Abstract:
Somaclonal variation and recalcitrance nature were the major bottlenecks of
agrobacterium mediated genetic transformation in Bhendi (
Abelmoschus esculentus (L.). Therefore,
Agrobacterium mediated in planta transformation protocols was standardized using sprouted seed explants of bhendi cv. MDU-1, an important vegetable.
agrobacterium tumifaciens-mediated transformation was performed using EHA105 strain harboring the binary vector pBinAR
carrying
cry2Aa 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-
cry2Aa) 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
cry2Aa transgenes into the bhendi genome was confirmed by polymerase chain reaction (PCR). Work is in progress to grow the T
0 seeds for molecular characterization of the inserted transgene using southern and western blot methods.
R Anandan, Rohini Srivastava, S Murugan and M Prakash. Sonication and vacuum infiltration assisted genetic transformation in bhendi [Abelmoschus esculentus (L.) Moench] using cry2Aa gene for pest resistance. J Pharmacogn Phytochem 2019;8(3):4746-4749.