基于转录组测序解析草地贪夜蛾对溴氰虫酰胺的解毒代谢分子机制  

Transcriptome sequencing reveals the molecular mechanism of detoxification metabolism induced by cyantraniliprole in fall armyworm Spodoptera frugiperda

作　　者：杨媛雪 王爱玉 张芸 张开军[2] 薛超[1] 刘怀[2] 赵鸣[1] 张建华[1] Yang Yuanxue;Wang Aiyu;Zhang Yun;Zhang Kaijun;Xue Chao;Liu Huai;Zhao Ming;Zhang Jianhua(Institute of Industrial Crops,Shandong Academy of Agricultural Sciences,Jinan 250100,Shandong Province,China;College of Plant Protection,Southwest University,Chongqing 400715,China)

机构地区：[1]山东省农业科学院经济作物研究所,济南250100 [2]西南大学植物保护学院,重庆400715

出　　处：《植物保护学报》2023年第3期800-811,共12页Journal of Plant Protection

基　　金：山东省科技成果转移转化补助(鲁渝科技协作)项目(2022LYXZ026)。

摘　　要：为阐明草地贪夜蛾Spodoptera frugiperda对溴氰虫酰胺的解毒代谢分子机制,通过LC50的溴氰虫酰胺诱导草地贪夜蛾3龄幼虫后,利用酶活测定和转录组测序鉴定解毒代谢相关基因,并采用实时荧光定量PCR技术对细胞色素P450单加氧酶(cytochrome P450 monooxygenase,P450)基因进行验证分析。结果表明,经LC50的溴氰虫酰胺处理后,草地贪夜蛾3龄幼虫体内3种解毒代谢酶活性较对照均有所升高,但仅P450活性较对照显著升高,而谷胱甘肽S-转移酶和羧酸酯酶与对照无显著差异。经LC50的溴氰虫酰胺处理后草地贪夜蛾3龄幼虫转录组中共筛选到1408个差异表达基因,其中上调表达的基因有935个,下调表达的基因有473个。药物代谢-细胞色素P450通路、药物代谢-其他酶通路及细胞色素P450对异生物质的代谢通路中有超过20个基因存在差异表达。在草地贪夜蛾转录组中筛选鉴定到121个P450基因,其中,属于CYP2、CYP3、CYP4以及Mito家簇的基因分别有9、45、58和9个,而经LC50的溴氰虫酰胺诱导后有8个P450基因上调表达,实时荧光定量PCR验证结果显示其中有6个P450基因CYP321A8、CYP6B2、CYP6B7、CYP6B5、CYP4C1及CYP4D1表达量显著上调,这些都属于与害虫抗药性相关的CYP3和CYP4家簇基因。表明P450基因CYP321A8、CYP6B2、CYP6B7、CYP6B5、CYP4C1及CYP4D1可能在草地贪夜蛾对溴氰虫酰胺的解毒代谢中发挥着重要作用。In order to clarify the molecular mechanism of metabolic detoxification of cyantraniliprole in fall armyworm Spodoptera frugiperda,the 3rd instar larvae was induced by cyantraniliprole at the dose of LC50 to screen and identify the relevant genes related to metabolic detoxification through the enzyme activity assay and transcriptome sequencing analysis,and the relative expression levels of cytochrome P450 monooxygenase(P450)genes were validated using quantitative real-time PCR(qPCR).The results showed that the activities of the three detoxification metabolic enzymes were increased after treatment with cyantraniliprole,but only the activity of P450 was significantly increased,while the activities of glu-tathione S-transferase and carboxylesterase were not significantly increased compared with the control.A total of 1408 genes were differentially expressed induced by cyantraniliprole at LC50 in S.frugiperda,of which 935 genes were up-regulated and 473 genes were down-regulated.More than 20 genes in the pathway of drug metabolism-cytochrome P450,drug metabolism-other enzymes,and metabolism of xe-nobiotics by cytochrome P450 showed differential expression.Based on transcriptome sequencing,a to-tal of 121 P450 genes(nine in CYP2 clade,45 in CYP3 clade,58 in CYP4 clade,and nine in Mito clade)were identified in S.frugiperda.Among them,the expression of eight P450 genes was up-regulated af-ter induced by cyantraniliprole at LC50 dose,and it was found that the relative expression levels of CYP321A8,CYP6B2,CYP6B7,CYP6B5,CYP4C1 and CYP4D1 were significantly up-regulated through qPCR analysis.The six P450 genes belonged to the CYP3 Clade and CYP4 Clade,which were associat-ed with insecticide resistance.It indicated that P450 including CYP321A8,CYP6B2,CYP6B7,CYP6B5,CYP4C1 and CYP4D1 might play important roles in metabolizing cyantraniliprole in S.frugiperda.

关 键 词：草地贪夜蛾 溴氰虫酰胺 转录组 解毒代谢 细胞色素P450单加氧酶 

分 类 号：S433.4[农业科学—农业昆虫与害虫防治]