PLANT TISSUE CULTURE.

Plant research often involves growing new plants in a controlled environment.

These may be plants that we have genetically altered in some way or may be plants of which we
need many copies all exactly alike.

These things can be accomplished through tissue culture of small tissue pieces from the plant of
interest. These small pieces may come from a single mother plant or they may be the result of
genetic transformation of single plant cells which are then encouraged to grow and to ultimately
develop into a whole plant.

Tissue culture techniques are often used for commercial production of plants as well as for plant
research. Tissue culture involves the use of small pieces of plant tissue (explants) which are cultured
in a nutrient medium under sterile conditions.

Using theappropriate growing conditions for each explant type, plants can be induced to rapidly
produce new shoots, and, with the addition of suitable hormones new roots.

These plantlets can also be divided, usually at the shoot stage, to produce large numbers of new
plantlets. The new plants can then be placed in soil and grown in the normal manner.

Many types of plants are suitable for use in the classroom. Cauliflower, rosecuttings, African violet
leaves and carnation stems will all easily produce clones (exact genetic copies) through tissue
culture.

Cauliflower florets in particular give excellent results since they can be grown into a complete plant
in the basic tissue culture media, without the need for additional growth or root hormones.

Green shoots are generally observable within three weeks, and roots develop within six weeks. The
most important part of this activity, however, is to maintain as sterile an environment as possible.

Even one fungal spore or bacterial cell that comes into contact with the growth media will rapidly
reproduce and soon completely overwhelm the small plant piece that you are trying to clone.

Objectives

1. To understand a procedure that is often used to propagate many plants of

The same genetic background.

2. To understand the importance of sterile techniques.

USE OF TISSUE CULTURE IN VEGETABLE CROPS

For most vegetables, the use of plants derived from tissue culture for field production of an edible
product is not a feasible alternative when compared to current plant production systems.

This is due in part to the fact that most vegetable crops are seed-propagated. Seeds of these
vegetables are usually produced in abundance and are therefore relatively inexpensive to produce
when compared to the costs of plants from micropropagation; in addition, seeds are durable,
compact, self-sufficient, easily handled and relatively stable genetically.

Even ill vegetables where asexual propagation is the established mode of propagation (e.g.
potato,)Tissue Culture as a Plant Production System for Horticullllral Crops.

1986MartinusNijhofJ Publishers, Dordrechr. 260 sweet potato), micropropagation does not

represent an economic alternative because of the ease of obtaining a large number of plants
through bedding or the use of seed pieces.

Where micropropagation of vegetable crops may show the most promise at the moment are
situations where it serves as an adjunct to existing seed technology (e.g. in the production of specific
parental genotypes for hybrid seed production) and in specific cases where difficulties are involved
in traditional propagation methods for a given species.

PARENTS FOR HYBRID SEED PRODUCTION

Genotypes incapable of self-pollination

Many vegetable seed companies have recently established tissue culture facilities at their main field
installations. While these in part were built to interface with programs currently under way in
affiliated biotechnological companies, these laboratories are also being used to investigate the
feasibility of producing parental plants for hybrid seed production.

This approach appears to be most feasible in situations where the parental genotype is incapable of
self-pollination, whether through male sterility, gynoecy or self-incompatibility, or in cases where
the quality of resulting seed is extremely poor.

Male sterility and gynoecy are useful traits in hybrid production in that costs concomitant with hand
emasculation are avoided. Also, in species where the natural mode of pollination is through natural
vectors, these phenotypes eliminate the costs involved in hand pollination.

Where micropropagation is most useful in producing male sterile plants is in species where an
established cytoplasmic sterility system does not exist. For example, in tomato
(Lycopersiconesculentum) a number of nuclear genes exist that convey male sterility (54). Since a
cytoplasmic male sterile factor has not yet been identified, these male sterile lines must be
maintained in a heterozygous situation and male fertile sergeants’ must be rogued out each
generation in order to insure true hybrid seed production from the male sterile.

Tissue culture production of tomato plants allows for rapid propagation of these genotypes with
fidelity to the phenotypes. Another interesting situation exists in asparagus (Asparagus officinalis), a
dioecious vegetable where male plants exhibit superior survival and establishment in the field and
also tend have superior vigour
Morphogenesis means the generation of form, and usually in the context of developmental biology
where it means the generation of tissue organization and shape in animal and plant embryos (it also
covers the generation of internal organization in complex single-cell. Morphogenesis therefore deals
with apparently straightforward problems such as: how epithelial ducts branch in glands how nerves
migrate to and recognize their targets, how mesenchymal cells come together to form pre-muscle
and pre-bone condensations, how tendons link to the appropriate bones, and how cells change their
shapes.

Morphogenesis is important"

 It is responsible for tissue organization and hence for much of an organism's anatomy,
physiology and behavior.
 Mutations that affect morphogenesis underpin many human congenital abnormalities.
 Mutations that alter shape alter the fitness of a species under selection pressure and so drive
evolutionary change.

"Morphogenesis is difficult to study": Current knowledge about the morphogenesis of complex

tissues is limited for three reasons:

1. Many of the key events take place during early development when organ rudiments are
small and difficult to study, although genetic manipulation is now allowing morphogenesis
to be investigated in organisms such as Drosophila with very small embryos.
Most tissues will not develop much of their form in vitro and so are inaccessible to standard
experimental manipulation.

2. The intrinsic complexity of morphogenesis makes experimentation difficult.