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Lecture 6

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98 views25 pages

Lecture 6

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abdo01123808428
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© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Lecture 6

Micropropagation
Techniques in Plant Tissue
Culture
Prepared and presented by :
Dr. Abdelaziz Mohamed Nasr
Lesson Objectives
• Understand the concept and significance of micropropagation.

• Learn various micropropagation techniques.

• Discuss the applications and advantages of micropropagation.


Introduction
• Plants can be propagated through their two developmental life cycles; the
sexual, or the asexual.

• In the sexual cycle new plants arise after fusion of the parental gametes and
develop from zygotic embryos contained within seeds or fruits.

• By contrast, in the vegetative (asexual) cycle the unique characteristics of any


individual plant selected for propagation (termed the mother plant, stock
plant) are usually perpetuated.

• A group of such asexually reproduced plants are termed clones.


Seed propagation advantages
• They are often produced in large numbers so that the plants
regenerated from them are individually inexpensive.

• Many may usually be stored for long periods without loss of


viability.

• They are easily distributed.

• Most often plants grown from seed are without most of the pests
and diseases which may have afflicted their parents.
Seed propagation disadvantages

• Some plants do not produce viable seeds.

• Some plants produce seeds only after a long juvenile


period.

• Some seeds lose their viability after long storage.


Micropropagation techniques
• Micropropagation is the mass vegetative production of plants in vitro for the
purpose of commercial plant production.

• The propagation could happen through terminal or axillary buds, or by the


propagation of adventitious shoots or embryos from somatic cells.
Micropropagation advantages
• Cultures are started with very small pieces of plants (explants), so only a small
amount of space is required to maintain plants or to greatly increase their
number.

• Methods are available to free plants from specific virus diseases, and certified
virus-tested plants can be produced in large numbers.

• A more flexible adjustment of factors influencing vegetative regeneration is


possible.

• It may be possible to produce clones of some kinds of plants that are


Micropropagation advantages
• Plants may acquire a new temporary characteristic through micropropagation
which makes them more desirable to the grower than conventionally-raised
stock. A bushy habit (in ornamental pot plants) and increased runner
formation (strawberries) are two examples.

• Production can be continued all the year round.

• Plant material needs little attention between subcultures.


Micropropagation disadvantages
• A specialized and expensive production facility is needed.

• Explants and cultures have to be grown on a medium containing sucrose or


some other carbon source.

• As they are raised within glass or plastic vessels in a high relative humidity,
and are not usually photosynthetically self-sufficient, the young plantlets are
more susceptible to water loss in an external environment.
Stages of Micropropagation

• Stage 0: Mother plant selection and preparation.

• They must be typical of the variety or species, and free


from any symptoms of disease.

• It may be advantageous to treat the chosen plant (or


parts of it) in some way to make in vitro culture
successful.
Stages of Micropropagation
• Stage I: Establishing an aseptic culture

• The customary second step in the micropropagation process is to obtain an


aseptic culture of the selected plant material.

• Success at this stage firstly requires that explants should be transferred to the
cultural environment, free from obvious microbial contaminants; and that this
should be followed by some kind of growth (growth of a shoot tip, or formation
of callus).

• Usually, a batch of explants is transferred to culture at the same time. After a


short period of incubation, any container found to have contaminated explants
Stages of Micropropagation
• Stage II: The production of suitable propagules

• The production of new plant outgrowths or propagules, which, when


separated from the culture are capable of giving rise to complete
plants.

• Newly derived axillary or adventitious shoots, somatic embryos.

• They can also be used as the basis for further cycles of


multiplication.
Stages of Micropropagation

• Stage III: Preparation for growth in the natural


environment

• Steps are taken to grow individual or clusters of


plantlets, capable of carrying out photosynthesis, and
survival without an artificial supply of carbohydrates.

• It includes the in vitro rooting of shoots prior to their


Stages of Micropropagation
• Stage IV: Transfer to the natural environment

• Plantlets are transferred from the in vitro to the ex vitro external environment
is extremely important. If not carried out carefully, the transfer can result in a
significant loss of propagated material.

• Shoots developed in culture have often been produced in high humidity and a
low light ‘intensity’. This results in there being less leaf epicuticular wax or
wax with an altered chemical composition, than on plants raised in growth
chambers or greenhouses.

• In some plants, the stomata of leaves produced in vitro may also be atypical
Stages of Micropropagation
• When supplied with sucrose (or some other carbohydrate) and kept in low-
light conditions, micropropagated plantlets are not fully dependent on their
own photosynthesis.

• They need to change to be fully capable of producing their own requirements


of carbon and reduced nitrogen.

• In practice, plantlets are removed from their Stage III containers, then
transplanted into an adequate rooting medium (such as a peat:sand compost)
and kept for several days in high humidity and reduced light intensity.

• The change only occurs after the plants have spent a period of several days ex
Shoot (or shoot tip) culture
• The term shoot culture is now preferred for cultures started from explants
bearing an intact shoot meristem, whose purpose is shoot multiplication by
the repeated formation of axillary branches.

Establishment of shoot tip culture of male P. vera


after 3 days of culture on the CIM containing 1.0
mgl -1 BA
Shoot culture
• Shoot cultures are conventionally started from the apices of lateral or main
shoots, up to 20 mm in length, dissected from actively-growing shoots or
dormant buds.

• Larger explants are also sometimes used with advantage:

• Better survive the transfer to in vitro conditions.

• More rapidly commence growth.

• Contain more axillary buds.

• However, the greater the size of the explant, the more difficult it may be to
Shoot culture
• The shoot tip used are usually macerated from shoots originating from
meristem tip culture.

• According to some researchers, there is a competition between cell


proliferation, and the formation of the virus particles in meristem region of
plant. Nucleic acid production capacity in meristematic tissue during cell
division is used for cell division and this situation prevents the reproduction of
virus.

• According to other researchers, transportation of viruses to the meristem


region of the plant is prevented due to lack of transport system in meristem.
Shoot culture

• The growth and proliferation of axillary shoots in shoot


cultures is usually promoted by incorporating growth
regulators (usually cytokinins) into the growth medium.

• In some plants, pinching out the main shoot axis is used


as an alternative, or an adjunct, to the use of growth
regulators for decreasing apical dominance.
Current applications

• Conventional shoot culture continues to be the most


important method of micropropagation, although node
culture is gaining in importance.

• It is very widely used by commercial tissue culture


laboratories for the propagation of many herbaceous
ornamentals and woody plants.
What is the take-home message
from today’s lecture.

Let’s discuss.
Any more questions?

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