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利用RNA干扰在烟粉虱(Bemisia tabaci (Gennadius))中实现组织特异性基因沉默

Tissue-specific gene silencing by RNA interference in the whitefly Bemisia tabaci (Gennadius).

作者信息

Ghanim Murad, Kontsedalov Svetlana, Czosnek Henryk

机构信息

Department of Entomology, Institute of Plant Protection, Agricultural Research Organization, The Volcani Center, P.O. Box 6, Bet Dagan 50250, Israel.

出版信息

Insect Biochem Mol Biol. 2007 Jul;37(7):732-8. doi: 10.1016/j.ibmb.2007.04.006. Epub 2007 Apr 25.

Abstract

The hemipteran whitefly Bemisia tabaci (Gennadius) species complex and the plant viruses they transmit pose major constraints to vegetable and fiber production, worldwide. The whitefly tissue- and developmental-specific gene expression has not been exhaustively studied despite its economic importance. In 2002, a functional genomic project was initiated, which generated several thousands expressed sequence tags (ESTs) and their sequence. This project provides the basic information to design experiments aimed at understanding and manipulating whitefly gene expression. In this communication, for the first time we provide evidence that the RNA interference mechanism discovered in many organisms, including in Hemiptera, is active in B. tabaci. By injecting into the body cavity long double-stranded RNA (dsRNA) molecules, specifically directed against genes uniquely expressed in the midgut and salivary glands, we were able to significantly inhibit the expression of the targeted mRNA in the different organs. Gene expression levels in RNAi-silenced whiteflies were reduced up to 70% compared to whiteflies injected with buffer or with a green fluorescent protein (GFP)-specific dsRNA. Phenotypic effects were observed in B. tabaci ovaries following dsRNA targeting the whitefly Drosophila chickadee homologue. Disruption of whitefly gene expression opens the door to new strategies aimed at curbing down the deleterious effects of this insect pest to agriculture.

摘要

半翅目粉虱烟粉虱(Bemisia tabaci,Gennadius)复合种及其传播的植物病毒对全球蔬菜和纤维生产构成了重大限制。尽管烟粉虱具有经济重要性,但其组织和发育特异性基因表达尚未得到详尽研究。2002年启动了一个功能基因组项目,该项目产生了数千个表达序列标签(EST)及其序列。该项目为设计旨在理解和操纵烟粉虱基因表达的实验提供了基础信息。在本通讯中,我们首次提供证据表明,在包括半翅目在内的许多生物中发现的RNA干扰机制在烟粉虱中是活跃的。通过向体腔注射特异性针对在中肠和唾液腺中独特表达的基因的长双链RNA(dsRNA)分子,我们能够显著抑制不同器官中靶向mRNA的表达。与注射缓冲液或绿色荧光蛋白(GFP)特异性dsRNA的粉虱相比,RNAi沉默粉虱中的基因表达水平降低了70%。在用靶向粉虱果蝇山雀同源物的dsRNA处理后,在烟粉虱卵巢中观察到了表型效应。粉虱基因表达的破坏为旨在抑制这种害虫对农业的有害影响的新策略打开了大门。

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