Genetic transformation of lowland rice variety GR11 for drought tolerance and its ratification for upland paddy cultivation

Abstract

In the current investigation experiments were performed to produce high yielding transgenic rice plants, suitable for upland paddy cultivation in Gujarat. DREB 2A gene isolated from drought tolerant rice variety was placed in a binary vector driven by 35S promoter of cauliflower mosaic virus (CaMV) and a 5’non-coding region (5’-UTR) of rice alcohol dehydrogenase (ADH) gene acting as translational enhancer. Three to four weeks old proliferating calli of GR 11 variety of rice were used for transformation with Agrobacterium strains LB 4404 harbouring pRION_DREB 2A plasmid. NPT II selectable marker gene in putatively transformed plants confers resistance to kanamycin which is used for selection of positively transformed GR 11 callus. In vitro regeneration of the transformed callus produced T0 lines of rice harbouring DREB 2A cassette. The frequency of transformation was 10%. Stable integration of transgene conferring tolerance to drought in T0 rice plants were verified using PCR, Reverse transcription PCR (RT PCR), Real Time PCR (absolute and relative quantification). The transgenic copy number in T0 cisgenic lines were calculated as one to three copies of stably integrated DREB 2A gene per copy of genome of GR 11 rice variety.