PJB-2007-145
IMPROVEMENT OF BASMATI RICE AGAINST FUNGAL INFECTION THROUGH GENE TRANSFER TECHNOLOGY
AYESHA ASGHAR1, HAMID RASHID2, M. ASHRAF1, M. HAROON KHAN2 AND ZUBEDA CHAUDHRY2
Abstract
In this study, gene for fungal resistance (RCC2) has been introduced into Basmati 385 by Agrobacterium mediated transformation. Maximum callus induction (88%) was achieved on MS medium with 2.0 mg/l 2,4-D. Different combinations and concentration of growth regulators (NAA and BAP) were used to develop an efficient culture environment for higher regeneration frequencies. Maximum plant regeneration (80%) from calli was achieved on RM5 (NAA 1.0 mg/l+ BAP 5.0 mg/l). Hygromycin was used as selectable agent and at concentration of 50 mg/l proved to be lethal for scutellum derived calli. Calli of more than 5 mm in size were infected with Agrobacterium strain EHA101. Rice chitinase gene-RCC2 with vector pB1333-EN4 was introduced under the control of enhanced CaMV 35S promoter. Transformation efficiency proved to be highest when 21-24 days old calli were used with co-cultivation period of 2-3 days. Selection of the calli was carried out with hygromycin (50 mg/l) in addition to cefotaxime (1000 mg/l). After two weeks of selection, calli were transferred to RM5 containing hygromycin 50 mg/l + cefatoxime 1000 mg/l. A significant regeneration frequency of transformed plants was attained which was 10-11%.
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