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褐飞虱(半翅目:稻虱科)Dicer-2基因的分子特征及基因功能分析

Molecular characterization and gene functional analysis of Dicer-2 gene from Nilaparvata lugens (Hemiptera: Geometroidea).

作者信息

Zhang Xiao-Yi, Lu Kai, Zhou Jia-Liang, Zhou Qiang

机构信息

State Key Laboratory of Biological Control and Institute of Entomology, Sun Yat-sen University, Guangzhou, China.

出版信息

Insect Sci. 2013 Feb;20(1):61-8. doi: 10.1111/j.1744-7917.2012.01539.x. Epub 2012 Jul 31.

Abstract

Nilaparvata lugens (Stål) (Hemiptera: Geometroidea), a serious rice pest in many countries of Asia, causes a great loss in rice production every year. RNA interference (RNAi) is a powerful technology for gene function study in insects and a potential tool for pest control. As a core component of RNAi pathway, Dicer-2 (Dcr-2) protein determines the production of small interfering RNA (siRNA) and is crucial for the efficiency of RNAi. In this study, the full-length complementary DNA (cDNA) of N. lugens Dcr-2 (NlDcr-2) was first cloned and analyzed, and then the RNAi experiment was conducted to explore the function of NlDcr-2 gene. The complete Dcr-2 cDNA of N. lugens was 4 971 bp in length with an open reading frame (ORF) of 1,656 amino acids. Phylogenetic and protein domain analysis showed that the predicted NlDcr-2 protein was similar to Tribolium castaneum. In the RNAi experiment, the messenger RNA level of NlDcr-2 was significantly reduced by NlDcr-2 double-stranded RNA (dsRNA) (dsDcr-2). Fifty-five per cent decrease of NlDcr-2 was found after 4 days of unremitting feeding. No significant effect was observed on the development of N. lugens after dsRNA ingestion.

摘要

褐飞虱(Nilaparvata lugens (Stål),半翅目:稻虱科)是亚洲许多国家水稻的一种严重害虫,每年都会给水稻生产造成巨大损失。RNA干扰(RNAi)是昆虫基因功能研究的一项强大技术,也是一种潜在的害虫防治工具。作为RNAi途径的核心组成部分,Dicer-2(Dcr-2)蛋白决定小干扰RNA(siRNA)的产生,对RNAi效率至关重要。在本研究中,首次克隆并分析了褐飞虱Dcr-2(NlDcr-2)的全长互补DNA(cDNA),然后进行RNAi实验以探究NlDcr-2基因的功能。褐飞虱完整的Dcr-2 cDNA长度为4971 bp,开放阅读框(ORF)为1656个氨基酸。系统发育和蛋白结构域分析表明,预测的NlDcr-2蛋白与赤拟谷盗的相似。在RNAi实验中,NlDcr-2双链RNA(dsRNA)(dsDcr-2)显著降低了NlDcr-2的信使RNA水平。持续喂食4天后,发现NlDcr-2降低了55%。摄入dsRNA后,未观察到对褐飞虱发育有显著影响。

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