Canola (Brassica napus L.) regeneration and transformation via hypocotyl and hypocotyl derived calli.

Canola (Brassica napus L.) is a leading vegetable oil and protein rich meal crop in the world. Like any other crop plant it is also a target for genetic manipulation. During this study a system of plant tissue culture was established for efficient regeneration and genetic transformation of three cultivars (Wester, Con-1, and Pakola). In vitro regeneration from hypocotyls derived callus and directly from hypocotyls of different cultivars was monitored. A procedure for callus induction, regeneration and selection of explants was also investigated. In case of callus induction more efficient response was observed in Wester and Con-1 compared to Pakola (63.8%). Similarly, high regeneration was noted in Wester (87.6) and Con-1(84.4) while low in Pakola (54.2%). Two Agrobacterium (LBA4404) strains harboring plasmid pCAMBIA2301 and pCAMBIA1391Z (harboring promoter P2F1: GUS) were used for transformation in canola plants using hypocotyls of all three cultivars. PCR and transient GUS expression was used as the basis for the confirmation of transformation.