Somatic Hybridization: Development of hybrid plants through the fusion of somatic protoplasts of two different plant species/varieties is called somatic hybridization.
Somatic hybridization technique involved following steps
Isolation of protoplast
Fusion of the protoplasts of desired species/varieties
Identification and Selection of somatic hybrid cells
Culture of the hybrid cells
Regeneration of hybrid plants
Isolation of Protoplast (Separation of protoplasts from plant tissue) Can be done using two methods
Mechanical Method
Enzymatic Method
Mechanical Method: Used for vacuolated cells like onion bulb scale, radish and beet root tissues
Low yield of protoplast
Laborious and tedious process
Low protoplast viability
Enzymatic Method: Used for variety of tissues and organs including leaves, petioles, fruits, roots, coleoptiles, hypocotyls, stem, shoot apices, embryo microspores.
Mesophyll tissue - most suitable source
High yield of protoplast
Easy to perform
More protoplast viability
Protoplast Fusion: Two ways
Spontaneous Fusion
Induced Fusion
1) Chemofusion : fusion induced by chemicals
2) Mechanical Fusion: Physical fusion of protoplasts under microscope by using micromanipulator and perfusion micropipette.
3) Electrofusion : Fusion induced by electrical stimulation.
Identification and Selection of somatic hybrid cells: Hybrid identification - Based on difference between the parental cells and hybrid cell with respect to
Pigmentation
Cytoplasmic markers
Presence of chloroplast
Nuclear staining
Culture of the hybrid cells: Hybrid cells are cultured on suitable medium provided with the appropriate culture conditions.
Advantages of somatic hybridization: Production of novel interspecific and intergenic hybrid Pomato (Hybrid of potato and tomato)
Production of fertile diploids and polypoids from sexually sterile haploids, triploids and aneuploids
Transfer gene for disease resistance, abiotic stress resistance, herbicide resistance and many other quality characters
Production of heterozygous lines in the single species which cannot be propagated by vegetative means.
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#Somatichybridization
#PTC
#Plantregeneration
19 сен 2024
