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Agrobacterium-mediated transformation; Brassica oleracea var.botrytis; cryIA(b) gene; GUS; Plutella xylostella

A number of factors that are known to influence genetic transformation were evaluated to optimizeAgrobacterium-mediated transformation of hypocotyl explants of cauliflower variety Pusa Snowball K-1. The binary vector p35SGUSINT mobilized intoAgrobacterium strain GV2260 was used for transformation and transient GUS expression was used as the basis for identifying the most appropriate conditions for transformation. Explant age, preculture period, bacterial strain and density were found to be critical determinants of transformation efficiency. Using the optimized protocol, the syntheticcryIA(b) gene was mobilized into cauliflower. Molecular analyses of transgenics established the integration and expression of the transgene. Insect bioassays indicated the effectiveness of the transgene against infestation by diamondback moth (Plutella xylostella) larvae.

R Chakrabarty¹ N Viswakarma¹ S R Bhat¹ P B Kirti^{1 2} B D Singh^{1 3} V L Chopra¹

1. National Research Centre on Plant Biotechnology, Indian Agricultural Research Institute, New Delhi - 110 012, India
2. Department of Plant Sciences, University of Hyderabad, Hyderabad - 500 046, India
3. School of Biotechnology, Banaras Hindu University, Varanasi - 221 005, India

Published

September 2002

Manuscript received

13 March 2002

Accepted

16 July 2002