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Asian Journal of Agricultural Research
Year: 2015  |  Volume: 9  |  Issue: 4  |  Page No.: 139 - 154

Development of Transgenic Rice (Oryza sativa L.) Plant Using Cadmium Tolerance Gene (YCFI) through Agrobacterium Mediated Transformation for Phytoremediation

Md. Mahmudul Islam and Md. Khalekuzzaman    

Abstract: Cadmium has adverse effects on plant development and ultimately reduces production. The investigation was undertaken for successful gene transfer and phytoremediation through an efficient Agrobacterium-mediated genetic transformation method using cadmium tolerance gene (YCF1). Embryogenic calli induced after 20 days of highly regenerating rice cultivar BRRI dhan29 and Agrobacterium strain GV 3101 was transformed with binary vector pCAMBIA 1303-YCF1 which contained the hygromycin phosphotransferase (HPT) gene as a selectable marker and the yeast cadmium factor 1 (YCF1) gene, were used for genetic transformation in the experiment. The transformed colonies were selected on 15 mg L-1 hygromycin and 50 mg L-1 rifampicin to select hygromycin resistant shoots. Hygromycin-resistant shoots were subsequently rooted on root induction medium. Rooted plantlets were transferred to pot-soil, hardened and grown in a greenhouse until maturity and stable integration, expression of YCF1 gene was also confirmed by using PCR analysis. The maximum transformation efficiency of 22% was obtained using 500 mg L-1 cefotaxime as a bacteriostatic agent to inhibit growth of Agrobacterium and 100 mM acetosyringone in co-cultivation medium. Southern blot analysis was performed to confirm that transgenes (HPT and YCF1) were stably integrated into the plant genome. All transgenic plants showed single-copy of transgene integration in the host genome. This transgenic rice plant will uptake cadmium from soil and will protect rice grain from cadmium and store into cell vacuoles of rice plants. As a result soil will be free from cadmium through phytoremediation process.

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