Agrobacterium- mediated Genetic Transformation of oilseed Brassica juncea(L.)

Abstract

Brassica is an important oilseed crop in Bangladesh. The available Brassica varieties cultivated in Bangladesh are susceptible to many diseases and pests specially, with fungal diseases. In this context, an alternative to trait improvement especially disease resistant variety would be evolved by applying biotechnological approaches like genetic transformation. So, the present investigation was carried out for the development of Agrobacterium-mediated genetic transformation system for oilseed Brassica grown in Bangladesh. Prior to the transformation an efficient in vitro regeneration protocol was developed for two varieties of Brassica juncea namely, BARI Sarisha-11 and BARI Sharisha-16. The best shoot regeneration was obtained from hypocotyl explants using MS medium supplemented with 2.0 mg/l BAP and 0.2 mg/l NAA. MS medium hormone was found to be effective in inducing roots. Three gene construct (LBA4404/pBI121, LBA4404/ pCAMBIA2300polyA AFPFP, EHA105/pGII-pSOUP-VST-N-CHITIN) of Agrobacterium were used for transformation experiments. Transformation frequency regarding the recovery of shoots for BARI Sarisha-11 and BAR Sarisha-16 using  gene construct I was 0.84% and 0.96% respectively, while that was 0.63% and 0.78% respectively in case of construct III. The stable integration of the putatively transformed shoots developed by using EHA105/pGII-pSOUP-VST-N-CHITIN (containing bar and chitinase gene) was confirmed through PCR analysis.

Keywords: Oilseed Brassica; In vitro Regeneration; Genetic Transformation Bangladesh.