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
DOI:
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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