Genetic Transferring AtMYB44Gene Into Rice Mediated By Agrobacterium Tumefaciences

Rice is one of the most important crops. Plant transgenic technology is a useful method to breed stress-resistant rice. In recent years，plenty of genes conferring to stress resistantence have been isolated and widely applied. AtMYB44is a new effective stress-resistant gene from Arabidopsis thaliana. Based on optimizing the transgenic rice technical system, we transformed AtMYB44gene into Japonica rice Wanjing97through Agrobacterium tumefaciences. The main results were as follows:1. To improve the Agrobacterium-mediated transformation frequency of Japonica rice Wanjing97, we optimized the factors which may influence rice genetic transformation, and analyzed the impacts of innoculum OD660, coculture temperature and coculture duration on transformation using gfp gene by monitoring fluorescence of GFP. Our results showed that the optimal conditions for Agrobacterium-mediated transformation of Wanjing97were determined to be innoculum OD660of0.1, coculture at21℃~23℃dark for72h. Under this condition, the rate of gfp-positive calli was up to70.9%. The results suggest that gfp could be used as an effective reporter gene to improve rice genetic transformation.2. To get the stress-resistant rice, we transformed AtMYB44gene into Japonica rice Wanjing97through Agrobacterium tumefaciences and selection with proper concentration of basta (4mg/L), a total of680resistant calli were gained, the frequency of transformation is63.79%, and352bar-resistant plants which contain AtMYB44gene were obtained. PCR assay using AtMYB44-specific primers and bar-specific primers showed that160were positive. Southern blot analysis indicated that target gene has integrated into rice genome.