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Biotechnology
Year: 2008  |  Volume: 7  |  Issue: 3  |  Page No.: 530 - 536

GUS Gene Transformation in Rice (Oryza sativa L.) Variety BRRI Dhan-30 Mediated by Agrobacterium tumefaciens

M. Asaduzzaman, M.A. Bari, M. Rahman, M. Minami, K. Matsushima and K. Nemoto    

Abstract: Agrobacterium mediated transformation has already been reported in rice but under the present investigation efforts has been made to establish the transformation protocol in Bangladeshi rice variety BRRI dhan-30. High percentage of callus induction at 97.3% was obtained when seeds of rice (Oryza sativa L.) cv. BRRI dhan-30 were cultured on modified N6 medium supplemented with 3% sucrose, 2.0 mg L-1 2, 4-D and 0.8% agar under dark condition. Maximum  88.28%  calli  induced  shoots  and  were  obtained from BRRI dhan-30 in the suitable regeneration medium was MS medium supplemented with 3% (w/v) sucrose, 2.0 mg L-1 BAP+0.05 mg L-1 NAA+3% sorbitol and 0.8% agar. Agrobacterium mediated gene transfer protocol for rice variety BRRI dhan-30 were performed using Agrobacterium tumefaciens strain LBA4404, which harbored the plasmid pCL3 containing genes for β-glucuronidase (GUS) and kanamycin resistance. It was found that kanamycin concentration up 50 μg mL-1 were effective for selection of transformants. Putatives transformants as  indicated  by  kanamycin  test  were  subjected  to GUS assay. GUS activities were found in rice calli after co-cultivation.

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