Protoplast fusion

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. PROTOPLAST FUSION 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 .

3. METHODS OF 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 hybrid.

4. Ito (1973) has devised cavity slide method for the fusion of protoplast of members of L iliacae. CHEMICAL METHOD: C hemical 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. POLYETHYLENE GLYCOL 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 IN DUCED 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.

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