以pmi为筛选标记的水稻转基因研究

Development of Rice GeneticTransformation Using pmi Gene as A Selectable Maker

[目的]建立一种快速、高效的甘露糖安全筛选体系。[方法]以绿色荧光蛋白(GFP)编码基因为报告基因,用农杆菌介导的方法将磷酸甘露糖异构酶基因(pmi)转入水稻品种皖粳97,并对转化和筛选条件进行优化。[结果]受体品种皖粳97,在32℃光照条件下诱导愈伤的出愈率可达90.3%,且生长较快;培养基中甘露糖筛选压力以5g/L蔗糖+15g/L甘露糖较为适宜,在该条件下筛选21d,进而分化成苗,转化频率可达20.6%。[结论]该研究确立的甘露糖筛选体系,能明显缩短筛选周期、提高转化效率,有利于转基因水稻的安全生产。

[ Objective ] The research aimed to establish a rapid, effective and safety mannose screening system. ~ Method ] The phospho- mannose isomerase gene （pmi） was transformed into Japonica rice variety Wanjing 97 with green fluorescent protein （GFP） gene as repo.rter gene. Further, the transformation and selection conditions were optimized. [ Result] The results showed that callus emerging rate could be up to 90.3% and the growth rate under continuous illumination at 32 ~C was much higher than the other conditions. The pressure of 5 g/L sucrose plus ＋ 15 g/L mannose was the best condition for selective medium. Under this optimal condition, the transgenic plants could be generated af- ter 21 d, and the transformation efficiency could up to 20.6%. [ Conclusion] The study developed a short-time consuming selection system with high transformation efficiency which facilitates to improve the safety of transgenic rice production.