A de novo transcriptome of European pollen beetle populations and its analysis, with special reference to insecticide action and resistance
A de novo transcriptome of European pollen beetle populations and its analysis, with special reference to insecticide action and resistance
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DOI：
10.1111/imb.12099
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发表时间：
2014-08-01
影响因子：
2.6
通讯作者：
Nauen, R.
中科院分区：
文献类型：
--
作者：
Zimmer, C. T.;Maiwald, F.;Nauen, R.
花粉甲虫Meligethes aeneus是欧洲油菜种植中最重要的鞘翅目害虫，每年侵扰数百万公顷，如果不保持在经济损失阈值以下，则造成重大产量损失。该物种主要用杀虫剂控制，但最近对拟除虫菊酯类产生了高度抗性。本研究的目的是提供一个转录组资源，以调查耐药机制。在Roche（Indianapolis，IN，USA）和Illumina（LGC Genomics，柏林，德国）平台上对cDNA进行测序，得到总共相似的53 m读段，其组装成43 396个表达序列标签（EST）。人工注释揭示了编码杀虫剂靶位点和解毒酶的基因的良好覆盖。共鉴定出77个非冗余细胞色素P450基因。Illumina RNAseq序列（来自敏感和拟除虫菊酯抗性菌株）针对参考转录组的定位鉴定了在拟除虫菊酯抗性菌株中高度过表达的细胞色素P450（CYP6BQ23）。单核苷酸多态性分析证实了一个耐药菌株的电压门控钠通道中存在靶位点耐药突变（L1014 F）。我们的研究结果提供了新的见解与拟除虫菊酯抗性的重要基因在M。埃涅阿斯。为进一步研究杀虫剂的作用方式和花粉甲虫的抗性机制提供了一个完整的EST资源。
The pollen beetle Meligethes aeneus is the most important coleopteran pest in European oilseed rape cultivation, annually infesting millions of hectares and responsible for substantial yield losses if not kept under economic damage thresholds. This species is primarily controlled with insecticides but has recently developed high levels of resistance to the pyrethroid class. The aim of the present study was to provide a transcriptomic resource to investigate mechanisms of resistance. cDNA was sequenced on both Roche (Indianapolis, IN, USA) and Illumina (LGC Genomics, Berlin, Germany) platforms, resulting in a total of similar to 53 m reads which assembled into 43 396 expressed sequence tags (ESTs). Manual annotation revealed good coverage of genes encoding insecticide target sites and detoxification enzymes. A total of 77 nonredundant cytochrome P450 genes were identified. Mapping of Illumina RNAseq sequences (from susceptible and pyrethroid-resistant strains) against the reference transcriptome identified a cytochrome P450 (CYP6BQ23) as highly overexpressed in pyrethroid resistance strains. Single-nucleotide polymorphism analysis confirmed the presence of a target-site resistance mutation (L1014F) in the voltage-gated sodium channel of one resistant strain. Our results provide new insights into the important genes associated with pyrethroid resistance in M. aeneus. Furthermore, a comprehensive EST resource is provided for future studies on insecticide modes of action and resistance mechanisms in pollen beetle.