Optimisation of plant tissue culture conditions in a popular semi-dwarf indica rice cultivar ADT 39 for effective Agrobacterium-mediated transformation
Abstract
The in vitro culture conditions plays a critical role in promoting callus and shoot induction in rice during Agrobacterium-mediated transformation. In this research, an attempt was made to optimize the key factors affecting the efficiency of Agrobacterium-mediated transformation in ADT 39 rice cultivar. Among the different combinations of the NB-Asmedia tried with varying concentrations of casein hydrolysate from 0.25 g l-1 to 1.25 g l-1, the media containing 0.5 g l-1 casein hydrolysate supplemented with 2 mg l-1 2,4-Dichlorophenoxyacetic acid (2,4-D), 1 mg l-16-Benzylaminopurie (6-BAP) and 1mg l-1 Naphthalene Acetic Acid (NAA) was found to be the best with callus induction frequency of 94.44 per cent. Incubation in NB-As media after infection for 20 min. considerably reduced the Agrobacterium overgrowth after co-cultivation. For shoot induction, regeneration media RNMH30 containing varied concentrations of 6-BAP from 2 mg l-1 to 4 mg l-1 was tried, the media containing 3.0 mg l-1 6-BAP added with 1 mg l-1 NAA recorded the highest shoot induction frequency of 71.11 per cent. The overall transformation efficiency in ADT 39 with the optimized culture conditions was found to be 7.96 per cent.
Key words: ADT 39, Agrobacterium-mediated transformation, Callus induction frequency, Shoot induction frequency, Transformation efficiency.
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