PLANT PROPAGATION

Laboratory Manual

Copyright © 2000, 2001, 2002, 2003,

2004, 2006
Amended Fall 2006
- All Rights Reserved -
James B Calkins,
Ph.D.
CopyrCopyt © 2000, 2001, 2002, 2003, 2004, 2006
Emily E. Hoover,
– All Rights Reserved –

James B. Calkins, Ph.D.

Emily E. Hoover, Ph.D.

Fall Semester 2009

Department of Horticultural Science
College of Food, Agricultural, and Natural Resource
Sciences
University of Minnesota
 Table of Contents
I. Introductory Information
Page

A. Plant Growth Regulators & Other Chemical Compounds Used

in Plant
Propagation: General Information & Formulations.....................4

B. Sexual Propagation
1. Seed Propagation Techniques: Seed Collection, Handling, Viability,
Germination,
and Germination Requirements.........................................................7

2. Propagation By Seed: Description of Seeds, Collection, Cleaning,

Viability Tests, Storage, Germination, Dormancy, Overcoming
Dormancy, Environmental Factors That Influence Germination, Seed
Sowing, & Transplanting Seedlings....................................................9

C. Asexual (Vegetative) Propagation

1. Stem Cuttings / Herbaceous Species: Herbaceous Stem Cuttings and
Their
Rooting Requirements....................................................................15

2. Stem Cuttings / Woody Species: Softwood, Semi-Hardwood, and

Hardwood
Cuttings and Their Rooting Requirements......................................16

3. Leaf & Leaf-Bud Cuttings / Herbaceous & Woody Species: Leaf-Piece,

Leaf-Blade, Leaf-Petiole, and Leaf-Bud Cuttings and Their
Propagation Requirements...............................................................19

4. Root Cuttings / Herbaceous & Woody Species: Root Cuttings and

Their Cultural
Requirements.................................................................................20

5. Vegetative Propagation Using Specialized Structures: Bulbs, Corms,

Tubers,
Tuberous Roots & Stems, Rhizomes, Stolons, Runners, Layers, and
Crowns.............................................................................................21

6. Grafting and Budding: Whip & Tongue Grafts, Side Veneer Grafts,
Cleft Grafts,
 Approach Grafts, and Budding........................................................23

D. Propagation Ferns and Fern Allies: Spores and Their Cultural

Requirements.....................................................................................26

E. Media Effects on Propagation: Importance and Measurement

of
Growing Medium Porosity and Bulk Density Characteristics ......28
F. Experimental Design.......................................................................30
II. Laboratory Exercises (E = Experiment; P = Project; T = Technique)
Page Starting Date Report Due
#

Individual Reports [E]

Introduction to Experimentation: 32
Herbaceous Stem Cuttings [E]
Sexual Propagation: Using Scarification 35
to Overcome Hard Seed Coat Dormancy
[E]
Asexual Propagation: Adventitious Shoot 37
Initiation in Response to Treatment with
BA [E]
Asexual Propagation: Propagation of 39
Plants Using Leaf Cuttings [E]
Sexual Propagation: Effects of Seed Coat 42
Removal, Treatment with GA, Seed
Drying, and Cold Stratification on the
Germination of Apple Seeds [E]
Sexual Propagation: Effects of Growing 47
Medium on Seed Germination and
Seedling Performance [E]
Projects
Asexual Propagation: Propagation of 50
Herbaceous Garden and Landscape
Perennials Using Herbaceous Stem
Cuttings
Asexual Propagation: Designing an 53
Experiment to Maximize Rooting of Leaf-
Bud Cuttings
Sexual Propagation: Manipulation and 55
Germination of Seeds of Annual,
Herbaceous Perennial, and Woody
Perennial Plants
Let’s Go Grocery Shopping: Propagation 58
Produce
Techniques [T]
Sexual Propagation: Seed Morphology, 61
Viability Testing, Germination testing,
and Epigeal vs. Hypogeal Germination
[T]
Asexual Propagation: Whip & Tongue 64
Grafting [T]
Asexual Propagation : Unique Vegetative 69
Structures [T]
Asexual Propagation: T-Budding [T] 72
Asexual Propagation: Approach Grafting - 76
Spudmatoes [T]
Asexual Propagation: Air Layering [T] 79

HORT 1001 - Plant Propagation

Plant Growth Regulators & Other

Chemical Compounds Used In Plant
Propagation
General Information & Formulations

The term plant hormone is very specific and often misused. Plant hormones are
organic compounds, which are naturally produced in plants (endogenous) and
transported from their site of synthesis to their site of action where they regulate
physiological processes at low concentrations. Hormone biosynthesis itself (timing,
quantity, and physiological purpose including relationships to other growth
substances) is influenced by internal genetic factors and the environment. They
function to regulate plant growth and development by controlling the growth and
differentiation of cells, tissues, and organs. As a group, plant hormones are a
specific subcategory of a broader classification of plant growth regulatory
compounds called plant growth regulators (PGR's). In contrast to plant hormones,
plant growth regulators are applied to plants; they do not have to be translocated,
may require higher concentrations to elicit a response, and may be either naturally
or artificially synthesized. For this reason, when compounds having hormonal
effects are applied to plants exogenously they should be referred to as plant growth
regulators rather than hormones.

Plant growth regulators are widely used in plant propagation and in many cases
have revolutionized our ability to successfully propagate plants by increasing the
success in rooting cuttings and allowing greater manipulation of plant tissues in
micro propagation (tissue culture) systems. There are five primary types of plant
hormones and all are used as exogenously applied plant growth substances:
auxins, gibberellins, cytokinins, ethylene, and abscisic acid.
Auxins
Discovered in 1935, auxin compounds are commonly used in plant propagation to
induce the production of adventitious roots on cuttings and to control
morphogenesis in micro propagation systems (tissue culture). Along with the
development of mist systems, which slow respiration and reduce water loss
(transpiration) by cooling the cuttings and increasing humidity, the discovery of
auxin as a root promoter is one of the primary advances in the ability to propagate
plants from cuttings. Although a number of auxins are synthesized by plants,
indole-3-acetic acid (IAA) is the auxin most commonly produced. Auxins are
synthesized in leaf primordia, young leaves, and developing seeds and move
downward in plants. They are active in cell elongation, cell division, and cell
differentiation. The stimulation of cell elongation by auxin is involved in
phototropism (growth toward light) and etiolation (stretching in the absence of
light). Auxins are also implicated in the regulation of apical dominance (inhibition of
subordinate buds), cambial growth, flower initiation and development, fruit
development, and the promotion and inhibition of leaf, flower, and fruit abscission
(formation of abscission layers). Although also implicated in other processes such
as the induction of embryos in cell cultures, enhanced pollen tube growth, and tuber
and bulb development, the most important function of auxin in propagation systems
is its ability to promote the initiation of adventitious roots on cuttings.

Although it has the benefit of being soluble in water, indoleacetic acid (IAA) is
unstable and breaks down quickly when prepared and applied exogenously as a
plant growth regulator. It is destroyed when exposed to light (photooxidation) and
is broken down by enzyme systems within plant tissues. For these reasons,
synthetic auxins, such as indole-3-butyric acid (IBA) and naphthalene acetic acid
(NAA), which are more stable, are more commonly used in plant propagation. Like
IAA, IBA is also produced naturally in plants.

Auxins are available in both dry and liquid formulations. Dry formulations are talc
based and liquid formulations may be water or alcohol based. In general, both are
effective and easy to apply. The talc formulations are generally cheaper; they are
also easy to see on the ends of the treated cuttings. In comparison, liquid
formulations are easier to work with and tend to be more effective than talc
formulations, however, the potential for toxic effects is increased for some species
when alcohol based formulations are used.

Cytokinins
As a group, the cytokinins are believed to be involved in plant growth by stimulating
cell division, cell enlargement, and cell differentiation. In addition to kinetin, which
was the first cytokinin to be identified, a number of natural and synthetic cytokinins,
including zeatin, isopentenyladenine, and benzyladenine (BA), are known.
Cytokinins appear to interact with auxins, and this interaction is widely used in plant
propagation; a high auxin/cytokinin ratio promotes rooting, a high cytokinin/auxin
ratio promotes shoot development, and high levels of both at the same time results
in callus development. Cytokinins are also thought to slow plant senescence, inhibit
the breakdown of chlorophyll and proteins, and to help overcome apical dominance
and seed dormancy. The various responses associated with cytokinins likely involve
interactions with auxins and gibberellins.

Gibberellins
Gibberellins are believed to have many functions in plants. They are found at
relatively high concentrations in stem apices (especially in the leaf primordia),
roots, fruits, and tubers and are known to promote shoot growth through cell
division and elongation. A lack of gibberellin may be involved in dwarfed plants.
Gibberellins are also thought to be involved in the development and germination
(dormancy regulation) of seeds and the promotion of flowering. Over 100
gibberellins have been identified in plants. GA 3 and GA4+7 are the most common
gibberellins used in horticulture.

Abscisic Acid
The role of abscisic acid (ABA) as it affects plant growth is mixed. ABA has been
shown to have both stimulatory and inhibitory effects, which seem to be
concentration dependent. ABA is thought to be involved in the dormancy of buds
and seeds as well as plant responses to stress (especially drought). ABA is also
thought to be involved in stomatal regulation and water and ion uptake by roots. It
may also influence leaf and fruit abscission, but may not play the primary role.

Ethylene
Unlike the four groups of compounds previously discussed, ethylene is a gas under
standard conditions. It is thought to be involved in a variety of plant responses
including epinasty, leaf and fruit senescence and abscission, promotion of flowering,
and overcoming apical dominance (stimulation of lateral buds). Horticulturally, it is
used to promote fruit ripening, defoliation, fruit thinning, and flowering. Ethylene
has also been used to promote the initiation of adventitious roots and to overcome
seed and bud dormancy.

We will be using a variety of plant growth substances and other

compounds in class. Formulations and recipes for the preparations
most commonly used follow:

IBA (indole butyric acid) will be available as a root promoter in both dry (talc or
powder) and liquid) formulations. IBA will be available in a commercially prepared
talc formulation at concentrations of 1000, 3000, 8000, and 16,000 ppm (Hormex 1,
3, 8, & 16, respectively). IBA will also be available as a liquid formulation in
concentrations of 1000, 3000, 8000, and 16,000 ppm. For liquids, a quick-dip
method of application is used wherein cuttings are dipped for only 3 to 5 seconds.
Stock solutions are stable and can be kept indefinitely if they are kept clean; they
should also be kept tightly sealed to prevent evaporation and associated increases
in concentration.

NAA (naphthalene acetic acid) will be available as a liquid formulation in

concentrations of 1000, 3000, 8000, and 16,000 ppm. Although NAA is sometimes
more effective than IBA in promoting root development, it is also more likely to
cause injury to the cuttings; these effects are species specific. Stock solution
characteristics, storage requirements and cutting treatment procedures are the
same as for IBA.

NAD (napthylacetamide) is a synthetic auxin and will be available as a liquid

formulation. NAD does not induce as strong a reaction in plants as NAA. Although
NAD can sometimes cause injury to plant material, it tends to be more species
specific than NAA. Stock solution characteristics, storage requirements, and cutting
treatment procedures are the same as for IBA.

BA (benzyladenine) will be available as a liquid formulation at a concentration of

150 and 1500 ppm. Remember, benzyladenine is the most commonly used
cytokinin in the field of plant propagation and is used to promote the development
of adventitious shoots.

GA (gibberellic acid) will be available as a potential seed treatment for overcoming

dormancy in some seeds. A commercially prepared stock solution of GA 3 (Pro-Gibb)
will be diluted (stock = 4% = 40,000 ppm) to a concentration of 400 ppm for use in
class. Higher concentrations may also be used.

WRS (willow rooting substance) is simply a water-based extract of willow stems.

Although the mode of action is unknown, it can enhance the rooting of cuttings and
may be responsible for the ease of rooting common for willow. As a rooting
promoter, it remains unquantified and effectiveness may be variable from batch to
batch. The extract is stable for long periods if refrigerated. When using WRS,
cuttings are generally allowed to take-up the extract for some time prior to sticking.

TTC (2,3,5-triphenyl-2H-tetrazolium chloride), simply referred to as tetrazolium

chloride, is often used to test the viability of seeds. When metabolized by living
tissue, this colorless material is converted to an insoluble, red compound
(formazan). When applied to seeds and other plant tissues, this color change
indicates the presence of living tissue and is used as an indicator of viability.
Tetrazolium chloride solutions are prepared by dissolving TTC in water. A 1%
(10,000 ppm) solution is commonly used. Seeds are soaked for 2 to 24 hours.

H2SO4 (sulfuric acid) may be used as a pre-germination seed treatment to

scarify seeds having hard, impermeable seed coats and thereby promote
germination.

Note: ppm = parts per million; a commonly used unit of measure for concentration. It is the same as
milligrams per liter (mg/l); in other words, 1ppm = 1mg/l. Also, percent (%) multiplied by 10,000 =
ppm.
Always keep petri dishes containing liquid formulations of plant growth regulators covered when not in
immediate use to prevent evaporation and subsequent increases in the concentration of the growth
regulator.

HORT 1001 - Plant Propagation

Seed Propagation Techniques:

Seed Collection, Handling, Viability,
Germination Testing & Germination
Requirements

Seeds, reproductive organs composed of a protective covering, storage tissue, and

an embryo, are the most common form of reproduction for plants in the wild and a
common method of propagation for cultivated plants. In most, but not all, cases,
the propagation of plants from seeds is by definition sexual propagation since the
formation of most seeds involves the transfer of pollen from the male parent to the
female parent (pollination) and the subsequent union of male and female gametes
(fertilization) to form a single cell called a zygote. Fertilization occurs within
structures called ovules located within the female ovary. The zygote begins to
divide and eventually develops into an embryo (embryogenesis). As the embryo
develops, stored food reserves accumulate within the developing seed leaf or leaves
(cotyledons) or separately in the form of a tissue called endosperm. The developing
seedling utilizes the stored reserves during germination and establishment. At the
same time, the ovary, which may contain multiple ovules, develops into what is
botanically called the fruit. For some plants, seeds can develop in the absence of
fertilization from cells of the female parent; when this occurs, the process is called
apomixis; resulting seedlings are genetically identical to the mother plant.

In general, producing plants from seed is relatively cheap compared to other

methods of propagation. This is perhaps the greatest advantage of propagating
plants from seed. Seeds also provide a convenient and effective way to store plants
for a long period of time. Many seeds can be stored so they are on hand when
needed such as during times of the year when seed production and availability is
limited. Depending on your objectives as a propagator, there are other advantages
and disadvantages associated with the propagation of plants from seed.

One very important attribute associated with the propagation of plants from seed is
the genetic diversity, or variability, within seedling populations. This genetic
diversity is the cornerstone of natural selection in the wild and the improvement
(increased vigor, higher yield, increased nutritional value, greater environmental
adaptability, enhanced pest resistance, etc.) of domesticated plants for human use
through plant breeding and selection. From a grower’s standpoint, genetic diversity
and, therefore, plant variability may be looked upon as being both desirable and
disadvantageous depending on perspective. From a production standpoint,
uniformity is desirable as it simplifies the mechanics of production. If, however, the
plants being produced are destined for native plantings, native genetic diversity is
preferable regarding performance and survival.

Whether you are collecting seed yourself or purchasing seed from a supplier, a
reliable source of high quality seed is an important factor when propagating plants
from seed. So, too, is the handling of the seed following its collection or
procurement and during the period of germination. When purchasing seed, knowing
the geographic location (latitude, longitude, and elevation) where the seed was
collected or produced is important. The geographic locality from which the seed
was obtained is called provenance. If available, additional information about the
growing environment (soil type, pH, light, moisture conditions, etc.) is also of
interest. This information will tell you something about the potential for the
resulting plants to survive in a given environment. Knowing the source and
understanding the handling and germination requirements of specific types of seed
are critical to propagation success and the long-term performance of the plants
produced.

Seed germination is a remarkable and complicated process. You will find that seeds
differ greatly when it comes to their specific germination requirements. Some seeds
will germinate immediately without special treatment while others may require
manipulation and/or specific environmental conditions for germination to occur. For
example, seeds of most annuals, including vegetable seeds, will germinate quickly
when placed in a favorable environment (moisture, temperature, aeration, and
sometimes light). Such seeds are said to be quiescent. Seeds that are viable, but
fail to germinate when exposed to conditions favorable for germination are said to
be dormant. The seeds of many woody plants and some herbaceous perennials
exhibit some type of dormancy. Seed dormancy may be caused by a variety of
factors and is an evolutionary characteristic important in the survival of many plant
species. Overcoming dormancy, which may be physically and/or physiologically
induced, is often a challenge when propagating plants from seed. Pre-germination
treatments, such as scarification (breaking seed coats) and stratification (cool,
moist conditions), are often critical to success when propagating plants from seed.
In addition, providing conditions that are optimal for the germination of specific
seeds can often determine success. Research and/or hands-on experience are
important in determining the proper handling and germination requirements of
specific species. Even so, germinating some seeds can remain a significant
challenge even for the experienced propagator.

Some additional things to ponder and watch for include:

 When does germination begin?

 How long does it take for all of the seeds planted to germinate (at least those that
appear likely to do so)?

 When a seed germinates, what emerges first – the shoot or root?

 Is germination epigeous/epigeal or hypogeus/hypogeal?

 Is the plant to which the seed belongs a monocot (monocotyledenous) or a dicot

(dicotyledenous)?

 How does the final germination percentage compare with the viability test
results?

 What might be some reasons why certain seeds or groups of seeds did not
germinate?

Propagation by Seed
Description of Seeds, Collection, Cleaning, Viability Tests,
Storage, Germination, Dormancy, Overcoming Dormancy,
Environmental Factors That Influence Germination, Seed
Sowing, & Transplanting Seedlings

Excerpted from

Landscape Plant Propagation Workbook: Unit III. Propagation by

Seed
Dewayne L. Ingram
Sexual (seed) propagation is an important means of reproducing crop plants,
including landscape plants. Many seeds can usually be harvested from a plant; thus,
many plants can be propagated from a single mature plant. An orchid seedpod can
contain over 3 million seeds but 500,000 to 1 million seeds per pod is common in
many plant species. Annuals and biennials are most commonly propagated from
seed. Sometimes seedage is the only practical means of propagating a plant,
because vegetative means have not been explored or have been unsuccessful.

Seed propagation offers genetic variability; thus, the offspring may not have the
exact characteristics of the parent plant. This allows for the selection of plants with
unique features and systematic breeding for identified characteristics. Genetic
variability is a disadvantage if the goal of the producer is to grow a uniform crop.
Seedling variation is quite high in some plants, while other plants are more true to
type. Techniques using genetically stable lines or line hybrids are used to produce
many of the flowering annuals and vegetables with minimal variability.

Growers find that it takes longer to produce some plants from seed than from
cuttings. Seedlings may remain in a juvenile stage of growth for a longer period of
time. Juvenile growth does not produce flowers or fruit and such growth often has a
different appearance than mature growth.

Description of Seeds: A seed is composed of three basic parts: the embryo,

food storage tissue and the seed covering. The embryo is a new plant resulting from
union of male and female gamete during fertilization. Cotyledons, or seed leaves,
are attached to the embryo. Seeds are classified by the number of cotyledons.
Monocotyledons (monocots) such as grasses and palms have one cotyledon, while
dicotyledons (dicots) such as bean and peach have two. Plants such as pines are
gymnosperms and their seeds may contain as many as 15 cotyledons. A mature
viable seed contains enough stored food or energy source for seed germination and
early seedling growth. The cotyledons of dicots usually contain this food reserve,
while some seeds have a mass of food reserve called endosperm surrounding or in
contact with the embryo (e.g., corn). Seed coverings can consist of the seed coat
and parts of the fruit or seedpod. These structures protect the embryo and food
reserve inside the seed. They can also inhibit germination until conditions are
suitable for germination and seedling development.

Seed Collection

Seeds should be collected when ripe and just before they fall to the ground. When
seeds are on the ground, the likelihood of disease and insect infestation is greatly
increased. Some seeds loose viability (ability to

germinate) soon after they are ripe and should not be collected from the ground.
Several of the tropical plants have seeds with short periods of viability. Generally,
palm seeds fit into this category.
Seed maturity is often difficult to determine. Generally there is a color change of the
fruit from a green to an orange, black, purple or red, and the fruit may become soft.
Winged seed, seedpods or fruit without fleshy pulp may turn a brownish color as
they mature and become drier. This water loss or drying coincides with seed
maturity and is a state in which the seed may be stored or distributed by wind or
water.

Seed Cleaning

The pulp from a fleshy fruit may contain germination inhibitors and should be
removed before seeds are sown. The seed would probably germinate after the fruit
decomposes, but the propagator may not want to wait several weeks and the
process may reduce germination. Seeds are usually removed from dried capsules
before treated or germinated.

Seeds may be cleaned individually or in bulk. Seeds may be harvested from dried
capsules, or the capsules can be crushed, if the seed is sufficiently hard, and sown
with the seeds. The flesh around a seed can be removed by hand or by some
mechanical means. A procedure for cleaning several seeds at one time has been
developed by Dr. Bijan Dehgan in the Department of Environmental Horticulture at
the University of Florida. The procedure was developed for cleaning cycad seeds but
will work with many seeds. The general procedure is as follows: The fleshy coat is
removed by using a long-stemmed, circular wire brush attached to a variable speed
drill. First, seeds should be soaked in water for 24 hours. Next, place seeds in large
wide-mouth jar or a large coffee can with a plastic top in place, through which the
stem of the wire brush is passed. Add a small amount of 20-mesh sand to the seeds
and enough water to cover the sand and seeds. Turn on the drill at a slow speed and
gradually increase the speed until all seed coats are removed. Wash the seeds
thoroughly. A lot of 300 to 400 seeds can be cleaned in approximately 10 to 15
minutes.

Seed Viability Tests

It is often difficult to determine from the appearance of a seed if it is alive and

would germinate under the proper conditions. There are methods of determining
seed viability or potential for seed germination. Stains can be used to determine if
the seed embryo is alive. Living tissue respires in order to maintain its integrity and
tetrazolium chloride turns red when it comes into contact with respiring tissue. The
seed is cut open and the stain applied to the exposed embryo. If the embryo is
alive, it will turn reddish in color.

Viable and dead seeds in some plant species can be separated by floating the seed
in water. When the seed is filled with the embryo and endosperm to support
germination and early seedling growth, the seed will sink in water. When a seed is
empty or contains a relatively large amount of air, the seed will float. This is a good
means of identifying dead oak and pindo palm seeds, but it does not work with
Zamia floridana, Cycas cercinalis, coconut, fish poison tree and several other plants.
Seed Storage

Seeds of many plants can be stored and germinated weeks, months or years later.
Factors affecting the viability of seeds during storage include (1) the inherent
longevity of the plant species, (2) seed moisture content, (3) the temperature and
(4) the relative humidity. The length of time seeds can be stored differs with the
plant species. Magnolia, wax myrtle and elms have short-lived seeds and may be
stored only for a few weeks or maybe up to a year without losing viability.
Koelreuteria, Calendula, Petunia and Zinnia are examples of seeds that can be
stored for 2 to 15 years, and Acacia and Elaeagnus can be stored for 15 to 20 years.
Research is being conducted with freezing techniques that could increase the length
of time seeds can be stored.

The moisture content of the seed should be reduced to 20 to 30 percent of seed

weight before storage. This can be accomplished by placing them in the sun or in a
drying oven. Obviously, if the seed becomes too dry, the embryo will die.

Many seeds will store best if a temperature 35 to 45F is provided. However, some
of the tropical plants will not tolerate temperatures below 40F. Generally, seeds
from tropical plants do not store well.

The relative humidity in the storage container should be 20 to 30 percent. The

moisture in the air and the seed will become equal over time in the container;
therefore, the environment in the container should be relatively dry and the seeds
should be dried somewhat before storage. Plastic bags or bottles work well as
storage containers.

Seed Germination

Although the specific characteristics of seeds differ with plant species, the general
germination process is the same. The germination process can be described in three
phases: activation, digestion and cell division, and elongation. The process begins
with the imbibition or uptake of water. This may take a few hours or several days,
and the seeds will usually swell or enlarge due to the increased volume. Water
uptake triggers the activation of certain biochemical processes that result in the
synthesis of the building blocks (nucleic acids, amino acids, enzymes, etc.) for
growth and development. The endosperm is digested by enzymes to produce
energy-rich compounds and nutrients that are transported to the embryo. Cell
division and elongation occurs and the developing embryo expands. The radical or
root emerges from the seed by breaking through the seed coat.

Dormancies

Many seeds are ready to germinate when the fruit is ripe or the capsule is dried.
Seeds from tropical plants usually are not dormant when mature, but many of the
plants from temperate climates are dormant when collected. Dormancies are
protective mechanisms that have evolved to allow a seed to germinate at the
appropriate time. For example, dogwood seed mature in late fall but are dormant at
that time. These seeds require a cold period to satisfy some factor inside each seed
before it will germinate. If dogwood seeds were germinated in the fall, the tender
seedling would probably not survive the winter. Therefore, the cold requirement
facilitates natural germination in the spring at the beginning of the growing season.

Seed dormancy can be caused by a hard seed coat that is impermeable to water or
gases, or resistant to embryo expansion. Zamia, camellia and redbud are examples
of seed dormancy due to an impermeable seed coat. Seeds from many of the stone
fruits, such as peaches, are dormant due to physical restriction of seed expansion.

The fleshy pulp, the endosperm or the seed coat can contain chemical inhibitors of
germination. Some of these inhibitors are water-soluble and can be leached out of
the seed, but many cannot be washed out and must be broken down by some
chemical process. Okra seeds contain a water-soluble inhibitor that can be leached
by soaking the seeds in water for 12 to 24 hours.

The embryo may be immature when the fruit is ripe or the capsule is dry. A period of
time in the proper environment is required before germination is possible. The
embryo will often increase in size during this period and will develop the properties
necessary for germination. Orchid seeds are dormant when released by the plant,
due to an immature embryo.

Some seeds have double or multiple dormancy: more than one type of dormancy. An
immature embryo may prevent germination of a seed, even though a dormancy due
to a hard seed coat has been overcome. A seed may require a cold period to break a
dormancy due to the seed coat and then require a warm period for development of
an immature embryo. Nandina seeds require a cold-warm-cold sequence of
temperatures before germination.

Even though some seeds will germinate without a preconditioning period, the
germination rate and uniformity might be enhanced by some treatment. This
condition is referred to as a physiologically intermediate dormancy.

Overcoming Seed Dormancies

Scarification and stratification are the two most common means of breaking seed
dormancy, but hot water soaks and plant growth regulator treatments are also used.
Scarification involves the breaking of the impermeable or hard seed coat. This may
be accomplished by mechanical means or by acid treatments. Scratching the seed
coat with sand paper and cutting the seed coat with a knife are suitable for some
seeds. A hammer or pliers can be used to break the seed coat in some cases. Care
must be taken in any scarification procedure not to injure the seed embryo. If a
specific treatment for a particular plant species is not known, sample seed lots
treated with various techniques followed by germination tests is recommended.

Acid scarification has been used successfully in some plants to break the seed coat.
Such scarification occurs in nature when the seed coat is partially digested by fungi
or bacteria or enzymes in the digestive tract of birds and animals. Sulfuric acid is
usually used for scarification. Seeds may be soaked for 5 to 60 minutes, depending
upon the toughness of the seed coat and the sensitivity of the embryo. Dry, clean
seeds are placed in a nonmetal, noncorrosive, acid-resistant container, and the acid
is added slowly in a ratio of one part seed to two parts acid. The mixture should be
gently stirred intermittently during the soak to ensure uniform results. Seeds soaked
in acid should be washed thoroughly under running water for 10 minutes to dilute
the acid in contact or inside the seeds before they are sown.

Acids are extremely dangerous and proper handling procedures must be followed.
Never pour water into acids, because tremendous heat can be generated in a
violent reaction. Dilute acids by pouring the acid into water very slowly. Acid-
resistant clothing including gloves should be worn when handling acids. Acids
should be disposed of properly as hazardous material and should never be poured
down the sink.

Stratification is the technique of providing a moist chilling treatment to seeds to

overcome a dormancy. This technique has been used to overcome dormancies due
to an impermeable seed coat or chemical inhibition. Stratification could reduce the
concentration of germination inhibitors or increase the concentration of growth-
promoting hormones. Seeds should be soaked for 12 to 24 hours and then placed in
a moist medium. Coarse vermiculite, sphagnum peat moss, equal volumes of peat
and perlite, and coarse sand have proven to be good stratification media that retain
moisture yet allow aeration. Seeds should be mixed with one to three times their
volume of medium and stored in a container that provides a barrier to moisture loss,
such as a polyethylene bag. Seeds can be stratified naturally by placing them
outdoors in an area or container protected from rodents, but the temperatures
outdoors may not be uniformly low enough in Florida to provide optimum results.

Seeds are usually stratified at 32 to 40 F (0 to 2 C) for a period of 1 to 4 months.

Seeds are then separated from the stratification medium and germinated at 70 to
80 F (17 to 22 C). Germination at higher temperatures (>90 F; >27 C) may
induce a secondary dormancy.

Soaking in water will often soften seed coats and leach water-soluble inhibitors from
the seed to reduce germination time. Best results are obtained when hot water is
used, but the temperature sensitivity of the embryo differs with species. Generally,
water at 170 to 212 F (67 to 88 C) is poured over seeds in a ratio of one part seed
to four or five parts water and allowed to cover the seed and cool for 12 to 24
hours. Changing the water periodically during prolonged soaking is imperative.
Seeds should not be allowed to dry after the treatment but should be sown
immediately.

Environmental Factors

An ample supply of high quality water must be applied during seed germination and
seedling development. There must be a proper balance between water and air in
the propagation medium. Waterlogged media cannot supply the oxygen necessary
for germination and seedling growth. Monitoring the moisture level in the medium is
important. Water with excessive dissolved salts can result in poor seed germination
and seedlings growth. Seeds and seedlings differ in their sensitivity to salt levels,
but generally the seedlings are less tolerant than mature plants.

The maximum, minimum and optimum temperatures for seed germination and
seedling growth differ with plant species and maybe even cultivars within species. If
the optimum temperature for a seed is not known, the range of 75 to 80 F would be
optimum for many plants and should be considered. Keep in mind that plants native
to warmer climates or those that flourish during the warmer months may have
higher optimum temperatures than plants that flourish in cooler temperatures.
Reducing night temperatures 5 to 10 F (3 to 5 C) has proven beneficial in
germination of some plants native to temperate climates.

Seeds can be grouped according to their requirement of light for germination. Many
woody plants do not require light for seed germination, but most epiphytes such as
mistletoe and strangler fig require light. Other species have a light requirement for
germination, but this can be overcome by chilling or chemical treatments as in
lettuce, tobacco and many native weed seeds. Allium, Amaranthus and Phlox are
examples of seeds whose germination is inhibited by light. Some species such as
Tsuga and Betula are sensitive to day length, but sometimes this can be overcome
by temperature treatments.

Emerging roots from a seed can absorb nutrients. The presence of nutrients in low
to moderate concentrations at this time will result in more rapid seedling growth.
Care should be taken to avoid excessive dissolved salt levels, which will injure such
tender root tissue. Application of soluble fertilizers on a periodic basis is
recommended over incorporation of fertilizers in the propagation medium before
the seeds are sown.

Seed Sowing

Germinating seeds in a controlled environment such as a greenhouse and in a

sterile medium will result in optimum seed germination and early seedling
development. Problems with weed seeds; nematodes, insects and diseases can be
greatly reduced with such procedures. Plastic, wood or metal flats are common
containers for seed germination, although individual containers, celled flats and
preformed peat pellets are also used. Seeds may be germinated in a flat and then
the most vigorous seedlings can be transplanted to a larger production container,
or, under certain circumstances, the seeds may be sown in the production container
in which they will be sold.

The germination medium should be well drained but should hold sufficient moisture
to maintain optimum moisture in the seed. The particle size of the propagation
medium components is important, because this primarily determines moisture and
aeration characteristics of the medium. The particle size must also be considered in
relation to the size of the seed to be sown in the medium. There must be sufficient
contact between the seed and the particle for exchange of moisture. A large seed
can be germinated in a medium with relatively large particles, but a small seed
would settle toward the bottom of such a medium. A small seed should be
germinated in a medium with relatively small particles to provide an appropriate
contact between the seed and the particle.

The proper planting depth differs with seed. Seeds that require light for germination
obviously cannot be planted deeply or may not be covered at all. Generally, seed
should not be planted deeper than two or three times their diameter. Many large
seed, especially tropical species like palms, may only be inserted into the medium
surface where they will remain moist.

Transplanting Seedlings

Seedlings should be transplanted before they over-grow a container and their

growth habit or form is altered. Seedlings produced in flats at a high density
obviously must be transplanted earlier than seedlings produced in larger containers
at a low density. Seedlings must be hardened before transplanting. Hardening refers
to a gradual change in the environment so the seedlings can adapt to withstand
more stressful conditions than those provided in the propagation phase. The
irrigation frequency is usually reduced and the light level and fertilization may be
increased. Proper hardening will ensure that the seedlings are established in the
production environment at an optimum rate.

This document is excerpted from Circular 725, Florida Cooperative

Extension Service, Institute of Food and Agricultural Sciences,
University of Florida. Reviewed: April 1993. Dewayne L. Ingram,
former Professor, Environmental Horticulture, Cooperative Extension
Service, Institute of Food and Agricultural Sciences, University of
Florida, Gainesville FL 32611.

Copyright Information: This document is copyrighted by the University of

Florida, Institute of Food and Agricultural Sciences (UF/IFAS) for the people of
the State of Florida. UF/IFAS retains all rights under all conventions, but
permits free reproduction by all agents and offices of the Cooperative
Extension Service and the people of the State of Florida. Permission is
granted to others to use these materials in part or in full for educational
purposes, provided that full credit is given to the UF/IFAS, citing the
publication, its source, and date of publication.
 HORT 1001 - Plant Propagation

Stem Cuttings / Herbaceous

Species:
Herbaceous Stem Cuttings & Their Rooting
Requirements

Herbaceous stem cuttings are commonly used to propagate a wide variety of

succulent, non-woody (herbaceous) plants. In general, they are easy to root. Many
species of house plants, bedding plants, and native and introduced herbaceous
landscape perennials are easily propagated using herbaceous stem cuttings.
Examples of species that are commonly propagated from herbaceous stem cuttings
include geranium (Pelargonium spp.), begonia (Begonia spp.), chrysanthemum
(Dendranthema  grandiflora), garden phlox (Phlox paniculata), dieffenbachia
(Dieffenbachia spp.), and dracaena (Dracaena spp.).

As always, when preparing herbaceous stem cuttings, care should be taken to select
healthy, well-conditioned plants. Stock plants are often cut back to force increased
numbers of new shoots to be used as cuttings. Cuttings should be kept cool and
prevented from drying during transit and prior to sticking. Herbaceous stem
cuttings are typically 5-15 cm (2-6") long and the leaves are typically removed from
the lower portion of the cutting prior to sticking. Cuttings are typically collected by
making a cut immediately subtending (below) a node. While treatment with auxins
is generally not required for rooting and usually does not increase rooting
percentages for herbaceous stem cuttings, they can improve rooting uniformity for
some species. During rooting the cuttings are maintained under humid conditions
(mist or fog) to reduce transpiration and prevent drying until the cuttings are well
rooted. Cuttings from some herbaceous species will, however, root under less
controlled conditions; e.g., remember Grandma rooting cuttings in water in the
kitchen window? Most herbaceous stem cuttings are collected from terminal shoots,
but cuttings may be collected from both terminal and lower portions of the stems.
Flowers that are present or appear later should be removed so that all energy
(photosynthate) is exclusively utilized for the initiation and production of roots.

Polarity is another consideration when preparing herbaceous stem cuttings or, for
that matter, any type of cutting. Polarity has to do with maintaining the orientation
of the cutting relative to its orientation on the source plant; in other words, up vs.
down (distal vs. proximal). Polarity is an inherent condition wherein the cutting
responds differently on one end than the other. For example, stem cuttings tend to
form roots on the basal end relative to their orientation when collected from the
source plant. When cuttings are stuck upside down, they still tend to form roots on
what was the proximal end even though polarity has been reversed.

20
 HORT 1001 - Plant Propagation

Stem Cuttings / Woody Species:

Softwood, Semi-Hardwood, and Hardwood Cuttings

&
Their Rooting Requirements

Softwood & Semi-Hardwood Cuttings

Softwood cuttings, also called summer softwood or greenwood cuttings, are
collected during late spring and early summer when the new growth of woody
plants is still "soft" or unlignified. New wood remains "soft" for a period of 2 to 8
weeks depending on species and environmental conditions. For softwood cuttings,
the source plants are still actively growing and shoots have not set terminal buds.
Leaves near the tips of the growing shoots are not completely expanded. Timing is
important; the wood must not be too succulent or too woody. Cuttings collected
when the wood is too soft will often rot while those collected too late may root
poorly or not at all. For some species, timing is very critical regarding success with
softwood cuttings. For example, lilac (Syringa spp.) cuttings must be collected and
stuck within a very short time window just prior to terminal bud set to root reliably
and in sufficient percentages to be profitable. Softwood cuttings generally root
easier and quicker than semi-hardwood and hardwood cuttings, but, since the
cuttings are soft and tender, they also require more care during collection, handling,
and rooting. Softwood cuttings are widely used in the propagation of many
deciduous landscape shrubs and some trees and a variety of woody tropical and
subtropical plants some of which are used as houseplants. Examples of species
typically rooted from softwood cuttings include honeysuckle (Lonicera spp.), lilac
(Syringa spp.), rose (Rosa spp.), spirea (Spiraea spp.), and viburnum (Viburnum
spp.).

Semi-hardwood cuttings are collected during late summer and early fall and, owing
to the presence of leaves, are similar in appearance to softwood cuttings. As the
name implies, however, the condition of the wood (soft vs. hard or mature) falls
somewhere between that of softwood and hardwood cuttings. At the time of
collection, stock plants are still in leaf although terminal buds have generally been
set and the leaves along the full length of the stems are fully expanded. The wood
has not completely matured, but is no longer soft and pliable as with softwood
cuttings. Many broadleaf evergreens, and several woody tropical and subtropical
species, are propagated using semi-hardwood cuttings. Although less common, a
number of deciduous species can also be propagated from semi-hardwood cuttings.
Examples of species typically rooted from semi-hardwood cuttings include
euonymus (Euonymus spp.), evergreen azaleas (Rhododendron spp.), evergreen
hollies (Ilex spp.), cliff green (Paxistima canbyi), and boxwood (Buxus spp.).

When preparing softwood and semi-hardwood cuttings, care should be taken to

21
select healthy, well-conditioned plants. Cuttings should be kept cool and moist
during transit and prior to sticking. Since the cuttings have leaves, special care
must be taken to prevent drying out during transit and storage and the cuttings
must be rooted under humid conditions (mist or fog) to reduce transpiration and
prevent drying until the cuttings are well rooted. For some species, polyethylene
tents with occasional sprinkling are sufficient to control transpiration. Bottom heat
can be helpful in the spring if temperatures are cool and treatment with plant
growth substances may be helpful in promoting rooting. Cuttings are typically
collected from terminal growth, should be 8 to 15 cm (3-6") long and should have 2
or more nodes. The basal cut is usually made just below a node. Leaves should be
stripped from the basal portion of each cutting prior to sticking. If flowers or flower
buds are present, they should be removed so that all energy (photosynthate) is
exclusively utilized for the initiation and production of roots.
Several labs will investigate the difference between softwood and semi-hardwood
cuttings. The objective of these labs is the production of entire plants from
softwood and semi-hardwood cuttings and to compare rooting for these two types of
cuttings. By the time classes begin in the fall, we are way beyond the optimal time
to collect and stick softwood and most semi-hardwood cuttings. We will, therefore
be using container grown plants that have been maintained under conditions that
will keep them from going dormant. In this way we can provide plants with new
growth suitable for use as softwood cuttings and semi-hardwood cuttings. Observe
your cuttings at least weekly and record your observations.

Hardwood Cuttings
Hardwood cuttings can be successfully used to propagate a number of woody trees,
shrubs, and vines. The wood used is completely mature or woody and the cuttings
are typically collected from dormant plants in late fall, winter, or early spring.
Exactly when the cuttings are collected may influence how the cuttings are handled
as an artificial cold treatment may be needed to initiate shoot growth for cuttings
collected prior to exposure to sufficient chilling (a period of exposure to
temperatures just above freezing required to overcome dormancy). Many
deciduous and narrow-leaved evergreen species and some broad-leaved evergreens
are propagated using hardwood cuttings. Examples of species typically propagated
from hardwood cuttings include honeysuckle (Lonicera spp.), spirea (Spiraea spp.),
forsythia (Forsythia spp.), grape (Vitis spp.), currants and gooseberries (Ribes spp.),
juniper (Juniperus spp.), white cedar (Thuja spp.), false cypress (Chamaecyparis
spp.) and yew (Taxus spp.).

As with any propagation technique, when preparing hardwood cuttings care should
be taken to select healthy, well-conditioned plants. Although drying-out is of
considerably less concern than with other, more succulent, types of cuttings,
hardwood cuttings are best kept cool and should be prevented from drying during
transit and prior to sticking. Cuttings that are stuck in early spring may be collected
just prior to sticking or they may be collected in the fall and stored outdoors or
indoors under more controlled conditions (cool temperatures and moist conditions
to prevent drying). In areas with severe winters, collection and storage of cuttings
in the fall or early winter, prior to the onset of potentially lethal winter temperatures
is advisable. This is especially true for species that are not reliably cold hardy.

For most species native to north temperate regions, exposure to a period of cool
22
(34-38F) temperatures is required to overcome dormancy and initiate bud break
and the resumption of normal growth in the spring. For such species, the wood
used to prepare hardwood cuttings must also be exposed to such conditions. This
cold treatment occurs naturally during the winter for cuttings collected in the late
winter or early spring or is provided artificially through refrigerated storage for
cuttings collected in the fall or early winter. In some cases, prepared cuttings are
stored under relatively warm conditions for a time to induce callus formation prior to
sticking to hasten rooting. Remember that depending on species and the time of
collection, such cuttings may also require a period of chilling, either before or after
callusing, to overcome dormancy. For some species, hardwood cuttings may be
collected and stuck directly in outdoor fields or beds in the fall or early spring.
While the practice is used to a limited extent in northern climates such as
Minnesota, it is quite common in warmer areas such as the central and southern
U.S.

Hardwood cuttings are typically rooted under humid conditions (mist or fog) to
reduce transpiration and prevent drying until the cuttings are well rooted. In some
cases, shading the cuttings, humidity tents, regular sprinkling, or a combination of
these methods can provide sufficient reduction in water loss. Bottom heat,
wounding, and treatment with plant growth substances are often helpful in
promoting rooting of hardwood cuttings. The concentrations of rooting promoters
used are typically higher than those used for softwood and semi-hardwood cuttings.
Cuttings may be collected from both terminal and lower potions of the stems. Age
of the stem may affect rooting success. Cuttings that include a short section of
stem from older wood are called mallet cuttings while those that include only a
small piece of older wood torn from the adjacent stem are called heel cuttings.
Hardwood cuttings usually have 2 or more nodes and may be anywhere between 10
and 76 cm (4-30") long. The basal cut is usually made just below a node. In the
case of hardwood cuttings, which initially are void of leaves, much of the success is
dependant on the stored food reserves, mainly carbohydrates, present in the
cutting. If flower buds are present and are recognizable, they should be removed,
along with any flowers that appear following bud break, so that all energy (stored
carbohydrate and new photosynthate) is exclusively utilized for rooting.

Cutting orientation is very important with hardwood cuttings. Because the leaves
are not present, make sure to observe the buds. In all cases, the buds should be
pointing upright to assure correct polarity (up vs. down relative to the orientation of
the cutting on the stock plant).

23
 HORT 1001 - Plant Propagation

Leaf & Leaf-Bud Cuttings /

Herbaceous &
Woody Species:
Leaf-Piece, Leaf-Blade, Leaf-Petiole, and
Leaf-Bud Cuttings & Their Propagation
Requirements

Herbaceous Species: Leaf-Piece, Leaf-Blade, and Leaf-

Petiole Cuttings
Some plants can be propagated using leaf cuttings. Several different types of leaf
cuttings are used as determined by the amount of leaf tissue used. They include
leaf-blade cuttings, which consist of the leaf blade only, leaf-petiole cuttings, which
include the leaf blade and the attached petiole, and leaf-piece cuttings, which are
comprised of only pieces of leaves. Although somewhat species dependent, the
potential to produce many new daughter plants from a limited number of stock
plants, or even a single stock plant, is very high when leaf cuttings are used owing
to the small size of the propagules involved. Since no buds or roots are present on
the cuttings, successful propagation from leaf cuttings requires the development of
both adventitious roots and shoots. The portions of the leaf used for propagation
purposes generally do not remain as a permanent part of the new plants produced.
Depending on species, plant growth substances have had mixed results on
propagation success with leaf-piece, leaf-blade, and leaf-petiole cuttings. Many
species, mostly herbaceous, that are used as houseplants or bedding plants can be
propagated from leaf cuttings. Some examples of species that can be propagated
from various types of leaf cuttings include begonia (Begonia spp.), Cape primrose
(Streptocarpus spp.), African violet (Saintpaulia spp.), snake plant (Sansevieria
spp.), and peperomia (Peperomia spp.).

Herbaceous & Woody Species: Leaf-Bud Cuttings

While they may root, some plants are unable to initiate adventitious shoots from
leaf-piece, leaf-blade, or leaf-petiole cuttings and the rooted cuttings, therefore,
never develop into new plants. Many of these plants can, however, be propagated
easily from cuttings if the propagule includes a preformed shoot meristem. They
can, for example, typically be propagated from stem cuttings. For some of these
plants, a cutting that includes the node portion of the stem along with the attached
leaf is all that is needed to be successful. Such cuttings, consisting of the leaf
blade, the petiole, the node, and the associated axillary bud, are called leaf-bud
cuttings or single-node cuttings. The bud portion of the cutting contains the new
24
shoot primordia and only roots need to be initiated to produce a new plant. Many
species, including most herbaceous species that are used as house or bedding
plants, can be propagated from leaf-bud cuttings. Leaf-bud cuttings are also used to
commercially propagate a number of woody species. Some examples of species
that can be propagated from leaf-bud cuttings include peperomia (Peperomia spp.),
geranium (Pelargonium spp.), rhododendron (Rhododendron spp.), red maple (Acer
rubrum), clematis (Clematis spp.), and blackberry (Rubus alleghaniensis).

As always, leaf-piece, leaf-blade, leaf-petiole, and leaf-bud cuttings should only be

collected from healthy, well-conditioned plants. The cuttings should be kept cool
and prevented from drying during transit and prior to sticking. The cuttings are
maintained under high humidity (usually under mist or in fog houses) to reduce
transpiration until the requisite roots and shoots are produced.

25
 HORT 1001 - Plant Propagation

Root Cuttings / Herbaceous &

Woody Species:

Root Cuttings & Their Cultural Requirements

For some plants, root pieces can be used to vegetatively propagate new plants.
True root cuttings must be distinguished from rhizomes, which, although they are
often mistaken as roots and can certainly be used as propagules, are by definition
modified, underground stems. The common spice we know as ginger (Zingiber
officinale) is a good example. Although it is called ginger root, it is a rhizomatous,
tropical herb and a good example of a plant that can be easily propagated from
rhizomes, but not true roots. Rhizomes have nodes and associated bud initials.
Root cuttings do not have nodes and must initiate adventitious shoots if they are to
be used as a method of propagation.

When compared to most other methods of propagation, propagation using root

cuttings is relatively uncommon. The list of plants that have the capacity to initiate
shoots from root tissue is limited and, because roots are located below ground, root
cuttings are more difficult to collect. For most plants, other, more effective and
economical propagation methods are available. In certain cases and under certain
circumstances, however, root cuttings are the method of choice.

Depending on species, root cuttings may be collected at various times during the
year. Most are collected during late summer and early fall. While root cuttings can
also be collected during the winter, unless the soil has been prevented from
freezing by mulching heavily, frozen ground poses a problem in colder regions. As a
general rule, any plant that suckers from the roots is a likely candidate for
propagation from root cuttings. Some examples of species that can be propagated
from root cuttings include sumac (Rhus spp.), garden phlox (Phlox paniculata),
devil's walking stick (Aralia spinosa), Oriental poppy (Papaver orientale),
horseradish (Armoracia rusticana), sweet fern (Comptonia peregrina), and red
raspberry (Rubus ideaus).

As with other types of cuttings, root cuttings should only be collected from healthy,
well-conditioned plants. The root cuttings should be kept cool and prevented from
drying during transit and prior to sticking. Unlike stem cuttings, where polarity
(direction of growth; up vs. down) can usually be easily determined from the
orientation of the buds, when working with root cuttings it is important to indicate
proper polarity in some fashion at the time of collection. Knowing polarity is
generally of greater concern for plants with large, fleshy roots than those with thin,
delicate roots. To this end, distal ends of root cuttings are typically cut on the bias
while the proximal ends are given a flush cut. Cuttings are typically 5 to 20 cm (2-
8") long and will vary greatly in thickness depending on species (actual length is

26
often correlated with diameter; shorter cuttings for thicker roots and vice versa).
Cuttings from plants with large fleshy roots are either stuck upright or inclined on
their sides, proximal end up, in shallow trenches cut into the rooting medium. The
cuttings are then covered with medium such that their tops are just below the
surface. Cuttings from plants with thin, fibrous roots are typically planted
horizontally just below the surface. The cuttings are kept moist and maintained
under optimal conditions until shoots and additional roots are produced.

27
 HORT 1001 - Plant Propagation

Vegetative Propagation Using

Specialized
Vegetative Structures
Bulbs, Corms, Tubers, Tuberous Roots & Stems, Rhizomes,
Stolons, Runners, Layers, and Crowns

Specialized vegetative structures such as bulbs, corms, tubers, tuberous roots and
stems, and pseudobulbs are mainly associated with vegetative propagation of
herbaceous species. Rhizomes, stolons, runners, layers, and crowns may be used
to propagate both herbaceous and woody species. Some of these structures,
including bulbs, corms, tuberous roots and stems, and some rhizomes, function
primarily in food storage during dormant periods in addition to serving a
reproductive function. Rhizomes (modified, underground stems) and stolons
(horizontal stems that creep across the ground) root and produce shoots at the
nodes and can serve both storage and reproductive functions. These vegetative
structures can be quite short, resulting in clumps, or distinctly elongated resulting in
rather large colonies. A runner is a specialized type of stolon, which roots and forms
a new plant at its tip when it contacts the soil. In addition to serving a storage
function, bulbs (tulip, lily) and corms (crocus, gladiolus), produce naturally
detachable offshoots called offsets, which become or can be artificially separated
from the parent structure (a form of division). This natural form of reproduction can
also be exploited commercially by inducing the formation of offsets, which are then
removed from the parent structure and grown on to produce new plants.

Scooping, Scoring, Chipping, Scaling, & Twin Scaling

Bulbs are produced by monocotyledonous plants and function in storage and
reproduction. Their structure is defined by a short stem terminated by a growing
point or flower primordia enclosed by thick, fleshy, modified leaves or scales. Bulbs
and may be classified as being either tunicate (also referred to as laminate) or non-
tunicate (also referred to as scaly). Tunicate bulbs are protected by a layer of dry,
membranous bulb scales called the tunic. The inner scales are arranged in tight,
contiguous layers or lamina resulting in a rather solid structure. A basal plate can
be found at the base of each bulb and it is from here that new roots develop
annually. Onion is perhaps the most recognizable example of a tunicate bulb. Tulip
and daffodil are also examples of tunicate bulbs. Non-tunicate, or scaly, bulbs lack
the outer layer of dry scales and the fleshy scales are clearly visible and distinctly
individual. Having a less solid structure, and lacking the tunic, scaly bulbs are more
easily damaged and susceptible to drying compared to tunicate bulbs. Roots are
also formed at the base of scaly bulbs each year, but they live for two seasons.
Lilies are the best examples of scaly bulbs.

28
Scooping, scoring, chipping, and scaling are common methods used in the
propagation of plants that produce bulbs. These methods induce the production of
offsets or, in this case, bulblets. Scooping involves the mechanical removal of the
basal plate, which exposes the bases of the bulb scales inside the bulb. The bulbs
are callused in the open or in a dry medium for a few days at 21C (70F). This
reduces the chances of infection and decay. The bulbs are then incubated at higher
humidity by covering them with a moist substrate, such as moistened peat, to
encourage the development of adventitious bulblets at the bases of the exposed
bulb scales. The bulblets are then removed, planted, and grown-on to larger size.

Scoring is the practice of making cuts through the basal plate of the bulb. As with
scooping, the bulbs are allowed to callus for a few days and then incubated at
higher humidity. Bulblets are initiated in the axils of the bulb scales along the cuts.
The process involves making three cuts, in the pattern of an asterisk, through the
basal plate using a sharp knife. Once again the bulblets are removed and grown-on
to a larger size.

Scaling involves the peeling-off of individual bulb scales from the mother bulb for
use in propagating new plants. The bulb scales may be treated with a fungicide and
are then planted or incubated in moist peat moss or sand at temperatures of 18-
21C (65-70F) for a period of a few weeks or more depending on species and
treatment. The individual scales will root and grow. In addition bulblets will be
initiated at the base of the scales. Most lilies can be propagated by scaling. When
two or sometimes more scales are used in propagation, by dividing the bulb into
several slabs or smaller sections each with a piece of basal plate, the process is
called twin scaling or chipping, respectively. Grape hyacinth are easily propagated
by twin scaling and narcissus by chipping.

Some bulbous plants also produce small "bulbs" along their stems. When produced
above ground they are called bulbils; when produced below ground they are called
bulblets. Both may be produced naturally or induced by plant manipulation. All
bulbous species naturally produce bulblets. Lily is probably the best example of a
plant capable of producing bulbils.

Layering
Layering is another method of vegetative or asexual propagation that occurs
naturally or under controlled conditions. Layering is the rooting of shoots while they
are still attached to the parent plant. Layers may be natural or induced and can be
used to propagate both herbaceous and woody plants. In cultivation, the process
involves covering a portion of the stem with moist soil or some other medium in
order to promote rooting. Wounding and/or treatment with root promoters are often
involved. Air layering, tip layering, mound layering or stooling, trench layering, and
serpentine layering are the most common types. Stolons, runners, and rhizomes
might be considered naturally rooted layers. Rooted suckers produced from stem
tissues are also essentially natural layers.

Division
Division is perhaps the simplest form of asexual propagation. It is simply the

29
division of the parent or mother plant into two or more viable sections, which are
then planted. The removal of offsets from bulbs can be considered a type of
division. Other examples of plants propagated by division include plants having
tubers (caladium, potato), tuberous roots (dahlia, sweet potato), tuberous stems
(tuberous begonia, cyclamen), rhizomes (lily-of-the-valley and iris), stolons
(bugleweed, club mosses), runners (strawberry, spider plant, boston fern), or
branched crowns (most herbaceous perennials and some woody shrubs). The
crowns of some plants may also produce offshoots, which are similarly divided into
separate plants. Some plants can also be propagated from naturally occurring
suckers (adventitious shoots arising from below ground) most often from root
tissues (sumac, raspberry). Finally, cormaceous plants, such as gayfeather, can
also be propagated by division.

30
 HORT 1001 - Plant Propagation

Grafting & Budding

Whip & Tongue Grafts, Side-Veneer Grafts, Cleft Grafts,
Approach Grafts, and Budding

Grafting / Graftage
Grafting is the art and science of combining two or more pieces of plant tissue such
that they knit together to form a complete and viable plant. Grafting takes its cue
from and is an adaptation of the ability of some plants to naturally form graft unions
in the wild. Grafting is used to propagate selected landscape plants and fruit trees
that are otherwise difficult to propagate by other, more economical, means such as
the use of cuttings. Grafting is also used in cases where it is desired and useful to
combine two or more genotypes to produce superior plants. Grafting a desired top
onto a specific rootstock can confer special characteristics to the finished plant such
as dwarfing, enhanced vigor, precocious flowering, enhanced mineral nutrition, or
tolerance to specific soil conditions. In addition, grafting can be used to influence
plant form, as when a weeping form is grafted onto a standard. Grafting can also be
used to repair damage to existing plants that are valuable in the landscape or a
production setting. Finally, graftage can be employed in the creation of unique
plants or multiple varieties (e.g., 5 in 1 apple) which are produced mainly as
curiosities.

The plant tissues most often used in grafting include a scion piece and a rootstock.
The scion consists of a portion of the crown of an existing plant; it may be as small
as a single bud. The rootstock may consist of an entire root system or simply pieces
of roots depending on the type of graft and its purpose. If the graft is successful,
the scion is destined to become the growing shoot while the rootstock will serve as
the resulting plant's root system. Sometimes additional pieces of stem are inserted
between the primary scion and rootstock; they are called interstocks and serve to
introduce additional characteristics (such as dwarfing) into the finished product.
Interstocks are most commonly used in the development of improved fruit trees.

It is the ability of the cambial tissues within the scion, interstock, and rootstock to
produce callus tissue which differentiates into functional xylem and phloem and
leads to the development of a viable, continuous connection between the cambial
tissues of the grafted parts that enables grafting to serve as a practical means of
propagation. To this end, it is very important that the stock and scion pieces fit
closely together and be held securely in place until the graft is sufficiently healed.

In most instances, the scion is grafted onto a stock that has an existing root system.
Such rootstocks may be rooted cuttings, seedlings, transplants, or even much older,
larger plants. Either the entire root system or just parts of the root system (e.g.,
piece rooting) from the stock plant may become part of the finished graft. The
technique whereby a scion is grafted onto an unrooted cutting, and the cutting is
rooted at the same time as the graft heals, is called stenting. Although we will be

31
using this technique in class, and it does have some specific commercial uses,
stenting is a relatively uncommon practice in the real world. Grafting is labor
intensive and, from a commercial perspective, success rates must be in the range of
90% or more to be economical. Grafting is, therefore, limited to high value plants
that cannot be propagated using less expensive methods. The rooting of cuttings is
also a more costly venture than growing plants from seed. Only very high success
rates or the production of a grafted plant with superior rootstock can, therefore,
justify stenting as a method of producing grafted plants. Although it is a viable
technique in certain instances (e.g., nurse grafting), these considerations explain
the relatively uncommon commercial use of stenting; the costs involved require that
the rooting success for the grafted cuttings be essentially 100% since the grafting
costs are typically already limiting.

Compatibility between the scion and rootstock is of paramount importance in the

success of any graft. In general, the more closely related the two parts (scion &
rootstock), the more likely the graft will be successful. Therefore, grafts between
different plants within a species are typically compatible, grafts made between
different species within the same genus are also frequently successful, and,
although much less common, unions between different genera within the same
family are also sometimes successful. Grafts between plants separated by more
than a family relationship will almost always fail completely. The relationship
between grafting success and plant taxonomy is further illustrated by the following
examples: A specific variety or cultivar of green ash (Fraxinus pennsylvanica;
Oleaceae, Olive Family) budded onto a seedling green ash rootstock (= same
species) will be successful; white ash (Fraxinus americana) grafted onto a green ash
rootstock (= same genus) will also typically be successful; and common privet
(Ligustrum vulgare; Oleaceae, Olive Family) grafted onto a green ash rootstock (=
same family) will also usually be successful. Green ash grafted onto a sugar maple
(Acer saccharum; Aceraceae, Maple Family) rootstock (= different families) will,
however, prove unsuccessful.

In addition to compatibility between the species to be grafted, the inherent ability of

the individual species being grafted to produce callus will also influence grafting
success. Other factors, including the selection of healthy stock plants, relatively
warm temperatures, good aeration, and adequate moisture (high humidity) are all
required for grafting success. Except for the expertise of the grafter in preparing
the stock and scion, these factors are most often responsible for the success or
failure of grafted plants.

Budding
Budding is a specialized form of grafting wherein the scion has been reduced to a
single bud from the plant to be propagated. The technique is widely used in the
production of landscape plants; mainly named tree cultivars (e.g., Acer platanoides
`Deborah', Malus `Spring Snow'). Budding is also commonly used as a method of
propagating various fruit cultivars. In most cases, the single-bud scion is typically
budded onto a seedling rootstock grown specifically for that purpose. The stems
that provide the scion buds are called bud sticks or bud wood. In most cases, the
bud is grafted onto the stock near the soil line and the top of the rootstock is
removed once the grafted bud begins to grow. Reducing the size of the scion to a
single bud greatly increases the efficiency of propagating the parent plant.

32
Although roses are also commonly budded, the practice is becoming less common
in favor of cuttings and the subsequent production of plants on their own roots.

We will investigate two budding techniques in class – T-budding (also called shield
budding) and chip budding. The main difference between the two is the condition of
the understock, which is essentially determined by the time of year that the
budding operation is performed. T-budding is done when the bark is said to be
slipping - a time when cambial growth is active and the bark is, therefore, easily
separated from the underlying wood. Chip-budding is used when these conditions
are lacking and the bark is not easily separated from the wood. Although there are
other forms of budding, T-budding and chip-budding are the primary budding
techniques used.

Cleft Grafting

An infrequently used, but at times very valuable type of graft, mostly used in fruit
production is cleft grafting. Cleft grafting is often referred to as ‘top—working’.
Sometimes growers are faced with a recently planted block of trees of a cultivar
that is under performing. As opposed to pushing the whole block out and
replanting, some growers will opt to use the established planting to support the
growth of new more desirable cultivar. By ‘top-working’ the existing trees they save
the price of removing the existing orchard, purchasing and planting new trees. In
addition, they save considerable time in getting the new cultivars into production.
We will be cleft grafting apple scion wood onto simulated apple rootstocks.

Spudmatoes?!
Although relatively uncommon in practice, many herbaceous species can also be
grafted. As with woody species, various types of grafts can be used and the more
closely related the stock and scion the more likely a graft will be successful. For
example, various types of squash, say acorn squash, or butternut squash, or
pumpkin (all Cucurbita spp.; Cucurbitaceae, Gourd Family), can be grafted to one
another successfully. Such grafts can be used to create novelty plant combinations,
impart disease resistance, or to improve the ability of plants to grow under specific
conditions.

33
 HORT 1001 - Plant Propagation

Propagating Ferns & Fern Allies:

Spores & Their Cultural Requirements

Ferns are unique and often treasured plants both in the wild and in cultivated
landscape and garden settings. All true ferns were at one time included in the same
family, the Polypodiaceae, but have since been separated into several families by
some recent authorities. Family classifications are, therefore and unfortunately,
somewhat confused in the literature. Most references still list most true ferns as
members of the Family Polypodiaceae. The club mosses (Lycopodiaceae; also called
ground pines and running pines or cedars) and horsetails (Equisetaceae), also
common in the wild and sometimes planted in the landscape, are often grouped
together with ferns and referred to as the fern allies. Although perhaps not as
closely related as once thought, they are also non-flowering plants and have life
cycles that are similar to that observed for ferns.

Classified as non-flowering plants, ferns and their allies do not produce flowers or
seeds. They are, however, classified as vascular plants owing to the presence of
conducting tissues for the transport of water and minerals (xylem) and
photosynthates (phloem). As a group, these interesting plants typically posses true
roots and leaves (in the case of ferns, fronds) both of which originate from a
perennial rhizome. The rhizome may grow horizontally or vertically and may be
found below or above ground depending on species. The “leaf”, or frond, is divided
into two main parts – the stipe (the leaf stalk or petiole) and the blade (the leafy,
expanded portion of the frond). In addition, the portion of the stipe to which the
leaflets are attached is sometimes referred to as the axis or rachis. Depending on
species, the blade can be quite variable; it may be simply lobed or divided into
smaller sections (or leaflets) called pinnae, pinnules, or pinnulets depending on the
degree of division.

The life cycle of ferns and other non-flowering plants makes them unique, but has
served well for hundreds of millions of years. In the wild, reproduction occurs by
both vegetative and sexual means. The complete life cycle of the ferns and fern
allies is characterized by two, independent, distinctly different, alternating
generations of plants, one of which produces sex cells while the other produces
spores. The familiar, spore-bearing plants we call ferns are examples of the
sporophytic or spore-producing generation. The sporophyte can also propagate
itself vegetatively by means of its rhizome. Spores are comprised of a single cell
containing nucleated protoplasm and are produced in miniature, stalked
(sporangiophore) spore sacs called sporangia, which are produced on either the
underside of the fronds, or on separate spore-bearing fronds. Fronds that bear
spores are called fertile fronds or sporophylls. When produced on the undersides of
the leaves, the sporangia are typically clustered in covered structures called sori.
Each sorus looks like a small dot and as a group the sori are typically arranged in an
organized pattern on the undersides of mature fronds. Millions of spores are
produced, but few will land in a spot suitable for growth. Under favorable conditions,
spores germinate to produce small, free-living plants called prothallia, which
comprise the sexual generation and look nothing like what we think of when we
think of ferns. Each prothallium (also prothallus), or gametophyte, consists of a
small, inconspicuous, green, heart-shaped structure and grows flat on the soil
surface. Separate male and female sex organs (antheridia and archegonia,
respectively), which produce eggs and sperm (also called spermatozoids or
antherozoids), are produced on the underside of the prothallium. In the presence of
moisture, the motile sperm swim about and unite with an egg produced by the
archegonium. The resulting zygote then begins to divide and develops into the
familiar spore-bearing plant (sporophyte) we call a fern. The entire process is
relatively slow and generally requires a period of several years.

In cultivation, ferns may be propagated by a variety of means depending on the

goals of the propagator and the species of fern involved. Division of clumps or the
rhizome is the most common for plants grown in the garden and where plants that
are identical to the parents are desired. When large numbers of plants are desired
and genetic variability is of less concern or desired, spores are used. Some ferns
naturally produce or ban be stimulated to produce bulbils in the axils of their leaves
which can be used to propagate new plants. As with plants propagated by division,
the plants produced are genetically identical to the parent. Time is also of concern;
division will produce full size plants the quickest, followed by bulbils, followed by
spores, which will require several years.

The first step in propagating ferns from spores is the collection of the spores. When
the spores are ripe and ready to be released, fertile fronds are collected and placed
in a clean dry container. They are allowed to dry for several days and then lightly
tapped or shaken to dislodge the spores. Fern spores are often viable for only a
short period of time (days) and should, therefore, be sown as soon after harvest as
possible. If immediate sowing is not possible, most spores can be stored and
viability maintained under refrigeration. Sowing is accomplished by filling a small
container with moist, sterile, growing medium and carefully sprinkling the spores
onto the surface of the medium. The culture must be kept in a moist, sterile
environment in indirect light. Temperatures of 60 to 70ºF (15 to 20ºC) are
recommended. Following germination of the spores, the developing prothalli will
appear as green film on the surface of the medium. When small fronds appear, the
young ferns are transplanted and gradually hardened off.

The American fern Society is an excellent source of information about ferns. Visit
their web page for more information about ferns (http://www.amerfernsoc.org).
 HORT 1001
Plant Propagation

Media Effects on
Propagation:
Importance & Measurement of
Growing Medium Porosity & Bulk
Density Characteristics

A carefully planned growing medium, specifically suited to the plants being grown,
is one of the critical aspects of successful plant propagation and production in
general. The characteristics of a growing medium become even more critical for
plants grown in containers where the volume of the container limits the growing
environment on many levels. Special care must, therefore, be focused on all
aspects of production to optimize plant growth and performance in containers.
Media porosity and bulk density are just two of the many factors that must be
considered when selecting a growing medium.

Porosity
Growing medium porosity characteristics are but a single, yet highly important,
group of production factors that influence plant performance or may even
determine the success or failure of an entire crop. When choosing a growing
medium our goal is to strike a balance between aeration and moisture holding
capacity such that the medium provides sufficient oxygen and moisture for plant
roots without requiring excessively frequent moisture replenishment (rainfall or
irrigation). It should be noted that additional considerations such as fertility, pH,
and weight, are also involved in designing a proper growing medium.

The total porosity of a container medium is a measure of the space within the
container that is not occupied by the solid medium components under standard
moisture and planting or growing conditions. In other words it is the space within
the medium that is occupied by either water (water-retention porosity) or air
(aeration porosity) at the time of measurement. Porosity measurements are
standardized such that they are measured for a growing medium at field or, in this
case, container capacity. Field or container capacity is when the medium contains
as much water as it can possibly hold just after it has been fully saturated and then
drained.

Desired Values:

Media porosity is a dynamic characteristic; it is influenced by the characteristics of

the growing medium and container depth. The porosity characteristics of a growing
medium also change over time in response to settling and the degradation of
organic components. As a growing medium ages in a container, aeration porosity
typically decreases while water retention porosity increases. The porosity
characteristics of a given medium are directly related to the depth of the containers
in which it is used; a growing medium will always hold less air and more water in a
shallow container compared to a deeper container of the same volume. When it
comes to plant growth and performance, porosity requirements are also species
specific since plants differ in the capacity of their roots to tolerate low oxygen, high
moisture conditions.
In general:

Aeration Porosity (should be at least 10%) – 15-30% (perhaps higher; up to

45%)
Water-Retention Porosity – 35-50%
Total Porosity – 50-80% (sometimes up to 95%)

Bulk Density
Bulk density is the mass or weight per unit volume (in situ) of oven-dry growing
medium and is commonly expressed as g/cm 3. It is an important characteristic of
any growing medium whether it be soil or artificial. Bulk density is determined by a
variety of factors including the type and size of the particles that make up the
medium, how tightly the particles fit or are packed together, as well as organic
matter content, moisture content, and biological activity which influences the
aggregation of particles and ultimately determines the structure of the growing
medium (how the particles are held together and arranged). These characteristics
in turn influence medium porosity and subsequently aeration and soil moisture
characteristics. Bulk density also influences water infiltration and thereby mediates
erosion potential and soil moisture water recharge. The relationships between bulk
density and porosity, and their effects on plant performance, are complex.

Typical & Desired Values:

The bulk density of a growing medium is dependent on the texture of the

various medium components and the structure of the medium (i.e., how the
medium components are organized). Bulk density is also dependent on the
degree of compaction and the effects of soil moisture on shrinkage and
contraction of the growing medium. The effects of bulk density on plant
performance are also species specific as plants differ in their capacity to
tolerate compacted soils.

A bulk density of:

0.1 to 0.6 - is typical of organic soils

1.0 to 1.3 - is typical of granulated clay surface soils
1.3 to 1.8 - is typical of course textured (sandy) surface soils

In general:

A bulk density of 0.80 to 1.20 g/cm3 would be considered acceptable for

most growing media. Values considerably lower than these are
commonly encountered with, artificial, soilless growing media, and are
just fine when it comes to plant growth.

Bulk density values of 1.25 - 1.65 g/cm3 have been shown to reduce root
growth.
38
39
 HORT 1001
Plant Propagation

Experimental Design
During the semester, you will be carrying out experiments, and reporting on them in written
form. Each experiment begins with experimental variables. What is an experimental variable and
how are they chosen? An experimental variable is defined as any individual characteristic that
has been chosen to be different from one group of individuals to another group within the
experiment. What does that mean for you in plant propagation?

For example, when you are attempting to root herbaceous stem cuttings, some examples of
variables that will be used during the semester are:

 Effect of plant growth regulator application on inducing rooting - one group of cuttings
will be treated with one concentration of IBA and another group would not be treated.
 Effect of treatment with different concentrations of a plant growth regulator - each group
of cuttings will be treated with different concentrations e.g. IBA at 1000, 3000, and 5000
ppm.
 Effect of the carrier on plant growth regulator action - a group of cuttings treated with
IBA in talc vs IBA dissolved in alcohol.
 Effect of leaf area on rooting success of cuttings - compare the effect of number of leaves
on rooting.

This list of possible variables could go on, limited only by time, space and supplies. However,
the variables could change depending on the type of plant material you were using. In
experimental design (at least for plant propagation class), there are 2 important rules to
remember when thinking about experiments: always use a control (plant material that remains
untreated); and always repeat your treatments.

What is a control?

A control is a group of cuttings that remain untreated. For example, if you were interested in
treating 5 cuttings of Spirea japonica with 1000 ppm IBA in talc, you would also leave 5 cuttings
untreated. This allows you, at the end of the experiment, to conclude whether IBA at 1000 ppm
in talc increased root initiation or increased the speed roots appeared. If you treated all of the
cuttings with 1000 ppm IBA you would have nothing to compare them to. The same goes for all
of the variables listed above. If you are going to treat a group of cuttings in a certain way, you
must leave the same number of cuttings untreated for the control.

What is a treatment?

A treatment is defined as the combination of variables applied to experimental units (each

individual cutting is an experimental unit) in the study. For single variable experiments, the
variable and the treatment are the same. For example, if you were comparing whether 1000 ppm
IBA enhanced rooting of Echniacea purpurea, the variable and treatment are the same – IBA

40
treatment. In an experiment comparing seed scarification treatments, the variable might be
scarification methods including the treatments mechanical, acid, and hot water scarification. All
other factors need to remain the same. So after treatment with different scarification methods, all
seeds would be planted at the same time, in the same media, placed side by side in the same
greenhouse. The only difference in the seeds would be the treatment that you imposed earlier.

You must take care when treatments involve combinations of several variables since the effects
can be confounded and make it impossible to make conclusions from the experiment. For this
reason, we will be doing very few multiple variable experiments.

Designing experiments

For each experiment designed, you begin by choosing your variables. How were the variable
chosen for the experiments that we are doing in Plant Propagation? We chose them to
demonstrate different methods to effect rooting in asexual propagation. We are experimenting
with plant material that we don’t know how successful some of the treatments will be. We
looked through the text and web sources to determine plant material that might work for the
treatments that we are imposing. The next step was to determine the number of experimental
units that will receive each treatment. For our example of herbaceous cuttings, each cutting
would be an experimental unit or for the scarification example each seed would be an
experimental unit. In this class, 5 experimental units should be sufficient but each treatment
should be repeated on no fewer than 3 experimental units. In a few experiments, each student
will only be doing one set of treatments and we will combine data from 5 or more students for
the final analysis.

Next, we thought about what kind of data should be collected. In the seed scarification
experiment you might collect data on number of seeds germinated and calculate the %
germination for each treatment; number of days to emergence and calculate the average for each
treatment. Other examples might be, number of roots, root length, days to rooting, number of
plantlets formed, etc.

Implementing your experiment

Collect and prepare the necessary plant materials and apply your treatments. Observe your
experiments frequently; at least once a week. Collect data as necessary. Carefully record exactly
what you did from beginning to end. Make sure you record dates in your lab book that you
started and ended the experiment as well as the date that you collected data.

Be sure everything is labeled! Labels must include the scientific name of the plant. Each
treatment needs to have its own label. You can use codes to identify treatments but be sure to
make notes confirming the meaning of the code. You will be doing lots of experiments and so
codes that were clear when you did them in September, might not be so clear after 12 more
experiments and 8 weeks!

When the experiment is completed and all data has been collected, transplant material you wish
to keep. Every pot that goes into the greenhouse must have a label or it will be thrown away. We
recommend taking these plants home regularly as we have limited greenhouse space and plants
don't do well being transported outside in December
41
 Introduction to Experimentation:
Herbaceous Stem Cuttings
[E; individual; 1 species, 2 treatments; 5 replications/treatment = 10
cuttings/student]

Objectives:
 To introduce experimentation including application of treatments (in this
case treatment with IBA) and data collection.

 To learn how to take cuttings.

 To produce uniformly rooted cuttings.

To Do Before Class:
Read p. 15 in your lab manual and pp. 154-156 in your lab text (Plant
Propagation).

Plant Material:
Solenostemon scutellarioides [syn. Coleus  hybridus] (Coleus; Lamiaceae, Mint
Family)

Treatments:
1. Control
2. 1000 ppm IBA in talc

Materials Provided:
 Pruners
 Razor blades
 Labels
 Permanent marking pen (black Sharpie)

Methods:
1. Label two colored plastic stakes with the date, treatment and the plant
material (Coleus).

2. Take 10 cuttings of the plants provided, with each cutting having 2-3
nodes. Take each cutting just below a node. The cuttings should all come from
the same plant and be as similar as possible, including size, and the location on
the plant from which they were collected. Be sure to keep track of polarity –
which end was up – for each of your cuttings.

3. Wrap the cuttings in moist paper toweling.

42
 4. Finish preparing your cuttings by removing the leaves from the bottom
node of each cutting.

5. Once your cuttings have been prepared dip the bases of five (5) cuttings in 1000
ppm IBA in talc; the remaining five (5) cuttings will be the controls (non-treated).

6. After treatment, take all 10 cuttings (keeping the treatments separate) and stick
them into your mist bench plot with the appropriate labels.

7. Check on your cuttings each lab period and record your observations. Begin by
shaking off the sand and wrap the cuttings in moist paper toweling, keeping
treatments separate. Count the number of cuttings that have produced roots.
Then evaluate the cuttings on a 0-5 visual rating scale: 0 = no roots, 5 = roots
abundant. In general, rooted cuttings are ready for harvest when they have a
root system that is about 33% to 50% the size of the top. The sample tables
provided may be used to record your data.

9. When you are finished collecting your data, pot up all your rooted coleus
cuttings; place them in the designated location in the greenhouse for later use.

10. As always, be sure to clean up all work areas when you are finished. Good
sanitation is extremely important in greenhouse production systems so please
be sure to do your part in keeping work and growing areas clean throughout
the semester.

Results – Data and Observations:

 Final data will be collected after 3 weeks. For this and subsequent reports
your data should be summarized for each date of observation and
presented in the form of a bar graph or table.

Discussion – Using your results and observations, discuss

your findings and answer the following questions:
 Where did the roots emerge from - between the node, at the node, or at the
bottom of the cutting? What is the significance of where the roots emerge?

 Did the application of 1000 ppm IBA aid in rooting?

 If you could change one thing about this experiment, what would it be and
why?

Recommendation:
43
 Based on your findings, provide a one-sentence recommendation outlining
the best method of propagating coleus using stem cuttings.

44
Example of Data Table – use one such table for each date data is taken
Title of Experiment: Introduction to Experimentation:
Herbaceous Stem Cuttings
Date data taken: _______________

For the control:

# of cuttings started __________________ # of cuttings rooted
__________________

Cuttings treated with 1000ppm IBA

# of cuttings started __________________ # of cuttings rooted
__________________

Root Rating
Cutting Number
Treatm Average
ent 1 2 3 4 5

Control

1000
ppm IBA

Note: This table is also provided as an example of the

format that might be used when collecting and reporting data for future
experiment

Root ratings are an example of one type of data that will be collected for this
and other experiments. Use the diagrams below (Thanks to Ohio State
University) as a guide to rate the rooting success of your treatments. Assign
a 1 to 5 rating to each cutting and use the data to calculate an average root
rating for each treatment.

45
46
 Sexual Propagation: Using Scarification to
Overcome
Hard Seed Coat Dormancy
[E; Individual; 2 species, 5 treatments, 2 replications = 20 seeds per student]

Objectives:
 To experiment with different methods to overcome hard seed coat dormancy

To Do Before Class:
Read pp. 9-14 in your lab manual and pp. 19-20 (up to “Conditions needed for
Germination”) in your text.

Plant Material:
Gleditsia triacanthos (Common Honey Locust; Fabaceae, Bean Family)

Gymnocladus dioica (Kentucky Coffee Tree; Fabaceae, Bean Family)

Materials to Bring To Class:

 Permanent marking pens
 Lab notebooks

Materials Provided:
 Files
 Hammers
 Labels
 Pots
 Potting medium

Treatments:
1. Control (non-treated; seed coat not broken)
2. Mechanical scarification hammer
3. Mechanical scarification file
4. Hot water treatment
5. Acid scarification

Methods:
1. Make labels for each of the five (5) treatments – the two (2) species tested for
each treatment will be planted together in the same pot (= 4 seeds/pot).

2. Partially fill five (5) of the pots provided with growing medium. Each
treatment will be planted in a separate pot.

47
 3. Plant two (2) control seeds for both species in one pot and label. The seeds
should be planted such that they are 1 to 2 times as deep as their diameter.

4. For the sake of timing and safety, seeds that have been soaked in boiling
water and acid scarified will be provided. Plant two (2) seeds of each species
from each of these treatments together in separate containers by treatment
and label.

5. For one of the mechanical scarification treatments, scarify the seed coats of
two (2) seeds of each species with a file, making sure you file in the center of
the seeds so you do not damage the embryos. Once the seed coat is broken,
stop so you do not damage the cotyledons. Plant seeds and label.

6. For the second mechanical scarification treatment, scarify the seed coats with
a hammer by wrapping the seed securely in paper (this prevents the seed
from shooting away when it is hit with the hammer) and tapping until the
seed coat is cracked. Ideally, pound hard enough to crack the seed coat, but
not so hard that you damage the seed. Plant the seeds and label.

7. Transfer the pots to the designated location in the greenhouse.

8. Check for germination each lab period. When you believe all the seeds that
are going to germinate have done so, you may want to dig up those seeds
that did not germinate in an attempt to find out why they failed to do so.

Results – Data and Observations:

 Observations that may be taken for each treatment include (but are not
limited to): % germination, number of days to first emergence, type of
germination (hypogeal or epigeal).

Discussion – Using your results and observations, discuss

your findings and answer the following questions:
 What differences, if any, did you observe between treatments?

 What are the advantages and disadvantages of using one scarification

technique over another?

Recommendation:
 Based on your findings, provide a one-sentence recommendation outlining
the best method of propagating these two species from seed.

48
 Asexual Propagation: Adventitious Shoot
Initiation in Response to Treatment with BA
[E; individual; 1 species, 3 treatments, 5 replications/treatment
= 15 two-segmented phyllocad cuttings/student]

Objectives:
 To better understand the techniques involved in the propagation of new
plants from pieces of the parent plant; in this case phyllocads (a flattened
stem or branch that functions as a leaf).

 To investigate whether treatment with BA influences the formation of

adventitious shoot meristems on phyllocad cuttings.

 To produce entire plants from phyllocad cuttings.

To Do Before Class:
Read pp. 4-6 and 15 (Herbaceous Stem Cuttings section) in your lab manual.

Plant Material:
Schlumbergera bridgesii (Christmas Cactus; Cactaceae, Cactus Family)

Materials To Bring To Class:

 Permanent marking pens
 Lab notebooks

Materials Provided:
 Plant material
 Razor blades
 Label stakes
 Newspaper
 Benzyladenine solution (150 ppm in EtOH)
 Benzyladenine solution (1500 ppm in EtOH)

Treatments:
1. Control (no treatment)
2. 150 ppm BA in 50% EtOH
3. 1500 ppm BA in 50% EtOH

49
Methods:
1. Select a plant from which to collect your cuttings.

2. Using a clean, sharp razor blade, collect fifteen (15) phyllocad cuttings (two
phyllocad segments per cutting) of similar size, age, and quality. Be sure to
keep track of the polarity of your cuttings.

3. Divide your cuttings into three (3) groups of five (5) and prepare labels for
each group indicating treatment and date. One group will serve as a control.
Treat the next two groups of cuttings in the with BA (150 ppm & 1500ppm) by
dipping the whole cutting into the solution provided. Be sure to re-cover the
solutions when not in immediate use to limit evaporation and associated
increases in concentration.

4. Stick your cuttings in your assigned plot in the mist bench along with the
appropriate labels.

5. Observe your cuttings on a regular basis and record your observations.

6. The plants produced are yours to keep; when you are done, pot them up and
take them home if you like. If you have good quality plants and wish to
donate any of them for use by students in future classes, we would be happy
to accept them. Please label all plants to be donated with their full scientific
and common names and the date.

7. As always, be sure to clean up all work areas when you are finished.

Results – Data & Observations:

 Data and observations that may be taken for each treatment include (but are
not limited to): root initiation, shoot initiation, subsequent root and shoot
development, number of phyllocads produced, etc.

Discussion – Using your results, discuss your findings and

answer the following questions:
 Which comes first, roots or shoots?

 From where do the roots and shoots arise?

 Are there differences between propagation by phyllocads and standard stem

cuttings

 What effects, if any, did your treatments have on root and shoot
development?

Recommendation:

50
Based on your findings, provide a one-sentence recommendation outlining the
best treatment for propagating from Christmas cactus using phyllocad
cuttings.

51
 Asexual Propagation: Propagation of Plants
Using
Leaf Cuttings

[E; Group; 1 species, 6 treatments, 3 replications/treatment =18

cuttings/student]

Objectives:
 To observe effects of benzyl adenine (BA) on shoot meristem formation.

 To produce entire plants from leaf cuttings.

To Do Before Class:
Read pp. 19 in your lab manual and p. 157 in your text.

Plant Material:
Saintpaulia ionantha (African Violet; Gesneriaceae, Gesneria Family)

Materials To Bring To Class:

 Permanent marking pens
 Lab notebooks

Materials Provided:
 Plant material
 Razor blades
 Label stakes
 Paper toweling
 1500 ppm benzyladenine (BA)

Treatments:
Leaf-petiole cuttings

52
 1. Control
2. 150 ppm (BA)

Leaf-blade cuttings
1. Control
2. 150 ppm (BA)

Leaf-piece cuttings
1. Control
2. 150 ppm (BA)

Methods:
1. Remove 15 leaves from an African violet plant; 6 with petioles; 6 without
petioles, and three that will be cut down the midrib vein. You will have a total
of 15 leaves. All cuttings should come from the same plant and be as close
to the same age and size as possible. You may want to arrange to exchange
with other students when the experiment is completed so you have a variety
of African violets to take home.

2. Keep leaves wrapped in a moist paper towel while working. Be gentle.

3. Treat the bases of three cuttings with 1500 ppm BA and leave 3 cuttings
untreated for each treatment. For the leaf-piece cuttings treat the midrib of 3
cuttings with the BA and leave three untreated.

4. Stick the cuttings in your assigned plot in the mist bench. The cuttings
should be oriented with just the bases of either their petioles or leaf-blades
stuck into the rooting media. For the leaf-piece cuttings, the midrib should be
in contact with the sand.

5. Observe your cuttings on a regular basis and record your observations. Lift
the cuttings gently and by lifting from beneath to avoid breaking any roots.

6. As your cuttings become well rooted, remove the cuttings from the mist
bench, collect data, and pot them up. Be sure to keep your plants labeled
and continue to follow shoot development after potting.

7. As always, be sure to clean up all work areas when you are finished.

Results – Data & Observations:

 Data and observations that may be taken for each treatment include (but
are not limited to): callus formation, root initiation and development,
shoot initiation and development, time to harvest and transplantation,
plant (roots and shoots) quality, etc.

53
Discussion – Using your results and observations discuss
your findings and answer the following questions:
 Did roots and/or shoots initiate? If so, where?

 Which came first, roots or shoots?

 Were there any differences in root and shoot development between the
treatments or among the different types of cuttings?

 Why might a plant propagator use leaf-type cuttings compared to other

types of cuttings (e.g., stem cuttings)?

 What effects, if any, did your treatments have on root and shoot
development?

 If you could change one thing about this experiment, what would it be and
why?

Recommendation:
 Provide a one-sentence recommendation outlining the best treatment from
your data for propagating African violet using leaf cuttings.

54
 Sexual Propagation: Effects of Seed Coat
Removal, Treatment with GA3, and Cold
Stratification on the Germination of Apple
Seeds
[GE; groups of 4; 1 species, 8 treatments, 5 seeds/treatment = 40
seeds/group]

Objectives:

 To investigate the effects of handling and various pre-plant seed

treatments on the germination of apple seeds.

To Do Before Class:
Read pp. 4-6, 7-14 in your lab manual and pp. 10 & 19-20 (up to “Conditions
needed for Germination”) in your text.

Plant Material:
Malus domestica (Apple; Rosaceae, Rose Family) – cultivar announced in class.

Materials to Bring to Class:

 Razor blades
 Permanent marking pens
 Ziploc bags
 Tweezers
 Lab notebooks

Materials Provided:

 Label stakes
 Paper toweling
 Pots
 Growing medium
 GA

Treatments:
1. Stratify, Seed coats intact, No GA3
2. Stratify, Seed coats intact, GA3

55
 3. Stratify, Seed coats removed, No GA3
4. Stratify, Seed coats removed, GA3
5. Direct Sow, Seed coats intact, No GA3
6. Direct Sow, Seed coats intact, GA3
7. Direct Sow, Seed coats removed, No GA3
8. Direct Sow, Seed coats removed, GA 3

Methods (see flow chart; Figure 1):

1. For this experiment you will be working in groups of four (4).

2. Prepare label stakes for each of the treatments to be done today – Treatments
1-4. Be sure to write the date and the names of your group members on
each stake.

3. Fill five (4) pots or, if plug trays are used, four (4) rows of plugs with slightly
moistened growing medium.

4. Extract 25 apple seeds keeping them moist AT ALL TIMES by wrapping

them in a moist paper towel. Each treatment will be replicated five (5) times
(5 seeds per treatment) for a total of 20 seeds. We are having you extract 25
seeds because one of the procedures, removing the seed coat, may be
difficult to do. Having extras is important!

5. Remove seed coats from 10 seeds – KEEP THEM MOIST!

6. Sow 5 seeds with seed coats in a pot or row of plugs and 5 without seed
coats in another pot or set of plugs (Treatments 1 and 3). Seeds should be
planted at a uniform depth and should be no more than 1 cm deep. Place the
appropriate label stake in the growing container.

7. Place 5 seeds with seed coats and 5 without seed coats in a Petri dish
filled with 400 ppm GA3, and allow the seeds to soak for 1 hour (Treatments
2 and 4). Then sow the 5 GA3-treated seeds with seed coats and the 5 GA 3-
treated seeds without seed coats into separate pots or set of plugs in the
plug tray. Place the appropriate stake in each pot or row.

8. Water carefully and place all pots or the plug tray in a large plastic bag with
your group number and names of each group member as well as the date of
this portion of the experiment written with a Sharpie. Place the bags into
cold stratification (cool, moist conditions).

CONTINUING ACTIVITIES: Approximately 6 weeks after the

seeds were placed into conditions of stratification remove
the seeds from stratification:
1. Remove seeds prepared in the first half of the experiment from stratification.

56
 2. Prepare label stakes for each of the treatments to be done today
(Treatments 5-8). Be sure to write the date and your group members
name on the stakes.
3. Extract 25 apple seeds keeping them moist AT ALL TIMES. Each
treatment will be replicated 5 times (5 seeds per treatment) for a total of 20
seeds. Again, we are having you extract 25 seeds because it may be difficult
to remove the seed coats.

4. Remove seed coats from 10 seeds – KEEP THEM MOIST!

5. Sow 5 seeds with seed coats and 5 seeds without seed coats into
separate pots or rows of the plug tray (Treatments 5 and 7). As before, pay
close attention to depth of planting. Label with the appropriate stakes.

6. Place 5 seeds with seed coats and 5 seeds without seed coats in a Petri
dish filled with 400 ppm GA3, and allow the seeds to soak for 1 hour
(Treatments 6 and 8). Then sow seeds into 2 separate pots or rows of the
plug tray. Label with appropriate stakes.

7. You will have 8 pots or 8 rows of a plug tray total (each with one treatment).
Place the pots or plug trays in the greenhouse.

8. Observations on germination must be taken and recorded each lab period.

You may make your observations as a group or you can designate one
member to collect the data and report to the group.

Results – Data and Observations:

 Data and observations that may be taken for each treatment include (but are
not limited to): # days to emergence, # of seeds rotted, germination %
(# seeds germinated/total number of seeds), seedling size and vigor. You
should discuss data collection as a group to standardize your efforts. Also, at
the end of these lab directions there is a data sheet that we will be using
in the lecture portion of this class. Information for the data sheet should
come from lab observations.

Discussion – Using your results and observations discuss

your findings and answer the following questions:
 How did the treatments compare with respect to germination percentage?

 Is there a difference in germination if seeds are stratified vs. direct sow?

 What effect does stratification have on germination?

57
  Is removing the seed coat analogous to scarification?

 Are there inhibitors to seed germination in the seed coat? How might you
test your ideas?

 What other treatments might be used to overcome dormancy requirements?

 What recommendations would you give to someone who wanted to know

about overcoming dormancy of woody species?

Recommendation:
 Based on your findings, provide a one-sentence recommendation outlining
the best treatment for germinating apple seeds.

58
Apple Seed Experiments - Data Sheet: Date:

Treatment # of Seeds Germinated % Germination

Stratified

1. With seed coat

2. With seed coat, GA3 treated

3. Without seed coat

4. Without seed coat, GA3 treated

Not Stratified

5. With Seed coat

6. With seed coat, GA3 treated

7. Without seed coat

8. Without seed coat, GA3 treated

Comments:

59
Figure 1. Flowchart of Apple Seed Experiments

with GA
Treatments 2 & 4

Stratification
(with & without seed coat)
without GA
Treatments 1 & 3

with GA
Treatments 6 & 8

No Stratification
(with & without seed coat)
without GA
Treatments 5 & 7

60
 Sexual Propagation: Effects of Growing
Medium on Seed Germination and Seedling
Performance
[E; 1 species, 7 treatments 3 replications (3 containers of each
medium; 4 seeds/container) = 21 containers
and 84 seeds]

Objectives:
 To demonstrate the effects of various growing media on seed germination
and seedling performance.

 To highlight the importance of selecting a suitable growing medium based on

the plant being grown and the container used.

To Do Before Class:
Read pp. 28-29 in your lab manual and pp. 32-35 in your text. On web: Growing
media in container production in greenhouse and nursery. Part 1: Components and Mixes
http://www.uaex.edu/Other_Areas/publications/PDF/FSA-6097.pdf &
http://www.uaex.edu/Other_Areas/publications/PDF/FSA-6098.pdf

Plant Material:
Tagetes sp. (Marigold; Asteraceae, Aster/Sunflower Family)

Materials To Bring To Class:

 Permanent marking pens
 Lab notebooks

Materials Provided:
 Containers
 Various growing media (use the same media that were used for the porosity/bulk
density experiment)
 Marigold seeds
 Label stakes

Treatments:
1. Growing Medium #1
2. Growing Medium #2

61
 3. Growing Medium #3
4. Growing Medium #4
5. Growing Medium #5
6. Growing Medium #6
7. Growing Medium #7
Methods:
1. This experiment will be done in groups; the same groups as for the
porosity/bulk density experiment.

2. Fill three (3) containers to 1/4" below the brim with each of the seven (7)
growing media. for a total of 21 containers filled with growing media.

3. Each of us will fill a container with medium differently. For example, I might
pack the medium more tightly and have a different understanding of what full
means than someone else does. To reduce the effects such differences in the
method of filling containers might have on the experimental results, the same
person should fill all the containers with the appropriate media.

4. Label each container indicating your group name, the date, and the medium.

5. Plant 4 marigold seeds on the surface of the medium in each container.

6. Cover the seed with additional medium filling each container to the brim
resulting in a planting depth of 1/4". Be sure to cover the seed with the same
medium as is already in the container and once again, have the same person
perform this task (this does not have to be the same person who initially filled
the containers).

7. Carefully water your seeds; wet the medium completely being very careful
not to splash medium from the containers or disturb your seeds.

8. Place your containers in the flats provided and carefully transfer them to the
designated location in the greenhouse.

9. Check your seeds and follow the germination and growth of your seedlings
each lab period until final data is collected. Water your containers carefully
as needed.

10. As always, be sure to clean up all work areas when you are finished.

Results – Data & Observations:

 Data and observations that may be taken for each treatment include (but are
not limited to): time to emergence of first and last seedlings, % germination,
seedling vigor, seedling appearance, and % survival.

Discussion – Using your results, discuss your findings and

answer the following questions:
62
  Did your different media influence seed germination? If so, why?

 Was the growth of your seedlings influenced by the various media? If so,
how?

 Which medium was the best choice for growing marigolds from seed to the
seedling stage in the containers used?

 How do your results match up with your earlier hypotheses about the
suitability of each medium for growing plants based on porosity values?

Recommendation:
 Based on your findings, provide a one sentence recommendation outlining
the best medium for growing marigolds in the containers used

63
Asexual Propagation: Rooting of Herbaceous
Stem Cuttings
in Response to Type of Auxin,
Concentration of Auxin,
Different Carriers of Auxin

[P; 1 species, each student will do one experiment, with 4-6

treatments per experiment,
5 replications/treatment = 20-30 cuttings/student; shared data for
the final report]

Objectives:
 To successfully propagate plants from stem cuttings.

 To observe how treatment with auxin influences rooting in different plants.

To Do Before Class:
Read pp. 4-6 and 15 in your lab manual and pp. 154-155 in your text.

Plant Material:
The plant material used will be determined by availability in the horticulture
garden.

Materials To Bring To Class:

 Pruners
 Permanent marking pens
 Polyethylene bags
 Lab notebooks

Materials Provided:
 Label stakes
 Paper toweling
 Treatment compounds

64
Treatments:
Experiment 1 - Rooting in Response to Different Types of Auxin

1. Control
2. 50% ethanol
3. 1000 ppm IBA in 50% ethanol
4. 1000 ppm NAA in 50% ethanol
5. 1000 ppm IAA in 50% ethanol

Experiment 2 - Rooting in Response to Different Concentrations of an Auxin

1. Control
2. 1000 ppm (0.1%) IBA in talc
3. 3000 ppm (0.3%) IBA in talc
4. 8000 ppm (0.8%) IBA in talc
5. 16,000 ppm (1.6%) IBA in talc

Experiment 3 - Rooting in Response to Different Carriers of an Auxin

1. Control
2. 50% ethanol
3. 1000 ppm IBA in 50% ethanol
4. Talc
5. 1000 ppm IBA in talc
6. 1000 ppm IBA (K-IBA) in water

Methods:
1. Experiments will be assigned by the instructor. Each student will do one
experiment, selecting one species for their cuttings. Be sure to record the
genus, species, and common name of the plant.

2. Each student should prepare labels for each of the treatments included in
their experiment. The stakes should also be labeled with the date, the name
of the plant, and a brief description of the experiment (i.e., type,
concentration, carrier).

3. Each student should collect the required number of cuttings needed for their

65
 experiment from the plant species chosen. In all cases, the cuttings should
have four (4) nodes with fully expanded leaves (ask if you are unsure how to
do this). As you collect your cuttings, be sure to place them in polyethylene
bags with moist paper toweling to prevent them from drying out.

4. Return to the classroom and finish preparing the cuttings by removing the
leaves from the bases of the cuttings. Keep the cuttings covered with moist
paper towel to keep them from drying out. Divide the cuttings into groups of
five with one group of five cuttings for each of the treatments in your
experiment. Treat the cuttings in each group with one of the treatments
included in your experiment.

5. After treatment, carefully transfer the cuttings to the mist house and stick
them in your plot in the mist bench with the corresponding label. Keep your
cuttings separated and labeled by treatment at all times.

6. As always, be sure to clean up all work areas when you are finished.

7. Observe the cuttings for each of the experiments at least weekly and record
your observations.

Results – Data & Observations:

 Data and observations that may be taken for each treatment in each
experiment include (but are not limited to): callus formation, rooting (time to
first root emergence, time to harvest, % rooting, and root system quality and
symmetry), signs of necrosis at the site of application, length of longest root,
# of roots, and performance of the cutting growth.

When taking data, pay close attention to where roots are forming.

Discussion – Using your results and observations, discuss

your findings and answer the following questions:
 Where along the stem were roots initiated?

 Did rooting continue to improve over time?

 What effects, if any, did your treatments have on rooting?

 Which treatment(s) resulted in the best rooting?

 If you could change one thing about this experiment, what would it be and
why?

Recommendation:

66
 Discuss your observations with other members of your group and, as a group,
provide a one-sentence recommendation for each experiment outlining the
best treatment for propagating your specific herbaceous plant using stem
cuttings.

67
 Asexual Propagation: Designing an
Experiment to Maximize Rooting of Leaf-
Petiole and Leaf-Bud Cuttings
[P; individual; 1 species; experiments designed individually by each
student using
a minimum of 20 experimental units]

Objectives:
 To review the scientific method by providing an opportunity to design an
experiment involving leaf-bud and leaf-petiole cuttings.

 To compare the effects of various treatments on leaf-bud and leaf-petiole

cuttings.

To Do Before Class:
Design your experiment by clearly identifying objectives, treatments to be
done, and data to be taken. You need to use at least 20 experimental units. You
must choose the variables. Each of the resulting treatment combinations
should be replicated five (5) times. You must have your experimental design
approved by the instructor prior to initiating your experiment.

Read pp. 4-6, pp. 19 and 30-31 in your lab manual.

Plant Material:
Peperomia scandens ‘Variegata’ (Variegated False Philodendron; Piperaceae,
Pepper (the spice) Family)

Materials To Bring To Class:

 Pruners
 Permanent marking pens
 Lab notebooks

Materials Provided:
 Plant material
 Label stakes
 IBA in talc (1000, 3000, 8000, 16000 ppm)

68
  IBA in water (1000 ppm)
 IBA in 50% ethanol (1000 ppm)
 NAA in 50% ethanol (1000 ppm)
 BA in 50% ethanol (150ppm & 1500 ppm )
 Distilled water
 talc
 50% ethaol (ETOH)

Treatments:
To be determined by student – leaf-bud and Leaf-Petiole cuttings combined with
other experimental variables selected by the student.

Methods:
To be determined by student.

Results – Data & Observations:

 To be determined by the student - data and observations that may be
taken for each treatment might include (but are not limited to): root
initiation (time to rooting and root number), root quality and percentage,
shoot initiation (time to shoot initiation and shoot number), subsequent
aspects of root and shoot development, etc.

Discussion – Using your results, discuss your findings and

answer the following questions:
 Which came first roots or shoots?

 Where did the roots and shoots arise?

 Were multiple or single plants produced?

 Were there differences between the treatments when compared?

 Were there differences between types of cuttings?

Recommendation:
 Based on your findings, provide a one-sentence recommendation outlining
the best method for propagating Peperomia scandens ‘Variegata’ from
leaf-bud cuttings and a one-sentence recommendation outlining the best
method for propagating Peperomia scandens ‘Variegata’ from leaf-petiole

69
cuttings?

70
 Sexual Propagation: Manipulation and
Germination of Seeds from an Herbaceous
Perennial, or Woody
Perennial Plant

[P; individual; 1 species of choice, treatments and replication

numbers to
be determined by the student]

Objectives:

 To determine the requirements for successfully germinating seeds from a

variety of plants based on research and experimentation.

 To experiment with different techniques used to break seed dormancy.

 To design your own experiments to answer specific, testable research

questions pertaining to seed handling and germination.

To Do Before Class:
Read the Introductory Information section on seeds in the lab manual (pp. 7-14).

You will need to do some research to hopefully learn about the germination
requirements of the plant you are working with before you set up the experiments.
Don’t put this off! A little research will go a long way when it come to success or
failure with this project. There are many books and research papers specific to the
germination of seeds from various plants. The internet can also be a valuable
source of information. One website that has information on many garden plants is
the Plantfacts site at http://plantfacts.osu.edu/. And then, of course, you can always
visit the library.

Plant Material:

Seeds of one type of plant material (1 herbaceous perennial or 1 woody perennial)

to be distributed by the lab instructor.
Materials To Bring To Class:

71
 Permanent marking pens
 Lab notebooks

Materials Provided:

 Label stakes
 Paper toweling
 TTC (for viability testing)
 Gibberellic acid (400 ppm)
 Containers
 Growing media
 Scarification and stratification facilities

Treatments:

To be determined by each student depending on the plant material chosen.

Methods:

To be determined by each student depending on the plant material chosen. This

may require several attempts and experimentation to achieve successful
results.

Results – Data and Observations:

 Observations that may be taken for each treatment may include (but are not
limited to): number of days to germination, % germination, % viable seed,
type of germination (epigeal or hypogeal), monocot or dicot, type of
dormancy, etc.

Discussion – Using your results and observations discuss

your findings in the following format and answer the
following questions:

 Don’t worry if you are not successful in germinating the seeds you have

72
 been given; the point is that you make a reasonable attempt. If you have a
problem getting seeds to germinate, you should consider and develop a
hypothesis as to why you think you were unsuccessful. Again, research into
the germination requirements of your seeds is necessary in this regard.

! Based on the information gained while attempting to germinate your seeds and
research on your selected plants, you should design a seed packet for each
species of plant that you collected and researched. As an example, think of the
vegetable and flower seed packets you have seen offered for sale. Be creative
in your seed packet designs; remember marketing is an important
aspect of

any successful horticultural enterprise. What type of design and

information would catch your attention and persuade you to purchase a packet
of seeds?

At a minimum, each seed packet should include the following information:

1.)A picture or drawing of the mature plant and a young seedling.

2.) Family relationship (family name), scientific/botanical name (Genus

species, and variety if applicable), and common name(s).

3.) Classification based on lifecycle (herbaceous perennial, woody

perennial tree or shrub) and descriptive information (mature height,
habit, etc.).

4.) Geographic range of the species (Where is it native?).

5.) Cultural information – planting directions, light requirements

(sun/shade), moisture requirements, soil preference (texture, pH),
fertility requirements, preferred habitat, potential pests, etc.

6.) Specific information gleaned from your research and experiments –

number of days from sowing to emergence, % viable seed, %
germination, germination requirements (light, temperature
requirements), treatments needed to overcome dormancy, etc.

7.) Price – How much are your seeds worth; how many seeds/packet and
how much would you charge?

8.) References used (at least 3 references should be used – books,

periodicals, and the internet –the internet cannot be the only source for
references).

Recommendation:

73
  Based on your findings, provide a one-sentence recommendation outlining
the best method for germinating the seeds for each of the species you
investigated.

Lets Go Grocery Shopping: Propagating

Produce

[P; Individual; at least 3 species; 5 propagation methods; at least 2

experimental treatments/ technique]

Although you may have never thought of it as such, your neighborhood grocery
store may serve as a rich source of propagules for the production of new plants at
home. For this project, your objective will be to successfully propagate new plants
from produce purchased in local grocery stores. Proceed by selecting the produce
you intend to work with and then formulating a strategy to propagate new plants
from your purchases. Experiment with methods that are different than the standard
practice; in fact for many types of produce, based on what is available in stores, you
will have no choice but to use a different method. For example, cole crops such as
cabbage or brussel sprouts are commercially propagated from seed, however, you
would need to determine how to propagate them from the heads of cabbage or the
individual sprouts or stalks sold in stores. Design experiments to determine the
best method of propagating your material (e.g., different types of propagules or
stem cuttings with and without auxin treatment) and/or trying similar techniques on
a variety of crops. Be creative and don't be afraid to take some chances just for the
fun of it.

74
Your goal is to produce multiple plants from one source. When considering
and choosing propagation techniques, remember this goal!

Some propagation techniques and potential treatments to consider

include:

 Seeds (but not prepackaged vegetable or flower seeds; sorry! Seeds count as
one technique); treatments might include various treatment to enhance
germination.

 Scooping, Scoring, Chipping, or Scaling of bulbs to produce bulblets;

treatments might include method (e.g., scooping vs. scoring) or treatment
with BA (Note – this can be a tricky one; be sure what you think is a bulb, is really a
bulb.).

 Leaf cuttings (leaf-piece, leaf-blade, leaf-petiole, or leaf-bud cuttings);

treatments might include type of cutting or growth regulator treatments.

 Stem Cuttings (herbaceous stem or softwood, semi-hardwood, or hardwood

cuttings); treatments might include size of cutting, wounding, or treatment
with growth regulators.

 Root Cuttings; treatments might include size of cutting or treatment with

growth regulators (Note – this can also be a tricky one; be sure what you think is a
root really is root tissue.).

 Division (tubers, tuberous roots, tuberous stems, rhizomes); treatments

might include different growing media or size of cutting.

 Grafting

 Other Techniques; check with the instructor if you have other ideas.

To satisfy the requirements of this laboratory exercise,

you are required to:

1. Experiment with at least five (5) different propagation techniques.

2. Apply at least two experimental treatments per technique; one

treatment should be a control. If you attempt grafting as a technique, you
will not be required to apply treatments in that case.

3. Successfully propagate completely new plants (roots and shoots)

with at least one (1) technique.

4. Work with at least three (3) different species of produce. The five

75
 techniques you choose may be applied to the same or different varieties of
produce (e.g., both leaf and stem cuttings from the same species or leaf
cuttings from one species and stem cuttings from another). Remember,
seeds count as one technique so propagating three species from seed
doesn’t count as three techniques. Once again, be creative in your plant
selections and methodology. Anyone can easily propagate potatoes from
the tubers available in stores, and potatoes are certainly a fine choice as
one selection; however, you are encouraged to try more unusual produce
selections in your trials.

When making your produce selections, be sure to think about what

techniques might apply. Not all choices will work; bananas are a good
example – Why?

5. Cite at least three references, one of which must be from a source that
is not web based.

In addition to achieving at least a modicum of success, to receive full credit, your

methods must be clearly deliberative and appropriate to the produce you have
selected. For example, dried bay leaves are obviously dead, and no matter what
the methods used, an unsuccessful outcome is predetermined. Also, growth
regulator treatments should be appropriate for the technique used.

Remember, propagation entails the efficient and economical production of many

plants using propagules collected from a group of existing plants or perhaps even
from a single mother plant – in other words propagation involves multiplication. It is
not simply growing a single onion plant from an onion bulb, which is neither efficient
nor economical. Multiplication - optimal increases in the number of quality plants in
production is key to any successful propagation strategy.

Your efforts to propagate your produce selections will be presented in poster form.
Posters are a common and concise method used by researchers to present research
findings at scientific meetings. Feel free to be creative as you design your poster. It
should be complete, yet brief and to the point. The poster should have a title. Be
sure the species, techniques, and treatments used are clearly identified; you should
essentially present a mini lab report for each of the experiments performed. As a
general outline, you might consider using your produce selections as the primary
sections followed by the experiments performed including the techniques and
treatments used. For each produce selection, the following information should be
presented:

1. Objective(s).

2. Plant material used – identify the produce selections using their correct
botanical (Genus, species, and cultivar if known) and common names. You
should also indicate what plant families your produce selections belong to.

3. Methods used – describe the techniques used and the treatments applied.

4. Results – success or failure; if you were unsuccessful, indicate why you think

76
 your efforts failed.

5. A description of how your produce selections are typically propagated

commercially.

6. A one sentence recommendation – based on your efforts, what was the best
way found to propagate your produce selections; if you were not successful,
indicate what you might recommend trying next.

7. Literature cited – list the references from which you obtained information
about your produce selections.

Posters will be presented during a show-and-tell session at the end of the semester.
Samples of your successes should be part of your presentation.

77
 Sexual Propagation: Seed Morphology,
Viability Testing, Germination
Testing, and Epigeal/Epigeous vs.
Hypogeal/Hypogeous Germination
[T; individual; Seed morphology 4 species, 2 replications per species = 8
imbibed seeds/student,
Seed viability 4 species, 5 replications per species = 20 imbibed
seeds/student
Hypogeal vs. Epigeal, 4 species, 5 replications per species = 20 dry
seeds/student]

Objectives:
 To test viability of different seeds.

 To use different methods for determining germination percentage.

 To observe different types of germination.

To Do Before Class:
Read pp. 7-14 in your lab manual and p. 20 in your text - “How a Seed
Germinates”.

Review the information on cut tests and tetrazolium staining at the following web
site: http://msucares.com/pubs/publications/p1978.htm

Plant Material:
Allium cepa (Onion; Liliaceae, Lily Family)

Phaseolus vulgaris (Bean; Fabaceae, Bean Family)

Pisum sativum (Pea; Fabaceae, Bean Family)

Zea mays (Corn; Poaceae, Grass Family)

Materials To Bring To Class:

 Permanent marking pens
 Polyethylene bags
 Lab notebooks

78
Materials Provided:
 Labels
 TTC
 Growing medium
 Pots
 Presoaked seeds
 Dry seeds

Methods:
I. Seed Morphology

a. Obtain two (2) presoaked (imbibed) seeds and one (1) dry seed for each of
the four species provided. The presoaked seed have been soaked in water
overnight to allow them to absorb water. The uptake of water by living
seeds is called imbibition and the seeds are said to be imbibed. Imbibition
is the first step in seed germination. First, observe the differences between
the seeds of the different species. Then compare the dry seed to the
imbibed seeds for each species. Diagram and label your seeds and make
notes about your observations. When you have finished observing your
seeds from the outside, carefully disect them open and diagram your
observations. Microscopes will be available.

II. Viability Testing (% viable seed)

a.Cut tests allow you to look inside the seed to observe the health of
the embryo.

i. Select five (5) presoaked seeds for each of the four species for a total of
20 seeds.

ii. Work on moist a paper towel, cut the corn and onion seeds in half with a
sharp razor blade. Remove the seed coats from the beans and peas
and pull the halves apart.

iii. Observe the condition of the seed and the color of the embryo for each
seed. As a general rule, if the embryo is white in color, it is probably
viable.

iv. Record your observations.

b. Tetrazolium tests (TTC) utilize an enzyme reaction in which live tissues stain
red, and dead tissues do not stain. TTC tests can be used to estimate seed
germination and vigor and can be a useful tool in determining seed quality

79
 and viability during harvesting, conditioning, storage, and distribution.

i. Place the seeds that you cut open in a petri dish, cover with tetrazolium
(TTC) solution, and let sit for up to 1 hour checking staining intensity
every 15 minutes.

ii. After you have reached the desired level of staining, drain the tetrazolium
solution and rinse the seeds two to three times in cold tap water and
evaluate immediately.

iii.Evaluate the stained seeds under magnification and good light. The most
desirable color for seeds is a dark pink to a light red. Darker red seeds
may also be viable. Viable seeds may be completely stained, or the seeds
may have slight, small dead areas over the cotyledon or the chalazal end,
or have dead or weak tissue over less than one-third of the cotyledonary
area. The radicle tip in a good seed may be quite dark, because this is an
area of high-metabolic activity, and the small amount of tissue allows
deeper penetration of the tetrazolium solution, resulting in a dark
appearance.

III. Determining type of germination epigeal/epigeous vs.

hypogeal/hypogeous germination

Epigeal germination = hypocotyl elongates and the cotyledons rise

above the soil
surface

Hypogeal germination = epicotyl elongates and the cotyledons

remain below the soil
surface

a) Plant five (5) seeds of each species in a pot filled with growing medium.
You will have 4 pots, one for each species. Label the pot with a stake
(marked with the species, your name, and the date) and place in the
greenhouse. Also wrap (5) seeds of each species in a damp but not wet
paper towel. Place the paper towel with the seeds in a plastic bag and
store at room temperature.

b) Observe these seeds each lab period. As they germinate, compare your
observations to those for seeds germinated in rag dolls to confirm your
conclusions. When it comes to determining whether germination is epigeal
or hypogeal, it is sometimes difficult to visualize what is happening from
seedlings germinated in rag dolls. It can be difficult to discern what the
orientation and relationship of the seedling to the soil would be had it
actually been planted.

c) After your seeds in the rag dolls have germinated, carefully examine and
diagram your seedlings. Compare your observations to those made earlier.
Based on your observations, determine germination percentage, whether

80
 germination is hypogeal or epigeal, and whether the plant is a monocot or
a dicot, for each type of seed.

d) Follow the emergence of the seeds planted in pots and compare to the
seeds germinated in rag dolls to better understand the difference
between epigeal and hypogeal germination.

Observations:
 Observations for each species, should include (but not be limited to): %
germination, rate of germination, type of germination (hypogeal or epigeal),
and whether the species is a monocot or a dicot.

Some Questions for Thought:

 What environmental factors may have influenced the rate of germination?

 Which technique for determining viability is the easiest? Fastest? Gives the
most information?

 If you were setting up a seed testing laboratory, which of these techniques

might it be important to include? Why?

81
 Asexual Propagation: Whip & Tongue Grafting
[T; individual; 5 grafts/student]

Objectives:
 To better understand the theory behind and demonstrate the techniques
involved in whip & tongue grafting.

 To successfully propagate plants using whip & tongue grafts or at least come
to appreciate the skills involved in grafting as a method of propagation.

 To successfully complete four whip & tongue grafts without cutting oneself;
please be careful!

To Do Before Class:

Read pp. 23-25 (up to “Budding”) in your lab manual and the sections on
grafting in general and whip & tongue grafting in your text (pp. 58-59).

Plant Materials:
Cornus sericea (Red-twig or Red-Osier Dogwood; Cornaceae, Dogwood Family) =
stock or scion

Cornus sericea 'Flaviramea' (Yellow-twig Dogwood; Cornaceae, Dogwood Family)

= stock or scion

Materials To Bring To Class:

 Pruners
 Lab notebooks

Materials Provided:
 Budding rubbers
 Parafilm
 Label stakes
 Paper toweling
 Peat moss (moistened)
 Rooting promoter (3000 ppm IBA talc)

82
Methods:
 We will be using red-twig and yellow-twig dogwood stems to create our whip
& tongue grafts. By grafting these two dogwood selections with different
colored stems together, we will be attempting to create a novel or specialty
plant that has both red and yellow winter stems. Grafting together two plants
that look very different will also serve to visualize and illustrate the grafting
process.

1. Prepare the stock:

a. We will be using unrooted, hardwood cuttings for our rootstocks. The

scion piece will be grafted onto the unrooted cutting. The cutting will then
be rooted at the same time the graft heals. Once again this technique is
called stenting.

b. Select enough woody stems of either red-twig or yellow-twig dogwood to

prepare four (4) cuttings. Stems that are pencil-sized or slightly larger are
best. You can use all red-twig or all yellow-twig for your rootstocks or you
might want to try using a couple of each as stock pieces. Each cutting
should have at least one (1) node at its base and a relatively long section
of internode above. Depending on the length of the internodes for the
plant material available, your rootstock cuttings may have more than one
(1) node.

Note: You may want to collect your scion wood at the same time to ensure
that you have stems of equal diameter (see 2. a. below).

c. Make the grafting cuts (see Figure 1). The cuts required for the whip &
tongue graft are fairly simple and straightforward. Using a clean sharp
razor blade, begin by making a slanting cut 1 to 1.5 inches (4cm) long
within the internode section at the top of each rootstock cutting. Start at a
point about ½ the distance between the pith and the outer edge of the
bark, on the upper side of the slanting cut, make a second cut straight
downward about half as long as the first cut to form the “tongue”.

d. To insure a good fit and close contact between the stock and the scion
pieces, all cuts should be straight and clean; remember, too, that
hemoglobin is not effective as a plant growth regulator. Don’t let your
wood dry out; keep your stock pieces wrapped in moist paper toweling in
a polyethylene bag when you are not working with them directly and be
aware that the wood will dry out as you are making your graft cuts.

2. Prepare the scion:

a. Select just enough woody stems to make four (4) scions having opposite
stem colors as the stock pieces already prepared. It is important that the

83
 wood used for the scions be equal in diameter to the stock pieces. Each
scion piece should have one (1) or two (2) pairs of buds (nodes) at the top
and a relatively long section of internode at its base.

b. Make the grafting cuts (see Figure 1). Holding the scion piece upside
down, make exactly the same cuts in the base of the scion piece as were
made in the stock; the length and angle of the cuts on the scion piece
must be identical to those on the stock to insure a good match. Again,
keep your scion pieces from drying out.

3. Join the stock and scion pieces:

a. Slide the stock and scion together. It is very important that the stock and
scion pieces fit together tightly (See Figure 1). Hold the grafts up to the
light to look for gaps and make adjustments as needed. Once again, keep
your stock and scion pieces, especially the cut surfaces that will be joined
together, from drying out.

b. Wrap the joined pieces with parafilm followed by a budding rubber. Wrap
the budding rubber from the bottom up. Both the parafilm and the
budding rubber should extend a half inch or so above and below the cut
surfaces of the graft to provide sufficient support. A budding rubber may
be used alone, however, the parafilm is easier to apply and helps reduce
frustration by holding the pieces of the graft in place while the budding
rubber is applied. Do not use parafilm alone as it breaks down and does
not last long enough.

c. Don't get frustrated. Remember, learning to graft takes practice

and patience. Your technique should improve with each attempt.

4. Treat the bases of your grafted cuttings with 3000 ppm IBA in talc.

5. Stick two (2) of your completed grafts directly into your plot in the mist
bench. Be sure to label them. Pre-callus the remaining two (2) grafts for two
weeks prior to sticking by packing them in moist peat moss in a polyethylene
bag; label the bag with your name and the date and place in the box
provided.

6. Observe your grafts on a regular basis and record your observations. Avoid
the temptation to remove the budding rubber and parafilm to "sneak a peek"
for at least five (5) weeks.

7. When your grafted rootstocks are well rooted, and the grafts have healed, pot
them up. The grafts will continue to heal in the greenhouse. If your grafts
should show signs of wilting after potting, enclosing the potted graft in a
polyethylene bag for a time and then gradually hardening-off may be
helpful.

8. As always, be sure to clean up all work areas when you are finished.
84
Observations:
 Observations should include (but not be limited to): callus formation, bud
break, root development and success or failure of your grafts.

Some Questions To Ponder:

 Were your grafts successful or not? If not, why not?

 What factors might have influenced the success or failure of your whip &
tongue grafts?

 Based on this experience, what might you do differently in subsequent

attempts at whip & tongue grafting?
 Why is it important for the stock and scion to be the same diameter? What if
they were not similar?

 Why is grafting done? Why do you suppose grafting is a more expensive

means of propagation?

 Which factor would you say is most important in grafting success?

85
Figure 1. Making a Whip and Tongue Graft.

Preparing Preparing Joining the stock and

the scion the stock the scion

 Asexual Propagation: Unique Vegetative
Structures
[T; individual]

Objectives:
 To become familiar with storage structures

 To better understand and demonstrate several techniques commonly used to

propagate plants with unique vegetative storage structures.

To Do Before Class:
 Read 21-22 in your lab manual, 25-26 (pictures on p. 27) in your text
 Scaling p 258 in your text
 Rhizomes pp. 149, 169, 184, 191, and 288 in text
 Stolons p162-163 in text
 Bulbils p.273 in text
 Tubers p. 235 in text

Types of structures and plant material to be used:

Scaly Bulbs: Lilium sp. (Asiatic Lily; Liliaceae, Lily Family)

Tunicate Bulbs: Tulipa Sp. (Tulip; Liliaceae, Lily Family)

Rhizomes: to be determined

Stolons: spider plant, strawberries

Bulbils: Tiger lily (Lilium lancifolium, Liliaceae, Lily Family)

Tubers: Potato

Materials To Bring To Class:

 Pruners
 Latex gloves (if you think you might be sensitive to plant exudates)
 Lab notebooks

Materials Provided:
 Pots
 Labels
  Incubation medium – 50:50 milled sphagnum moss:perlite

Methods:
For the purpose of comparison, and to better understand their structure, observe
and diagram the dissected tunicate (tulip and grape hyacinth) and scaly (lily) bulbs
provided. Other types of “bulbs” (corms, tubers, tuberous roots, etc.) may also be
provided for the purpose of observation. Be sure to record your observations and
label your diagrams.

Scaly Bulbs
1. Obtain structures for propagating:
 10 lily scales
 3 pieces of rhizome
 3 pieces of stolon
 2 bulbils
 1 tuber

2. The shared lily bulb will be used to demonstrate scaling (see p. 258 in your
text). Simply separate 10 individual scales from the basal plate of your lily
bulb. The scales should be plump and in good condition.

3. Place your bulb scales into a clean polyethylene bag together with three to
four times their volume of the moistened incubation medium provided.

4. Be sure to label the bag with your name, the date, the species, and the
method used. Place your bag in the boxes provided; they will be incubated
in a warm, dark place.

5. Your bulb scales will be available for observation during subsequent lab
periods during the semester; observe them on a regular basis for bulblet
development and record your observations. Remove any materials showing
signs of decay; if you are not sure, ask.

Rhizomes: To be determined

1. Remove plants from pots if necessary. Using a sharp razor blade cut off a
piece of the rhizome (see pictures p. 288 in your text).

2. Each cutting needs to have at least one node on it.

3. Plant the pieces of rhizome together in a pot, cover with soil. Label your pot
with name, date, type of structure and place it in the greenhouse.

4. In subsequent lab periods, take observations on above ground growth.

Stolons: spider plant, strawberries

1. Remove plants from pots if necessary. Using a sharp razor blade, cut off a
piece of the stolon (see pictures p. 197 in your text, they are called
runners).
 2. Each cutting needs one node.

3. Plant the pieces of stolon together in a pot, cover with soil. Label your pot
with name, date, type of structure and place it in the greenhouse.

4. In subsequent lab periods, take observations on above ground growth.

Bulbils: Tiger lily (Lilium lancifolium, Liliaceae, Lily Family)

1. Remove 2 bulbils from plants (see pictures p. 273 in your text).

2. Plant the 2 bulbils in a pot, cover with soil. Label your pot with name, date,
type of structure and place it in the greenhouse.

3. In subsequent lab periods, take observations on above ground growth.

Tubers
1. Remove plants from pots if necessary. Using a sharp razor blade, divide tuber
(see pictures p. 235 in your text).

2. Each tuber cutting needs to have at least one node on it.

3. Plant the pieces of tuber together in a pot, cover with soil. Label your pot with
name, date, type of structure and place it in the greenhouse.

4. In subsequent lab periods, take observations on above ground growth.

Wash your hands and, as always, be sure to clean up all work areas when you are
finished.

Observations:
 Observations should include (but not be limited to): labeled diagrams of the
various bulbs provided, the success or failure of each technique, time to
bulblet initiation, and the number and relative size of the bulblets produced.

Some Questions to Ponder:

 For each species, which method of propagation, scooping, scoring, chipping
or scaling, worked the best?

 In each case, where did bulblets form and how many were produced?

 If you were unsuccessful with a specific technique, what might be some

reasons why you were unsuccessful?

 If you were to try these propagation methods again, what might you do
differently?
 Asexual Propagation: T-Budding
[T; individual; 5 grafts/student]

Objectives:
 To better understand and demonstrate the techniques involved in T-budding.

 To successfully propagate a novelty plant using T-budding.

To Do Before Class:
Read pp. 23-25 in your lab manual and the sections on T-budding in your text
(pp. 62, 114-115).

Plant Materials:
Hibiscus rosa-sinensis; named cultivars (Various selections of Chinese Hibiscus;
Malvaceae, Mallow Family) = scions (stock plants) & stocks (potted plants)

Materials To Bring To Class:

 Pruners
 Permanent marking pens
 Polyethylene bags
 Lab notebooks

Materials Provided:
 Budding rubbers
 Parafilm
 Label stakes
 Paper toweling

Methods:
1. Prepare the stock:

a. For this grafting exercise you will be using a plant that already has a root
system as the stock. Select a healthy, vigorous hibiscus plant from the
assortment provided; the plants we will be using are named cultivars and
were propagated from softwood cuttings.

91
 b. Starting near the base of the plant, and working up, you will be grafting
five (5) buds, each from a different hibiscus cultivar, onto the rootstock
plant. Try and choose locations along the stem that are relatively straight
and evenly spaced along the lower portion of the stem.

c. Make the grafting cuts (See Figure 1). For each bud-graft, begin by making
a 1 inch (2.5 cm) long, vertical cut through the bark using a clean, sharp
razor blade. Make a second, horizontal cut centered on and just
intersecting the top of the first cut to form a "T". Make sure your cuts
connect and go all the way through the bark (it is thicker than you think),
yet not cut into the underlying wood.

d. Using the corner of your razor blade, peel back the resulting flaps at the
top of the "T" as if you were opening a book, being careful to include the
green cambium layer with the flap. Take care to avoid contaminating the
inner surfaces of the wound. The scion bud-piece will be inserted into the
resulting opening in the bark.

2. Prepare the scion:

a. When T-budding, the scion basically consists of a single bud with some
attached bark and wood. An assortment of hibiscus plants, again named
cultivars, will serve as our source of budwood. Choose the varieties you
want to work with – you will need to collect five (5) buds; each from a
different variety than your rootstock plant. If all five buds are successful
you will have created a six-in-one hibiscus plant.

b. To collect the individual bud scions, choose healthy-looking, mature buds.

Remove the leaf blade from the subtending leaf, but leave the petiole
intact to serve as a handle. To remove the bud, undercut the bud starting
your cut about 3/4 of an inch (1 cm) below the bud and continuing up and
beyond the bud for about 1 inch (2.5 cm).

3. Join the stock and scion to complete the graft:

a. Using the petiole as a handle, and the bud pointing up, insert the bud scion
into the T-shaped cut in the bark of the rootstock plant. Remove the
excess bark above the bud by cutting it off horizontally such that the scion
fits flush under the edge of the bark at the top of the "T".

b. Starting below the graft, wrap the graft with a parafilm covering all the cut
surfaces but not the bud itself. Trim the petiole such that only a short stub
remains.

c. Throughout the process, prevent excessive drying of the stock and bud
scions. To this end, work quickly and complete each graft separately. Buds
may be temporarily wrapped in clean, moist paper toweling in a
polyethylene bag to prevent them from drying out.

92
 4. Observe your grafts on a regular basis and record your observations. Avoid
the temptation to remove the parafilm to "sneak a peek" for at least several
weeks. If the petiole remains green and eventually abscises naturally the
bud is probably in good shape. If, however, the petiole dries-up and remains
firmly attached, the graft has probably failed.

5. As always, be sure to clean up all work areas when you are finished.

Observations:
 Observations should include (but not be limited to): callus formation,
condition of the petiole stub, appearance of the bud, bud break, and success
or failure of your grafts.

Some Questions to Ponder:

 Were your grafts successful or not? If not, Why not?

 Why would plant propagators use T-budding?

 What environmental conditions are important for successful T-budding?

93
Figure 1. T-Budding.

Preparing the stock Preparing the scion

Joining the stock and scion

94
 Asexual Propagation: Approach Grafting –
Spudmatoes

[T; individual; 2 grafts/student]

Objectives:
 To better understand and demonstrate the techniques involved in approach
grafting.

 To successfully propagate plants using approach grafting.

To Do Before Lab:
Read pp. 23-25 in your lab manual and p. 303 in your text.

Plant Materials:
Lycopersicon esculentum (Tomato; Solanaceae, Potato Family) = scion

Solanum tuberosum (Potato; Solanaceae, Potato Family) = stock

Materials To Bring To Class:

 Permanent marking pens
 Lab notebooks

Materials Provided:
 Budding rubbers
 Parafilm
 Paper toweling
 Bamboo stakes
 Twist ties

Methods:
1.To create your spudmatoes, a grafted plant which will produce both potatoes
and tomatoes, the potato will need to serve as the rootstock since the
potatoes are produced below ground. The tomato plant will become the
scion.

95
 2.Select two (2) healthy potato plants and two (2) healthy tomato plants from
the plug-grown plants provided. Be sure you can tell them apart (note that
the tomatoes have a distinctly different smell!).

3.Repot the two plants as close as possible to each other in the center of one of
the containers provided.

4.Once planted, gently bring the stems of the two plants together to determine
where the stems naturally come into contact without undue force or
contortion. Using a razor blade, remove any leaves that are attached to the
stems in this region.

5.Using a clean, sharp razor blade, carefully shave a thin (1/32 to 1/16 of an
inch or about 1 mm thick) strip from each stem in the region where the
stems came in contact when pushed together in Step 4. The wounded area
should be 1 to 1.5 inches (2.5 to 4 cm) long. Be very careful; the stems are
delicate and it is very easy to cut right through them.

6.Push the stems together again such that the wounded areas are in solid
contact. Carefully wrap the graft with a strip of parafilm making sure the
stems remain in solid contact (See Figure 1).

7.When you have finished wrapping the graft, stake the grafted plant using a
bamboo stake and a couple of twist ties.

8.Repeat the procedure with your remaining potato and tomato plants. Label
your plants with your name and the date.

9.Water your grafted plants and transfer them to the designated location in the
greenhouse.

10.Observe your grafts on a regular basis and record your observations. Avoid
removing the wrapping to "sneak a peek" for at least five (5) weeks.

11.When the grafts have healed you can complete the process by severing the
root system of the tomato plant just below the graft union and the top of the
potato plant just above the graft union. Be careful to know for sure which
stems you are cutting before you make your cuts. Keep the plant staked to
help reduce the chances of breaking the graft and use a razor blade to cut
the stems.

12.If the top of your grafted plant (tomato) does not wilt, it is a good sign that
your graft has been successful.

13.As always, be sure to clean up all work areas when you are finished.

Observations:

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  Observations should include (but not be limited to): callus formation and
success or failure of your grafts.

Some Questions to Ponder:

 Were your grafts successful or not? If not, why not?

 What is the primary difference between the approach graft and the other
grafts we have studied so far? What, then, is the advantage of the approach
graft?

Figure 1. Making an approach graft.

Preparing the stock and the scion Joining the stock and
the scion

97
 Vegetative Propagation: Air Layering
[T; individual; 2 air layers/student]

Objectives:
 To better understand and demonstrate the techniques involved in air
layering.

 To successfully propagate a house plant by air layering.

To Do Before Class:
Read the section on “Layering” in your lab manual (p. 22) and the section on air
layering in your text (pp.24-25 “Layering”, 64, and 105-107).

Plant Material:
Schefflera actinophylla (Umbrella Tree; Araliaceae, Aralin Family)

and/or Perhaps Others

Materials To Bring To Class:

 Permanent marking pens
 Polyethylene bags
 Lab notebooks

Materials Provided:
 Label stakes
 Bamboo stakes
 Sphagnum moss (moistened)
 Twist ties or string

Methods:
Go and get one (1) potted Schefflera plant.

1. Select a location approximately midway up the stem on each plant for your
air-layers.

2. Carefully remove any shoots or leaves from the portion of the stem to be

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 layered.

3. Wound the stem. This may be done by making a slanting cut upward into
the stem or by girdling the stem. You may each of the plants in the same
fashion or try a different technique with each plant.

4. If a slanting cut is used, carefully use a razor blade to make the cut such that
it extends into the stem about 1/3 its diameter and is about 1 inch long. (Do
not cut halfway into the stem or it could easily break off.)

5. If girdling is chosen, girdle the stem by removing a ring of stem tissue about
1/4 inch (6 mm) wide and 1/16 inch (2 mm) deep.

6. Cover the wounded area, and an additional inch or so on either side, with
moistened sphagnum moss. If the stem was wounded with a slanting cut,
hold the cut open by inserting some of the sphagnum into the open cut.

7.Cut open one of your polyethylene bags to create a flat sheet. Use this sheet
of polyethylene film to wrap the layer. The polyethylene wrapping should
extend far enough beyond the moss to allow it to be secured with string or
twist ties.

8. Using label stakes, label your layers with your name and the date and place
them in the designated location in the greenhouse. Water your plants.

9. Observe your plants at least weekly and record your observations. If the
sphagnum moss appears to be drying out, carefully open the plastic sleeve
and remoisten it. When you observe numerous roots growing into the moss,
remove the plastic sleeve and cut the stem below the root mass. Carefully
remove the excess moss and spread the roots, taking care to keep the roots
moist at all times. Plant the rooted layer in the planting media and pots
provided. Water the new plant thoroughly after potting.

10. As always, be sure to clean up all work areas when you are finished .

Observations:
 Observations should include (but not be limited to): success or failure.

Some Questions to Ponder:

 Where do new roots appear?

 Do you think using rooting promoters would improve success?

 If you were propagating your house plant on a commercial basis, how might
you manipulate the parent plant to increase propagation efficiency?

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Air-Layer

1.) Girdled Stem

2.) Peat Moss around girdled section
3.) Plastic fastened around Peat Moss and
girdled area