Browsing by Author "BALACHANDRAN, S.M"
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ThesisItem Open Access DEVELOPMENT OF TRANSGENIC HYBRID RICE PARENTAL LINE, APMS6B WITH Bt FUSION GENE, RESISTANT TO YELLOW STEM BORER (Scirpophaga incertulas)(PROFESSOR JAYASHANKAR TELANGANA STATE AGRICULTURAL UNIVERSITY RAJENDRANAGAR, HYDERABAD, 2015) AMARNATH, D; BALACHANDRAN, S.MIn the present study, the genotype APMS6B which is a maintainer line for hybrid seed production was used for tissue culture and transformation experiments with cry1Aabc gene which codes toxic proteins for lepidopteron pest, yellow stem borer. As callus is used for transformation, callus induction potential from seeds was studied using 2,4-D of 1.5 mgL-1, 2.0mgL-1, 2.5mgL-1, 3.0mgL-1. Maximum callus induction was observed at concentration of 2.5mgL-1 of 2,4-D. In order to assess the regeneration potential calli, were cultured on media, with 0.3mgL-1, 0.5mgL-1, 1.0mgL-1 and 1.5mgL-1concentrations of NAA, where 0.5mgL-1 concentration of NAA showed marginal advantage of regeneration over other concentrations. Calli of 21 days old were used for Agrobacterium mediated transformation under 40mgL-1 concentration of Acetosyrongone and kept for co-cultivation for 3 days. Co-cultivated calli were washed with carbencellin treated water of 250mgL-1 concentration and cultured on selection media containing hygromycin 50mgL-1. Calli survived on selection media were transferred to regeneration media, then to rooting media and later hardened using yoshida media in a biosafety glasshouse. Thirty one putative transgenic plants were obtained, out of which 27 were analysed for the presence of transgene using hygromycin primers. Out of these only two plants were confirmed to be positive.