Abstract

V. V. Singh , Vandana Verma, Aniruddh K. Pareek1 , Monika Mathur , Rajbir Yadav , Poonam Goyal , Ajay Kumar Thakur , Y. P. Singh , K. R. Koundal , K. C. Bansal, A. K. Mishra, Arvind Kumar and Sandeep Kumar

A simple and easy protocol for regeneration and genetic transformation of Brassica juncea (L.) Czern and Coss cv. Pusa bold has been developed. The optimum regeneration was found on combination of BAP (3.0 mg/l) and IAA (0.2 mg/l). The genetic transformation protocol has been standardized using Agrobacterium tumefaciens (gv3101). Cotyledonary petioles used as explant were pre-cultured for 48 h and than co-cultivated with Agrobacterium suspension for 2 days. Putative transformants were selected from selection medium containing 20 mg/l Kanamycine and 250 mg/l cefotaxime. Complete plant formed after placing shoots on rooting medium (0.5 mg/l IBA). Confirmation of integration of transgene was done by PCR using nptII gene primer.