Transformation Of Rapid-cycle Brassica Napus And The Generation Of T-DNA Tagging Lines

T-DNA transfer mediated by Agrobacterium tumefaciens,the preferred method for large-scale transformation,is widely used in the model plant for the construction of mutant library. Arabidopsis thaliana genome project was completed in 2000 and genome-scale database of insertion mutant also have been built. Mutant population of A.thaliana SM lines (48 000 lines), SALK lines (150 000 lines), GABI-Kat population (42 603 lines) and Syngenta CS lines (120 000 lines) is widely used in A.thaliana functional genomics research .Rapeseed (Brassica napus L. AACC, 2n = 38) is allotetraploid and large genome size (1132Mb) ,about 9 times larger than A.thaliana genome (125Mb),indispensable vernalization ,long growth cycle, limited the breeding process and the B.napus functional genomics research.Rapid-cycle Brassica napus(RCB) was transformed by binary vector pCambia2031, and T-DNA insertion mutants was identified by PCR. Through tissue culture, and dipped flowers obtained T-DNA. Fifty one insertional mutants were recovered by tissue culture and floral dip of RCB. Among them, twenty-eight were generated from tissue culture and twenty three from floral dip, the transformation rate was 2.69%,1.04% respectively. Of the TAIL-PCR analysis 11 T-DNA tagged lines, 6 was isolated effectively.In addition, in order to shorten the time for rape genetic transformation and eliminate selectable marker genes, twin T-DNA binary vector pSB130-SG1 for Gastrodia antifungal protein(GAFP) was constructed and could be transformed into RCB to obtain genetic modified rapeseed which is resistant to fungal disease without selectable marker gene.