CRY 1AB trangenic cowpea obtained by nodal electroporation

Abstract

Electroporation-mediated genetic transformation was used to introduce Cry 1 Ab insecticidal gene into cowpea. Nodal buds were electroporated in planta with a plasmid carrying the Cry 1Ab and antibiotic resistance npt II genes driven by a 35S CaMV promoter. T1 seeds derived from electroporated branches were selected in vitro on a medium containing geneticin. PCR and Southern blot analyses confirmed the stable integration of the Cry 1Ab gene into genome of transgenic T1 cowpea plants. Copy numbers of the gene in cowpea were estimated to be between one to three per genome. However transgene integration occurred at high molecular weights and corresponded to the hybridisation bands obtained for plasmid control. When tested for resistance to Maruca vitrata, T1 plant progenies reduced larval survival to as low as 11% three days after infestation (DAI). T2 progenies of Cry 1Ab transgenic lines also positively hybridised with npt II probes when subjected to southern analyses. T3 progenies significantly reduced larval survival and larval weight and inhibited M. vitrata feeding on cowpea leaves.