Transformation of Cotton (Gossypium hirsutum L.) With Insect Resistant Gene by Particle Bombardment and Agrobacterium

A combination of two techniques i.e. particle bombardment and Agrobacterium method was applied to transfer marker NPT-II and insecticidal cry1A(b) gene into zygotic embryos of cotton variety MNH-93. Wounding in embryos produced by bombardment of tungsten particles increased the efficiency of Agrobacterium mediated transformation. Primary transformants selected on selection medium containing Kanamycin (100 g/ml) for 6-8 week showed the presence and expression of cry1A(b) gene in the primary transformants. Insect Bioassays with 2nd instar larvae of of American bollworm (Heliothis armigera), PCR amplification of the coding region of cry1A(b) gene in plant genome and ELISA with the antibodies against cry1A(c) confirmed the transformation.