小麦TaNADP-ME2基因重组植物表达载体构建及对水稻的遗传转化

The Construction of Plant Overexpression Vector of Wheat TaNADP-ME2 Gene and Genetic Transformation of Rice

小麦TaNADP-ME2是一光反应基因,并对干旱、盐、低温等非生物胁迫作出响应。构建TaNADP-ME2基因的重组植物表达载体,并转化水稻获得转基因植株。利用重组技术构建植物表达载体,冻融法转化农杆菌,农杆菌介导方法转化水稻成熟胚诱导的愈伤组织,潮霉素筛选和PCR鉴定转基因植株。成功构建了重组植物表达质粒pSUE2,并将pSUE2转入农杆菌EHA105,潮霉素磷酸转移酶(HPT)基因和TaNADP-ME2基因PCR鉴定获得16棵转基因阳性水稻植株。为进一步确定TaNADP-ME2基因的水分利用效率功能奠定了基础。

The wheat TaNADP-ME2 was not only responsive to light but also to drought,salt and low temperature.The recombinant plant overexpression vector was constructed,transformed into rice and acquired transgenic rice plant.The vector was constructed by recombinant technology.The agrobacterium was transformed by freeze-thaw method.The callus induced from the mature embryo of rice was transformed by agrobacterium-mediated method.The transgenic rice was identified by hygromycin and PCR method.The results revealed that recombinant plant overexpression vector pSUE2 has been constucted succeedingly and transformed into EHA105.The 16 transgenic rice positive plants were acquired by the PCR detection from HPT and TaNADP-ME2 gene.This research laid a foundation for identification the function of TaNADP-ME2 gene in enhancing water use efficiency in drought tolerance.