To improve the Agrobacterium-mediated transformation frequency of Japonica rice Wanjing 97, an orthogonal experiment L9 (33 ) was designed to optimize the factors regulating rice genetic transformation, and the impacts of innoculum OD600 , coculture temperature and coculture duration on transformation using gfp gene to monitor the expression of target gene were analyzed. Results showed that the optimal conditions for Agrobacterium-mediated transformation of Wanjing 97 were determined to be innoculum OD660 of 0. 1, cocultured at 21℃ -23 t dark for 72 h. Under this condition, the rate of gfp-positive calli was up to 70. 9% . The results suggested that gfp could be used as an effective reporter gene to improve rice genetic transformation.%为提高农杆菌介导的水稻遗传转化效率,以晚粳97为转化材料,绿色荧光蛋白gfp基因为报告基因,采用正交试验L9(33)对影响农杆菌介导水稻的遗传转化因子进行优化.通过观察愈伤组织荧光表达情况,分析菌液浓度、共培养温度与共培养时间对农杆菌转化水稻的影响.结果表明,在OD660值为0.1、共培养21℃～23℃黑暗条件下,农杆菌与水稻愈伤共培养72 h,最有利于水稻的遗传转化,该条件下晚粳97 愈伤组织荧光表达率达到70.9%.
展开▼
