Cellular differentiation and totipotency
Plant consist of different organs and each organ is
composed of different tissues, which in turn are made
up of different individual cells.
The process of the growth accompanied by
morpho-physiological specialization of the cells
produced by the meristems is called differentiation.
In plant tissue culture, differentiation is used to
describe the formation of different cell types and
organs in the callus or cell culture.
The tissues after differentiation
which loose meristematic activity are
called permanent tissues.
These highly mature and
differentiated cells possess the ability
to grow, divide and redifferentiate
under appropriate experimental
conditions.
All the living cells in a plant body are
capable of giving rise to whole plants
irrespective of their specialization. This
property of cells is called cellular
totipotency.
Plants specialized cells perform
different functions known as
cytodifferentiation.
In plant tissue culture, explant is used
to initiate growth in culture. Non-
dividing, differentiated, quiescent cells
of the explant when grown on a nutrient
medium first undergo changes to
achieve the meristematic state.
The phenomenon of mature cells reverting to a
meristematic state and forming undifferentiated
callus tissue is termed as dedifferentiation.
The ability of dedifferentiated cells to form a
whole plant or plant organ is called
redifferentiation.
The phenomenon of dedifferentiation and
redifferentiation are inherent capacity of a plant
cell and thus giving rise to a whole plant known
cellular totipotency.
Clonal propagation
Clonal propagation is the multiplication of
genetically identical individuals by asexual
reproduction, and clonal propagation in vitro
is called micropropagation.
Micropropagation provides significant
advantage over traditional method like it can
be executed in a relatively short span of time
and space.
It is feasible to obtain a number of plants
from a single explant within a very short
time.
Suitable explants from vascular plants
including angiosperms, gymnosperms and
pteridophyte can be cultured in vitro and
induced to form adventitious buds, shoot
or whole plants.
Murashige outlined three major stages involved in
micropropagation (1974) :
Stage-I: Selection of suitable explants, their sterilization and
transfer to nutrient media for establishment i.e. initiation
of a sterile culture of explants.
Stage-II: Proliferation or multiplication of shoots from the explant
on medium
Stage-III Transfer of shoots to a rooting medium followed on
by its transfer into soil
Stages of micropropagation
Micropropagation and plant regeneration
can be grouped into the following categories:
1. Axilliary bud proliferation
2. Organogenesis
3. Somatic embryogenesis
Axillary bud proliferation
It is the stimulation of axilliary buds, which
are present in the axil of each leaf, to
develop into a shoot.
Due to presence of cytokinin in a culture
medium, the shoots develop from axilliary
buds.
The shoots are then separated and rooted
to produce plants and could be used as
propagules for further propagation.
This approach can be subdivided into
a) Meristem and shoot tip culture
b) Bud culture
Meristem and shoot tip culture
Morel and Martin (1952) developed the
technique of meristem culture for in vitro virus
eradication.
Explants of Dahlia meristem tips between
0.2-0.5 mm are used to produce virus free
plants and this method is popularly referred to
as meristem tip culture.
Schematic representation of shoot tip culture
Bud culture
Bud culture is of two types:
Single node culture and
Axillary bud method
Single node culture
Generally a bud is isolated along with a piece of
stem with the purpose of forming a shoot by
allowing the bud to develop.
The bud found in the axil of a leaf is similar to
that one in the stem tip. Bud is isolated and
cultured on a nutrient medium for in vitro
development.
The buds in the axil of a newly formed leaves can
be further subcultured and allowed to develop into
plantlet.
Once enough shoot have been obtained these
must be rooted and then transferred to soil.
Schematic representation of vegetative propagation by single node technique
Axillary bud method
In this method a shoot tip is isolated, from
which the axillary buds in the axils of leaves
develop under the influence of a relatively high
concentration of cytokinin.
High concentration of cytokinin suppresses
the effect of apical dominance and allows the
axillary buds to develop.
Advantages of micropropagation
A small amount of plant tissue is needed as the
initial explant for regeneration of millions of
clone plants in a short time.
Production of disease free plants by meristem
culture because shoot apices of virus infected
plants are frequently devoid of viruses eg.
Dianthus, potato
Germplasm storage
Formation of artificial seeds through somatic
embryogenesis.
