RNAi-mediated resistance to rice black-streaked dwarf virus in transgenic rice
RNAi-mediated resistance to rice black-streaked dwarf virus in transgenic rice
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RNAi介导的转基因水稻对水稻黑条矮缩病毒的抗性
DOI：
10.1007/s11248-016-9999-4
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发表时间：
2017-04-01
影响因子：
3
通讯作者：
Yu, Hengxiu
中科院分区：
文献类型：
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作者：
Ahmed, Mohamed M. S.;Bian, Shiquan;Yu, Hengxiu
水稻黑条矮缩病毒（Rice black-streaked dwarf virus，RBSDV）是呼肠孤病毒科（Reoviridae）斐济病毒属（Fijivirus）的成员，在我国和亚洲许多国家的水稻生产中造成严重的经济损失。利用常规育种方法培育抗RBSDV的品种受到限制，目前尚未发现高水平抗RBSDV的种质资源，而利用RNA干扰（RNAi）技术培育抗RBSDV的品种是最有前途的方法之一。RBSDV非结构蛋白P7-2由S7 - 2基因编码，是一种潜在的F-box蛋白，通过泛素化途径参与植物与病毒的互作。P8是由S8基因编码的具有转录抑制活性的次要核心蛋白。本研究利用含有RBSDV基因S7 - 2或S8的RNAi构建体的微型双T-DNA载体转化水稻愈伤组织，获得了含有目的基因构建体和选择标记基因潮霉素磷酸转移酶（HPT）的植株。从这些转基因植株的后代中，我们获得了无选择标记（HPT基因）的植株。携带S7-2-RNAi或S8-RNAi的T5纯合转基因株系在田间感染的情况下对RBSDV表现出高水平的抗性。因此，我们的研究结果表明，RNA干扰S7 - 2或S8基因的表达可能是诱导水稻对RBSD病高水平抗性的有效途径。
Rice black-streaked dwarf virus (RBSDV), a member of the genusFijivirusin the familyReoviridae, causes significant economic losses in rice production in China and many other Asian countries. Development of resistant varieties by using conventional breeding methods is limited, as germplasm with high level of resistance to RBSDV have not yet been found. One of the most promising methods to confer resistance against RBSDV is the use of RNA interference (RNAi) technology. RBSDV non-structural protein P7-2, encoded byS7-2gene, is a potential F-box protein and involved in the plant-virus interaction through the ubiquitination pathway. P8, encoded byS8gene, is the minor core protein that possesses potent active transcriptional repression activity. In this study, we transformed rice calli using a mini-twin T-DNA vector harboring RNAi constructs of the RBSDV genesS7-2orS8, and obtained plants harboring the target gene constructs and the selectable marker gene,hygromycin phosphotransferase(HPT). From the offspring of these transgenic plants, we obtained selectable marker (HPTgene)-free plants. Homozygous T5transgenic lines which harbored either S7-2-RNAi or S8-RNAi exhibited high level resistance against RBSDV under field infection pressure from indigenous viruliferous small brown planthoppers. Thus, our results showed that RNA interference with the expression ofS7-2orS8genes seemed an effective way to induce high level resistance in rice against RBSD disease.