褐飞虱安全高效RNAi靶基因的筛选及Snf7同源基因的控害效果  

Screening of safe and efficient RNAi target genes and the control effects of a Snf7 homologous gene in the brown planthopper

作　　者：黎丹 毛婕 李灿 张文庆[1] LI Dan;MAO Jie;LI Can;ZHANG Wenqing(State Key Laboratory of Biocontrol/School of Life Sciences,Sun Yat-sen University,Guangzhou 510275,China;Guizhou Provincial Key Laboratory of Rare Mountain Animals and Economic Insects/School of Biology and Environmental Engineering,Guiyang University,Guiyang 550005,China)

机构地区：[1]有害生物控制与资源利用国家重点实验室/中山大学生命科学学院,广东广州510275 [2]贵州省山地珍稀动物与经济昆虫重点实验室/贵阳学院生物与环境工程学院,贵州贵阳550005

出　　处：《中山大学学报（自然科学版）》2020年第6期1-11,共11页Acta Scientiarum Naturalium Universitatis Sunyatseni

基　　金：国家重点研发计划项目(2017YFD0200900)。

摘　　要：褐飞虱(Nilaparvata lugens)是一种具有高繁殖力、强适应性和迁飞性的重要水稻害虫。为更有效地获得安全高效的褐飞虱RNAi靶标基因,建立了一套安全高效RNAi靶基因的筛选流程,获得了32个安全高效致死候选基因,并检测了其中12个基因的RNAi效率。进一步对其中的转运必需内吞体分选复合物Ⅲ亚基Snf7同源基因(简称为NlSnf7)的控害效果进行了研究。NlSnf7基因在褐飞虱各个时期和组织均有表达。通过注射法和饲喂法RNAi技术对NlSnf7进行低剂量干扰,结果表明干扰后NlSnf7的mRNA表达水平显著降低,褐飞虱的校正死亡率在每头注射50 ng后第13天可达70%,饲喂50 ng/μL后第9天可达48%;注射dsNlSnf7后第5天NlSnf7的蛋白水平也显著降低。此外,选择无连续21 bp相同序列的亚洲玉米螟和白背飞虱进行dsNlSnf7注射,发现与对照组相比,实验组均无显著变化。这些结果表明dsNlSnf7片段是一个高效安全的RNAi靶标分子。The brown planthopper(BPH,Nilaparvata lugens)is an important rice pest with high fecundity,strong adaptability and migratory ability.In order to obtain safe and efficient RNAi target genes,we established a set of process for RNAi target gene screening,and 32 candidate target genes were obtained,of which 12 were tested for RNAi efficiency.Furthermore,we investigated the control effects of a endosomal sorting complex which was required for transport Ⅲ subunit Snf7(NlSnf7).NlSnf7 gene was expressed in various stages and tissues of BPH.After RNA interference of NlSnf7,its mRNA expression level was significantly reduced,and the corrected mortality rate of BPH reached 70%on the 13th day after injection of dsNlSnf7 at 50 ng/head,and 48%on the 9th day after feeding of dsNlSnf7 at 50 ng/μL.NlSnf7 protein was also significantly reduced on the 5th day after injection of dsNlSnf7.In addition,when dsNlSnf7 was injected into Ostrinia furnacalis and Sogatella furcifera in which no 21 bp continuously identical sequence was found,there were no significant changes between the experimental group and the control group.These results indicated that dsNlSnf7 fragment is an efficient and safe RNAi target molecule.

关 键 词：褐飞虱(Nilaparvata lugens) RNAi靶基因 转运必需内吞体分选复合物Ⅲ亚基Snf7 

分 类 号：S43[农业科学—农业昆虫与害虫防治]