杀虫剂处理的梨小食心虫转录组测序分析及差异表达解毒酶基因筛选  

Sequencing of Transcriptome and Screening of Differentially Expressed Detoxifying Enzyme Genes of Grapholita molesta Treated with Insecticides

作　　者：刘晓庆 庞钦玮 冀佳悦 杨春[1] 高玲玲 郭艳琼[1] LIU Xiaoqing;PANG Qinwei;JI Jiayue;YANG Chun;GAO Linging;GUO Yanqiong(College of Plant Protection,Shanxi Agricultural University,Taigu 030801,China;Institute of Pomology,Shanxi Agricultural University,Taigu 030815,China;Agriculture and Food,Commonwealth Scientific and Industrial Research Organization(CSIRO),Wembley 6913,Australia)

机构地区：[1]山西农业大学植物保护学院,山西太谷030801 [2]山西农业大学果树研究所,山西太谷030815 [3]澳大利亚联邦科学与工业研究组织农业与食品部,西澳文布利6913

出　　处：《山西农业科学》2023年第11期1233-1244,共12页Journal of Shanxi Agricultural Sciences

基　　金：山西省自然科学基金面上项目(202103021224153);国家现代农业产业技术体系(CARS-28-19)。

摘　　要：试验采用药膜法测定阿维菌素、高效氯氟氰菊酯、吡虫啉对梨小食心虫的室内毒性,旨在挖掘与杀虫剂解毒代谢相关基因,为进一步研究梨小食心虫解毒代谢机制奠定基础。根据生物测定结果,分别将3种杀虫剂原药用丙酮稀释至LC_(10)、LC_(30)、LC_(50)浓度,并将健康成虫放入带有杀虫剂薄膜的玻璃瓶24 h,收集存活的成虫,-80℃保存备用。采用Illumina HiSeq 2000进行高通量测序,获得的非重复序列基因(Unigene),使用NCBI比对工具BLAST分别与NR、NT、SWISS-PROT、Pfam、GO、KOG、KEGG数据库进行比对,获得相应的注释信息;进一步采用生物信息学方法筛选差异表达的解毒酶基因。结果表明,从转录组中共获得101873条Unigenes,平均长度为1992 bp,在NR、NT、SWISS-PROT、Pfam、KEGG、KOG、GO数据库比对,分别有57209、39186、43749、46168、46678、42247、27486条Unigene获得注释。阿维菌素、吡虫啉、高效氯氟氰菊酯处理梨小食心虫后差异表达基因分别有825、641、1391个,其中筛选到20个解毒代谢相关基因,包括4个羧酸酯酶基因、5个谷胱甘肽硫转移酶基因和11个细胞色素P450基因。In order to dig out genes related to pesticide detoxification metabolism,lay a foundation for further research on detoxification metabolism mechanism of Grapholita molesta.In this study,the indoor toxicity of avermectin,lambda-cyhalothrin,and imidacloprid to G.molesta was determined by drug film method.According to the bioassay results,the three insecticide technical drugs were diluted with acetone to the concentration of LC_(10),LC_(30),and LC_(50) respectively,and the healthy adults were put into the glass bottle with insecticide film for 24 hours.The surviving adults were collected and stored at-80℃for standby.High-throughput sequencing was performed by Illumina Hiseq 2000 and Unigenes were obtained.The Unigenes were compared by comparison tool BLAST from NCBI in databases such as NR,STRING,SWISS-PROT,Pfam,GO,KOG,KEGG in order to obtain the corresponding annotation information.In the next step,the bioinformatics method was used to screen the differentially expressed detoxification enzyme genes.The results showed that a total of 101873 Unigenes were obtained with an average length of 1992 bp.There were 57209,39186,43749,46168,46678,42247,27486 Unigenes annotations in NR,NT,SWISS-PROT,Pfam,KEGG,KOG,and GO databases,respectively.There were 825,641,and 1391 differentially expressed genes after avermectin,imidacloprid,and lambda-cyhalothrin at sublethal concentration.20 detoxification metabolism related genes were screened and identified,including 4 CarEs genes,5 GSTs genes,and 11 P450 genes.

关 键 词：梨小食心虫 解毒代谢相关基因 Illumina HiSeq 2000 转录组 

分 类 号：S482.3[农业科学—农药学] S433.4[农业科学—植物保护]