
in initial invasion by algae, colonization and establishement of symbioses
in mature precoralloids, and continued invasion of active coralloid apices
would be interesting. There is also urgent need for a detailed study of the
various dermal tissues of precoralloids, coralloids and normal lateral roots
in order to determine the identity of the papillose sheath (root cap or epidermis)
and the algal zone (epidermis or specialized cortical tissues).
Symbiosis between Nz-fixing Cyanobacteria and cereal crops has been
attempted as a possible alternative to reliance on fertilizers (see Ahem and
Staff 1994). Further investigation of the process of invasion and establishment
of symbiosis in cycad coralloid roots may provide significant results.
Reproductive Biology. The study of the reproductive biology of

gymnosperms requires a good deal of attention; only a few taxa have been
Concluding Remarks 459
studied. Detailed investigations are necessary to fill up the gaps in our

knowledge of anatomy, reproductive biology, cytology, genetics and breeding
behaviour. Physiological, biochemical, cell and nuclear studies have been
more or less completely ignored and deserve immediate attention. Significant
knowledge (so far unreported because of the exclusive use of the light
microscope) will emerge by employing new techniques-histo- and
cytochemistry, electron (TEM, SEM), fluorescence and interference
microscopy.
Pollen Biology. Intensive investigations (using modem tools and techniques)

on pollination biology-the role of the pollination drop, anemophily/
entomophily, pollen viability, origin and role of pollen tube, wall/alveoli
formation in female gametophyte, archegonium etc.-are necessary.
Life History. The life history of several gymnosperms and their temporal
correlations in the reproductive cycle needs to be investigated. Wild gene
resources can be identified and superior genetic variation can be multiplied
or bred under control conditions through the knowledge of reproductive
biology. It can be used for tree improvement programmes.
An ethnobotanical survey for the use of gymnosperms would be of great
use (Sharma 1994).
Finally, our understanding and knowledge of the gymnosperms is far
from complete. Detailed investigations (structural, physiological and
biochemical) have so far been confined to only a few taxa. Therefore, there
is urgent need to intensify these studies.
The possibilities of studying this fascinating group of plants-
Gymnosperms: the last group of archaegoniates-are endless.
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Plant Index
(*Extinct)
Abies 10, 18, 146, 441, 446, 450 Biota 17, 203, 204, 206, 208, 210, 211,
Acmopyle 7, 221 212, 214, 215, 216, 219, 429, 439
Actinostrobus 203 Boletaceae 147
Adiantum 102 Boletus 147
Agaricaceae 147 Botrychium 9
Agathis 7, 18, 251, 257, 262, 314, 439, Bowenia 4, 48, 49, 54, 55
445, 446 Bryum 51
Agrobacterium 436 Butomopsis 11
Allicospermum* 100
Amentotaxus 8, 20, 292 Calamopityaceae 2, 3
Ameylon* 127, 129 Callistophytaceae* 2
Anabaena 48, 59 Callistophytales* 22
Androstrobus* 47 Callitris 7, 14, 18, 203, 441, 447
Aneurophytales 2, 3, 22, 23 Callixylon 22, 24, 143
Araucaria 7, 18, 137, 142, 145, 251, Calothrix 48
252,253,254,255,256,257,258,2 59, Calocedrus 203
260,262,263,264,266,314,439, 445 Cardiocarpus* 133, 135
Araucariaceae 7, 144, 146, 222, 251,257, Carnoconites* 94, 96
441 Casuarina 405
Araucarioxylon* 88, 128 Cathaya 146
Arberia* 91 Caytonanthus* 82, 83, 84, 85
Arber.iella* 88 Caytonia* 11, 82, 84, 85
Arceuthos 203 Caytoniaceae 82
Archaeopteridales * 2, 3, 22, 23 Caytoniales 11, 21, 22, 23, 82
Archaeopteris* 24, 25 Cedrus 12, 14, 18, 145, 146, 430, 442,
Arctobaeira* 98, 99 450
Artisia* 131 Cephalotaxaceae 7, 144, 265, 290, 310
Athrotaxis 7, 14, 18, 188 Cephalotaxus 7, 12, 14, 18, 19, 20, 265,
Austrobaileya 406 267,268,269,272,273,274,275,2 77,
Austrocedrus 203 279,280,2&1,285,288,289,309,4 40,
Austrotaxus 8, 20, 278, 292, 455 451
Azolla 10 Ceratozamia 4, 47, 54, 55, 414, 415
Chamaecyparis 7, 14, 145, 203, 439, 441
Baenia* 46, 47 Cheirolepidaceae 291
Baiera* 98, 99 Chigua 4, 48, 50
Bennettitales* 20, 21, 37 Classostrobus* 291
Bijuvia* 45 Classopolis* 291
492 Index
Coniferales 4, 6, 20, 21, 22, 23, 144, Fitzroya 203

290, 309 Fokienia 203
Cordaianthus* 131, 133, 134, 135, 137, Frenelopis* 291
139,142,143,290 Fusarium 449
Cordaitaceae* 127, 142
Cordaitales* 20, 21, 22, 23, 127. 136, Gangamopteris* 88, 92
142, 290, 291 Ginkgo 4, 5, 9, 10, 14, 15, 17, 18, 19, 71,
Cordaites* 127, 131, 132, 138 98, 99, 100, 105, 106, 108, 109, 110,
Corystospermaceae* 21 111, 112, 114, 115, 118, 119, 122, 133,
Corystospermales* 82 247,401,410,415,423
Crossotheca* 33 Ginkgoaceae 100, 444
Cryptomeria 7, 14, 145, 188, 189, 190, Ginkgoales 3, 4, 20, 22, 23, 98
191,193,195,198,199,202,314,439 Ginkgoidium* 99
Cunningham!a 7, 188 Ginkgoites* 98, 99, 100
Cupressaceae 7, 144, 202, 203, 310,441 Glossopteridaceae* 86
Cupressus 7, 14, 192, 202, 203, 408, Glossopteridales 1, 21, 22, 23, 86
410, 411, 439, 441 Glossopteris* 86, 87, 88, 89, 92
Cycadales 4, 20, 21, 22, 23, 44, 97 Glossotheca* 89
Cycadaceae 56 Glyptolepis* 141, 142. 143
Cycadeoidaceae* 38 Glyptostrobus 7, 188
Cycadeoidales* 21, 22, 23, 36, 95, 97 Gnetaceae 366
Cycadeoidea* 38, 39, 41, 404 Gnetales 4, 8, 21, 311, 366
Cycadofilicales* 21 Gnetum 8, 9, 10, 11, 12, 13, 14, 16, 17,
Cycas 4, 18,48,49, 50, 51, 54, 55,56, 19, 22, 36, 247, 301, 311, 314, 349,
59, 61, 63, 65, 66, 71, 73, 74, 75, 76, 366,368,369,371,374,378,404,405,
79, 80, 247, 401, 412, 414, 439, 451, 406,451
457
Cydonia 406 Hebeloma 147
Czekanowakiales* 2, 21, 22, 23
lsoetes 9, 10
Dacrydium 7, 144, 145, 221, 440, 450
Denkania* 89, 90, 91, 92 Juniperus 7, 14, 145, 203,409, 439, 441,
Dictyopteridium* 91, 92 450
Dictyozamites* 38
Dioon 4,44,50,51,52,54, 73,74,455 Kaloxylon* 30
Diselma 203 Karkenia* 100
Doratophyllum* 46 Keteleeria 146
Encephalartos 4, 47, 48, 50, 54, 55, 71, Laccaria 147

73,81,410,414,452,455 Lagenostoma* 34
Ephedra 8, 9, 10, 14, 17, 18, 22, 311, Laru 12, 18,145,420,440,445,449
312,313,316,319,320,321,324,327, Lebachia* 135, 137, 138, 140, 143,
329,331,333,337,340,341,342,349, Lebachiaceae* 139, 140, 141, 142
393, 404, 410, 411, 416, 453 Lepidozamia 4, 50
Ephedraceae 312 Leptocycas* 44
Ephedrales 4, 21, 311, 312 Libocedrus 7, 203
Ephedripites* 404 Lidgettonia* 89, 90, 92
Equisetum 50, 51 Lyginopteridaceae* 29
Eretomonia* 89 Lyginopteridales* 21
Ernestiodendron* 137, 138, 139, 140, Lyginopteris* 29, 33
143, 265 Lyssoxylon* 44
Index 493
Macrozamia 4, 47, 48, 50, 54, 73,451,455 Podocarpaceae 7, 144, 146, 221, 310,
Marchantia 51 443
Marsilea 10 Podocarpus 7, 14, 18, 145, 222, 223,
Medullosaceae* 94, 97 225,227,228,230,232,234,237,2 38,
Medullosales* 21 240,243,245,247,249,250,251, 440
Mesoxylon* 128, 129, 131, 135 Polypodiaceae 4
Metasequoia 7, 145, 188 Polypodium 86
Microcachrys 14, 221 .Polytrichum 51
Microcycas 4, 49, 50, 53, 55, 81 Protopityales* 22, 23
Microstrobus 7 Pseudoctenis* 44
Miterospermum* 135 Pseudofrenelopis* 291
Pseudolarix 145, 146
Neocallitropsis 203 Pseudotaxus 292, 309
Nilssonia* 37, 46 Pseudotsuga 146, 409, 423, 428, 437,
Nilssoniale~* 37 445,449
Nipaniophyllum* 94, 95 Pseudovoltzia* 140, 141
Nostoc 48, 59 Psilotum 10
Nothotaxus 20 Pteridium 51, 52
Pteridospermales 21, 22, 23
Ortiseia* 135 Pterophyllum 37
Oscillatoria 59 Ptilophyllum* 37, 38
Ottozamites* 38
Ottokaria* 91, 92 Regnellidium 10
Paeonia 11, 406 Sagenopteris* 82, 8'3

Palaeocycas* 46 Sahnia* 94, 96
Palaeotaxus* 292, 309 Salvinia 10
Palissaya* 265 Saxegothea 7, 221, 262, 278
Papuacedrus 7, 203 Sciadopitys 7, 16, 188, 439
Pennsylvanioxylon* 128, 129, 13t Scutum* 91, 92
Peltaspermaceae* 82 Selaginella 10
Pel~spermales* 21, 22 Senotheca* 91
Pentoxylales* 2, 21, 22, 23, 94 Sequoia 6, 11, 144, 188, 407, 445
Pentoxylon 94, 95, 96 Sequoiadenodron 6, 144, 188
Pherosphera 14, 221 Sphenobaiera* 98, 99, 100
Phyllocladus 7, 145, 450 Sphenopteris 30, 33
Picea· 12, 146, 407, 408, 423, 433, 445, Stamnostoma* 30
450, 456 Stangeria 4, 9, 48, 50, 54, 66
Pilgerodendron 203 Stangeriaceae 56
Pilularia 10 Stylites 10
Pinaceae 6, 144,146,290,293,310,442
Pinus 12, 14, 18, 145, 146, 147, 149, Taiwania 7, 188
150,151,152,153,154,155,156,1 57, Tamaxellia* 291
160, 161, 162, 163, 164, 165, 166, 167, Taxaceae 292
168, 169, 170, 171, 172, 173, 175, 176, Taxales 4, 8, 20, 22, 23, 292, 309
177, 179, 180, 181, 182, 184, 185, 187, Taxodiaceae 6, 144, 188, 293, 310, 444
188,222,240,349,393,409,410,4 16, Taxodium 6, 14, 145, 188, 314, 440
423,425,428,429,433,439,445,4 47, Taxus 8, 14, 16, 18, 20, 192, 268, 278,
452, 457 292, 293, 294,1295, 296, 297, 298, 299,
Pistacia 448 300,301,302,303,304,307,309,4 10,
Pitys* 22 411, 440, 444, 453
494 Index
Tetraclinis 447 Walchia* 137

Thuja 7, 202, 203, 411, 439, 445 Walchiostrobus* 139, 140, 143
Thujopsis 203, 439 Welwitschia 8, 9, 10, 11, 12, 13, 14, 16,
Torreya 8, 10, 14, 16, 20, 278,292, 309, 19, 22, 301, 311, 343, 344, 345, 346,
411, 451 349, 351, 352, 354, 356, 357, 358,
Trichopitys* 100, 101, 125, 126 359,360,361,362,363,364,365,404,
Tsuga 10, 18, 146, 262, 423, 445, 449, 430
450 Welwitschiaceae 343
Welwistschiales 4, 8, 311, 343
Ullmania* 141, 142, 143 Widdringtonia 203
Wielandiella* 38
Vertebraria* 86, 88 Williamsonia* 38
Vitis 406 Williamsoniella* 38
Voltzia 141
Voltziaceae* 139, 140, 141, 143 Zamia 4, 9, 18, 47, 48, 49, 50, 51, 52,
Voltziales* 22, 23, 137, 138, 139, 142, 53,55, 66,73, 74,81,410,412,423
290 Zamiaceae 56
Voltziopsis* 142, 143 Zamites* 38

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The Gymnosperms

Springer-Verlag Berlin Heidelberg GmbH

Dr. Cbhaya Biswas

Copyright © I 997 Springer-Verlag Berlin Heidelberg

All rights reserved. No part of this publication may be reproduced,

Exclusive distribution in North America (including Canada and Mexico),

ISBN 978-3-662-13166-4 ISBN 978-3-662-13164-0 (eBook)

Professor Panchanan Maheshwari

The Gymnosperms is a well-illustrated comprehensive account of living

For the preparation of this book we are grateful to:

2. Seed Development 12-19

Gymnosperm opsida-Cycad ophytes

10. Glossopteridales 86-93

11. Pentoxylales 94-97

12. Ginkgoales 98-126

14. Voltziales 137-143

15. Coniferales 144-146

15.2 Taxodiaceae 188-202

15.3 Cupressaceae 202-221

15.4 Podocarpaceae 221-251

15.5 Araucariaceae 251-265

15.6 Cephalotaxaceae 265-290

15.7 Phylogenetic Considerations of Coniferales 290-291

16. Taxales 292-310

17. Gnetopsida 311

18. Ephedrales 312-342

19. Welwitschiales 343-365

20. Gnetales 366-404

21. Phylogenetic Considerations: Ephedra,

22. In Vitro Experimental Studies 408-439

23. Economic Importance 440-456

Paper and Board 445

24. Concluding Remarks 457-459

Indubitable seeds first appeared in the Devonian (395-359 my B.P. =million

Antiquity and Fossil History

angiosperms appeared and diversified rapidly. They began to replace the

Fig.1.3 A, B. World distribution. A Living cycad genera. B Gnetum. (A After Schuster

Cupressaceae. The largest family of the conifers includes ca. 19 genera,

Podocarpaceae. Podocarpaceae is the most important family of the Southern

Araucariaceae. An extremely old family with fossil record extending

Cephalotaxacae. A single taxon, Cephalotaxus, is restricted to eastern

Ephedrales. The Ephedrales comprise a single mono generic family,

Welwitschiales. Welwitschia is a monotypic genus of limited distribution.

Some of the other features .typical of gymnosperms are: There are no

Gymnosperms and Pteridophytes

Gymnosperms and Angiosperms

In most gymnosperms, cytokinesis following meiosis is simultaneous

The male gametes may be flagellate spermatozoids, as in cycads

Megaspore Wall. Pettitt ( 1966) has reinterpreted the structure of the

detritus (containing sporopollenin and lipids) which accumulates against

Archegonia. The archegonia develop in all gymnosperms, except Gnetum

Pollination and Fertilization

Pollination. In different gymnosperms the ovules receive pollen at various

Fertilization. The release· of male gametes varies in different taxa of

One of the objectives of systematists is to discover a phylogenetic system