miRNA在水稻抗感黑条矮缩病品种中的表达差异和调控作用

项目名称： miRNA在水稻抗感黑条矮缩病品种中的表达差异和调控作用

项目编号： No.31201496

项目类型： 青年科学基金项目

立项/批准年度： 2013

项目学科： 植物保护学、园艺学与植物营养学

项目作者： 兰莹

作者单位： 江苏省农业科学院

项目金额： 23万元

中文摘要： 灰飞虱传播引起的水稻黑条矮缩病近年来在我国华东华北稻区爆发流行，利用植物自身抗性培育抗病品种是防治该病毒病的根本策略。RNA沉默机制是植物抵御病毒侵染的重要途径，miRNA是其中的关键组分。申请者利用课题组已建立的RBSDV人工接种体系结合田间鉴定筛选出具有高耐（抗）病水平且专一针对病毒抗性的水稻品种苏69，以高感品种淮稻5号为对照，分别构建两者RBSDV侵染前后的小RNA文库，拟通过高通量测序，获取抗感品种中差异表达的miRNA，并利用定量PCR, RNA杂交和RACE检测等技术，验证候选miRNA的表达水平及剪切功能；并利用RNA病毒载体构建miRNA瞬时表达系统，通过转化水稻株系对调控功能进行验证，以进一步检验miRNA在抗病性分化中的作用，深化对病毒致病及植物抗病机制的认识，为抗病品种的遗传改良提供理论依据和技术支持。

中文关键词： 水稻黑条矮缩病；水稻；miRNA；降解组；病毒小RNA

英文摘要： Rice black streaked dwarf virus is becoming one of the most serious problems in affecting rice production in East and North China. Using host defense resistance to cultivate resistant varieties is the most basic fundamental strategy. Plants can activate RNA silencing to against virus infection through regulating resistance genes and silencing of viral genes. miRNA is one of the key components in RNAi mechanism. We use artificial inoculation system of rice black-streaked dwarf virus and field identification to select anti-virus resistance rice variety Su69 and another high susceptible variety HuaiDao 5 as control. Through high-throughput sequencing, qRT-PCR and Northern Blot technologies, we built four small RNA libraries before and after RBSDV infection on susceptible rice varieties and resistant rice varieties respectively. RACE analysis is also used to identify the predicted target genes. To further identify the effects of miRNAs in the rice resistance pathways, we use RNA virus vectors to construct miRNA transient expression system in transformed rice lines.This study may provide a theoretical basis and technical support for the genetic improvement of disease-resistant varieties.

英文关键词： Rice black streaked dwarf virus；rice；miRNA；Degradome；Virus-derived small interfering RNAs