中间偃麦草SGT1基因的克隆及其抗病功能的分析

Isolation and Preliminarily Functional Analysis of SGT1 Gene of Thinopyrum intermedium

为了探索中间偃麦草SGT1基因在小麦抗病反应中的应用潜力,采用RT-PCR和RACE方法克隆出中间偃麦草SGT1基因,命名为TiSGT1。该基因编码蛋白具有SGT1蛋白典型的功能域结构,与大麦SGT1序列高度同源。通过基因重组技术构建了TiSGT1的高效组成型表达载体,通过基因枪法将该载体转化到小麦品种扬麦12基因组中。对转基因植株PCR、Southern杂交与RT-PCR分析表明,TiSGT1基因可在T0、T1和T2代转基因小麦中遗传和表达。黄矮病、白粉病抗性鉴定结果表明,SGT1的超量表达可以提高小麦对黄矮病、白粉病的抗性,TiSGT1可以作为小麦广谱抗病性改良的潜在基因资源。

To understand the role of SGT1 gene in wheat（Triticum aestivum L.）diseases resistance,we used RT-PCR and RACE methods to isolate SGT1 genes from Thinopyrum intermedium.The SGT1 was named TiSGT1.The deduced TiSGT1 protein possesses the typical domains of known SGT1 proteins and shows highly homology with barley SGT1 protein.A highly efficiency expression vector of TiSGT1 was constructed by genetic recombination,and transformed into the wheat cultivar ＇Yangmai 12＇ by biolistic particle method.Transformation plants were analyzed by PCR,Southern hybridization,and RT-PCR methods.The results showed that the TiSGT1 gene could be inherited from T0 to T2 generations,and could be expressed.The results of disease resistance test indicated that the over-expression of SGT1 in the transgenic wheat enhanced resistance to barley yellow dwarf virus and powdery mildew,and TiSGT1 could be used as a potential gene resource for improving broad spectrum resistance of wheat.