2,4-D RESISTANT TRANSGENIC COTTON PLANTS PRODUCED BY AGROBACTERIUM-MEDIATED GENE TRANSFER
Clen Zhixian;Danny J.Llewllyn;Fan Yunliu;Li Shujun;Guo Sanduei;Jiao gaili;Zhao Junxia(1.Institute of Cotton, Shanxi Academy of Agricultural Sciences,Yuncheng,Shanxi,044000,China)(2.CSIRO Division of Piant Industry,Canberra,ACT2601,Australia)(3.Biotechnolo
Hypocotyl segments from 5-6 day old seedlings of Gossypium hirsutum cv.Jin 7 were co-cultivated with Agrobacterium tumefaciens strain AGLI17-2 containing a binary vector pGA470(355 -GUS-nos3'/35S-tfdA -Nos3'). Transformed kanamycin-resistant calli were selected on MS medium containing 0.1mg/L 2,4-D, 0.1mg/L kinetin.50mg/L kanamycin and 500mg/L cefotaxime for 2 months,then numbered and tested for GUS activity after 3 months. Selected GUS positive calli were cultured on embryo induction medium (MS+1.9g/L KNO_3) until globular embryos developed and germinated. Plantlets were developed from these embryos over the next 2-3 months. Plantlets were analyzed for NPT Ⅱ and GUS activity.Eighty-seven percent of plantlets examined expressed the NPTⅡand GUS. Histochemical assays revealed strong GUS reactions in roots,stems and leaves of transgenic plants.Southern hybridization confirmed the presence of the tfdA gene in the genome of the transgenic plants.R1 cotton plants were sprayed with different concentrations (25-800ppm) of 2,4-D.All GUS positive plants proved to be resistant to 2,4-D.
【CateGory Index】： S562.034