Department of Biotechnology, College of Agriculture, University of Agricultural Sciences, Dharwad-580005, India.
Two explants viz., shoot tip and immature zygotic embryos were used for transformation as they showed good elongation and rooting. Shoot tip of about 5-6mm size from in vitro grown seedlings were co-cultured for 48 hrs. Putative transformants were selected on MS medium with 100mg/l kanamycin. In addition, immature embryos were vacuum infiltrated in the presence of Agrobacterium culture. Optimum period for co-cultivation was 48 hrs for shoots and 24 hrs for zygotic embryos. Five cry1Ac positive plants were obtained and of which only three were positive for the expression of the transgene.
