1Department of Pathology and Microbiology, College of Biotechnology, S V P University of Agriculture and Technology, Meerut, 250110, India.
Department of Biotechnology, College of Agriculture, S V P University of Agriculture and Technology, Meerut-250110, India.
*Corresponding author nikita.biotech@gmail.com
The aim of present study is to establish an efficient callus induction, regeneration and transformation system for basmati rice varieties. Seeds of four basmati rice varieties viz. Basmati-370, Pusa Sugandha-4, Vallabh Basmati-22 and Taroari were sterilized and incubated on MS medium with different concentrations of 2,4-D (1.5, 2.0, 2.5 mgL-1). The best response of callus induction and proliferation for all the four basmati rice varieties was observed when 2 mg/l of 2, 4-D was used in the medium. The callus induction frequency was 94.1% for PS-4, 91.7% for B-370, 83.5% for VB-22 and 70.5% for Taroari. Twenty one day old maintained calli when transferred to regeneration media containing basic MS salts and vitamins with NAA 1 mgL -1, BAP 2 mgL-1 and Kinetin 2 mgL-1 showed 42.5% for B-370, 27.5% for PS-4, 12.5% for VB-22 and 15% for Taroari. The embryogenic calli derived from seed scutellum of two Basmati rice varieties i.e. B-370 and PS-4 were used for transformation by Agrobacterium tumefaciens strain EHA105 harboring pCAM-BIA1305 with GUS (β-glucuronidase) and hpt (hygromycin phosphotransferase) genes for studying the transformation efficiency. From GUS histochemical analysis it was found that B-370 exhibited a transformation frequency of 6.6% whereas transformation frequency of PS-4 was found to be 5.3%.
Agrobacterium tumefaciens, Callus, Regeneration, Transformation, GUS
