小麦WZY2基因RNA干扰表达载体的构建及遗传转化

Construction of RNAi expression vector of WZY2 gene from wheat and its genetic transformation

【目的】利用基因枪将WZY2基因RNA干扰表达载体pAHC-WZY2-Ri导入"郑引1号"小麦,获得WZY2基因表达缺失型小麦,为深入分析WZY2基因功能奠定基础。【方法】以植物表达载体pAHC25为基础,构建含有反向重复序列的RNA干扰表达载体pAHC-WZY2-Ri,利用基因枪将其转入"郑引1号"小麦幼胚愈伤组织中,经过筛选、PCR检测得到阳性植株。【结果】从转化的1 500个愈伤组织中获得27株再生植株。利用PCR对再生植株进行检测,获得阳性植株3株,转化率为0.2%。【结论】构建了WZY2基因的RNA干扰表达载体,成功地将WZY2基因RNA干扰表达载体导入"郑引1号"小麦,获得阳性植株。

【Objective】 The study aimed to obtain the transgenic wheat with the gene of WZY2 silenced by transforming WZY2 into the RNAi expression vector,and provide information for further investigation of WZY2.【Method】 Using pAHC25 as a basic vector,the RNAi expression vector of WZY2 named pAHC-WZY2-Ri was constructed and introduced into wheat variety ＂Zhengyin No.1＂ by biolistic particle.Then the transgenic plants were identified by PCR.【Result】 The 1 500 embryos of ＂Zhengyin No.1＂ were bombarded by biolistic particle,and 27 regenerated plants were obtained.Three of them were identified positive by PCR and the transformational frequency was 0.2%.【Conclusion】 In this study,the RNAi expression vector of WZY2 was constructed,and was successfully transformed into ＂Zhengyin No.1＂.