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甜菜夜蛾转录组分析:从头组装及与溴氰菊酯抗性相关基因的检测。

Transcriptome analysis of Mythimna separata: De novo assembly and detection of genes related to beta-cypermethrin resistance.

机构信息

Department of Biology, Taiyuan Normal University, Jinzhong, Shanxi, China.

Department of Agricultural Entomology, College of Plant Protection, Shanxi Agricultural University, Taigu, Shanxi, China.

出版信息

Arch Insect Biochem Physiol. 2022 Sep;111(1):e21921. doi: 10.1002/arch.21921. Epub 2022 May 30.

Abstract

The oriental armyworm Mythimna separata (Walker) is a devastating pest of cereal crops mainly in Asia and Oceania and recently become resistant to beta-cypermethrin (beta-CP). However, molecular biological studies of its response to beta-CP are scarce, and related genomic information is not available. In this study, we sequenced and de novo assembled the transcriptome of beta-CP susceptible M. separata (MsS-whole, abbr. MsS-W). A total of 30,486 unigenes were obtained, with an N50 length of 506 bp. A number of 21,051 unigenes were matched to public databases, of which 10,107 were classified into 59 gene ontology subcategories, 5792 were assigned into 25 clusters of orthologous groups of proteins subgroups and 12,123 were assigned to 257 Kyoto Encyclopedia of Genes and Genomes pathways. A total of 729 simple sequence repeats were detected. In addition, a total of 323 cytochrome P450-associated sequences from nine lepidopterous species, of which 130 were from M. separata, were analyzed using the maximum likelihood method and Bayesian inference. Among the 130 cytochrome P450-associated sequences from M. separata, 60 were dropped into CYP3 clan, which is associated with metabolizing xenobiotics and plant natural compounds. Furthermore, the beta-CP susceptible (MsS-2) and resistant (MsR-2) M. separata population transcriptomes were sequenced. Certain critical genes involved in beta-CP detoxification were detected and verified by quantitative real-time polymerase chain reaction. Collectively, our results provided a basis for further studies of the molecular mechanism of insecticide resistance in M. separata.

摘要

东方粘虫(Mythimna separata)是一种主要分布在亚洲和大洋洲的毁灭性粮食作物害虫,最近对β-氯氰菊酯(β-CP)产生了抗药性。然而,其对β-CP 反应的分子生物学研究很少,相关的基因组信息也不可用。在本研究中,我们对β-CP 敏感的东方粘虫(MsS-whole,简称 MsS-W)进行了测序和从头组装转录组。共获得 30486 条 unigenes,N50 长度为 506bp。有 21051 条 unigenes与公共数据库匹配,其中 10107 条被分类为 59 个基因本体论亚类,5792 条被分配到 25 个同源蛋白簇,12123 条被分配到 257 个京都基因与基因组百科全书途径。共检测到 729 个简单序列重复。此外,利用最大似然法和贝叶斯推断对来自 9 种鳞翅目物种的共 323 个细胞色素 P450 相关序列进行了分析,其中 130 个来自东方粘虫。在来自东方粘虫的 130 个细胞色素 P450 相关序列中,有 60 个归入 CYP3 族,该族与代谢外来物质和植物天然化合物有关。此外,还对β-CP 敏感(MsS-2)和抗性(MsR-2)东方粘虫种群转录组进行了测序。通过定量实时聚合酶链反应检测并验证了参与β-CP 解毒的某些关键基因。总之,我们的研究结果为进一步研究东方粘虫对杀虫剂的抗性分子机制提供了依据。

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