抗水稻黑条矮缩病转基因材料的鉴定分析  

Identification and analysis of transgenic lines against rice black-streaked dwarf virus

作　　者：尹鲜思 王少岭[1] 崔益平[1] 陈秋红[1] 王国梁[1,2] 王志龙[1] Yin Xiansi;Wang Shaoling;Cui Yiping;Chen Qiuhong;Wang Guoliang;Wang Zhilong(Hunan Provincial Key Laboratory of Crop Germplasm Innovation and Utilization, Changsha 410128, China;College of Agronomy, Hunan Agricultural University, Changsha 410128, China; 3.Department of Plant Pathology, The Ohio State University, Columbus, Ohio 43210, USA)

机构地区：[1]湖南农业大学农学院,湖南长沙410128 [2]美国俄亥俄州立大学植物病理系,俄亥俄哥伦布43210

出　　处：《湖南农业大学学报（自然科学版）》2016年第4期380-385,共6页Journal of Hunan Agricultural University(Natural Sciences)

基　　金：转基因生物新品种培育重大专项(2012ZX08009001);教育部创新团队发展计划项目(IRT1239);湖南农业大学大学生创新性实验计划项目(XCX15145)

摘　　要：针对水稻黑条矮缩病毒(RBSDV)的核心粒子外壳蛋白基因(S8)、毒质与非结构蛋白基因(S9)及其嵌合基因(S8&S9)构建了6个RNAi载体,经农杆菌介导转化武陵粳1号获得转基因植株。通过潮霉素溶液浸泡法对转基因植株进行初步筛选,剔除假阳性植株;再通过PCR从DNA水平进行转基因阳性鉴定;进一步通过q RT–PCR从RNA水平进行转基因表达量分析。选取q RT–PCR表达量高的单拷贝纯系S8后代进行灰飞虱传毒试验,结果表明,含有S8 RNAi片段的转基因水稻材料对RBSDV具有一定抗性。Genome segments S8 (encoding minor core capsid protein), S9 (encoding virplasm and non-structual protein)and their chimeric sequences S8&S9 of RBSDV were used to construct six RNAi vectors and transferred into rice cultivarWulingjing 1 to generate transgenic plants by Agrobacterium-mediated transformation. Transgenic plants were firstscreened by immersion with hygromycin solution to eliminate the false-positive plants, then identified by PCRgenotyping analysis from the DNA level,followed by qRT–PCR analysis of candidate genes at the RNA level.Single-copy transgenic lines with high expression of S8 detected by qRT–PCR were picked up for Laodelphaxstriatellus-mediated virus transmission test, and the results showed that transgenic lines containing S8 RNAi vectordisplayed resistance to RBSDV.

关 键 词：水稻 水稻黑条矮缩病 转基因株系 RNA 干扰 荧光定量PCR 

分 类 号：Q786[生物学—分子生物学]