根癌农杆菌介导的柑桔遗传转化技术

The embryoids of Jincheng orange and Xinhui orange ( Citrus sinensis Osbeck) were induced from nucellar embryo, and the mature shoots were obtained from 20-year-old plants of Beibei447 orange and Navel orange . After they were inoculated with agrobacterium strain LBA4404 harboring vector that contained Shiva A and Cecropin B gene, the effects of various media, concentrations of kanamycin, places of shearing and parafilm on the shoot induction rate and transformation frequency were studied. PCR (polymerase chain reaction) and Southern blot analysis verified that the Shiva A and Cecropin B gene were transferred into the genome of citrus. Transgenic plants were grafted into 6-month- old Trifoliate orange in greenhouse.