Insect resistance of transgenic broccoli ('Pusa Broccoli KTS-1') expressing a synthetic cryIA(b) gene

摘要

Broccoli has no suitable genes for insect resistance within the gene pool for transfer through conventional breeding. A protocol was optimized for transformation of hypocotyl explants of broccoli based on transient GUS expression. A synthetic cryIA(b)gene coding for an insecticidal crystal protein (ICP) of Bacillus thuringiensis (Bt) was transferred to the broccoli cultfvar 'Pusa Broccoli KTS-1' by co-cultivating hypocotyls explants with Agrobacterium tumefaciens. Transformed plants resistant to kanamycin were regenerated. Molecular analysis demonstrated gene integration and expression. Immunoblot analysis revealed high-level expression of Bt toxin protein in the transgenic plants. In insect feeding bioassay, transgenics showed significant and effective resistance against the larvae of the diamondback moth (Plutella xylostella). Inheritance of the transgene was confirmed in T_1 seedlings.