Improvement of Agrobacterium-mediated genetic transformation of rice using gfp as a reporter gene

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℃ 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.