影响农杆菌介导的柑桔基因转化因素研究

The factors affectingAgrobacterium-mediated gene-transformation of epicotyls ofCitrus in vitroare studied in this paper. TransgenicPoncitrus trifoliatawith CTV-cp gene were obtained. The main results are as follows: As a selectable agent, concentration of Kanamycin is 20~30μg/mL forCitrusexcept P.trifoliata, which is 50μg/mL. The optimal cocultivation time of explants &Agrobacteriumis 3 days. Gus transient expression frequencywas promoted, when the concentration of acetosyingone is 70~100μmol/L. Gustransient expression frequency is higher in epicotyls from 20d seedling. Underthe same transformation condition , the rate of explantwith resistant shoot and the rate of explantwithGus positive shoot are different in differentCitrusspecies. Extral gene is proved to be intoP. trifoliataby southern blot examination with probe of plasmind pGA482GG cut by Nco IE.