Somaclonal variation

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• Larkin and Scowcroft (1980)the first time
coined the word ‘somaclonal variation’ and
defined it as the genetic variablity, which is
generated during tissue culture
• Evans and Sharp(1983) analysed somaclonal
variations in plants regenerated from leaf
explants of tomato

Somaclonal Variation
• Somaclonal variation is a general
phenomenon of all plant regeneration systems
that involve a callus phase
• There are two general types:
–Heritable, genetic changes (alter the DNA)
– Stable, but non‐heritable changes (alter gene
expression, epigenetic)
• With or without mutagen
Somaclonal variation
􀂾 Genetic variation is essential for any crop breeding
programs
􀂾 Plant cell cultures are potential sources of useful
genetic variation
􀂾 Although particular genotypes are cloned in tissue
culture, phenotypic variants are observed amongst
regenerated plants Caused by changes in
chromosome number and structure:
𐂾 the expression of chromosomal mosaicism or
genetic disorders in explant cells
􀂾 new irregularities brought about by culture
conditions through spontaneous mutations

Schemes for obtaining somaclonal variation
1. Without in vitro selection:
explant → explant derived callus → shoot regeneration → plant
→ transfer to the field → screening for desirable traits →
agronomic trials
􀂉 both dominant and homozygous recessive traits can be directly
selected
􀂉 progenies of heterozygous regenerants used for selection of
recessive traits
􀂉 time consuming, not effective
2. With in vitro selection:
􀂉 protoplasts, cell suspensions and callus cultures can be searched
for biochemical mutants selection for resistance: inhibitors such as
antibiotics, amino acid analogs, herbicides, pathotoxins

Factors influencing somaclonal variation
􀂉 Genotype: frequency of regeneration
and Somaclones
􀂉 Explant source
􀂉 Duration of cell culture: most longterm
cultures are chromosomally variable
􀂉 Culture conditions: 2,4‐D, NAA, BAP

Factors important during in vitro selection
of somaclonal variants
􀂉 Selective agent: a dose response curve with respect
to growth inhibition must be performed for each
selection agent of interest
􀂉 Selection technique
􀂉in vitro selection of cell lines against an inhibitor is
best carried out at concentration that kill 70‐90% of
wild type cells
􀂉 Regeneration of plants
􀂉 In vivo testing
􀂉 Agronomic analysis: testing agronomic value in the
first generation and also in the second generation
for stability

Genetic change
Change in ploidy number
in culture
• Endomitosis
– Sister chromatids separate, no spindle formation or
cytoplasmic separation
• Endoreduplication
– Chromosomes undergo extra duplications at interphase
• Spindle fusion
– Results in bi‐ or multinucleate cells

Application of somaclonal variation
􀂉Novel variants:
􀂉 improved scented Geranium variety ″Velvet Rose ″
􀂉 tornless blackberries (Rubus) ″Lincoln Logan″
􀂉Disease resistance
􀂉eye spot disease (Helminthosporium sacchari), downy
mildew (Sclerospora sacchari), Fiji virus disease in
sugarcane
􀂉Abiotic stress resistance
􀂉freezing tolerance in Norstar winter wheat (proline
accumulation)
􀂉salt and drought tolerance in alfalfa, maize, rice,
tobacco

Application of somaclonal variation
􀂉Herbicide resistance
􀂉tobacco, soybean wheat, maize
􀂉 resistance to glyphosate, sulfonylurea,
imidazolinones etc.
􀂉 Insect resistance
􀂉 Russian wheat aphid (kirva) in wheat
􀂉 Seed quality
􀂉 Lathyrus sativa in central India
􀂉 Alien gene introgression

Basis of somaclonal variation
􀂉Karyotype changes
𐂉polyploidy, commonly associated with reduced
fertility
􀂉 Changes in chromosome structure
􀂉translocations, deletions, duplications,
inversions, breaks, multinucleate cells, abnormal
anaphase etc.
􀂉Single‐gene mutations
􀂉 Cytoplasmic genetic changes
􀂉 variations in mitochondrial and chloroplastic
DNA
􀂉 Mitotic crossing over (MCO)

Basis of somaclonal variation
􀂉 Gene amplification and nuclear changes nuclear
genes are affected by tissue culture stress increased
amount of total DNA and an increased proportion of
highly repeated sequences (AT‐ and GC‐rich fractions in
tobacco) ribosomal DNA is known to alter directly in
response to environmental and cultural pressures:
methylation, structuralrearrangements
􀂉Transposable elements

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