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褐飞虱唾液蛋白质组的筛选与功能分析

Screening and Functional Analyses of Nilaparvata lugens Salivary Proteome.

作者信息

Huang Hai-Jian, Liu Cheng-Wen, Huang Xiao-Hui, Zhou Xiang, Zhuo Ji-Chong, Zhang Chuan-Xi, Bao Yan-Yuan

机构信息

State Key Laboratory of Rice Biology and Ministry of Agriculture Key Laboratory of Agricultural Entomology, Institute of Insect Sciences, Zhejiang University , Hangzhou 310058, China.

出版信息

J Proteome Res. 2016 Jun 3;15(6):1883-96. doi: 10.1021/acs.jproteome.6b00086. Epub 2016 May 12.

Abstract

Most phloem-feeding insects secrete gelling and watery saliva during the feeding process. However, the functions of salivary proteins are poorly understood. In this study, our purpose was to reveal the components and functions of saliva in a rice sap-sucking insect pest, Nilaparvata lugens. The accomplishment of the whole genome and transcriptome sequencing in N. lugens would be helpful for elucidating the gene information and expression specificity of the salivary proteins. In this study, we have, for the first time, identified the abundant protein components from gelling and watery saliva in a monophagous sap-sucking insect species through shotgun proteomic detection combined with the genomic and transcriptomic analysis. Eight unknown secreted proteins were limited to N. lugens, indicating species-specific saliva components. A group of annexin-like proteins first identified in the secreted saliva displayed different domain structure and expression specificity with typical insect annexins. Nineteen genes encoding five annexin-like proteins, six salivaps (salivary glands-specific proteins with unknown function), seven putative enzymes, and a mucin-like protein showed salivary gland-specific expression pattern, suggesting their importance in the physiological mechanisms of salivary gland and saliva in this insect species. RNA interference revealed that salivap-3 is a key protein factor in forming the salivary sheath, while annexin-like5 and carbonic anhydrase are indispensable for N. lugens survival. These novel findings will greatly help to clarify the detailed functions of salivary proteins in the physiological process of N. lugens and elucidate the interaction mechanisms between N. lugens and the rice plant, which could provide important targets for the future management of rice pests.

摘要

大多数取食韧皮部的昆虫在取食过程中会分泌凝胶状和水样唾液。然而,唾液蛋白的功能却鲜为人知。在本研究中,我们的目的是揭示吸食水稻汁液的害虫褐飞虱唾液的成分和功能。褐飞虱全基因组和转录组测序的完成将有助于阐明唾液蛋白的基因信息和表达特异性。在本研究中,我们首次通过鸟枪法蛋白质组检测结合基因组和转录组分析,从一种单食性吸食汁液昆虫的凝胶状和水样唾液中鉴定出丰富的蛋白质成分。8种未知分泌蛋白仅存在于褐飞虱中,表明其唾液成分具有物种特异性。首次在分泌唾液中鉴定出的一组类膜联蛋白与典型昆虫膜联蛋白具有不同的结构域结构和表达特异性。19个基因编码5种类膜联蛋白、6种唾液腺特异性蛋白(功能未知的唾液腺特异性蛋白)、7种假定酶和1种粘蛋白样蛋白,呈现唾液腺特异性表达模式,表明它们在该昆虫唾液腺和唾液生理机制中具有重要作用。RNA干扰显示,唾液腺特异性蛋白-3是形成唾液鞘的关键蛋白因子,而类膜联蛋白5和碳酸酐酶对褐飞虱的生存不可或缺。这些新发现将极大地有助于阐明唾液蛋白在褐飞虱生理过程中的详细功能,并阐明褐飞虱与水稻植株之间的相互作用机制,这可为未来水稻害虫的防治提供重要靶点。

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