亚致死浓度吡虫啉和毒死蜱对柑橘木虱成虫组织蛋白酶基因表达的影响  

Effects of sublethal doses of imidacloprid and chlorpyrifos on the expression profiles of cathepsins in Diaphorina citri Kuwayama

作　　者：舒本水 邹燕 张婉莹 冯杰 吴仲真[1] 林进添[1] SHU Ben-Shui;ZOU Yan;ZHANG Wan-Ying;FENG Jie;WU Zhong-Zhen;LIN Jin-Tian(Zhongkai University of Agriculture and Engineering,Guangzhou City Key Laboratory of Subtropical Fruit Tree Pest and Disease Outbreak Control,Guangzhou 510225,China)

机构地区：[1]仲恺农业工程学院/广州市亚热带果树重大疫情控制重点实验室,广州510225

出　　处：《环境昆虫学报》2022年第4期1027-1036,共10页Journal of Environmental Entomology

基　　金：广东省科技计划项目(2016B02020009);广州市民生科技攻关计划项目(201704020190,201803020009);广东省教育厅特色创新项目(2019KTSCX063);广州市亚热带果树重大疫情控制重点实验室项目(201805010008)。

摘　　要：昆虫组织蛋白酶属于多功能酶系,在昆虫生命活动中具有重要作用,但昆虫组织蛋白酶对化学农药的响应却鲜有报道。吡虫啉和毒死蜱是目前防治柑橘木虱Diaphorina citri常用的化学药剂。本研究旨在探究吡虫啉和毒死蜱亚致死浓度处理对柑橘木虱成虫组织蛋白酶基因表达的影响。本研究基于柑橘木虱的转录组数据库,同时结合NCBI数据库,通过Blast比对鉴定得到8个柑橘木虱组织蛋白酶基因,序列分析得出其均属于半胱氨酸蛋白酶家族且含有保守的组氨酸活性位点。研究采用浸叶法处理得到经吡虫啉和毒死蜱处理24 h对柑橘木虱成虫的致死中浓度LC_(50)分别为97.88 mg/L和47.94 mg/L。采用荧光定量PCR(qPCR)分析这两种药剂亚致死浓度(LC_(20)和LC_(50))胁迫下柑橘木虱成虫组织蛋白酶基因表达水平变化。结果表明吡虫啉胁迫显著下调柑橘木虱DcCath-B,DcCath-F,DcCath-L和DcCath-L 1基因表达量;毒死蜱LC_(20)浓度胁迫显著下调柑橘木虱DcCath-B,DcCath-L和DcCath-W基因表达量,LC_(50)浓度胁迫下仅DcCath-B表达量显著下调,其他基因表达无明显差异。以上结果表明吡虫啉和毒死蜱亚致死浓度胁迫下,柑橘木虱成虫下调表达体内部分组织蛋白酶基因以响应此逆境。本研究为探索亚致死浓度吡虫啉和毒死蜱对柑橘木虱的毒理机制提供了一定的理论依据。Insect cathepsins are multifunctional enzymes that play important roles in insect life activities.However,the response of cathepsins in insects to pesticides has rarely been reported.By blasting against the transcriptome data and searching the NCBI database,eight Diaphorina citri cathepsin genes with conserved histidine active sites were identified in this study.The LC 50 values of imidacloprid and chlorpyrifos against the adults of D.citri for 24 h were 47.94 mg/L and 97.88 mg/L,respectively.The qPCR results showed that the imidacloprid treatments at the concentrations of LC 20 and LC 50 significantly down-regulated the expressions of DcCath-B,DcCath-F,DcCath-L,and DcCath-L1.mRNA expression levels of DcCath-B,DcCath-L,and DcCath-W were significantly down-regulated in adults with chlorpyrifos treatment at the concentration of LC 20.After treated with chlorpyrifos at the concentration of LC 50,the expression level of DcCath-B was significantly down-regulated,while the other seven cathepsin genes were changed with no significance.The results showed that some cathepsins of D.citri down-regulated the mRNA expression in response to sublethal concentration treatments of imidacloprid and chlorpyrifos.This study provided a theoretical basis for exploring the toxicological mechanism of imidacloprid and chlorpyrifos on D.citri.

关 键 词：柑橘木虱 组织蛋白酶 吡虫啉 毒死蜱 亚致死浓度 基因表达量 

分 类 号：Q963[生物学—昆虫学] S433[农业科学—农业昆虫与害虫防治]