Plant Tissue Culture

in vitro propagation
Rapid (compared to outdoors)
Asexual

Used for
1. Micropropagation
2. Regeneration
1

Vegetative (asexual) Propagation Occurs in

Nature
Layering - a drooping lower branch
contacts the soil (pressed down by snow or
vegetation); roots form at point of soil
contact forming a new genetically identical
tree
When trees of some species are cut down,
new shoots emerge from the stump
strawberries spread through sending out
above-ground horizontal shoots called
runners, also called stolons.

I. Micropropagation - Many
genetically identical plants are produced from
ONE PLANT
in vitro propagation (in tubes/petri plates etc.)
Rapid
Asexual

Example: A Potato plant is superior because it has

good moisture and starch content. (good for french
fries!)

Micropropagation
A single node will produce
a shoot within 4-6 weeks
that has 4-6 nodes.
Each plantlet can be
"subcultured" to produce
another 4-6 plants each.

 Hundreds of thousands of plants could be

developed from one node
Since these are produced from axillary buds,
the plantlets will be clones of the mother
plant.

Micropropagation of almost all the

fruit crops and vegetables is possible
Some examples: dwarfing sweet cherry,
Shade trees, Ornamental shrubs, Roses,
Clematis, Lilacs, Saskatoon berries,
Nutraceutical Plants, Rhododendron,
Azalea, mustard, corn, soybeans, wheat,
rice, cotton, tomato, potato, citrus, turf,
legumes

Advantages of Micropropagation
economical in time and space
greater output -can produce millions of
uniformly flowering and yielding plants
African Biotechnologies - fruit crops banana
and indoor pot flowers- 6 million pieces per
year

disease free
elite plants with exceptional characteristics

Advantages Contd
facilitates safer movements of germplasm
across nations - In vitro germplasm assures
the exchange of pest and disease free
material
great for
vegetatively reproduced crops
crops which produce few seeds or highly
heterozygous seeds.

II. Regeneration - The process whereby a

part of a plant can be turned into a whole new
plant
African
violet leaf
sprouting
new
shoots

Regeneration is possible because plant cells

can be made totipotent using hormones.
differentiated tissue: stems, leaves, roots, etc.
undifferentiated (embryonic) cells are totipotent:
can become a whole new plant by differentiating
into a whole new plant.

Plant Hormones are naturally occurring chemicals that

influence plant growth.
Growth Regulators are synthetic versions of hormones.

Cytokinins induce the production of

shoots
Auxins induce the production of roots
The action of the different hormones / regulators is not consistent:

Different plants will respond to the same chemical

differently.
Different plant parts from the same plant can respond
differently

Steps involved in Tissue

Regeneration
tissue must be sterile - completely free of any
microoganisms; done using aseptic technique
1 Starting tissue is called an explant: differentiated
cells (these cells have developed to be part of
specialized tissue (root, leaf, stem, ovary, cotyledon,
etc.).

Explants are plated on a sterile petri dish containing hormones and

nutrients that promote the explant cells to develop into
Callus - a mass of undifferentiated cells.

Callus cells are totipotent

Tobacco Regeneration

3. Individual cells (or clumps of cells) of the callus are

transferred aseptically to a different petri dish containing
sterile medium that encourages the undifferentiated callus cells
to become shoots and roots.
One mass of callus cells can be divided and transferred to
many plates for development into shoots and roots.

4. Once shoots and roots have developed, they are transferred

to soil and grown to maturity.

Micropropagation vs. Regeneration

Plants that develop from existing terminal or
axillary buds are true clones of the "mother
plant".
Meristematic buds tend to be protected from
variations caused by environmental factors.
Little or no variation usually occurs when this
method of tissue culture is used.

Micropropagation vs. Regeneration (contd)

Regeneration using callus can produce variants of
the original plant
Non-meristematic cells are not protected from
variations caused by the environment.
When these cells experience a mutation, the plants
that are produced from them will also show a
difference from the mother plant.

Tissue Culture is also used when

plants are Genetically Engineered
Plants that receive a new gene are
called Transgenic plants or GMOs

A Genetically Engineered Plant has a New Gene

Squash gets coat proteins from Cucumber or zucchini
virus to prevent viral disease (to vaccinate the squash)
Corn gets a gene from bacteria that allows it to survive
in the presence of weedkiller (one that is less harmful
to environment that other herbicides).
Corn gets a gene from different bacteria that allows it
to produce its own pesticide (protects against insects).

After a new gene is given to a plant,

scientists must figure out how to
regenerate the transformed plant cell.