ISSN 2375-1231
African Journal of Crop Science ISSN 2375-1231 Vol. 5 (12), pp. 001-005, December, 2017. © International Scholars Journals
Full Length Research Paper
Optimization and development of regeneration and transformation protocol in Indian mustard using lectin gene from chickpea [Cicer arietinum (L.)]
V. V. Singh1, Vandana Verma1*, Aniruddh K. Pareek1, Monika Mathur1, Rajbir Yadav1, Poonam Goyal1, Ajay Kumar Thakur1, Y. P. Singh1, K. R. Koundal2, K. C. Bansal2, A. K. Mishra1, Arvind Kumar1 and Sandeep Kumar1
1Directorate of Rapeseed-Mustard Research, Bharatpur, Rajasthan 321 303, India.
2NRCPB, IARI, New Delhi, India.
Accepted 11 April, 2017
Abstract
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.
Key words: Indian mustard, Agrobacterium tumefaciens, chickpea lectin gene, cotyledonary petioles, transformation.