2. Definition of micropropagation
Clonal propagation ‘IN VITRO’
Micro-propagation is the production of
whole plant from small section of plant
such as a stem tip, node, meristem,
embryo, or even a seed.
3. STEPS OF MICRO-PROPAGATION
Establishment
Proliferation
Rooting and hardening
Sometimes one step is also included in cases
where establishment of plantlets in soil is
particularly elaborate
4. Selection of an elite mother plant
STEP 1- ESTABLISHMENT- In this
step, selection of suitable
plants, their sterilization and
transfer to nutrient media for
establishment, i.e. initiation of a
sterile culture of the explant.
STEP 2 – PROLIFERATION- Proliferation
or multiplication of shoot from the
explant on medium.
STEP 3 – ROOTING & HARDNING-
Transfer of shoots to a rooting medium
followed later by planting into soil
Explant
Surface sterilization and
washing
Establishment on growth
medium
Transfer to proliferation
medium
Shoot formation
Transfer of shoot or
plantlets to sterilized soil
or artificial medium by
various gradual
weaning processes
5. METHODS OF MICRO-PROPAGATION
AXILLARY BUD PROLIFERATION APPROACH
• Meristem and shoot tip culture
• Bud culture
ORGANOGENESIS
• INDIRECT
• DIRECT
EMBRYOGENESIS
6. AXILLARY BUD PROLIFERATION APPROACH
Meristem and shoot tip culture Morel and Martin (1952) develop the technique of
meristem culture for in vivo virus of Dahlia.
G. Morel (1965) was developed the technique shoot tip
culture for micro propagation of orchid Cymbidium.
This method is more successful in herbaceous plant.
Bud culture Buds contain active meristem depending upon the physiological
state of the plant. The various types used in bud culture,
a). Single node culture
b). Axillary bud method
7. ORGANOGENESIS
Indirect This pathway includes a callus stage.
Callus is undifferentiated tissue that develops on or around an
injured or cut plant surface.
Direct This pathway is bypasses a callus stage.
This method is particularly suitable to herbaceous species .
8. EMBRYOGENESIS
• The process of initiation and development of embryos and
embryo like structure from somatic cells.
• It usually involves a callus intermediate stage which can
result in variation among seedlings.
• Its not a common micro-propagation technique but
currently being used to produce superior pine seedlings.
a). Direct embryogenesis
b). Indirect embryogenesis
9.
10. HOW does micro-propagation work?
Plant cell have the ability to reproduce the whole plant
from a single cell. This is called totipotency.
Totipotency is the ability of single cell to express the full
genome in the cells to which it gives rise by cell division.
Why do we do it?
To regenerate plant from single cell or plant tissues.
To produce large quantities of identical plant.
To create new plant varieties.