Internal secretory spaces in thickened underground systems

of Asteraceae species
Graziela Cury
A
and Beatriz Appezzato-da-Glria
A,B
A
PO Box 09, 13418-900, Piracicaba, So Paulo, Brazil.
B
Corresponding author. Email: bagloria@esalq.usp.br
Abstract. Secretory structures are present in many vascular plants and have an important ecological role as a plant
defence mechanism against herbivors and pathogens. Internal secretory spaces of lipid substances are widespread
in the Asteraceae. However, information about the occurrence of these structures in thickened underground systems is
sparse, compared with what we know about aerial systems. The main objective of the present paper was to investigate
the occurrence, formation and chemical nature of the secretory structures in six Asteraceae species belonging to the
following tribes: Eupatorieae (Mikania cordifolia and M. sessilifolia), Mutisiae (Trixis nobilis), Plucheeae (Pterocaulon
alopecuroides) and Vernonieae (Vernonia elegans and V. megapotamica). The samples were collected in areas of Cerrado
(tropical savanna) in the state of So Paulo, Brazil. The secretory structures found were cortical canals in roots
(T. nobilis, P. alopecuroides, V. elegans and V. megapotamica), cortical cavities in roots (M. cordifolia, M. sessilifolia
and P. alopecuroides), cavities in the secondary phloem of roots (T. nobilis), cortical cavities in the xylopodium
(M. cordifolia, M. sessilifolia, P. alopecuroides and V. megapotamica) and in the underground stem (T. nobilis),
and canals in the secondary xylem in the xylopodium (M. cordifolia and M. sessilifolia). Histochemical tests showed
the presence of lipid substances in all structures.
Introduction
The occurrence of secretory structures in vascular plants has
already been reported by various authors and, according to Fahn
(1988), the main types are glands, nectaries, trichomes, oil cells,
ducts and cavities. Some authors (Solereder 1908; Metcalfe and
Chalk 1950, 1979; Robson 1977, 1981; Metcalfe 1983) have
pointed out the importance of these structures in taxonomic
studies, with the purpose of distinguishing orders, tribes, genera
and even species, because different secretory structures may be
found in different parts of the same plant or be conned to one
of its organs. Consequently, a better understanding and
denition of the function and the location of such structures
are necessary. The secretory structures also play an
environmental role. Increasing the capacity of plants to
survive in their habitat by the production of substances is
important for the interaction of plants with the biotic and
abiotic environment (Harbone 1993), as well as a mechanism
of plant defence against herbivores (Fahn 1979, 2002; Dussourd
and Denno 1991).
Internal secretory spaces of lipid substances are widely
distributed among the species of the Asteraceae (Fahn 1979),
occurring both in aerial and underground organs. However, the
role of these structures is not yet well understood. It is known
that lipid compounds may act against herbivorous insects
(Murphy 2001) and there is extensive literature on the aerial
organs, whereas studies on underground structures are scarce
(Appezzato-da-Glria et al. 2008b). The difculties in sampling
and processing underground material for analysis are probably
the main limiting factors in the study of underground organs.
However, studies of underground organs in Asteraceae are
important, because such organs can be useful tools in
taxonomic and even ecological studies.
Most studies on secretory structures have focussed on the
occurrence of ducts in non-thickened roots (Grotta 1944;
Williams 1947, 1954; Hoehne et al. 1952; Lersten and Curtis
1986, 1988; Joseph et al. 1988; Curtis and Lersten 1990; Poli
et al. 1995; Luque et al. 1997; Sacchetti et al. 1997; Duarte
and Estelita 1999; Pagni and Masini 1999; Melo-de-Pinna and
Menezes 2002, 2003; Pagni et al. 2003; Lotocka and Geszprych
2004; Luque-Arias 2004; Machado et al. 2004; Fonseca et al.
2006; Vilhalva and Appezzato-da-Glria 2006; Appezzato-da-
Glria et al. 2008b), with only few having studied the secretory
structures inthickened undergroundsystems (Hoehne et al. 1952;
Panizza and Grotta 1965; Ragonese 1988; Curtis and Lersten
1990; Lotocka and Geszprych 2004; Machado et al. 2004;
Vilhalva and Appezzato-da-Glria 2006; Appezzato-da-Glria
et al. 2008b).
Because internal secretory spaces, which are common in
Asteraceae, can be used as a tool in taxonomic studies, a more
detailed analysis of such structures, as well as correct and clear
terminology are needed. The presence of intermediary structures
between well dened types (Meira 1991), e.g. between duct and
cavity (Cutter 1978; Fahn 1979; Metcalfe 1983), can lead to
dubious classication. For example, Curtis and Lersten (1990)
pointed out these difculties concerning terminology while
studying rhizome of Solidago canadensis L. They observed
CSIRO PUBLISHING
www.publish.csiro.au/journals/ajb Australian Journal of Botany, 2009, 57, 229239
CSIRO 2009 10.1071/BT08139 0067-1924/09/030229
that the so-called canals found in this species were actually
cavities very close to each other whose septum broke during
the process of tuberisation. Therefore, they proposed the termoil
reservoir, which was also later adopted by Lotocka and
Geszprych (rhizome of Rhaponticum carthamoides (Willd.)
Iljin; 2004).
The present study aims to analyse the occurrence, formation
and the chemical nature of the internal secretory spaces of
six Asteraceae species belonging to the following tribes:
Eupatorieae (Mikania cordifolia L.f. Willd. and M. sessilifolia
DC), Mutisiae (Trixis nobilis (Vell.) Katinas), Plucheeae
(Pterocaulon alopecuroides (Lam.) DC) and Vernonieae
(Vernonia elegans Gardner and V. megapotamica Spreng.).
Materials and methods
Mikania cordifolia, M. sessilifolia, Trixis nobilis, Pterocaulon
alopecuroides, Vernonia elegans and V. megapotamica were
selected from the list of Almeida et al. (2005). The aim was to
compare the location and the formation process of secretory
structures in species of the same genera (in the case of
Mikania and Vernonia) and among genera belonging to distant
tribes (Eupatorieae, Mutisiae, Plucheeae and Vernonieae). Adult
individuals were collected in natural populations in areas of
Cerrado located in Botucatu (22

53
0
S, 8

29W) and Itirapina

(22

13
0
S, 47

54W), So Paulo State, Brazil, where Asteraceae

is well represented. The vouchers of the specimens (88 792,
88 791, 92 159, 88 790, 88 787 and 88 789, respectively) are
deposited in the ESA Herbarium, Brazil.
For anatomical analyses, whole underground systems of three
adult plants from each species were xed in formaldehyde :
glacial acetic acid: 50% ethanol (1: 1 : 18 , v/v, FAA 50) for
48 h (Johansen 1940) and dehydrated in a series of graded
ethanol (50100%) and embedded in plastic resin (Leica
Historesin: Leica, Wetzler, Germany). Cross- and longitudinal
serial sections, 510 mm thick, were cut with a rotary
microtome, then mounted on glass slides and stained with
toluidine blue (Sakai 1973). To determine the chemical nature of
the substances found in the secretory structures, hand-cut sections
of xed material were submitted to the following histochemical
tests: Sudan IV (Jensen 1962) for lipids, Nadi reagent (David
and Carde 1964) for terpenoids, ruthenium red (Johansen 1940)
for mucilage and ferric chloride (Johansen 1940) for phenolic
compounds. The images were digitally captured with a Leica
DMLB microscope by using a video camera plugged to a
computer utilising the IM50 software for image analysis.
Results
All species studied showed thickened underground organs, a
xylopodium (for denition see Alonso and Machado 2007 and
Appezzato-da-Glria et al. 2008a) anda stemtuber (Table 1). The
xylopodium of the stem structure and the stem tuber emitted
adventitious roots whereas the xylopodium of the radicular
structure emitted lateral roots. The presence and location of
secretory structures in these organs are shown in Table 1.
The secretion had a natural yellowish-orange colour, which
can be seen on the slides stained with toluidine blue (Figs 1A, C,
D, G, H, 2B, DH). The secretion stained positively only with
SudanIV(Fig. 3AC, E, G), withthe reddishcolour indicatingthe
presence of lipids. There was no reaction to the other stains and
reagents used.
The xylopodium of M. cordifolia, although showing a
secondary structure, had retained the cortex from the primary
growth. These organs had cavities and canals. The cavities
(150 mm diameter, 452 mm length) were observed in the cortex
near the phloem(Fig. 1A) and their epitheliumcomprised cells of
the endodermis with Casparian strips and cells of the cortical
parenchyma opposite the endodermis (Fig. 1B). Figure 1C
shows that this cavity originates from a schizogenesis process
followed by cell lysis, with the dissolution and later division of
epithelial cells to increase the lumen. The formation of a canal
(100 mm diameter, 1.7 mm length) can be observed in the
secondary xylem. It is originated by the separation of
parenchyma cells of the vascular ray, forming its epithelium,
which leads to an increase of the lumen after cell divisions
(Fig. 1D). On the adventitious roots of the xylopodium,
cavities (350 mm diameter, 1.3 mm length) occur on the cortex
opposite the phloem. In cross-section, these cavities in the
primary root structure show a diamond-shaped lumen
circumscribed by four epithelial cells. Two of the epithelial
cells are endodermic with visible Casparian strips and two are
from the adjacent cortical parenchyma, which, as seen in a
longitudinal section, suffers separation in different places,
originating small intercellular spaces (Fig. 1E) in the
secondary root structure. The union of these spaces, by
complete cellular separation, gives rise to elongated structures
(Fig. 1F). After the division and lysis of epithelial cells (Fig. 1G),
the lumen of these cavities becomes roundish and bigger,
as seen in the cross-section (Fig. 1H).
In the xylopodium of M. sessilifolia there are cortical cavities
(100 mm diameter, 434 mm length), originating through the
separation of cells from the endodermis with visible Casparian
strips, and cells from the cortical layer opposite the endodermis.
Table 1. Occurrence and types of secretory structures in the underground systems of six Asteraceae species
Species Occurrence and the type of the secretory structure
Mikania cordifolia Xylopodium (cortex and secondary xylem): cavities Roots (cortex): cavities
Mikania sessilifolia Xylopodium (cortex and secondary xylem): cavities Roots (cortex): cavities
Trixis nobilis Underground stem (cortex and secondary
phloem): cavities
Roots: canals in the cortex and cavities
in the secondary phloem
Pterocaulon alopecuroides Xylopodium (cortex): cavities Roots (cortex): canals and cavities
Vernonia elegans Xylopodium: secretory endodermis Roots (cortex): canals and secretory endodermis
Vernonia megapotamica Xylopodium (cortex): cavities and secretory
endodermis
Roots (cortex): idioblasts, canals and secretory
endodermis
230 Australian Journal of Botany G. Cury and B. Appezzato-da-Glria
(A)
(B)
(C) (D)
(E)
(F)
(G)
(H)
Fig. 1. Mikania cordifolia. (A, B, D) Cross-sections of the xylopodium. (A, B) Cortical cavity (
*
). (B) Detail of cortical cavity, showing the Casparian strips on
the endodermic cells (arrow). Inset shows a detail of an endodermic cell with Casparian strips (arrows). (C) Longitudinal section of the xylopodium, showing
cortical cavity (
*
) under cellular lysis (arrow). (D) Secondary xylem canal (). (E, F) Longitudinal section of an adventitious root, showing the formation
of cortical cavity (
*
). (G, H) Cross-sections of adventitious roots, showing cortical cavities ( ); cellular lysis (arrow in G) and Casparian strips (arrow in H)
are shown. C=cortex; SPH=secondaryphloem; SX=secondaryxylem; VR=vascular ray. Scale bars =200 mm(A), 10 mm(B), 50 mm(C, E), 100 mm(D, F) and
30 mm (G, H).
Internal secretory spaces in thickened underground systems Australian Journal of Botany 231
(A)
(B)
(C) (D)
(E) (F)
(G) (H)
Fig. 2. Pterocaulon alopecuroides. (A) Cross-section of the xylopodium, showing cortical cavities. (B) Longitudinal section of the xylopodium, showing
cortical cavities (
*
). (C) Cross-section of the lateral root in primary growth, with cortical canals (arrows). (D) Longitudinal section of the lateral root in
primary growth, showing the cortical canal ( ). (E, F) Cross-section of lateral roots in secondary growth, with cortical cavities (
*
). (F) The beginning of the
cavity formation by separation of the cortical cells. (G, H) Longitudinal sections of lateral root. (G) Close cavities and (H) cavity formed by septum rupture.
The arrow indicates cellular lysis. C=cortex; CA=cambium; PPH=primary phloem; SPH=secondary phloem; SX=secondary xylem. Scale bars =100 mm
(A, G), 30 mm (B, D, F), 10 mm (C) and 50 mm (E, H).
232 Australian Journal of Botany G. Cury and B. Appezzato-da-Glria
Such cells form the epithelium after separation. The growth
of the lumen is followed by divisions in the epithelial cells
(Fig. 4A). It is possible to observe canals (200 mm diameter,
2.5 mm length) in the vascular ray (Fig. 4B) in the secondary
structure of the xylopodium of M. sessilifolia. Such canals result
from the separation of vascular parenchyma cells along its
extension (Fig. 4CF), thus forming an elongated structure.
The division of epithelial cells increases the lumen (Fig. 4B).
The adventitious roots of this xylopodium, in the primary
structure, show cortical cavities (100 mm diameter, 1 mm
length) opposite the phloem, with a roundish lumen and an
epithelium formed by a layer of cells, namely two cells from
the endodermis and two from the cortical parenchyma opposite
the endodermis. In the secondary structure, the epitheliumshows
cellular division, followed by an increase in the lumen. The
presence of these cavities, as seen in the longitudinal plane,
(A) (B)
(C) (D)
(E) (F)
Fig. 3. Xylopodium of Vernonia megapotamica. (A) Cross-section of cortical (
*
) and endodermic cells (arrow) with lipid substances. (B) Longitudinal
section, showing a cortical cavity ( ) and lipid drops in the epithelium (arrows). (C, D) Cross-sections of V. elegans adventitious roots. (C) Cortical canals
(arrows) and endodermis with lipid substances secretion. (D) The cortical canals (
*
) and the Casparian strips in endodermic cells (thin arrow), and an
endodermic cell in division (wide arrow). (E, G) Adventitious roots of V. megapotamica. (E) Longitudinal section, showing the cortical canal (
*
) and
(G) cross-section with secretory idioblasts of lipid substance in the outer cortex (arrows). C=cortex; E=endodermis; P=pith. Scale bars =30 mm (AE) and
100 mm (G).
Internal secretory spaces in thickened underground systems Australian Journal of Botany 233
results in a sequential separation of the endodermic cells with
Casparian strips and the cortical parenchyma cells opposite the
endodermis. In a more advanced stage of development, as seen
in the longitudinal section, the cavities join in lines without
anastomosis between then, maintaining their length (Fig. 4G, H).
The underground stem of T. nobilis, although showing a
secondary structure, retained the structure from primary
growth. This stem has cavities (62 mm diameter, 400 mm
length) in the internal cortex, opposite the phloem (Fig. 5A),
originating by separation of common cortical cells, which then
form the epithelium. The epithelium later suffers cellular
divisions to increase the lumen. There are also cavities
(230 mm diameter, 600 mm length) present in the phloematic
ray (Fig. 5A), which, as seen in the cross-section, are
originated by a separation and later lysis of cells, with a
septum rupture between the spaces, leading to lumen
expansion (Fig. 5BD). In the longitudinal plane, cavities can
be seen as little elongated spaces with a determined lumen. In the
(A) (B)
(C) (D) (E) (F)
(G) (H)
Fig. 4. Mikania sessilifolia. (AF) Cross-sections of the xylopodium. (A) Cortical cavity ( ). (B) Secondary xylem canal (
*
). (CF) Formation of the
secondary xylem canal (
*
) by cellular separation of the vascular ray (C) near the pith, (D) average portion of secondary xylem (arrows), (E) near the
vascular cambium and (F) on the cortex. (G, H) Longitudinal sections of adventitious roots, showing cortical cavities ( ). (H) Detail of an endodermic cell
with Casparian strips (arrow). C= cortex; P =pith; SX=secondary xylem. Scale bars = 50 mm (A), 200 mm (B), 100 mm (CG) and 30 mm (H).
234 Australian Journal of Botany G. Cury and B. Appezzato-da-Glria
adventitious roots of T. nobilis, in the primary structure, there are
cortical canals (50 mm diameter, 625 mm length) opposite the
phloem. These canals show a diamond-shaped lumen, resulting
from the separation of cells of the endodermis and the cortical
cells of the parenchyma, opposite the endodermis. These cells
form the epithelium. In the secondary structure, the epithelial
(A) (B)
(C) (D)
(E) (F) (G) (H)
Fig. 5. Trixis nobilis. (AD) Cross-sections of the thickened underground stem, showing cortical cavities ( ) and the cavities in the phloem ray (
*
). (BD)
Formation of the cavities in the phloem ray (
*
) by septum rupture (arrows). (EH) Longitudinal sections of the adventitious roots, showing the formation of
the cavities in the phloematic ray by septum rupture (arrows). C=cortex; CA=cambium; PHR=phloem ray; SPH=secondary phloem. Scale bars =100 mm
(A, EH) and 30 mm (BD).
Internal secretory spaces in thickened underground systems Australian Journal of Botany 235
cells divide and the lumen canal becomes bigger and roundish.
In the phloematic ray, small cavities are formed as a result of
the separation of cells from the parenchyma. In the longitudinal
section, it can be observed that the isolated cavities after
septum rupture result in the formation of elongated structures
(70 mm diameter, 700 mm length) (Fig. 5EH).
The xylopodium of P. alopecuroides has cortical cavities
(30 mm diameter, 220 mm length) near the phloem (Fig. 2A),
originating through separation of cortex cells, which then
form the epithelium. Cellular divisions of the epithelium
results in the increase of the lumen (Fig. 2B). The lateral roots
of the xylopodium, in the primary structure, have cortical
canals (86 mm diameter, 1 mm length) with origin and location
similar to those described for T. nobilis (Fig. 2C, D). However,
their shape is irregular and the epithelium does not divide.
In the secondary structure, canals are not visible. Nevertheless,
the separation of the cortical cells of the parenchyma (Fig. 2F)
leads to the formation of new secretory cavities (Fig. 2E).
In the longitudinal section, we observed that these small
cavities are very close to each other (Fig. 2G). With the
development of the organ, cells between the cavities, which
form septa separation, suffer lysis, resulting in the formation
of elongated structures (50 mm diameter, 450 mm length) whose
epithelium divides itself following the growth of the lumen
(Fig. 2H).
The xylopodium of V. elegans and V. megapotamica has a
secretory endodermis of lipid substances. Cells from the
cortical parenchyma in V. megapotamica show lipid content
(Fig. 3A). In V. megapotamica, cortical cavities (30 mm
diameter, 50 mm length) can be observed, with the lumen
being formed by the separation of cortical cells. These cells
constitute the epithelium of the cavity (Fig. 3B). The
adventitious roots of these two species have a lipid substance-
secreting endodermis (Fig. 3C) and, in the primary growth,
cortical canals are observed (1550 mm diameter, 900 mm
length). Their origin, location and shape are similar to those
described for T. nobilis (Fig. 3CE). However, cellular divisions
in the epithelium were not observed. . In V. elegans, an anticlinal
division of a single cell in endodermis with visible
Casparian strips, results in two cells which are part of the
epithelium of two adjacent canals (Fig. 3D). In the outer
cortex of adventitious roots V. megapotamica, idioblasts
containing lipid substances can be observed (Fig. 3G).
Discussion
Col (1903) determined the difference between canals and
cavities for the rst time, with cavities being shorter and wider
than canals. In the present study, when analysing the thickened
underground systems and their adventitious and laterals roots,
canals and cavities of different shapes, origins and location
inside the organs were observed.
These structures originated from cellular separation only
(schizogenous), cellular dissolution (lysigenous), or had a
mixed origin, i.e. rst cellular separation, followed by lysis of
epithelial cells, leading to the increase of lumen in these spaces
(Evert 2006). In the present study, cavities formed during a
schizolysigen process were observed on the xylopodium as
well as on the adventitious roots of M. cordifolia, on the
lateral roots of P. alopecuroides (cortical cavities) and on the
underground stem and adventitious roots of T. nobilis (cavities
in the phloematic ray). All other secretory structures originated
from the schizogenous process alone.
The presence of cortical canals was veried in the adventitious
roots of T. nobilis, V. elegans and V. megapotamica and on the
lateral roots of P. alopecuroides. They originated from a
schizogenous process and are located opposite the primary
phloem poles in which the endodermis participated on the
formation of cortical canals. These are types of secretory
structures, which are quite common in Asteraceae roots
(Tetley 1925; Williams 1947, 1954; Metcalfe and Chalk 1950;
Lersten and Curtis 1986; Luque et al. 1997; Luque 2001;
Melo-de-Pinna and Menezes 2002, 2003; Lotocka and
Geszprych 2004; Appezzato-da-Glria et al. 2008b). The
cortical canals are elongated longitudinally and therefore
different from the cavities observed in the study species.
It is important note the occurrence of canals in the roots of
T. nobilis, according to Melo-de-Pinna and Menezes (2003),
because the only references to the presence of canals in the
Mutisieae tribe have been for the genera Ianthopappus
(Melo-de-Pinna and Menezes 2002) and Richterago (Melo-de-
Pinna and Menezes 2003). The occurrence of canals in T. nobilis
not only complements the information about the tribe, but
also veries the occurrence of the same structure as in the
roots of Mutisieae.
In the adventitious roots of M. cordifolia and M. sessilifolia,
cortical cavities were elongated structures, visibly shorter than
the canals, and not constituting elongated structures in the
whole extension of the root. Lersten and Curtis (1986)
observed these kinds of cavities in Eupatorium rugosum Houtt
root, also belonging to the Eupatorieae tribe, and called
them structures tubular cavities. These authors noticed that,
in their case, all the tubular cavities were schizogenous as in the
Mikania species studied here.
The occurrence of canals and cavities was also observed in
the adventitious roots of T. nobilis (Mutisieae tribe). The cavities
originated by a process similar to that described by Curtis
and Lersten (1990) for the rhizome of S. canadensis (Astereae
tribe). Curtis and Lersten (1990) called these structures oil
reservoir and veried that at the beginning of the formation of
these reservoirs, schizogeny occurred, as described in the present
study for T. nobilis.
The occurrence of secretory structures in thickened
underground organs could be veried for ve of the study
species. All of them showed cortical cavities originating from
cellular separation and, with the exception of V. megapotamica,
multiplication of epithelial cells for lumen growth was
observed. Multiplication of epithelial cells followed by cellular
lysis was veried only for M. cordifolia. In T. nobilis, in
addition to the cortical cavities, the underground stem also
had cavities in the secondary phloem due to the fusion of the
smaller cavities. This fusion may be occurring, so that these
structures can follow the growth of phloematic ray, as observed
by Luque-Arias (2004) in Coespeletia roots. For M. cordifolia
and M. sessilifolia, the presence of canals in the secondary
xylem, originated by cell separation in the vascular ray after
multiplication of epithelial cells, as well as cortical cavities, was
reported. The presence of secretory canals in the secondary xylem
236 Australian Journal of Botany G. Cury and B. Appezzato-da-Glria
of the xylopodium in Asteraceae has been reported only for
Isostigma megapotamicum (Speng) Sherff (tribe Heliantheae)
by Vilhalva and Appezzato-da-Glria (2006). However, in their
study the canal was originated from cells derived from the
vascular cambium.
Few studies of secretory structures in thickened underground
organs of the Asteraceae refer to the occurrence of cortical
secretory canals, e.g. in rhizomes of Calea pinnatida Banks
(tribe Heliantheae) (Hoehne et al. 1952), Solidago microglossa
DC. (tribe Anthemideae) (Panizza and Grotta 1965) and
Eupatorium inulaefolium H.B.K. (tribe Eupatorieae)
(Ragonese 1988), rhizophores of Smallanthus sonchifolius
(Poepp & Endl.) H.Robinson (tribe Heliantheae) (Machado
et al. 2004) and the xylopodium of Pterocaulon angustifolium
DC. (tribe Plucheeae) (Appezzato-da-Glria et al. 2008b). The
presence of cortical secretory reservoirs was reported for the
rhizomes of Solidago canadensis (Curtis and Lersten 1990) and
R. carthamoides (Lotocka and Geszprych 2004) and secretory
canals in the secondary phloem were reported for the
xylopodium of Calea verticillata (Klatt) Pruski (tribe
Heliantheae) (Vilhalva and Appezzato-da-Glria 2006).
However, only in the case of E. inulaefolium and
R. carthamoides, was the origin of these structures noted,
being a result of separation and multiplication of the
epithelium cells, as occurred for ve of the species studied here.
Within the Vernonieae, Vilhalva (2004) suggested that the
general lack of secretory canals on the leaves may be transferred
to underground systems, giving this observation a taxonomic
connotation. Actually, the species of the tribe presented
herein, also lack canals on the xylopodium, a fact that may
contribute to their identication. In the present study, it was
observed that underground organs in the species of Vernonieae
had common characteristics, which can be used as a common
trait for this tribe. The xylopodium of V. elegans and
V. megapotamica have a lipidic secretory endodermis. This
characteristic has already been described for rhizophores of
Vernonia herbacea (Vell.) Rusby and V. platensis (Spreng.)
Less. (Hayashi and Appezzato-da-Glria 2005) and for
tuberous roots of V. brevifolia Less. and V. grandiora Less.
(Hayashi and Appezzato-da-Glria 2007). Lipids were observed
in the cortical parenchyma of the xylopodium of
V. megapotamica, as has also been observed for the
rhizophores of V. herbacea, V. plantensis (Hayashi and
Appezzato-da-Glria 2005) and tuberous roots of V. brevifolia
(Hayashi and Appezzato-da-Glria 2007). Secretory idioblasts
in the radicular cortex of V. megapotamica have also been
reported for Chresta sphaerocephala DC. by Appezzato-da-
Glria et al. (2008b).
Among the study species, V. elegans and V. megapotamica,
whichbelongtothe Vernonieae, hadsecretorycells not onlyinthe
canals and cavities but also in the endodermis, conrming the
observations made by our research team in other studies on
Asteraceae (Hayashi and Appezzato-da-Glria 2005, 2007;
Appezzato-da-Glria et al. 2008b). Tetley (1925) and
Williams (1954) suggested that the secretion in the endodermis
is related with that the secretory structures, considering that the
canals complement the phloem in the transportation of organic
matter and the endodermis constitutes a passage for phloem
substances to the canal.
Although there is no literature explaining the functional
differences between canals and cavities, Lersten and Curtis
(1988) justied the use of the term reservoir because this
structure has preferably a storage function rather than a
transporting function. Moreover, in P. alopecuroides
(Plucheeae), the formation of cavities on the lateral roots
similar to the oil reservoir described by Lersten and Curtis
(1988) was veried. It is interesting that these cavities seem to
merge into the cortical canals formed during the primary
growth of the root, because they are not found in the
secondary growth. Lotocka and Geszprych (2004) afrmed
that, in underground organs, some reservoirs stop working and
disappear with time, possibly being obliterated, which could
explain why the cortical canals of the primary structure were
not observed in the secondary growth of the lateral roots of
P. alopecuroides.
The location of the cavities and canals opposite or near the
root phloem and in the thickened underground organs of
the species analysed in the present study, as well as the lipid
nature of the secretions, are probably related to defence processes
against herbivores as discussed by Appezzato-da-Glria et al.
(2008b). In roots of Santolina leucantha Bertol. (Asteraceae,
Anthemideae), the intense secretory activity of secondary
products into the canals might have an important
environmental role in preventing herbivores (Pagni and Masini
1999). Lersten and Curtis (1989) suggested that in leaves of
S. canadensis, the location of the secretory reservoirs near the
larger veins may provide protection to the phloem against
herbivory. Franceschi et al. (2005) reported the occurrence of
reservoirs of chemical substances (phenols, terpenoids and
alkaloids) distributed along various tissues of the bark, and
dened them as constitutive chemical defences in conifers,
protecting the phloem nutrients against the use by organisms
such as herbivores. Williams (1954) pointed out the importance
of secretory canals near the phloem, with the function in
transportation of photo-assimilates.
It is important to emphasise that there is a lack of literature
on the formation and occurrence of secretory structures in
thickened underground organs in other families of vascular
plants, with the exception of laticiferous structures, as e.g. in
rhizome of Cyclanthus bipartitus Poit. (Cyclanthaceae) (Wilder
and Harris 1982) and in tuberous roots of Mandevilla illustris
(Vell.) Woodson and M. velutina (Mart. ex Stadelm.)
(Apocynaceae) (Appezzato-da-Glria and Estelita 1997).
Laticiferous structures also play a role in the protection against
herbivory (Fahn 2002).
Neither the function of these structures, nor the nature of
secreted compounds are well dened, including the gum-
secreting canals of the secondary xylem of the tubers of
Vochysia thyrsoidea Pohl (Vochysiaceae) (Jeronymo and
Paviani 1992) and secretory cavities of lysigenous origin in
bulbs of Oxalis latifolia H.B.K. (Oxalidaceae) (Estelita-
Teixeira 1982). This is probably due to the fact that the
Asteraceae constitutes the largest group of angiosperms
(Bremer 1994) and within their different growth forms, many
species show thickened underground organs (Tertuliano and
Figueiredo-Ribeiro 1993). Therefore, more information on
secretory structures is of crucial importance. These structures
may have great economical potential (Cutter 1978), being
Internal secretory spaces in thickened underground systems Australian Journal of Botany 237
possible new sources of chemical compounds important for
medicinal purposes (Wagner 1977).
Acknowledgements
We are grateful to FAPESP (Process 00/12469-3) for the nancial support,
and CAPES and CNPq for the grants. We also thank Instituto de Botnica
and Instituto Florestal for giving us permission to collect plant material,
Professor Vincius C. Souza (USP) for identifying plant species
and Dr Alessandra T. Fidelis, Chair of Vegetation Ecology (Technische
Universitt Mnchen, Germany) for reviewing the English and for
comments.
References
Almeida AM, Fonseca CR, Prado PI, Almeida-Neto M, Diniz S, Kubota U,
Braun MR, Raimundo RLG, dos Anjos LA, Mendonca TG,
de Melo Futada S, Lewinsohn TM (2005) Diversidade e ocorrncia
de Asteraceae em cerrados de So Paulo. Biota Neotropica 5, 117.
doi: 10.1590/S1676-06032005000300003
Alonso AA, Machado SR (2007) Morphological and developmental
investigations of the underground system of Erythroxylum species
from Brazilian cerrado. Australian Journal of Botany 55, 749758.
doi: 10.1071/BT07060
Appezzato-da-Glria B, Estelita MEM (1997) Laticifer systems in
Mandevilla illustris and M. velutina, Apocynaceae. Acta Societatis
Botanicorum Poloniae 66, 301306.
Appezzato-da-Glria B, Cury G, Misaki-Soares MK, Hayashi AH, Rocha R
(2008a) Underground systems of Asteraceae species from the
Brazilian Cerrado. Journal of the Torrey Botanical Society 135, 103113.
doi: 10.3159/07-RA-043.1
Appezzato-da-Glria B, Hayashi AH, Cury G, Misaki-Soares MK, Rocha R
(2008b) Occurrence of secretory structures in underground systems
of seven Asteraceae species. Botanical Journal of the Linnean Society
157, 789796. doi: 10.1111/j.1095-8339.2008.00823.x
Bremer K (1994) Asteraceae: cladistic and classication. (Timber Press:
Portland, OR)
Col M (1903) Recherches sur lappareil scrteur interne des Composes.
Le Journal de Botanique 17, 288318.
Curtis JD, Lersten NR (1990) Oil reservoirs in stem, rhizome, and root
of Solidago canadensis (Asteraceae, tribe Astereae). Nordic Journal of
Botany 10, 443449. doi: 10.1111/j.1756-1051.1990.tb01787.x
Cutter EG (1978) Plant anatomy. Part I: cells and tissues. (Edward Arnold:
London)
DavidR, Carde JP(1964) Colorationdiffrentielle des inclusions lipidiques et
terpeniques des pseudophylles du pin maritime au moyen du reactif Nadi.
Comptes Rendus de lAcadmie des Sciences 258, 13381340.
Duarte MR, Estelita MEM(1999) Caracteres anatmicos de Bidens pilosa L.,
Asteraceae. Hoehnea 26, 1527.
Dussourd DE, Denno RF (1991) Deactivation of plant defense:
correspondence between insect behavior and secretory canal
architecture. Ecology 72, 13831396. doi: 10.2307/1941110
Estelita-Teixeira ME (1982) Shoot anatomy of three bulbous species of
Oxalis. Annals of Botany 49, 805813.
Evert RF (2006) Esaus plant anatomy: meristems, cells and tissues of the
plant body their structure, function and development. (John Wiley &
Sons Inc.: Hoboken, NJ)
Fahn A (1979) Secretory tissues in plants. (Academic Press Inc.: London)
Fahn A (1988) Secretory tissues in vascular plants. New Phytologist 108,
229257. doi: 10.1111/j.1469-8137.1988.tb04159.x
Fahn A (2002) Functions and location of secretory tissues in plants and
their possible evolutionary trends. Israel Journal of Plant Sciences 50,
5964. doi: 10.1560/LJUT-M857-TCB6-3FX5
Fonseca MCM, Meira RMSA, Casali VWD (2006) Anatomia dos rgos
vegetativos e histolocaliza co de compostos fenlicos e lipdicos em
Porophyllum ruderale (Asteraceae). Planta Daninha 24, 707713.
doi: 10.1590/S0100-83582006000400011
Franceschi VR, Krokene P, Christiansen E, Krekling T (2005) Anatomical
and chemical defenses of conifer bark against bark beetles and
other pests. New Phytologist 167, 353376.
doi: 10.1111/j.1469-8137.2005.01436.x
Grotta AS (1944) Contribui co ao estudo morfolgico e anatmico de
Spilanthes acmella L. (Compositae). Anais da Faculdade de Farmcia
e Odontologia da Universidade de So Paulo 4, 130164.
Harbone JB (1993) Introduction to ecological biochemistry. (Academic
Press: London)
Hayashi AH, Appezzato-da-Glria B (2005) The origin and anatomy of
rhizophores in Vernonia herbacea and V. platensis (Asteraceae) from
the Brazilian Cerrado. Australian Journal of Botany 53, 273279.
doi: 10.1071/BT04094
Hayashi AH, Appezzato-da-Glria B (2007) Anatomy of the underground
systeminVernoniagrandioraLess. andV. brevifoliaLess. (Asteraceae).
Brazilian Archives of Biology and Technology 50, 979988.
doi: 10.1590/S1516-89132007000700009
Hoehne W, Grotta AS, ScavoneO(1952) Contribui coaoestudomorfolgico
e anatmico de Calea pinnatida Banks. Anais da Faculdade de
Farmcia e Odontologia da Universidade de So Paulo 10, 933.
Jensen WA (1962) Botanical histochemistry: principles and practice.
(WH Freeman: San Francisco)
Jeronymo AS, Paviani TI (1992) Anatomia da tuberosidade de
Vochysia thyrsoidea Pohl (Vochysiaceae). In Anais 8

congresso.
pp. 8390. (SBPC Brazilian Society for the Progress of Science:
So Paulo)
Johansen DA (1940) Plant microtechnique. (Mc Graw & Hill Book
Company: New York)
Joseph JP, Shah JJ, Inamdar JA (1988) Distribution, development and
structure of resin ducts in guayule (Parthenium argentatum Gray).
Annals of Botany 61, 377387.
Lersten NR, Curtis JD (1986) Tubular cavities in white snakeroot,
Eupatorium rugosum (Asteraceae). American Journal of Botany 73,
10161021. doi: 10.2307/2444120
Lersten NR, Curtis JD (1988) Secretory reservoirs (ducts) of two kinds in
giant ragweed (Ambrosia trida; Asteraceae). American Journal of
Botany 75, 13131323. doi: 10.2307/2444454
Lersten NR, Curtis JD (1989) Foliar oil reservoir and distribution in
Solidago canadensis (Asteraceae, tribe Astereae). Nordic Journal of
Botany 9, 281287. doi: 10.1111/j.1756-1051.1989.tb01003.x
Lotocka B, Geszprych A (2004) Anatomy of the vegetative organs and
secretory structures of Rhaponticum carthamoides (Asteraceae).
Botanical Journal of the Linnean Society 144, 207233.
doi: 10.1111/j.1095-8339.2003.00251.x
Luque R(2001) Cavidades secretoras en la raiz de Espeletiinae (Asteraceae).
Plantula 3, 19.
Luque-Arias R (2004) Estructura primaria del sistema radical de
Coespeletia Cuatrec. Interciencia 29, 1318.
doi: S037818442004000100007
Luque R, Menezes NL, Semir J (1997) La funcin secretora de la endodermis
de la raz de espcies de Lychnophora Mart. (Asteraceae). Plantula 1,
221228.
Machado SR, Oliveira DMT, Dip MR, Menezes NL (2004)
Morfoanatomia do sistema subterrneo de Smallanthus sonchifolius
(Poepp. & Endl.) H.Robinson (Asteraceae). Revista Brasileira de
Botnica 27, 115123. doi: 10.1590/S0100-84042004000100013
Meira RMSA (1991) Levantamento dos tipos de estruturas secretoras
em folhas de espcies de Asteraceae em vegeta co de oresta
(Atibaia SP). MSc Thesis, Universidade Estadual de Campinas,
Campinas, Brazil.
238 Australian Journal of Botany G. Cury and B. Appezzato-da-Glria
Melo-de-Pinna GFA, Menezes NL (2002) Vegetative organ anatomy of
Ianthopappus corymbosus Roque & Hind (AsteraceaeMutisieae).
Revista Brasileira de Botnica 25, 505514.
doi: 10.1590/S0100-84042002012000014
Melo-de-Pinna GFA, Menezes NL (2003) Meristematic endodermis and
secretory structures in adventitious roots of Richterago Kuntze
(MutisieaeAsteraceae). Revista Brasileira de Botnica 26, 110.
doi: 10.1590/S0100-84042003000100002
Metcalfe CR (1983) Secretory structures: cells, cavities and canals in leaves
and stems. Laticifers and latex. In Anatomy of the dicotyledons.
Wood, structure and conclusion of the general introduction.
(Eds CR Metcalfe, L Chalk) pp. 6497. (Clarendon Press: Oxford)
Metcalfe CR, Chalk L (1950) Anatomy of the dicotyledons: leaves, stem
and wood in relation to taxonomy with notes on economic uses,
(Clarendon Press: Oxford)
Metcalfe CR, Chalk L (1979) Anatomy of the dicotyledons. Systematic
anatomy of leaf and stem, with a brief history of the subject. (Clarendon
Press: Oxford)
Murphy DJ (2001) The biogenesis and functions of lipid bodies in animals,
plants and microorganisms. Progress in Lipid Research 40, 325438.
doi: 10.1016/S0163-7827(01)00013-3
Pagni AM, Masini A (1999) Morphology, distribution, and histochemistry
of secretory structures in vegetative organs of Santolina leucantha
Bertol. (Asteraceae). Israel Journal of Plant Sciences 47, 257263.
doi: 10.1560/GGYH-4404-QC86-KRHV
Pagni AM, Orlando R, Masini A, Ciccarelli D (2003) Secretory structures
of Santolina ligustica Arrigoni (Asteraceae), an italian endemic species.
Israel Journal of Plant Sciences 51, 185192.
doi: 10.1560/GGYH-4404-QC86-KRHV
Panizza S, Grotta AS (1965) Contribui co ao estudo morfolgico e
anatmico de Solidago microglossa D.C. Compositae. Revista da
Faculdade de Farmcia e Bioqumica da Universidade de So
Paulo 3, 2750.
Poli F, Sacchetti G, Bruni A (1995) Distribution of internal secretory
structures in Tagetes patula (Asteraceae). Nordic Journal of Botany
15, 197205. doi: 10.1111/j.1756-1051.1995.tb00143.x
Ragonese AM (1988) Canales secretores en los rganos vegetativos de
Eupatorium inulaefolium H.B.K. (Compositae). Acta Farmacutica
Bonaerense 7, 161168.
Robson NKB (1977) Studies in the genus Hypericum L. (Guttiferae)
1. Infrageneric classication. Bulletin of the British Museum (Natural
History, Botany series) 5, 291355.
Robson NKB (1981) Studies in the genus Hypericum L. (Guttiferae)
2. Characters of the genus. Bulletin of the British Museum (Natural
History, Botany series) 8, 55226.
Sacchetti G, Romagnoli C, Ballero M, Tosi B, Poli F(1997) Internal secretory
structures and preliminary phytochemical investigation on avonoid
and coumarin content in Santolina insularis (Asteraceae). Phyton 37,
219228.
Sakai WS (1973) Simple method for differential staining of parafn
embedded plant material using toluidine blue O. Stain Technology 48,
247249.
Solereder H (1908) Systematic anatomy of the dicotyledons. A handbook
for laboratories of pure and applied botany. (Clarendon Press: Oxford)
Tertuliano MF, Figueiredo-Ribeiro RCL (1993) Distribution of fructose
polymers in herbaceous species of Asteraceae from the cerrado.
New Phytologist 123, 741749.
doi: 10.1111/j.1469-8137.1993.tb03785.x
Tetley U (1925) The secretory system of the roots of the Compositae.
New Phytologist 24, 138162.
doi: 10.1111/j.1469-8137.1925.tb06657.x
Vilhalva DAA (2004) Morfo-anatomia de sistemas subterrneos de trs
espcies de Asteraceae do Cerrado do Estado de So Paulo. MSc
Thesis, Universidade Estadual de Campinas, Campinas, Brazil.
Vilhalva DAA, Appezzato-da-Glria B (2006) Morfo-anatomia do sistema
subterrneo de Calea verticillata (Klatt) Pruski e Isostigma
megapotamicum (Spreng.) Sherff Asteraceae. Revista Brasileira de
Botnica 29, 3947. doi: 10.1590/S0100-84042006000100005
Wagner H (1977) Pharmaceutical and economic uses of the Compositae.
In The biology and chemistry of the Compositae. (Eds VH Heywood,
JB Harbone, BL Turner) pp. 411435. (Academic Press Inc.: London)
Wilder GJ, Harris DH (1982) Laticifers in Cyclanthus bipartitus Poit.
(Cyclanthaceae). Botanical Gazette 143, 8493. doi: 10.1086/337274
Williams BC (1947) The structure of the meristematic root tip and origin
of the primary tissues in the roots of vascular plants. American Journal
of Botany 34, 455462. doi: 10.2307/2437260
Williams BC (1954) Observations on intercellular canals in root tips with
special reference to the Compositae. American Journal of Botany 41,
104106. doi: 10.2307/2439313
Manuscript received 1 August 2008, accepted 7 May 2009
Internal secretory spaces in thickened underground systems Australian Journal of Botany 239
http://www.publish.csiro.au/journals/ajb