Transgenic cotton: Harboring broad term resistance against insect and weeds through incorporation of CEMB double Bt and cp4EPSPS genes

Introduction of multiple traits in crop plants for desirable character is an essential tool of Biotechnological application. To overcome problem of insect pests and weeds cotton variety FBS-37 was genetically modified by transformation of CEMB Cry1Ac+Cry2A and GTGene cloned in different plant expression vectors under CaMV35S Promoter and nopaline synthase Nos Terminator. CEMB optimized protocol of cotton gene transformation i.e. Agrobacterium mediated shoot apex cut method was applied for transformation of these genes constructs in cotton. Overall transformation efficiency was found to be 1.05%. Putative transgenic cotton plants were shifted to soil pots and acclimatized in the green house conditions. PCR with gene specific primers amplified 190bp for EPSPS (Glyphosate GTG) and 1000bp for each of Cry1Ac and Cry2A. Quantification of Cry1Ac, Cry2A and GTG protein through ELISA determined maximum of 0.8μg/g, 1μg/g and 0.9μg/g of tissue respectively. 100% mortality was obtained in 2^nd instar larvae of Heliothus armigera when compared with non-transgenic control cotton plants where maximum damaged was seen in leaf bioassay. Moreover, transgenic cotton plants successfully survived when sprayed with 1600ml/acre of glyphosate as compared to control where 100% mortality and necrosis occur after 7 days of spray assay in control conditions.