The Research Of Insect-resistant Transgene Of Chinese Cabbage

The intron of the pin â…¡ gene in plasmid pBBB-pinâ…¡ had been deleted by Xba I restriction enzyme,and the modified plasmid (pBBB-pinIIM) were transferred into Agrobacteriwn tumefaciens C58.A spider insecticidal protein (SlP)gene were isolated from plasmid DNA of pBPCVH3 and inserted into the plasmid DNA of PSHIC whichcaning gus and bar genes,Restriction enzyme digestion analysis showed recombinant pSHK-sIp were gotCotyledons with Petiole from aseptic seedlings were used as explants.13 genotypes of Chines cabbage (Brasscia campestris L. ssp. Pekinensis) were testd for their shoot regeneration ability on Ms medium with 2mg/LBA,lmg/LNAA 3mg/L AgNOs.Three genotypes (BeiJing 80 ,xiaoza 66,etc)Showed high and stable regeneration frepuency and BeiJing 80 was selected for the transformation works using Agrobacterium tumefaciens C58 with plasmid pBBB-p'n" earring Camv35s driving bar and Pin II genes.Five Basta tolerant regenerated plants were obtained The best suitable Basta concentration to selest the transgenic cells was 4 mg/L. PCR,PCR-Southern and genomic Southern blotting analyses showed that the bar and pin II gene was integrated into the Chinese cabbage genome DNA.Results of preliminary insect bioassay showed that the growth and development ofpieris rapae larvae were retared.Brassica campestris L. ssp. chinensis were transformed by vacuum infiltration method. 41 herbicide resistant plant was acquire by screening the TO generation which collected from four sorts treatment. PCR,PCR-Southem and genomic Southern blotting analysis showed that the target genes were integrated into the Plant genome.Results demonstrated that flowering or not of the buds had little influence on the transformation frequency,but the transformants frequency in the lower parts were three times more than that in the upper parts of the plants. It indicated that the alakastrum of treated plant was the main part of Agrobacterium tumefaciens. Transformation.