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1. SOMATIC HYBRIDIZATION
Presented by
‘ M.Phool Badshah ’

2. S
 Development of hybrid plants
through the fusion of somatic
protoplasts of two different plant
species/ varieties is called somatic
hybridization .
Definition

3. AWS

s
Somatic hybridization
techniques
1- Isolation of protoplast
2- Fusion of protoplast of desired varieties
3- Identification and selection of somatic hybrid cells
4- Culture of the hybrid cells 5- Regeneration of hybrid plants
Five
steps :

5.  a
Mechanical method : In
this method, large and highly
vacuolated cells of storage
tissues such as onion bulb
scales, radish root and beet
root tissue could be used for
isolation.
The cells are plasmolysed in
an iso-osmotic solution
resulting in the evacuation of
contents in the centre of cell.
Enzymatic method :
The tissue is first treated with
pectinase (macerozyme) to
separate cells by degrading
middle lamella. These free
cells are then exposed to
cellulose to release
protoplasts
1- Isolation of protoplasts

6.  B
2- Fusion of protoplast of desired
varieties
Spontaneous
fusion : ‘Protoplasts
during isolation often
fuse spontaneously.’
Induced fusion
Fusion of freely isolated
protoplasts from different
sources with the help of fusion
inducing chemical agents
Intraspecific
Intergeneric
Chemofusion
Mechanical
fusion
Electrofusion

7. N
 Hybrid identification based on difference between the
parental cells and hybrid cells with respect to :
o Pigmentation
o Cytoplasmic markers
o Fluorochromes like FITC ( FLUOROCINE ISOTHIO-CYANATE
) RITC (RHODAMINE ISOTHIOCYANATE ) are used for labeling of
hybrid cells.
o Presence of chloroplast .
o Nuclear staining .
3- Identification and selection of somatic hybrid cells :

8. A
 Specific amino acid
 Chromosomal analysis
 Herbicides
 Antibiotics
 Phytotoxins
 Auxin autotrophy
 Genetic complementation
 Auxotrophic and metabolic mutants.
Hybrid Selection
( Several markers are used )

9. B
 Hybrid cells are cultured on suitable
medium provided with the appropriate
culture conditions.
4- Culture of the hybrid cells

10. D
 Plants are induced to regenerate from
hybrid calli.
 These hybrid plants must be at least
partially fertile, in addition to having some
useful property, to be of any use in breeding
scheemes .
5- Regeneration of hybrid plants

11. S
 Transfer gene for disease resistance, abiotic stress
resistance, herbicide resistance, and many other
quality characters.
 Production of novel interspecific and intergenic
hybrid.
 Production of fertile diploids, and polyploids from
sexually sterile haploids, triploids and aneuploids .
 Pomato ( hybrid of potato and tomato )
Advantages of somatic
hybridization