Author(s):

R Anandan, Masoud Tohidfar, P Satheesh Kumar, A Kamaraj and N Vijayakumar

Abstract: In planta transformation is a novel approach to generate transgenic with a short period of time. Somaclonal variation is a major constraint in tissue cultured dependent genetic transformation in Bhendi (Abelmoschus esculentus (L.). Therefore, agrobacterium tumifaciens mediated in planta transformation protocols was standardized using sprouted seed explants of bhendi cv. MDU-1, an important vegetable. agrobacterium -mediated transformation was performed using LBA4404 strain harboring the binary vector pBinAR carrying cry3a gene under the control of CaMV35s promoter and npt II gene as a selectable marker. The transformation event consisted of sonicating the explants for 3 minutes, vacuum infiltration (750 mm of Hg) for 2 minutes in Agrobacterium (pBinAR-cry3a) and co-cultivation for 3 days in MS medium with acetosyringone (100µM) showed transformation efficiency of 12.5%. The presence and integration of npt II and cry3a transgenes into the bhendi genome was confirmed by polymerase chain reaction (PCR). Work is in progress to grow the T₀ seeds for molecular characterization of the inserted transgene using southern and western blot methods.

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