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以宝丰7228小麦幼苗叶片为材料,利用DDRT-PCR技术对正常与盐胁迫条件下小麦叶片基因表达的差异进行分析后检测到27条差异cDNA片段,mRNA点杂交分析结果显示SR07片段存在明显的胁迫诱导表达特征。对SR07进行了序列测定和同源性对比分析后发现,SR07片段与植物水孔蛋白中质膜内在蛋白(PIPs)有87%的序列同源性。将SR07克隆到植物转化载体pCAMBIA中CaMV35S启动子下游,构建表达载体pCAMBIA-SR07,利用根癌农杆菌(Agrobacterium tumefaciens)质粒介导的遗传转化系统转化烟草(Nicotiana tabacum),经筛选后获得3个烟草转化系。转化系的NaCl、PEG和甘露醇抗性实验结果表明,SR07转化株的抗盐性与对照株相比没有显著性差异(P>0.05),而PEG和甘露醇抗性与对照株相比有显著性差异(P<0.05),推测SR07表达蛋白可能在植物水分调节方面有重要作用。
In this study, 27 different cDNA fragments were detected by using DDRT-PCR to analyze the differences of gene expression in wheat leaves under normal and salt stress conditions. The results of mRNA dot blot analysis showed that the SR07 fragment was significantly Stress induced expression characteristics. After sequencing and homology comparison of SR07, it was found that there was 87% sequence homology between the SR07 fragment and the plasma membrane intrinsic protein (PIPs) of plant aquaporin. SR07 was cloned into the plant transformation vector pCAMBIA downstream of the CaMV35S promoter to construct the expression vector pCAMBIA-SR07. Nicotiana tabacum was transformed by Agrobacterium tumefaciens plasmid-mediated genetic transformation system, and after screening, 3 Tobacco transformation system. The results of NaCl, PEG and mannitol resistance experiments showed that the salt tolerance of SR07 transformants was not significantly different from that of the control (P> 0.05), but the resistance of PEG and mannitol was Significant difference (P <0.05), suggesting that SR07 protein expression may play an important role in plant water regulation.