OPTIMIZATION OF GFP GENE EXPRESSION AND PRODUCTION OF TRANSGENIC CALLUS LINES OF RUBBER (HEVEA BRASILIENSIS MUELL.ARG) BY USING GYGOTIC EMBRYO DERIVED CALLUS THROUGH AGROBACTERIUM- MEDIATED GENETIC TRANSFORMATION

A genetic transformation protocol for rubber (Hevea brasiliensis) was established in zygotic embryo derived callus tissue through gfp (green fluorescence protein) detection. Different parameters for transformation efficiency were identified and optimized. Agrobacterium strain LBA 4404 harboring pCAMBIA 1304 for 3 days co-cultivation produced optimum transformation in rubber. Best callus maintenance media was found MS+ 2 mg/L BAP + 2 mg/L 2,4 D + 0.5 mg/L NAA. Callus age and phenotype significantly influenced the transformation efficiency. Proliferated callus of 1-2 months old with pro-embryogenic massages produced maximum gfp expression. Transformed callus lines were established without antibiotic pressure. Integration of gfp in rubber callus with desired gene could be a useful tool for recombinant rubber plant.