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RNA干扰去除“万圣节”基因“无躯”表明其在防治褐飞虱和灰飞虱方面的潜在应用。

RNA interference depletion of the Halloween gene disembodied implies its potential application for management of planthopper Sogatella furcifera and Laodelphax striatellus.

作者信息

Wan Pin-Jun, Jia Shuang, Li Na, Fan Jin-Mei, Li Guo-Qing

机构信息

Education Ministry Key Laboratory of Integrated Management of Crop Diseases and Pests, College of Plant Protection, Nanjing Agricultural University, Nanjing, China.

出版信息

PLoS One. 2014 Jan 28;9(1):e86675. doi: 10.1371/journal.pone.0086675. eCollection 2014.

Abstract

Sogatella furcifera and Laodelphax striatellus are economically important rice pests in China by acting as vectors of several rice viruses, sucking the phloem sap and blocking the phloem vessels. Ecdysteroid hormone 20-hydroxyecdysone regulates insect development and reproduction. A cytochrome P450 monooxygenase CYP302A1 (22-hydroxylase), encoded by the Halloween gene disembodied (dib), plays a critical role in ecdysteroidogenesis. The objective of this study is to test whether dib genes are potential targets for RNA interference-based management of S. furcifera and L. striatellus. We cloned and characterized Sfdib and Lsdib. The open reading frame regions of dib genes were generated and used for designing and constructing dsRNA fragments. Experiments were conducted using oral delivery of dsdib to investigate the effectiveness of RNAi in S. furcifera and L. striatellus nymphs. Real-time quantitative reverse transcriptase-PCR analysis demonstrated that continuous ingestion of dsdib at the concentration of 0.01, 0.05 and 0.50 mg/ml diminished Sfdib expression levels by 35.9%, 45.1% and 66.2%, and ecdysone receptor (SfEcR) gene mRNA levels by 34.0%, 36.2% and 58.5% respectively in S. furcifera, and decreased Lsdib expression level by 18.8%, 35.8% and 56.7%, and LsEcR mRNA levels by 25.2%, 46.8% and 68.8% respectively in L. striatellus. The reduction in dib and EcR transcript abundance resulted in observable phenotypes. The development of nymphs was impaired and the survival was negatively affected. Our data will enable the development of new insect control strategies and functional analysis of vital genes in S. furcifera and L. striatellus nymphs.

摘要

白背飞虱和灰飞虱是中国重要的水稻害虫,它们作为多种水稻病毒的传播媒介,吸食韧皮部汁液并堵塞韧皮部导管。蜕皮甾体激素20-羟基蜕皮酮调节昆虫的发育和繁殖。由万圣节基因脱壳(dib)编码的细胞色素P450单加氧酶CYP302A1(22-羟化酶)在蜕皮甾体生成中起关键作用。本研究的目的是测试dib基因是否是基于RNA干扰防治白背飞虱和灰飞虱的潜在靶点。我们克隆并鉴定了Sfdib和Lsdib。生成了dib基因的开放阅读框区域,并用于设计和构建dsRNA片段。通过口服dsdib进行实验,以研究RNAi对白背飞虱和灰飞虱若虫的有效性。实时定量逆转录PCR分析表明,连续摄入浓度为0.01、0.05和0.50mg/ml的dsdib,分别使白背飞虱中Sfdib表达水平降低35.9%、45.1%和66.2%,蜕皮激素受体(SfEcR)基因mRNA水平降低34.0%、36.2%和58.5%;使灰飞虱中Lsdib表达水平降低18.8%、35.8%和56.7%,LsEcR mRNA水平降低25.2%、46.8%和68.8%。dib和EcR转录本丰度的降低导致了可观察到的表型。若虫的发育受到损害,存活率受到负面影响。我们的数据将有助于开发新的昆虫控制策略,并对白背飞虱和灰飞虱若虫中的关键基因进行功能分析。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e4ff/3904942/4fcd94866ddd/pone.0086675.g001.jpg

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