AGROBACTERIUM MEDIATED TRANSFORMATION TO BUILD RESISTANCE AGAINST BACTERIAL BLIGHT IN RICE
Mohammad Haroon Khan, Hamid Rashid, Zahoor Ahmad Swati, Zubeda Chaudhry
Abstract
Mohammad Haroon Khan, Hamid Rashid, Zahoor Ahmad Swati, Zubeda Chaudhry
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%.
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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%.
Key concepts: Agrobacterium, Callus, Transformation (genetics), Transformation efficiency, Biology, Regeneration (biology), Cultivar, Horticulture