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转录组挖掘:用于测试海虱Caligus rogercresseyi对杀虱药物反应的多基因检测板。

Transcriptome mining: Multigene panel to test delousing drug response in the sea louse Caligus rogercresseyi.

作者信息

Valenzuela-Muñoz V, Gallardo-Escárate C

机构信息

Laboratory of Biotechnology and Aquatic Genomics, Interdisciplinary Center for Aquaculture Research (INCAR), Department of Oceanography, University of Concepción, P.O. Box 160-C, Chile.

Laboratory of Biotechnology and Aquatic Genomics, Interdisciplinary Center for Aquaculture Research (INCAR), Department of Oceanography, University of Concepción, P.O. Box 160-C, Chile.

出版信息

Mar Genomics. 2016 Feb;25:103-113. doi: 10.1016/j.margen.2015.12.005. Epub 2015 Dec 23.

Abstract

Controlling infestations of copepodid ectoparasites in the salmon industry is increasingly problematic given higher instances of drug resistance or loss of sensitivity. Despite the importance of this issue, the molecular mechanisms and genes implicated in resistance/susceptibility are only scarcely understood. The objective of the present study was to identify and evaluate the expression levels of candidate genes associated with delousing drug response in the sea louse Caligus rogercresseyi. From RNA-seq data obtained for adult male and female sea lice, 62.48 M reads were assembled in 70,349 high-quality contigs. BLASTX analysis against UniprotKB/Swiss-Prot and the ESTs available for crustaceans in the NCBI database identified 870 transcripts previously related to genes associated with delousing drug response. Furthermore, 14 candidate genes were validated through RT-qPCR and were evaluated with deltamethrin and azamethiphos bioassays. The results evidenced an overregulation of genes involved in ion transport in salmon lice treated with deltamethrin, while those treated with azamethiphos evidenced an overregulation of genes such as cytochrome P450, Carboxylesterase, and acetylcholine receptors. The present study provides a multigene panel to test delousing drug response to pyrethroids and organophosphates in a highly prevalent pathogen of the Chilean salmon industry.

摘要

鉴于药物抗性或敏感性丧失的情况增多,控制鲑鱼养殖业中桡足幼体体外寄生虫的侵扰问题日益严重。尽管这个问题很重要,但与抗性/易感性相关的分子机制和基因却鲜为人知。本研究的目的是鉴定和评估与海虱Caligus rogercresseyi中除虱药物反应相关的候选基因的表达水平。从成年雄性和雌性海虱获得的RNA-seq数据中,6248万个读段被组装成70349个高质量的重叠群。针对UniprotKB/Swiss-Prot以及NCBI数据库中可获得的甲壳类动物的ESTs进行的BLASTX分析,鉴定出870个先前与除虱药物反应相关基因有关的转录本。此外,通过RT-qPCR验证了14个候选基因,并用溴氰菊酯和谷硫磷生物测定法对其进行了评估。结果表明,用溴氰菊酯处理的鲑鱼虱中参与离子转运的基因上调,而用谷硫磷处理的鲑鱼虱中细胞色素P450、羧酸酯酶和乙酰胆碱受体等基因上调。本研究提供了一个多基因组合,用于测试智利鲑鱼养殖业中一种高度流行的病原体对拟除虫菊酯和有机磷的除虱药物反应。

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