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African Journal of Biotechnology

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A highly efficient method for Agrobacterium mediated transformation in elite rice varieties (Oryza sativa L. spp. indica)

Praderm Wanichananan, Tharathorn Teerakathiti, Sittiruk Roytrakul, Chalermpol Kirdmanee, Surin Peyachoknagul

Abstract


An Agrobacterium mediated transformation method was developed for the Thai rice variety,  Pathumthani 1 (PT1), and the Indian rice variety, Pokkali (PKL). Various aspects of the transformation method, including callus induction, callus age, Agrobacterium concentration and co-cultivation period were examined, in order to improve transformation efficiency. Optimized transformation conditions were established using Agrobacterium strain EHA105, which carries a virulent plasmid, pCAMBIA1301.  A modified Murashige and Skoog (MS) medium supplemented with 1 mg/l 2, 4-D and 0.5 mg/l picloram was optimized for callus induction. Three week old calli were used to co-cultivate with 0.8 -1 OD600  Agrobacterium for 30 min and the culture was continued on agar medium without antibiotics for 2 days. This method can be used to induce high quality calli within three weeks. Based on GUS determination, it was demonstrated that the transformation method was improved significantly, with a high level of transformation efficiency.

Keywords: Agrobacterium tumefaciens, indica rice, mature seed-derived callus, rice transformation, transgenic rice

African Journal of Biotechnology Vol. 9(34), pp. 5488-5495, 23 August, 2010



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