ASEXUAL

PROPAGATION
Asexual Propagation

 Asexual propagation is used to reproduce or multiply many

horticultural plants. Plants that are propagated asexually
are genetically the same as the mother plant.
 It is multiplication without passage through the seed cycle,
and is the best way to maintain some species, particularly
an individual that best represent that species.
 Asexual propagation involves taking a part of one parent
plant and causing it to regenerate itself into a new plant.
 Clones are groups of plants that are identical to their one
parent and that can only be propagated asexually.
Reasons for Asexual Propagation

1. to preserve the genetic characteristics of a particular

plant;
2. to propagate plants that do not produce viable seeds
(bananas, pineapple, seedless grape, etc.);
3. to propagate plants that produce seed that is difficult to
germinate or has a very short storage life (cotoneaster,
willow);
4. to bypass the juvenile stage of plant growth when the
plants will not flower and bare fruit (apple).
Examples of Asexual Propagation

 The Bartlett pear (1770) and the Delicious apple (1870)

are two examples of clones that have been asexually
propagated for many years.
 One of the oldest clones in existence is Thompson seedless
grapes.
 The Kieffer pear and the Peace rose are two examples of
clones that have been asexually propagated for many
years.
Suitable Species for Asexual Propagation

 Some asexually propagated crops that are grown

extensively are: tree fruits, cane fruits, strawberries,
sugar cane, potatoes, sweet potatoes, cassava,
cranberries, and most herbaceous and woody ornamental
plants.
 Almost all the flower crops and green plants grown as
greenhouse crops are also propagated asexually.
 Many plants, such as ginger, onion, gladioli, and dahlia are
also propagated asexually.
Suitable Species for Asexual Propagation

 In some plants, such as the sweet potato, adventitious

roots or runners (stolons) can give rise to new plants.
 In Bryophyllum and kalanchoe, the leaves have small buds
on their margins. When these are detached from the
plant, they grow into independent plants; they may also
start growing into independent plants if the leaf touches
the soil.
Major Methods of Asexual Propagation

 Cutting – involves rooting a severed piece of the parent

plant
 Grafting and Budding – joining two plant parts from
different varieties
 Layering – involves rooting a part of the parent plant and
then severing it
 Marcotting – an old term regarding the method of
propagation now known as air layering
CUTTING
Cutting

 Stem cuttings are made by removing a small branch or twig

from the plant. This cutting will usually contain two or more
buds, one of which will grow into the top of the plant. With
proper treatment, adventitious roots will be produced on
the end of the cutting that was closest to the root of the
original plant.
 Root cuttings are made in a similar fashion, but produce an
adventitious stem on the end of the cutting that was nearest
to the stem of the original plant.
 Leaf cuttings produce both roots and stems when the leaf is
placed under proper conditions.
Cutting

 Propagation by stem cuttings is the most commonly used

method to propagate many woody ornamental plants.
Stem cuttings of many favorite shrubs are quite easy to
root. Typically, stem cuttings of tree species are more
difficult to root. However, cuttings from trees such as
crape myrtles, some elms, and birches can be rooted.
 The four main types of stem cuttings are herbaceous,
softwood, semi-hardwood, and hardwood. These terms
reflect the growth stage of the stock plant, which is one
of the most important factors influencing whether or not
cuttings will root.
Cutting

 A greenhouse is not necessary for successful propagation

by stem cuttings; however, maintaining high humidity
around the cutting is critical.

 If rooting only a few cuttings, you can

use a flower pot (Figure 1). Maintain
high humidity by covering the pot
with a bottomless milk jug or by
placing the pot into a clear plastic
bag.
Figure 1
Cutting

 Cuttings can also be placed in

plastic trays covered with clear
plastic stretched over a wire frame
(Figure 2). Trays must have holes in
the bottoms for drainage. The
plastic will help keep the humidity
Figure 2
high and reduce water loss from
the cuttings.
Four Main Types of Stem Cuttings

1. Herbaceous cuttings are made from non-woody, herbaceous plants such as

coleus, chrysanthemums, and dahlia. A 3- to 5-inch piece of stem is cut
from the parent plant. The leaves on the lower one-third to one-half of the
stem are removed.
2. Softwood cuttings are prepared from soft, succulent, new growth of
woody plants, just as it begins to harden (mature). Shoots are suitable for
making softwood cuttings when they can be snapped easily when bent and
when they still have a gradation of leaf size (oldest leaves are mature
while newest leaves are still small).
Four Main Types of Stem Cuttings

3. Semi-hardwood cuttings are usually prepared from partially mature wood

of the current season’s growth, just after a flush of growth. This type of
cutting normally is made from mid-July to early fall. The wood is
reasonably firm and the leaves of mature size. Many broadleaf evergreen
shrubs and some conifers are propagated by this method.
4. Hardwood cuttings are taken from dormant, mature stems in late fall,
winter, or early spring. Plants generally are fully dormant with no obvious
signs of active growth. The wood is firm and does not bend easily.
Hardwood cuttings are used most often for deciduous shrubs but can be
used for many evergreens. Examples of plants propagated at the hardwood
stage include forsythia, privet, fig, grape, and spirea.
Three Types of Hardwood Cuttings

 The three types of hardwood cuttings are straight, mallet, and

heel (Figure 3). A straight cutting is the most commonly used
stem cutting. Mallet and heel cuttings are used for plants that
might otherwise be more difficult to root. For the heel cutting,
a small section of older wood is included at the base of the
cutting. For the mallet cutting, an entire section of older stem
wood is included.
Figure 3. The three types of
hardwood cuttings are straight,
mallet, and heel.
Procedures for Rooting Stem Cuttings

1. Avoid material with flower buds if possible. Remove any flowers and flower buds
when preparing cuttings so the cutting’s energy can be used in producing new roots
rather than flowers. Take cuttings from healthy, disease-free plants, preferably
from the upper part of the plant.
2. Avoid taking cuttings from plants that show symptoms of mineral nutrient
deficiency. Conversely, plants that have been fertilized heavily, particularly with
nitrogen, may not root well. The stock plant should not be under moisture stress. In
general, cuttings taken from young plants root in higher percentages than cuttings
taken from older, more mature plants. Cuttings from lateral shoots often root
better than cuttings from terminal shoots.
Procedures for Rooting Stem Cuttings

3. Early morning is the best time to take cuttings, because the plant is
fully turgid. It is important to keep the cuttings cool and moist until
they are stuck. An ice chest or dark plastic bag with wet paper towels
may be used to store cuttings. If there will be a delay in sticking
cuttings, store them in a plastic bag in a refrigerator.
4. While terminal parts of the stem are best, a long shoot can be divided
into several cuttings. Cuttings are generally 4 to 6 inches long. Use a
sharp, thin-bladed pocket knife or sharp pruning shears. If necessary,
dip the cutting tool in rubbing alcohol or a mixture of 1 part bleach to 9
parts water to prevent transmitting diseases from infected plant parts
to healthy ones.
Procedures for Rooting Stem Cuttings

5. Remove the leaves from the lower one-

third to one-half of the cutting (Figure 4
). On large-leafed plants, the remaining
leaves may be cut in half to reduce
water loss and conserve space. Figure 4. Remove the leaves from the
lower one-third to one-half of the
cutting.
6. Treating cuttings with root-promoting compounds can be a valuable tool in
stimulating rooting of some plants that might otherwise be difficult to root.
Prevent possible contamination of the entire supply of rooting hormone by
putting some in a separate container before treating cuttings. Any material
that remains after treatment should be discarded and not returned to the
original container. Be sure to tap the cuttings to remove excess hormone
when using a powder formulation.
Procedures for Rooting Stem Cuttings

7. The rooting medium should be sterile, low in fertility, and well-drained to provide
sufficient aeration. It should also retain enough moisture so that watering does not have
to be done too frequently. Materials commonly used are coarse sand, a mixture of one
part peat and one part perlite (by volume), or one part peat and one part sand (by
volume). Vermiculite by itself is not recommended, because it compacts and tends to
hold too much moisture. Media should be watered while being used.
8. Insert the cuttings one-third to one-half their length into the medium. Maintain the
vertical orientation of the stem (do not insert the cuttings upside down). Make sure the
buds are pointed up. Space cuttings just far enough apart to allow all leaves to receive
sunlight. Water again after inserting the cuttings if the containers or frames are 3 or
more inches in depth.
Procedures for Rooting Stem Cuttings

9. Cover the cuttings with plastic and place in indirect light. Avoid direct sun.
Keep the medium moist until the cuttings have rooted. Rooting will be
improved if the cuttings are misted on a regular basis.
10. Rooting time varies with the type of cutting, the species being rooted, and
environmental conditions. Conifers require more time than broadleaf plants.
Late fall or early winter is a good time to root conifers. Once rooted, they
may be left in the rooting structure until spring.
11. Newly rooted cuttings should not be transplanted directly into the landscape.
Instead, transplant them into containers or into a bed. Growing them to a
larger size before transplanting to a permanent location will increase the
chances for survival.
GRAFTING & BUDDING
Grafting & Budding

 Grafting and budding are horticultural techniques used to join parts from
two or more plants so that they appear to grow as a single plant. In
grafting, the upper part (scion) of one plant grows on the root system
(rootstock) of another plant. In the budding process, a bud is taken from
one plant and grown on another.
 Since grafting and budding are asexual or vegetative methods of
propagation, the new plant that grows from the scion or bud will be
exactly like the plant it came from. These methods of plant reproduction
are usually chosen because cuttings from the desired plant root poorly (or
not at all). Also, these methods give the plant a certain characteristic of
the rootstock - for example, hardiness, drought tolerance, or disease
resistance.
Grafting & Budding

 Since both methods require extensive knowledge of nursery

crop species and their compatibility, grafting and budding
are two techniques that are usually practiced only by more
experienced nursery operators.
 Most woody nursery plants can be grafted or budded, but
both processes are labor intensive and require a great deal
of skill. For these reasons they can be expensive and come
with no guarantee of success. The nurseryman must
therefore see in them a marked advantage over more
convenient propagation techniques to justify the time and
cost.
Grafting & Budding

 Grafting and budding can be performed only at very

specific times when weather conditions and the
physiological stage of plant growth are both optimum. The
timing depends on the species and the technique used.
 For example, conditions are usually satisfactory in June
for budding peaches, but August and early September are
the best months to bud dogwoods. Conversely, flowering
pears can be grafted while they are dormant (in
December and January) or budded during July and August.
Reasons for Grafting and Budding
 Change varieties or cultivars. An older established orchard of fruiting
trees may become obsolete as newer varieties or cultivars are developed.
The newer varieties may offer improved insect or disease resistance,
better drought tolerance, or higher yields.
 Optimize cross-pollination and pollination. Certain fruit trees are not
self-pollinating; they require pollination by a second fruit tree, usually of
another variety. This process is known as cross-pollination. Portions of a
tree or entire trees may be pollinated with the second variety to ensure
fruit set. To ensure good fruit set on the female (pistillate) plant, a male
(staminate) plant must be growing nearby. Where this is not possible, the
chances that cross-pollination will occur can be increased by grafting a
scion from a male plant onto the female plant.
Reasons for Grafting and Budding
 Take advantage of particular rootstocks. Compared to the selected scion,
certain rootstocks have superior growth habits, disease and insect resistance,
and drought tolerance. For example, when used as rootstock for commercial
apple varieties, the French crabapple (Malus sylvestris, Mill.) can increase
resistance to crown gall and hairy root. Malling VIII and Malling IX are used as
dwarfing rootstocks for apple trees when full-sized trees are not desired, such
as in the home garden.
 Benefit from interstocks. An interstock can be particularly valuable when the
scion and rootstock are incompatible. In such cases, an interstock that is
compatible with both rootstock and scion is used. An interstock could increase
the disease resistance or cold hardiness of the scion. Plants also may be double
worked to impart dwarfness or influence flowering and fruiting of a scion.
Reasons for Grafting and Budding
 Perpetuate clones. Clones of numerous species of conifers cannot be
economically reproduced from vegetative cuttings because the percentage
of cuttings that root successfully is low. Many can be grafted, however,
onto seedling rootstocks. Numerous clones of Japanese maple (Acer
palmatum, Thunb.) That either root poorly or lack an extensive root system
are grafted onto seedling Acer palmatum rootstock.
 Produce certain plant forms. Numerous horticultural plants owe their
beauty to the fact that they are grafted or budded onto a standard,
especially those that have a weeping or cascading form. In most cases,
multiple scions are grafted or budded 3 feet or higher on the main stem of
the rootstock. When used this way, the rootstock is referred to as a
standard. It may require staking for several years until the standard is large
enough to support the cascading or weeping top.
Reasons for Grafting and Budding
 Repair damaged plants. Large trees or specimen plants can be damaged easily at
or slightly above the soil line. The damage can often be repaired by planting
several seedlings of the same species around the injured tree and grafting them
above the injury.
 Increase the growth rate of seedlings. The seedling progeny of many fruit and nut
breeding programs, if left to develop naturally, may require 8 to 12 years to
become fruitful. However, if these progeny are grafted onto established plants,
the time required for them to flower and fruit is reduced dramatically.
 Index viruses. Many plants carry viruses, although the symptoms may not always
be obvious or even visible. The presence or absence of the virus in the suspect
plant can be confirmed by grafting scions from the plant onto another plant that is
highly susceptible and will display prominent symptoms.
GRAFTING
When to Graft

 Unlike budding, which can be performed before or during the

growing season, most grafting is done during winter and early
spring while both scion and rootstock are still dormant.
 Containerized plants may be moved indoors during the actual
grafting process; after grafting, these plants are placed in
protected areas or in unheated overwintering houses. Field-
grown stock, of course, must be grafted in place
 Some deciduous trees are commonly grafted as bare
rootstock during the winter and stored until spring planting.
Indoor winter grafting is often referred to as bench grafting
because it is accomplished at a bench.
Selecting and Handling Scion Wood
 The best quality scion wood usually comes from shoots grown the previous
season. Scions should be severed with sharp, clean shears or knives and
placed immediately in moistened burlap or plastic bags. It is good practice
during the harvesting of scions and the making of grafts to clean the cutting
tools regularly. This may be done by flaming or immersing them in a
sterilizing solution. Isopropyl (rubbing) alcohol also works well as a sterilant,
although it evaporates quite readily.
 For best results, harvest only as much scion wood as can be used for grafting
during the same day. Select only healthy scion wood that is free from insect,
disease, or winter damage. Be sure the stock plants are of good quality,
healthy, and true to type. Scion wood that is frozen at harvest often knits
more slowly and in lower percentage.
Selecting and Handling Scion Wood

If large quantities of scion wood must be harvested at one time, follow these steps:
 Cut all scions to a uniform length, keep their basal ends together, and tie them in bundles of
known quantity (for example, 50 scions per bundle).
 Label them, recording the cultivar, date of harvest, and location of the stock plant.
 Wrap the base of the bundles in moistened burlap or sphagnum, place them in polyethylene
or waterproof paper bags, and seal the bags.
 Store the bundles for short periods, if necessary, either iced down in insulated coolers or in a
commercial storage unit at 32° to 34°F.
 Never store scions in refrigerated units where fruits or vegetables are currently kept or have
been stored recently. Stored fruits and vegetables release ethylene gas, which can cause
woody plant buds to abort, making the scions useless.
 Keep the scions from freezing during storage.
Types of Grafts

 Cleft Graft  Whip and Tongue Graft

 Bark Graft  Saddle Graft
 Side-Veneer Graft  Bridge Graft
 Splice Graft  Inarch Graft
Cleft Graft

 One of the simplest and most popular forms of grafting,

cleft grafting, is a method for top working both flowering
and fruiting trees (apples, cherries, pears, and peaches) in
order to change varieties. Cleft grafting is also used to
propagate varieties of camellias that are difficult to root.
This type of grafting is usually done during the winter and
early spring while both scion and rootstock are still
dormant. Cleft grafting may be performed on main stems
or on lateral or scaffold branches.
Cleft Graft
 Preparing the Rootstock. The stock should be sawed off with a clean, smooth cut perpendicular to
the main axis of the stem to be grafted. Using a clefting tool wedge and a mallet, make a split or
"cleft" through the center of the stock and down 2 to 3 inches. Remove the clefting tool wedge and
drive the pick end of the tool into the center of the newly made cleft so that the stock can be held
open while inserting the scion.
 Preparing the Scion. In cleft grafting, one scion is usually inserted at each end of the cleft, so
prepare two scions for each graft. Select scions that have three or four good buds. Using a sharp,
clean grafting knife, start near the base of the lowest bud and make two opposing smooth-tapered
cuts 1 to 2 inches long toward the basal end of the scion. Cut the side with the lowest bud slightly
thicker than the opposite side. Be sure the basal end of the scion gradually tapers off along both
sides.
 Inserting the Scion. Insert a scion on each end of the cleft, with the wider side of the wedge facing
outward. The cambium of each scion should contact the cambium of the rootstock.
 Securing the Graft. Remove the clefting tool from the cleft so that the rootstock can close.
Pressure from the rootstock will hold the scions in place. Thoroughly seal all cut surfaces with
grafting wax or grafting paint to keep out water and prevent drying. If both scions in the cleft
"take," one will usually grow more rapidly than the other. After the first growing season, choose the
stronger scion and prune out the weaker.
Cleft Graft
Bark Graft

 Bark grafting is used primarily to top work flowering and

fruiting trees. In contrast to cleft grafting, this technique
can be applied to rootstock of larger diameter (4 to 12
inches) and is done during early spring when the bark slips
easily from the wood but before major sap flow. The
rootstock is severed with a sharp saw, leaving a clean cut
as with cleft grafting.
Bark Graft
 Preparing the Stock. Start at the cut surface of the rootstock and make a vertical slit
through the bark where each scion can be inserted (2 inches long and spaced 1 inch
apart).
 Preparing the Scion. Since multiple scions are usually inserted around the cut surface of
the rootstock, prepare several scions for each graft. Cut the base of each scion to a 1 1⁄2-
to 2-inch tapered wedge on one side only.
 Inserting the Scion. Loosen the bark slightly and insert the scion so that the wedge-
shaped tapered surface of the scion is against the exposed wood under the flap of bark.
Push the scion firmly down into place behind the flap of bark, replace the bark flap, and
nail the scion in place by driving one or two wire brads through the bark and scion into the
rootstock. Insert a scion every 3 to 4 inches around the cut perimeter of the rootstock.
 Securing the Graft. Seal all exposed surfaces with grafting wax or grafting paint. Once
the scions have begun to grow, leave only the most vigorous one on each stub; prune out
all the others. Bark grafts tend to form weak unions and therefore usually require staking
or support during the first few years.
Bark Graft
Side-Veneer Graft

 At one time the side-veneer graft was a popular technique

for grafting varieties of camellias and rhododendrons that
are difficult to root. Currently, it is the most popular way
to graft conifers, especially those having a compact or
dwarf form. Side-veneer grafting is usually done on potted
rootstock
Side-Veneer Graft
 Preparing the Stock. Rootstock is grown in pots the season before grafting, allowed to go
dormant, and then stored as with other container nursery stock. After exposure to cold
weather for at least six weeks, the rootstock is brought into a cool greenhouse for a few days
before grafting takes place to encourage renewed root growth. The plant should not be
watered at this time.
Make a shallow downward cut about 3⁄4-inch to 1 inch long at the base of the stem on the
potted rootstock to expose a flap of bark with some wood still attached. Make an inward cut
at the base so that the flap of bark and wood can be removed from the rootstock.
 Preparing the Scion. Choose a scion with a diameter the same as or slightly smaller than the
rootstock. Make a sloping cut 3⁄4-inch to 1 inch long at the base of the scion. Use the bark
grafting technique.
 Inserting the Scion. Insert the cut surface of the scion against the cut surface of the
rootstock. Be certain that the cambia contact each other.
 Securing the Graft. Hold the scion in place using a rubber grafting strip, tape, or grafting
twine. Seal the entire graft area with warm grafting wax or grafting paint. Remove the
rubber or twine shortly after the union has healed. Never allow the binding material to girdle
the stem.
Side-Veneer Graft
Splice Graft

 Splice grafting is used to join a scion onto the stem of a

rootstock or onto an intact root piece. This simple method
is usually applied to herbaceous materials that callus or
"knit" easily, or it is used on plants with a stem diameter
of 1⁄2-inch or less. In splice grafting, both the stock and
scion must be of the same diameter.
Splice Graft
 Preparing the Stock and Scion. Cut off the rootstock using a diagonal
cut 3⁄4-inch to 1 inch long. Make the same type of cut at the base of
the scion.
 Inserting the Scion. Fit the scion to the stock. Wrap this junction
securely with a rubber grafting strip or twine.
 Securing the Graft. Seal the junction with grafting wax or grafting
paint. Water rootstock sparingly until the graft knits. Over watering
may cause sap to "drown" the scion. Be sure to remove the twine or
strip as soon as the graft has healed.
Splice Graft
Whip and Tongue Graft

 The whip and tongue technique is most commonly used to graft

nursery crops or woody ornamentals. Both the rootstock and scion
should be of equal size and preferably no more than 1⁄2-inch in
diameter. The technique is similar to splice grafting except that
the whip on the rootstock holds the tongue of the scion in place
(and vice versa). This leaves both hands free to wrap the joint.
 For the whip and tongue graft, make similar cuts on both the
stock and scion. These cuts should be made with a single draw of
the knife and should have a smooth surface so that the two can
develop a good graft union. Up to this point, rootstock and scion
are cut the same as for a splice graft.
Whip and Tongue Graft
 Preparing the Stock and Scion. Cut off the stock using a diagonal cut. The cut
should be four to five times longer than the diameter of the stock to be
grafted. Make the same kind of cut at the base of the scion.
Next, place the blade of the knife across the cut end of the stock, halfway
between the bark and pith (on the upper part of the cut surface). Use a single
knife stroke to draw the blade down at an angle through the wood and pith.
Stop at the base of the initial diagonal cut. This second cut must not follow the
grain of the wood but should run parallel to the first cut.
 Inserting the Scion. Prepare the scion in the same way. Fit the scion into the
rootstock so that they interlock whip and tongue. Be certain that the cambia
are aligned.
 Securing the Graft. Wrap the junction with a grafting strip or twine, and seal it
with grafting wax or grafting paint. Never allow the binding material to girdle
the stem.
Whip and Tongue Graft
Saddle Graft

 Saddle grafting is a relatively easy technique to learn and

once mastered can be performed quite rapidly. The stock
may be either field-grown or potted. Both rootstock and
scion should be the same diameter. For best results, use
saddle grafting on dormant stock in mid- to late winter.
Stock should not be more than 1 inch in diameter.
Saddle Graft
 Preparing the Stock. Using two opposing upward strokes of the grafting knife,
sever the top from the rootstock. The resulting cut should resemble an
inverted V, with the surface of the cuts ranging from 1⁄2-inch to 1 inch long.
 Preparing the Scion. Now reverse the technique to prepare the base of the
scion. These cuts on the rootstock and scion must be the same length and
have the same slope so that a maximum amount of cambial tissue will make
contact when the two halves are joined.
 Inserting the Scion. Place the V-notched scion onto the saddle of the
rootstock. If rootstock and scion are the same diameter, cambial alignment is
easier; otherwise adjust as needed.
 Securing the Graft. Wrap the graft with a grafting twine, tape, or strip, then
seal it with grafting wax or grafting paint.
Saddle Graft
Bridge Graft

 Bridge grafting is used to "bridge" a diseased or damaged

area of a plant, usually at or near the base of the trunk.
Such damage commonly results from contact with grading or
lawn maintenance equipment, or it may be caused by
rodents, cold temperatures, or disease organisms. The
bridge graft provides support as well as a pipeline that
allows water and nutrients to move across the damaged
area.
 Bridge grafts are usually done in early spring just before
active plant growth begins. They may be performed any time
the bark on the injured plant "slips."
Bridge Graft
 Preparing the Scion. Select scions that are straight and about twice as long as the damaged
area to be bridged. Make a 11⁄2- to 2-inch-long tapered cut on the same plane at each end of
the scion.
 Preparing the Stock. Remove any damaged tissue so the graft is on healthy stems. Cut a flap
in the bark on the rootstock the same width as the scion and below the injury to be
repaired. Gently fold the flap away from the stock, being careful not to tear the bark flap.
 Inserting the Scion. First, insert and secure the scion below the injury; push the scion under
the flap with the cut portion of the scion against the wood of the injured stem or trunk.
Then go back and insert and secure the scion above the injury following these same steps.
Push the scion firmly into place. Pull the flap over the scion and tack it into place as
described for bark grafting .
When grafting with young stems that may waver in the wind, insert the scions so that they
bow outward slightly. Bridge grafts should be spaced about 3 to 4 inches apart across the
damaged area.
 Securing the Graft. Secure all graft areas with warm grafting wax or grafting paint. During
and after the healing period, remove any buds or shoots that develop on the scions.
Bridge Graft
Inarch Graft

 Inarching, like bridge grafting, is used to bypass or support

a damaged or weakened area of a plant stem.Unlike
bridge grafting, the scion can be an existing shoot, sucker,
or watersprout that is already growing below and
extending above the injury. The scion may also be a shoot
of the same species as the injured plant growing on its
own root system next to the main trunk of the damaged
tree. With the inarching technique, the tip of the scion is
grafted in above the injury using the same method as for
bark or bridge grafting.
Inarch Graft
BUDDING
Budding
 Budding is a grafting technique in which a single bud from the desired
scion is used rather than an entire scion containing many buds. Most
budding is done just before or during the growing season. However
some species may be budded during the winter while they are dormant.
 Budding requires the same precautions as grafting. Be sure that the
scion and rootstock are compatible, that the scion has mature buds,
and that the cambia of the scion and rootstock match. Be especially
careful to prevent drying or contamination of grafting materials. With
practice, the speed with which the process can be performed and the
percentage of successful grafts those that "take" - should equal or
surpass those of other grafting techniques used on the same species.
Generally, deciduous fruit and shade trees are well suited to budding.
T-Budding

 T-budding is most commonly used for summer budding of

apples, crabapples, dogwoods, peaches, and pears. T-
budding must be one when the bark will "slip." Slipping
means that, when cut, the bark easily lifts or peels in one
uniform layer from the underlying wood without tearing.
The exact time when this condition occurs depends on soil
moisture, temperature, and time of year. It varies with
species and variety. Dry or excessively hot or cold weather
can shorten the period when bark slips. Irrigation can be
valuable in extending the T-budding season.
T-Budding
 Preparing the Stock. Budding knives usually have a curved tip (Figure 10),
making it easier to cut a T-shaped slit. First, insert the point of the knife and
use a single motion to cut the top of the T. Then without removing the point
of the knife, twist it perpendicularly to the original cut and rock the blade
horizontally down the stem to make the vertical slit of the T. If bark is
slipping properly, a slight twist of the knife at the end of this cut will pop
open the flaps of the cut and make it easier to insert the bud. In practice, the
top of the T is usually slanted slightly (Figure 11).
 This same type of cut can be made using two separate strokes, one vertical
and one horizontal, and then using the back of the budding knife tip to pry up
the flaps slightly. Although much slower, this technique may be easier.
T-Budding

Figure 10. Budding knives Figure 11. T-shaped cut on rootstock.

T-Budding
 Removing Buds from the Budstick. The bud to be inserted is often just a
shield of bark with a bud attached or a very thin layer of wood with both the
bark shield and bud attached (Figure 12). Various techniques can be used to
make these cuts, but the shape of the cut remains the same.
 Begin the first scion cut about 1⁄2-inch below the bud and draw the knife
upward just under the bark to a point at least 1⁄4-inch above the bud. Grasp
the petiole from the detached leaf between the thumb and forefinger of the
free hand. Make the second cut by rotating the knife blade straight across the
horizontal axis of the budstick and about 1⁄4 inch above the desired bud. This
cut should be deep enough to remove the bud, its shield of bark, and a thin
sliver of wood.
T-Budding

Figure 12. Removing the bark shield with the bud attached.
T-Budding
 Inserting the Bud. Insert the bud shield into the T flaps of the stock and slide
it down to ensure that it makes intimate contact with the rootstock (Figure 13
).
 Securing the Bud. Pull the cut together by winding a 4- or 5-inch long
budding rubber around the stem to hold the flaps tightly over the bud shield
and prevent drying (Figure 14). Secure the budding rubber by overlapping all
windings and tucking the end under the last turn. Do not cover the bud.
T-Budding

Figure 13. Bark shield with bud inserted into T cut. Figure 14. Wrapped bud.
Chip Budding

 Chip budding is a technique that may be used whenever

mature buds are available. Because the bark does not
have to "slip," the chip-budding season is longer than the
T-budding season. Species whose bark does not slip easily
without tearing - such as some maples - may be
propagated more successfully by chip budding than by T-
budding.
Chip Budding
 Preparing the Stock and the Scion Bud. Although all the basics in handling
budwood and stock are the same for chip budding and Tbudding, the cuts
made in chip budding differ radically. The first cut on both stock and scion is
made at a 45 to 60° downward angle to a depth of about 1⁄8-inch (Figure 15).
After making this cut on a smooth part of the rootstock, start the second cut
about 3⁄4-inch higher and draw the knife down to meet the first cut. (The
exact spacing between the cuts varies with species and the size of the buds.)
Then remove the chip.
 Cuts on both the scion (to remove the bud) and the rootstock (to insert the
bud) should be exactly the same (Figure 16). Although the exact location is
not essential, the bud is usually positioned one-third of the way down from
the beginning of the cut. If the bud shield is significantly narrower than the
rootstock cut, line up one side exactly.
 Chip Budding

Figure 15. Rootstock cut for T budding. Figure 16. Removing chip from budstick.
Chip Budding
 Securing the Bud. Wrapping is extremely important in chip budding. If all
exposed edges of the cut are not covered, the bud will dry out before it can
take. Chip budding has become more popular over the past 5 years because of
the availability of thin (2-mil) polyethylene tape as a wrapping material. This
tape is wrapped to overlap all of the injury, including the bud (Figure 17),
and forms a miniature plastic greenhouse over the healing graft.
Chip Budding

Figure 17. Chip bud wrapped with plastic tape.

Budding Aftercare
 When irrigation is available, apply water at normal rates for plants that bud
before August 1. Ornamental peaches and pears often will break bud and
grow the same year they are budded. Dogwoods and most other species
budded after August 1 should be irrigated at a normal rate for only two to
three weeks after budding except during extreme drought. Following these
irrigation practices will enable buds to heal completely with no bud break
before frost.
 To insure a top-quality plant, it is essential to remove unwanted sprouts.
These sprouts should be "rubbed" off as soon as they are visible so that they
do not reduce the growth and quality of the budded stock. If they are
removed regularly and early, large scars or "doglegs" can be avoided.
LAYERING
Layering

 Stems that are still attached to their parent plant may form roots
where they come in contact with a rooting medium. This method
of vegetative propagation is generally successful, because water
stress is minimized and carbohydrate and mineral nutrient levels
are high. The development of roots on a stem while the stem is
still attached to the parent plant is called layering. A layer is the
rooted stem following detachment (removal) from the parent
plant.
 Some plants propagate naturally by layering, but sometimes plant
propagators assist the process. Layering is enhanced by wounding
the stem where the roots are to form. The rooting medium should
always provide aeration and a constant supply of moisture
Simple Layering
 Simple layering can be accomplished by bending a low growing, flexible stem
to the ground. Cover part of it with soil, leaving the remaining 6 to 12 inches
above the soil. Bend the tip into a vertical position and stake in place (
Figure 1). The sharp bend will often induce rooting, but wounding the lower
side of the bent branch may help also. Simple layering can be done on most
plants with low-growing branches. Examples of plants propagated by simple
layering include climbing roses, forsythia, rhododendron, honeysuckle,
boxwood, azalea, and wax myrtle.
 Simple layering can be done in early spring using a dormant branch, or in late
summer using a mature branch. Periodically check for adequate moisture and
for the formation of roots. It may take one or more seasons before the layer is
ready to be removed for transplanting.
Simple Layering

Figure 1. Simple layering.

Tip Layering
 Tip layering is quite similar to simple layering. Dig a hole
3 to 4 inches deep. Insert the tip of a current season’s
shoot and cover it with soil. The tip grows downward first,
then bends sharply and grows upward. Roots form at the
bend. The re-curved tip becomes a new plant (Figure 2).
Remove the tip layer and plant it in late fall or early
spring. Examples of plants propagated by tip layering
include purple and black raspberries, and trailing
blackberries.
Tip Layering

Figure 2. Tip layering.

Compound (serpentine) Layering

 Compound (serpentine) layering is similar to simple

layering, but several layers can result from a single stem.
Bend the stem to the rooting medium as for simple
layering, but alternately cover and expose sections of the
stem. Each section should have at least one bud exposed
and one bud covered with soil. Wound the lower side of
each stem section to be covered (Figure 3). This method
works well for plants producing vine-like growth such as
heart-leaf philodendron, pothos, wisteria, clematis, and
grapes.
Compound (serpentine) Layering

Figure 3. Compound (serpentine) layering.

Mound (stool) Layering

 Mound (stool) layering is useful with heavy-stemmed,

closely branched shrubs and rootstocks of tree fruits. Cut
the plant back to 1 inch above the soil surface in the
dormant season. Dormant buds will produce new shoots in
the spring. Mound soil over the new shoots as they grow (
Figure 4). Roots will develop at the bases of the young
shoots. Remove the layers in the dormant season. Mound
layering works well on apple rootstocks, spirea, quince,
daphne, magnolia, and cotoneaster.
Mound (stool) Layering

Figure 4. Mound (stool) layering.

Air Layering

 Air layering can be used to propagate large, overgrown house

plants such as rubber plant, croton, or dieffenbachia that have
lost most of their lower leaves. Woody ornamentals such as
azalea, camellia, magnolia, oleander, and holly can also be
propagated by air layering. For optimum rooting, make air
layers in the spring on shoots produced during the previous
season or in mid to late summer on shoots from the current
season’s growth. For woody plants, stems of pencil size
diameter or larger are best. Choose an area just below a node
and remove leaves and twigs on the stem 3 to 4 inches above
and below this point. This is normally done on a stem about 1
foot from the tip.
Air Layering
 Air layering differs, depending on whether the plant is a monocot or a dicot.
For monocots, make an upward 1- to 11⁄2-inch cut about one-third through
the stem. The cut is held open with a toothpick or wooden match stick.
Surround the wound with moist, unmilled sphagnum moss (about a handful)
that has been soaked in water and squeezed to remove excess moisture. Wrap
the moss with plastic and hold in place with twist ties or electrician’s tape.
No moss should extend beyond the ends of the plastic. Fasten each end of the
plastic securely, to retain moisture and to prevent water from entering. If
exposed to the sun, the plastic should be covered. Aluminum foil can also be
used, as it does not require twist ties or tape to hold it in place.
Air Layering
 The process for dicots is similar, except a 1-inch ring of bark is removed from
the stem. With a sharp knife, make two parallel cuts about an inch apart
around the stem and through the bark and cambium layer (see Figure 5).
Connect the two parallel cuts with one long cut. Remove the ring of bark,
leaving the inner woody tissue exposed. Scrape the newly bared ring to
remove the cambial tissue to prevent a bridge of callus tissue from forming.
Application of a root-promoting substance to the exposed wound is sometimes
beneficial. Wrap and cover using the same procedure as that described for
monocots.
 After the rooting medium is filled with roots, sever the stem below the
medium and pot the layer. The new plant will usually require some
pampering until the root system becomes more developed. Provide shade and
adequate moisture until the plant is well established.
Air Layering

Figure 5. Air Layering.

Natural Forms of Layering
 Sometimes layering occurs naturally, without the assistance of a propagator. Runners
and offsets are specialized plant structures that facilitate propagation by layering.
 A runner produces new shoots where it touches the growing medium (Figure 6). Plants
that produce stolons or runners are propagated by severing the new plants from their
parent stems. Plantlets at the tips of runners may be rooted while still attached to
the parent or detached and placed in a rooting medium. Examples include strawberry
and spider plant.
 Plants with rosetted stems often reproduce by forming new shoots, called offshoots,
at their base or in the leaf axles. Sever the new shoots from the parent plant after
they have developed their own root systems. Unrooted offsets of some species may be
removed and placed in a rooting medium. Some of these must be cut off, whereas
others may simply be lifted from the parent stem. Examples include date palm,
bromeliads, and many cacti.
Natural Forms of Layering

Figure 6. Natural forms of layering.

MARCOTTING
Marcotting

 Marcotting or air layering, an asexual or vegetative method of

plant propagation, can be easily performed with less skill. Air
layering is just slightly different from other methods of layering
such as tip layering, simple layering, compound or serpentine
layering, etc. In all these methods, the induction of root
development is usually done by wounding the part of the plant
to be rooted.
 In this layering method, roots are induced to form on the part of
the plant while it remains aerial (aboveground), hence the term
air layering. But in other layering methods, the same plant part
is rooted on the ground usually by bending it downward.
Common Procedures in Marcotting
 Plant and Shoot Selection
Girdled stem ready for wrapping with rooting
medium. A shoot with plenty of leaves is chosen
from a healthy plant. The size of the stem at the
part to be rooted is generally about that of an
ordinary pencil, but this is not essential. Both the
thickness and length of the stem vary depending
on the plant part to be layered (trunk, branch or
twig), the intended size of the marcotted plant to
be produced, and the plant species.
In roses, the stems used in marcotting are
normally thinner. In comparison, in herbaceous
plants like aglaonema and dieffenbachia, the
stems are thick.
Common Procedures in Marcotting
 Girdling and Scraping
This is unique in marcotting. However, this procedure is skipped in bamboo
and herbaceous plants. For trees, shrubs and semi-woody plants, a strip of bark 
 is first removed from around the portion of the stem to be rooted. This involves
pressing of a sharp knife against the bark preferably as close as possible below a
node, moving the knife in circular motion around the stem. A similar cut is made
generally about 2 cm to 5 cm below the first cut, but it can be wider with larger
stems. The two cuts are then connected by a straight cut and the bark is pried
loose and removed.
The debarked portion of the stem is then scraped to remove the phloem and
cambium, that slippery coating on the wood, to prevent the wound from healing
and the upper and lower barks from reconnecting.
Common Procedures in Marcotting
 Slitting and Wedging
In herbaceous plants, an inward cut is made
starting from below a node and slightly upward. The
cut has to traverse the horizontal line that marks the
node at the point about halfway of the thickness of
the stem and terminate above the same node. In
other words, this slanting cut must be able to severe
the horizontal connection of the node.
Coir dust, sphagnum moss or a piece of wood or
any other suitable material is then inserted into the
wound to serve as wedge. The purpose of this wedge
is to keep the upper and lower cut surfaces apart and
prevent healing just like in girdling and scraping.
Common Procedures in Marcotting
 Placing and Securing the Rooting Medium
A slightly moistened sphagnum moss or coconut coir
dust is placed around the debarked stem and wrapped
with a piece of plastic sheet. A transparent plastic sheet
is preferred to be able to see later if roots have
developed. In many plant species, however, the stems
can be marcotted even with pure soil.
The rooting medium may be as thick as 1 inch (2.5
cm) from side to side or bigger depending on the
earliness to develop roots and size of the stem. The
longer is the time required to induce rooting and the
bigger is the stem, the thicker should be the rooting
medium.
A leaf was used to hold the rooting medium in
place.
Common Procedures in Marcotting
 Both ends of the plastic sheet are gathered and tied securely against the
stem, with one end just under the bottom part of the debarked stem (lower
cut) and the other a short distance above the upper part (upper cut). It is
important that the upper cut should be covered with the rooting medium
because it is from this cut that roots form.
 As an alternative, the plastic sheet may be placed first on the stem with one
end tied just below the lower cut. The rooting medium is then inserted
gradually and the upper end of the plastic wrapping is tied securely to the
stem. This marcotting technique is more convenient and applies with any
rooting medium which crumbles if not held by the hand.
 To prevent breaking of the stem with big and heavy rooting medium, it is tied
to another branch or to a stick attached to the parent plant.
Common Procedures in Marcotting
 In stems which are more or less erect, the rooting medium can be held by any
container such as broken or halved pots, cans or plastic cups with open top.
For big containers, a support is needed to prevent them from dropping.
 A container can be made also with a relatively thick plastic sheet with the
bottom gathered and tied just below the lower cut and the top is expanded to
form a shape like that of a funnel. The sides are overlapped and stapled or
fastened by piercing with a piece of thin stick.
 In plants which easily form roots like Ficus and croton or san francisco
(Codiaeum variegatum), this funnel-shaped container can be made out of
some thick leaves. The sides are secured in place by piercing with a stick. The
container is then filled with rooting medium which is kept moist by regular
watering.
Common Procedures in Marcotting

 Separation of the Marcot from the Parent Plant

The rooted shoots are severed from the parent plant when plenty of roots
have developed. At this time the rooting medium becomes hard and rough when
touched. New shoots will also have sprouted from the portion of the stem
immediately below the rooting medium. In many plant species this occurs at
least 15 days from marcotting.
 Potting
The marcotted shoot is immediately potted into suitable container. The
intensity of care that will ensure the successful establishment of the layers will
depend on various factors such as size of the shoot, size of the rooting medium,
and profuseness of roots. For maximum survival, the newly potted marcots are
kept under partial shade and high humidity
 PROPAGATION BY
SPECIALIZED STEMS AND
ROOTS
Bulbs

Bulb -a specialized underground organ consisting of a basal

plate, growing points, flower primordia, and fleshy scales
Bulblets -miniature bulbs produced on scales, stems
Bulbils -aerial bulblets
Stem bulblets -underground bulbils produced on stem
 Functions as food storage and reproductive structure
 Found in monocots
Types of Bulbs

1. Tunicate bulbs (Laminate bulbs)

 Outer bulb scales are dry and membranous
 the tunic or covering protects from drying and mechanical injury
 Fleshy scales are in continuous, concentric rings (lamina)
 Onions, garlic, daffodil, tulip, amaryllis, narcissus
2. Nontunicate bulbs
 Scaly bulbs with a basal plate, scales, shoot primordia
 Scales are modified leaves
 Easter lily, other Lilium species
 Propagated by scales (takes 3 years)
Propagation

1. Offsets
 Tulips, daffodils, lilies
 Bulbs grown from bulblets
2. Stem bulblets
 Lilies
 Underground bulblets
 Aerial bulblets (bulbils)
3. Scales
 Easter lily
4. Basal sections
 Use of scooping, scoring, or sectioning
Corms

A corm is the swollen base of a stem axis enclosed by dry,

scale-like leaves
 distinctive nodes and internodes
 cormels -stolon-like structures of miniature corm

Species
 gladiolus, crocus

Propagation
use of cormels
division of corms
 sectioning
Tubers

A tuber is a swollen, modified stem structure that functions as an

underground organ
Species
 Potato
 Caladium
 Jerusalem
 Artichoke
 yam
Propagation
 by division--- 'eyes' or nodal shoot buds must be present on the cut
sections
Tuberous roots
A. Enlarged secondary roots that are storage organs
B. Fleshy tuberous roots
 sweet potato
 Cassava
 dahlia

Tuberous stems
A. Tuberous begonia, cyclamen
 enlarged hypocotyl section of the seedling plant
Rhizomes
Horizontally grown underground stems
 pachymorph ----- thick, fleshy (iris, ginger)
 leptomorph ------ slender, long (lily-of-the-valley)
 mesomorph ------ intermediate

Pseudobulbs
Storage structure found in orchids
TISSUE CULTURE
What is Tissue Culture?

 In biological research, tissue culture refers to a method in which

fragments of a tissue (plant or animal tissue) are introduced into a new,
artificial environment, where they continue to function or grow. While
fragments of a tissue are often used, it is important to note that entire
organs are also used for tissue culture purposes. Here, such growth
media as broth and agar are used to facilitate the process.
 While the term tissue culture may be used for both plant and animal
tissues, plant tissue culture is the more specific term used for the
culture of plant tissues in tissue culture.
Types of Tissue Culture

Seed Culture
 Seed culture is the type of tissue culture that is primarily
used for plants such as orchids. For this method, explants
(tissue from the plant) are obtained from an in-vitro
derived plant and introduced in to an artificial
environment, where they get to proliferate. In the event
that a plant material is used directly for this process, then
it has to be sterilized to prevent tissue damage and ensure
optimum regeneration.
Types of Tissue Culture

Embryo Culture
 Embryo culture is the type of tissue culture that involves the isolation of an
embryo from a given organism for in vitro growth. Note, the term embryo
culture is used to refer to sexually produced zygotic embryo culture.
 Embryo culture may involve the use of a mature of immature embryo. Whereas
mature embryos for culture are essentially obtained from ripe seeds, immature
embryo (embryo rescue) involves the use of immature embryos from
unripe/hybrid seeds that failed to germinate. In doing so, the embryo is
ultimately able to produce a viable plant. 
 For embryo culture, the ovule, seed or fruit from which the embryo is to be
obtained is sterilized, and therefore the embryo does not have to be sterilized
again. Salt sucrose may be used to provide the embryo with nutrients. The
culture is enriched with organic or inorganic compounds, inorganic salts as well
as growth regulators.
Types of Tissue Culture

Callus Culture
 Callus - This is the term used to refer to unspecialized, unorganized and
a dividing mass of cells. A callus is produced when explants (cells) are
cultured in an appropriate medium - A good example of this is the tumor
tissue that grows out of the wounds of differentiated tissues/organs.
 In practice, callus culture involves the growth of a callus (composed of
differentiated and non- differentiated cells), which is the followed by a
procedure that induces organ differentiation.
 For this type of tissue culture, the culture is often sustained on a gel
medium, which is composed of agar and a mixture of given macro and
micronutrients depending on the type of cells. Different types of basal
salt mixtures such as murashige and skoog medium are also used in
addition to vitamins to enhance growth. 
Types of Tissue Culture

Organ Culture
 Organ culture is a type of tissue culture that involves isolating an
organ for in vitro growth. Here, any organ plant can be used as an
explant for the culture process (Shoot, root, leaf, and flower).
 With organ culture, or as is with their various tissue components, the
method is used for preserve their structure or functions, which allows
the organ to still resemble and retain the characteristics they would
have in vivo. Here, new growth (differentiated structures) continues
given that the organ retains its physiological features. As such, an
organ helps provide information on patterns of growth, differentiation
as well as development.
Types of Tissue Culture
Protoplast Culture  Cell cloning
 *Protoplast -cells without cell walls. A  Genetic transformations
protoplast is the term used to refer to cell  Membrane studies
(fungi, bacteria, plant cells etc) in which the
cell wall has been removed, which is why
 In protoplast culture, a number of phases
they are also referred to as naked cells. can be observed. These include;
Protoplasts may be cultured in the following  Development of a cell wall
ways;  Cell division
 Hanging-drop cultures  Continuous growth or regeneration to a whole
 Micro culture chambers plant
 Soft agars matrix
 Once a protoplast has regenerated a cell
wall, then it goes through the process of cell
division to form a callus, which may then be
subcultured for continued growth. Protoplast
culture is an important method that provides
numerous cells (single cells) that can be
Major Steps of Tissue Culture
(Plants)
Initiation Phase (Stage 1)
 The initiation phase is the first phase of tissue culture.
Here, the tissue of interest is obtained and introduced and
sterilized in order to prevent any microorganism from
negatively affecting the process. It is during this stage
that the tissue is initiated in to culture.
Major Steps of Tissue Culture
(Plants)
Multiplication Phase (Stage 2)
 The multiplication phase is the second step of tissue culture
where the in vitro plant material is re- divided and then
introduced in to the medium. Here, the medium is composed
of appropriate components for growth including regulators
and nutrients. These are responsible for the proliferation of
the tissue and the production of multiple shoots.
 *This step is often repeated several times in order to obtain
the desired number of plants
Major Steps of Tissue Culture
(Plants)
Root formation (Stage 3)
 It is at this phase that roots are formed. Here, hormones
are required in order to induce rooting, and consequently
complete plantlets.
The Process of Plant Tissue Culture

Medium preparation
 The appropriate mixture (such as the MS mixture) is mixed with distilled
water and stirred while adding the appropriate amount of sugar and sugar
mixture. Here, sodium hydroxide or hydrochloric acid is used to adjust the
pH - Contents used here will depend on the plant to be cultured and the
number of tissues to be cultured.
 Agar is added to the mixture, heat and stirred to dissolve
 After cooling, the warm medium is poured into polycarbonate tubes (to a
depth of about 4 cm)
 With lids sitting on the tubes, the tubes are placed in a pressure cooker and
sterilized for 20 minutes
The Process of Plant Tissue Culture

Plant preparation
 Cut the plant part in to small pieces (e.g. cauliflower can be cut to florets
of about 1cm across). On the other hand, such parts as the African violet
leaves can be used as a whole.
 Using detergent and water, wash the plant part for about 20 minutes
 Transfer the plant part in to sterilizing Clorox solution, shake for a minute
and leave to sock for 20 minutes
 Using a lid, gently discard the Clorox and retain the plant part in the
container and then cap the container
Transferring the plant material to a tissue culture medium
* 70 percent alcohol should be used for the sterilization of the equipment used
and containers
The Process of Plant Tissue Culture
 Open the container and pour sterile water to cover half the container
 Cover with a sterile lid again and shake the container for 2 to 3
minutes in order to wash the tissue and remove the bleach
 Pour the water and repeat this three times
 Using sterilized gloves, remove the plant part from the container and
on to a sterile Petri dish
 Using a sterile blade cut the plant material to smaller pieces of about
2 to 3 mm across avoiding the parts that have been damaged by
bleach
 Using sterile forceps, place a section of the plant in to the medium
The Process of Plant Tissue Culture

 *depending on the plant used, it is important to check and find out

how it should be placed in the medium
 Replace the lid/cap and close tightly
 This procedure will result in the development of a callus, which then
produces shoots after a few weeks. Once the shoots develop, then the
plant section may be placed in the right environment (well lit,
warmth etc) for further growth.
 * Plant materials should be sterilized so as to remove any bacteria or
spores that may be present.
The Process of Plant Tissue Culture

 For plants, the medium culture acts as a greenhouse that provides the
explant with the idea environment for optimum growth. This includes
being free of microorganisms, nutrients as well as the right balance of
chemicals and hormones. Such media as BAP, TDZ are used while such
hormones as IBA and IAA are used to induce growth. Some of the
major reasons tissue culture is used for plants include;
 To produce large quantities of a given plant
 To accelerate the production of new varieties of a plant
 To maintain a virus free stock of the plant of interest