基于转录组的苹小卷叶蛾杀虫剂靶标及解毒代谢相关基因分析

doi:10.16380/j.kcxb.2020.04.010

摘要/Abstract

摘要： 【目的】建立苹小卷叶蛾Adoxophyes orana转录组数据库，挖掘杀虫剂靶标及解毒代谢相关基因。【方法】采用Illumina HiSeq^TM 2000高通量测序技术对苹小卷叶蛾进行转录组测序，挖掘并分析杀虫剂靶标基因；利用qPCR检测6个杀虫剂靶标基因在苹小卷叶蛾卵、幼虫、蛹和成虫各不同发育阶段的表达；挖掘并分析苹小卷叶蛾转录组中解毒代谢相关基因的代谢通路及进化关系。【结果】通过组装有效序列共获得48 610条unigene(GenBank登录号: GGMW00000000)。挖掘鉴定到155个杀虫剂靶标unigene；qPCR结果显示，1个蜕皮激素受体(ecdysone receptor, ECR)、2个乙酰胆碱酯酶(acetylcholinesterase, AChE)、1个氯离子通道蛋白(chloride channel, CLC)、1个几丁质酶(chitinase, CS)和1个鱼尼丁受体(ryanodine receptor, RyR)基因在苹小卷叶蛾不同发育阶段均存在表达差异。挖掘鉴定到69个羧酸酯酶(carboxylesterase, CarE)unigene、66个谷胱甘肽S-转移酶(glutathion S-transferase, GST)unigene和205个细胞色素50(cytochrome P450)unigene等解毒代谢相关基因，共鉴定20个CarE unigene， 32个GST unigene和30个P450 unigene与有毒物质代谢相关的通路有关。基于氨基酸序列对具有完整ORF的unigene聚类分析结果显示：12个CarEs中9个为G类，即鳞翅目保幼激素类；分别有10个AoGSTs属于Delta和Epsilon亚家族；18个P450全部聚到CYP3集团。【结论】该研究有助于苹小卷叶蛾杀虫剂靶标基因的挖掘及抗药性的研究。

关键词: 苹小卷叶蛾, 转录组, 杀虫剂靶标基因, 解毒代谢相关基因, 抗药性

Abstract: 【Aim】 To establish the transcriptome database of Adoxophyes orana, and to explore insecticide target and detoxification related genes. 【Methods】 The transcriptome of A. orana was sequenced by Illumina HiSeq^TM 2000 sequencing platform to explore insecticide target genes. The expression levels of six insecticide target genes from A. ornan at different developmental stages including egg, larva, pupa and adult were identified and verified using qPCR. The metabolic pathway and evolutionary relationship of
detoxification related genes in the transcriptome of A. orana were analyzed. 【Results】 A total of 48 610 unigenes (GenBank accession no.: GGMW00000000) were identified by assembling valid sequences. One hundred and fifty-five insecticides target genes were identified. The qPCR results confirmed that there was difference in the expression levels of six insecticide target genes including one ecdysone receptor (ECR) gene, two acetylcholinesterase (AChE) genes, one chloride channel (CLC) gene, one chitinase (CS) gene
and one ryanodine receptor (RyR) gene at different developmental stages of A. ornan. Additionally, 69 carboxylesterase (CarE) unigenes, 66 glutathion S-transferase (GST) unigenes and 205 cytochrome P450 unigenes were identified. A total of 20 CarE, 32 GST and 30 P450 unigenes were classified into the metabolism categories related to toxic substance metabolism. The results of cluster analyses based on the amino acid sequences of unigenes with complete ORF showed that 9 of 12 CarEs were classified to G class (Lepidoptera neotenin), 10 AoGSTs into subfamily Delta and another 10 AoGSTs into subfamily Epsilon, and all of 18 P450s into CYP3 group. 【Conclusion】 This study contributes to the discovery of insecticide target genes and the research of pesticide resistance in A. orana.

Key words: Adoxophyes orana, transcriptome, insecticide target gene, detoxification related gene, insecticide resistance

孙丽娜, 张怀江, 刘孝贺, 仇贵生. 基于转录组的苹小卷叶蛾杀虫剂靶标及解毒代谢相关基因分析[J]. 昆虫学报, 2020, 63(4): 470-481.

SUN Li-Na, ZHANG Huai-Jiang, LIU Xiao-He, QIU Gui-Sheng. Transcriptomic analysis of insecticide target and detoxification related genes in Adoxophyes orana (Lepidoptera: Tortricidae)[J]. Acta Entomologica Sinica, 2020, 63(4): 470-481.

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基于转录组的苹小卷叶蛾杀虫剂靶标及解毒代谢相关基因分析

Transcriptomic analysis of insecticide target and detoxification related genes in Adoxophyes orana (Lepidoptera: Tortricidae)

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