啤酒花抗病毒双价RNAi植物表达载体的构建及遗传转化

摘要

To construct RNAi vector both resistant to Hop latent virus (HpLV) and Hop latent viroid (HpLVd),the pathogenic region of 202 bp in HpLV and the conserved sequence of 155 bp in HpLVd were selected as interference sequence through sequence alignment.The bivalent RNAi vector against HpLV and HpLVd were constructed based on interference vector pUCCRNAi,then its functional region was connected to plant expression vector pCAMBIA2300-35S-OCS through specific restriction sites.And the objective fragments were transformed into hops through Agrobacteriurn-mediated method.PCR identification showed that RNAi expression vector had been successfully transformed into hops.The construction of RNAi expression vector and Agrobacterium-mediated transformation in this study provided references to induce posttranscriptional gene silencing in transformed plants and obtain new multiple-virus-resistant plant materials.%为了构建对啤酒花潜隐病毒(HpLV)和啤酒花潜隐类病毒(HpLVd)有抗性的RNAi载体,本研究通过序列比对选取HpLV外壳蛋白基因致病区202 bp和HpLVd 155 bp的保守序列作为干扰序列,以干扰载体pUCCRNAi为基础,成功构建出针对HpLV和HpLVd的双价RNAi载体,通过特异性酶切位点将构建成功的RNAi载体的功能区域连接在植物表达载体pCAMBIA2300-35S-OCS上,并利用农杆菌介导法将目的基因导入啤酒花中.经PCR鉴定证明RNAi载体已成功转入啤酒花中.本研究基于RNAi技术的植物表达载体构建和农杆菌介导的遗传转化,为诱导转基因植物的转录后基因沉默、获得抗多种病毒病的植物新材料提供参考.