玉米转ABP9和ZmCBF3双基因植株的获得

目的]获得转双基因的植株,研究植物逆境诱导基因ABP9和ZmCBF3的协同表达对玉米抗逆性的影响.方法]通过农杆菌侵染的方法将ABP9和ZmCBF3基因同时转入玉米中,并对T0代植株进行分子鉴定.使用含有ABP9启动子驱动ABP9和ZmCBF3双基因载体的农杆菌侵染玉米幼穗,获得T0代种子;在田间通过对玉米苗喷洒草铵膦除草剂筛选获得具有草铵膦抗性的玉米植株.结果]挑选10株抗性玉米苗小量提取DNA并进行PCR扩增分析,其中5个植株出现特异条带;大量提取PCR阳性玉米的DNA进行Southem杂交分析,9TQ-5号植株在与bar探针和ABP9探针杂交时均出现特异条带.结论]9TQ-5号植株为转基因阳性植株,为后续试验提供了材料.

Generation of Maize Plants Transformed with ABP9 and ZmCBF3 Genes

Objective] In order to study the coordinated expression of plant stress induced gene ABP9 and ZmCBF3 effect on the resistance of maize,transgenic plants are desired.Method] In this study,ABP9 and ZmCBF3 genes were introduced into maize through Agrobacterium tumefaciens-mediated transformation,and transgenic plants at T0 generation were identified at molecular levels.We used the Agrobacterium stain containing an expression vector in which ABP9 and ZmCBF3 were driven by ABP9 promoter to infect the young ear of maize,and harvested T0 generation seeds.T0 plants tolerant to glufosinate were obtained by leaf-spray in the field.Result] 10 glufosinate tolerant plants were selected and 5 of them were identified positive by PCR.Southern hybridization with bar and ABP9 probes showed that 9TQ-5 had specific bands.Conclusion] These results identified 9TQ-5 as the transgenic plant,which will be used in further research.