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AGROBACTERIUM MEDIATED TRANSFORMATION TO BUILD RESISTANCE AGAINST BACTERIAL BLIGHT IN RICE

MOHAMMAD HAROON KHAN¹, HAMID RASHID^1*, ZAHOOR A. SWATI³ AND ZUBEDA CHAUDHRY²

Abstract: Agrobacterium mediated transformation and regeneration efficiency of rice cultivar IR-6 was studied. Various concentrations and combinations of growth regulators were used to establish efficient culture environment for enhanced regeneration efficiency. Maximum (70.96%) callus induction was achieved on MS medium containing 2,4-Dichlorophenoxyacetic acid with concentration of 2 mg/l. Maximum regeneration frequency (80%) was observed on regeneration media containing NAA 1.0 mg/l and BAP 5.0 mg/l. Calli of more than 5 mm size were infected with Agrobacterium containing the binary vector pTCL5. Age of the calli was also found to be a limiting factor in transformation efficiency. Older calli of over four weeks were less efficient in transformation whereas 22-25 days old calli were found to be highly efficient in transformation. Selection of the calli was carried out with Hygromycin (50 mg/l) in addition to Cefatoxime (500 mg/l) in combination with Carbenicillin (500 mg/l). Transgenic plants regeneration frequency was observed as 2.26%.

¹Agricultural Biotechnology Programme, NARC Islamabad, Pakistan

²Institute of Biotechnology and Genetic Engineering, NWFP Agricultural University Peshawar, Pakistan

³Depatment of Biotechnology, University of Malakand, Pakistan.