Micropropagation

“… the art and

science of
multiplying
plants in vitro.”
 Plant Propagation

the process of creating new plants

Sexual propagation asexual or vegetative

multiplication of plants propagation
from seed generating new plants from
some vegetative part of a plant
results in seedling variability
pass through an undesirable  Clonal propagation
juvenile phase
No uniform crop containing
chosen desirable
characteristics
 Clonal propagation
Clonal propagation refers to the process of asexual reproduction by
multiplication of genetically identical copies of individual plants.

The term clone is used to represent a plant population derived from

a single individual by asexual reproduction.

Advantages of Vegetative Propagation

Asexual (vegetative) propagation of plants has certain advantages
over sexual propagation.
Faster multiplication — large number of plants can be produced
from a single individual in a short period.
Possible to produce genetically identical plants.
 In Vitro Clonal Propagation:

 The in vivo clonal propagation of plants is tedious

 expensive
 frequently unsuccessful
 In vitro clonal propagation through tissue culture is
referred to as micro propagation
 Use of tissue culture technique for micro propagation was
first started by Morel (1960) for propagation of orchids
 now applied to several plants
 handy technique for rapid multiplication of plants.
Stages of micro propagation:
 Stages of micro propagation:

Stage 0:
 This is the initial step in micro- propagation
 involves the selection and growth of stock plants for about 3
months under controlled conditions.

Stage I:
 Selection of appropriate explants
 organs, shoot tips and axillary buds
 initiation and establishment of culture in a suitable medium

Stage II:
 Explants grown on defined culture medium
 involves multiplication of shoots or rapid embryo formation
from the explant
 Stages of micro propagation:

Stage III:
 transfer of shoots to a medium for rooting

Stage IV:
 Acclimatization
 establishment of plantlets in soil
 transferring the plantlets of stage III from the laboratory to the
greenhouse
 some plant species, stage III is skipped, and un-rooted stage II
shoots are planted in pots or in suitable compost mixture
 Types of micro propagation:

1. Multiplication by axillary buds/apical shoots

2. Multiplication by adventitious shoots
3. Organogenesis
4. Somatic embryogenesis
 Multiplication by axillary buds/apical shoots

• Cultivation of axillary or apical meristems is called

"Meristem culture".
• Meristem culture involves development of
already existing shoot meristem.
• Technique is used for quick vegetative
propagation of a large number of plant species in
a short period.
• Shoot tip culture produces virus free callus and
eventually plantlet.
A

Stages of apical meristem culture

B

Stages of axillary bud culture

 Cytokinin is not required

Multiplication by adventitious shoots
 Organogenesis

 process of morphogenesis
involving the formation of
plant organs i.e. shoots,
roots, flowers, buds from
explant or cultured plant
tissues.

 two types — direct

organogenesis and indirect
organogenesis.
 Somatic Embryogenesis
 The process of regeneration of
embryos from somatic cells,
tissues or organs is regarded
as somatic (or asexual)
embryogenesis.
 Somatic embryogenesis may
result in non-zygotic embryos
or somatic embryos
 Somatic embryos are
structurally similar to zygotic
(sexually formed) embryos,
and they can be excised from
the parent tissues and
induced to germinate in tissue
culture media.