以pmf为筛选标记的水稻转基因研究

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32oC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5∥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。

Development of Rice GeneticTransformation Using pmi Gene as A Selectable Maker

Objective] The research aimed to establish a rapid,effective and safety mannose screening system.Method] The phosphomannose isomerase gene(pmi) was transformed into Japonica rice variety Wanjing 97 with green fluorescent protein(GFP) gene as reporter 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 ℃ 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 after 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.