In this slide - protoplast fusion describes fusion concept of two different type of protoplast of two different plants. And it's also contain an major concept and methods and also advantages and main application. In modern technology some of the methods and technology were used to make an new concept of fusion .

1. SUBMITTED BY:
K. JAYALAKSHMI
I -M.SC., BIOTECHNOLOGY

2. DEFINITION
 The protoplast fusion refers to the fusion of isolated
protoplasts of two different plants.
 It is also known as somatic cell hybridization or
somatic hybridization or somatic hybrid protoplast.
 The plants raised from the hybrid protoplasts are
known as somatic hybrids.
Protoplast I
Protoplast II
Somatic
hybrids

3. METHODSOF FUSION
 There are two methods to induce protoplast fusion.
They are,
i) mechanical method
ii) chemical method
 MECHNICAL METHOD:
Two different plants varieties are sucked
in a micropipette whose mouth is partially blocked
and forced out quickly.
To create an mechanical force, as a result
the two protoplasts fuse together and form a

4.  Ito (1973) has devised cavity slide method for the fusion of
protoplast of members of Liliacae.
 CHEMICAL METHOD:
Chemical substances called fusagens are used to
induced fusion of protoplasts .
Following methods,
1) protoplast fusion by aggregation mixture.
2) polyethylene glycol treatment.
3) protoplast fusion by using Ca++ at high PH
4) fusion of protoplasts induced by electric field.

5. PROTOPLAST
FUSION BY AGGREGATION
 Protoplast protoplast
(Strain (A )) ( strain (B))
protoplast mixture ( strain A+B)
aggregation mixture treatment
10% sucrose and 5.5% sodium nitrate
( incubate at 35 c/5 m) ( plasmolysis)
plasmolysed cells washing with water
(A+B) hybrid protoplast deplasmolysis

6. POLYETHYLENEGLYCOL
TREATMENT(PEG)
Protoplast Protoplast
(strain A) (strain B)
protoplast mixture (strain A+B)
poly ethylene glycol treatment
sedimentations mixture of somatic hybrids
and unfused protoplast
hybrid protoplast (A+B) washing with water

7. PROTOPLAST FUSION
BY USING CA++ AT HIGH PH
 This method was devised by keller and melches in
1973.
 They isolated protoplasts from two strains of tobacco
and mixed together In the presence of a fusagens
solution containing 0.05M calcium chloride and 4M
mannitol, after adjusting the Ph to 10.5 the suspension
was incubated at 25c for 40 mins.
 After incubation protoplasts fused together and
formed hybrids.

8. FUSION OF PROTOPLASTS
INDUCED BY ELECTRIC FIELD
 Zimmerman et.al(1981) used electroporation for the
fusion of protoplasts.
 An electric field is generated between two electrodes
and a protoplast mixture is kept in the electric field
which create pores on the protoplast membrane.
 When the protoplasts come close together at the point
of contact within 20-50µ seconds.
 As the result protoplast undergo fusion.

9. IMPORTANCE
 Produce hybrids .
 It is an ideal tool to overcome male and female sterility
of plants in breeding programs,
 Useful to study the interaction between cytoplasmic
organelles from hybrids.
 It is used to overcome genetic Incompatibility .
 Stress tolerant cultivators.

10. THANKYOU