Abstract: In order to obtain a new germplasm material of Brassica napus with both insect-resistance and herbicide-resistance properties,the hypocotyls of Xiangyou 15 were used as explants.The synthetic insect-resistant cry1ab/ac fusion gene was inserted into the expression vector pFGC5941 to form pFGC-Bt. pFGC-Bt was transformed into Brassica napus cultivar Xiangyou 15 by Agrobacterium tumefaciens infection of hypocotyls.In order to obtain novel materials of Brassica napus with insect-resistance and herbicide-resistance,the synthetic insect-resistant cry1ab/ac fusion gene was inserted into the expression vector pFGC5941 to form pFGC-Bt. pFGC-Bt was transformed into Brassica napus cultivar Xiangyou 15 by Agrobacterium tumefaciens infection of hypocotyls. After the callus differentiation and plant regeneration,seven T₀ transgenic plants were obtained. The plants were harvested individually,and the seeds were then planted in pots,when seedlings were 4-leaf-stage,herbicide basta was applied. The surviving plants were then detected by PCR with bar gene primers. Eleven positive plants were selected for further test of Cry1Ab/Ac protein double antibody sandwich immunochromatographic assay. Cry1Ab/Ac protein was detected in all the eleven transgenic plants.The entomocidal activity was evaluated by insect feeding bioassays performed with larvae of Pieris rapae. The transgenic plants and non-transgenic control leaves were used in the insect bioassay. The Pieris rapae larvae that fed on transgenic plants were observed to stop growing and died after 7 days of feeding,whereas larvae fed on control plants grew normally.After 3 days, the leaves were basically eaten clean. The results of feeding identification indicated that the transgenic insect fusion gene cry1ab/ac was highly resistant to Pieris rapae. The results identified the transgenic plants had double resistance to herbicide basta and insect.

Key words: Brassica napus, cry1ab/ac gene, bar gene, Genetic transformation

CLC Number: 

• Q78

• S635.03