Genetic transformation of lowland rice variety GR11 for drought tolerance and its ratification for upland paddy cultivation
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Authors
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R. Sandeep Raj
Gujarat State Biotechnology Mission, Department of Science and Technology, Udyog Bhavan, Gandhinagar 382 011
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Chandrakant Singh
Department of Plant Physiology, Navsari Agricultural University, Navsari 396 450
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Arpan Modi
Department of Agricultural Biotechnology, Anand Agricultural University, Anand 388 110
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N. Subhash
Department of Agricultural Biotechnology, Anand Agricultural University, Anand 388 110
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 producedT0 lines of rice harbouring DREB 2A cassette.The frequency of transformation was 10%. Stable integration of transgene conferring tolerance to drought inT0rice plants were verified using PCR, Reverse transcription PCR (RT PCR), RealTime PCR (absolute and relative quantification). The transgenic copy number inT0 cisgenic lines were calculated as one to three copies of stably integrated DREB 2A gene per copy of
genome of GR 11 rice variety