Somaclonal Variation

Somaclonal Variation
• Definition: It is the term used to describe
the variation seen in plants that have
been produced by plant tissue culture.
• Plant tissue cultures isolated from single
explant, or even a single cell can show
variation after repeated subculture.
• Dedifferentiation : Callus formation
phase.
• Redifferentiation : Morphogenesis phase.
• Cellular totipotency : Phenomenons of
dedifferentiation and redifferentiation are
inherent in the capacity described as
cellular totipotency, a property found only
in plant cells and not in animal cells,
although totipotent stem cells have now
been found in animal tissues also. In other
words, while a differentiated plant cell
retains its capacity to give rise to a whole
plant, an animal cell loses this capacity of
regeneration after differentiation.
 Somaclones
• Meristem tip culture and
micropropagation produced plants with
few exceptions, are true to type.
• Regeneration of plants through callus
or cell suspension culture can induce
changes among cells, tissues and
organs.
• Calliclones, protoclones.
• Larkin and Scowcroft 1982:
Somaclones.
• Morphological Characters: Plant
height, leaf numbers, leaf size and
color.
• Physiological Characters: Biotic stress
like Disease resistance, or Abiotic
stress like temperature resistant,
drought resistant etc.
 Somaclonal Potential
• The amount of variation that can be
expected in vitro will vary with the
clone, age of the clone, use of
mutagenic agents, and use of selection
pressure applied to single cells for
stress conditions such as salt level,
herbicides, microorganisms or their
byproducts, and specific metabolites.
 Kinds of Variation

• Epigenetic change
• Cytogentic
• Genotypic change
• Phenotypic change
– Genotype X environment interation
Epigenetic change(Transient)
– Change in expression of the gene
– Often not permanent
– Not expressed in sexual progeny.
– Useless for crop improvement.
– Dwarfing, leaf shape, growth habits,
drug/stress resistance.
 Cytogenetic Changes
Structural DNA changes

• Chromosomal rearrangements
– Deletions
– Inversions
– Duplications
– Translocations
Structural Changes in Chromosomes

Deletion Inversion

Duplication Translocation
 Genotypic Changes
• The change of a single DNA base
sequence.(Point mutation).
• Inherit in mendelian proportion.
• Expressed in sexual progenies
 Causes of variation
• Variations pre-exist in the somatic cells:
Variable chromosome numbers (diploid,
triploid and polyploid) of mother plant.
• Tissues can be acquired from and part of the
plant.
• All part of the plant body do not have same
tissue system and different tissue systems
differ in composition and physiological
function
• Age of explant play vital role in variation.
• Different in original plant
• Ploidy numbers
apex
5 cm
2n - 50% tobacco
4n - 50%

4n - 70% 20 cm
2n - 9%
8n - 16%
Variability generated due to culture process

• Duration of culture.
Age of culture
Frequency of sub-culture
• Role of growth regulators (GR)
Both auxins and cytokinins
GR can trigger a high rate of cell division
Directly induce mutation
• Culture medium, Levels of KNO3, some micro salts
and metals destabilize the normal genetic make up.
• Undefined substances like coconut milk and yeast
extract may induce somaclonal variation.
 To Increase Somaclonal Variation

• Callus and suspension cultures for several

cycles
• Regeneration of large number of plants
from long-term cultures
• Testing of selected somaclones for genetic
stability
• Multiplication of genetically stable
somaclones for developing new cultivars.
 Isolation for Somaclones
• In vivo screening:
Morphological varaition: like intensity of
chlorophyll, leaf shape, size, height,
branching patteren and flower numbers,
shape and color.
• Physiological changes such as disease
resistance and herbicide resistance.
 In Vitro Screening
• Major application is the selection of mutant
• Provide the variation for crop improvement
• Assumption is made at cellular level that
resistant to particular set of cultural and
environmental condition is expressed by
intact plant.
 Strategy
• Normal media having selective agent like
(salt, herbicides, heavy metals or phytotoxin) at
broad range of concentrations.
• Optimize D90 which kills 90% of cultures
• Further batches TC are then exposed to same
concentration in the medium.
• Surviving cultures are identified and multiplied
on normal medium and finally regenerate.
 Studies concerning different aspects of somaclonal
variation are important for several reasons.

1. First, it is hailed as a novel source of genetic

variation. However, successful utilization of
somaclonal variation heavily depends upon its
systematic evaluation and judicious utilization in
breeding programmes. This necessitates appropriate
experimentation.
2. Second, somaclonal variation is of interest as a
basic genetic process, since it contradicts the
concept of clonal uniformity. The cells and tissues
which are expected to produce true to type plants
through the processes of de-differentiation, division
and re-differentiation, possibly perceive the whole
process as stress, as a result of which the genome,
known for its plasticity, restructures itself to
modulate the expression of genes as demanded by
the in vitro conditions.
3. Third, somaclonal variation is unwanted when
the objectives is micropropagation of elite
genotypes or genetic transformation which
partly involve tissue culture. Under such
circumstances, prevention or at least
minimization of variation is of utmost
importance. To achieve this, the frequency,
nature and magnitude of somaclonal variation in
relation to manipulation of media components,
explant source, culture conditions etc. should
essentially be understood.
 Somaclonal variation induced in
morphological traits in some crop plants
Crop Characters
Sugar cane diameter, stalk length and weight Cane yield, sugar yield, stalk
number, length diameter, volume, density the weight
Potato Growth habit, maturity period, tuber uniformity and skin color.
Photoperiod requirement, fruit production, Late blight.
Tobacco Yield, Days to flowering, plant height, stem diameter, leaf number,
leaf length, leaf width and yield Leaf Shape, type of inflorescence.
Rice Number of tillers per plant, number of fertile tillers per plant,
average panicle length, frequency of fertile seed, plant stature and
flag leaf length. Flowering period, plant height, seed fertility,
heading date and acquired resistance to tungro rice virus and leaf
hopper.
Oat Plant height, heading date, twin culms, yellow leaf stripe, awn
morphology and fertility.
Maize Twin stalks from a single node, reduced pollen fertility and male
sterility.
Crop Characters
Brassica Altered leaf way, multiple branching of the stem, precocious
flowering from the apex, stem or leaf, abnormal leaves, reduced
lamina in leaves, spontaneously absorbing vegetative buds, slow
growth, failure to flower, large pollen grains. Delayed flowering
altered growth habit and gross morphology
Pelargonium Leaf shape, size and form, flower morphology, plant height,
fasciation, pubescence and anthocyanin pigmentation.

Pineapple Leaf colour, foliage density, leaf width and leaf spine formation.

Tomato Male sterility, jointless pedicel, tangerine vifescent leaf, flower

and fruit colour.

Wheat
showing resistance to rust and resistance to high temperature
 Cons and Pros of somaclonal variation

• Disadvantages
• No control, may change in undesirable direction.
i.e –ve, may lose important characters.
• Variations are unpredictable in nature
– Any kind of variation may occurs
– Phenotypic or morphogenic
• May not occur for complex agronomic traits.
• May not be genetically stable
– Might bring new variations in up coming progenies
– Gentically different from somaclonal variant parent
• Require extensive field testing
– To become a new line need several field trails
– Vegetative propagation of selected clone
– Sexually reproducible characters are preferred
– These variants could then be field tested to
ascertain their genetic stability.
– Reciprocal crosses between desirable R1'
progeny and seed derived controls would further
stabilize the variants and help in seedset among
the promising lines.
 Advantages
• Changes can occur in agronomic traits
– Morphogenic and phenotypic.
– Appropriate technology for genetic manipulation of crops
with polygenic traits.
• Changes occur at a high frequency
– Rate of high cell division.
– Increase in chance of variation.
• Some novel changes
– May not occur by conventional breeding.
– Significant contribution in plant breeding.
– Although variation does effect all Characters, and not
always those that are agriculturally useful.
– It is an alternative tool to plant breeding for generation of
new varieties that could exhibit disease resistance and
improvement in quality and yield.
• In vitro selection helps in isolation of lines
tolerant to abiotic and biotic stresses.
• In vitro selection shortens time in
somaclonal isolation with desirable trait.
• somaclonal variation offers an opportunity to
uncover the natural variability in plants and
to use this genetic variability for new product
development.