小地老虎硫氧还蛋白基因AiTrx2的分子特性及其对杀虫剂胁迫的响应  

Molecular Characteristics of Thioredoxin Gene AiTrx2 from Agrotis ipsilon and Its Response to Insecticide Stress

作　　者：洪牛 陈诗琪 鄢敏 杨建[2] 苗治国 刘苏[1] 李茂业[1] 杨洋[2] HONG Niu;CHEN Shiqi;YAN Min;YANG Jian;MIAO Zhiguo;LIU Su;LI Maoye;YANG Yang(Key Laboratory of Biology and Sustainable Management of Plant Diseases and Pests of Anhui Higher Education Institutions,College of Plant Protection,Anhui Agricultural University,Hefei 230036,China;Yibin Branch of Sichuan Provincial Tobacco Corporation,Yibin Sichuan 644600,China)

机构地区：[1]安徽农业大学植物保护学院,植物病虫害生物学与绿色防控安徽普通高校重点实验室,合肥230036 [2]四川省烟草公司宜宾市公司,四川宜宾644600

出　　处：《西北农业学报》2024年第12期2399-2409,共11页Acta Agriculturae Boreali-occidentalis Sinica

基　　金：中国烟草总公司四川省公司科技项目(SCYC202112);安徽省教育厅优秀青年人才支持计划(gxyq2021167)。

摘　　要：鉴定小地老虎(Agrotis ipsilon)硫氧还蛋白基因(AiTrx2),明确其编码蛋白质的催化活性,阐明其在小地老虎不同发育期、不同组织及在氯虫苯甲酰胺、高效氯氟氰菊酯和辛硫磷胁迫下的表达模式。利用同源检索方法从小地老虎转录组中鉴定AiTrx2基因,利用大肠杆菌表达重组蛋白分析其活性,使用实时荧光定量PCR技术研究基因表达水平的变化。从小地老虎转录组中鉴定了AiTrx2的cDNA序列,其编码的蛋白具有硫氧还蛋白的典型特征,且与斜纹夜蛾亲缘关系最近。在大肠杆菌中成功表达了AiTrx2重组蛋白,测得该蛋白具有还原酶活性,过表达AiTrx2重组蛋白能够增强大肠杆菌对氧化胁迫的耐受性。在小地老虎不同组织与不同龄期中均检测出AiTrx2表达,分别以幼虫脂肪体和成虫阶段表达水平最高。使用LC_(50)浓度氯虫苯甲酰胺、高效氯氟氰菊酯和辛硫磷处理小地老虎幼虫后,AiTrx2表达水平显著上调。明确了小地老虎AiTrx2的分子特性和表达模式,发现氯虫苯甲酰胺、高效氯氟氰菊酯和辛硫磷处理能够诱导AiTrx2表达水平显著上调,说明AiTrx2可能在防御杀虫剂诱导的氧化胁迫中起重要作用。The purpose of this study is to identify the thioredoxin gene(AiTrx2)from Agrotis ipsilon,elucidate the catalytic activity of AiTrx2 protein,and unravel the expression profile of AiTrx2 at different developmental stages,in various larval tissues,and following exposure with chlorantraniliprole,lambda-cyhalothrin,and phoxim.These investigations will pave the way for unveiling the physiological function of AiTrx2 in A.ipsilon.A homology search method was used to identify AiTrx2,the recombinant AiTrx2 protein was expressed using an Escherichia coli expression system,and a real-time quantitative PCR technology was used to analyze the variation of AiTrx2 transcription levels.The cDNA sequence of AiTrx2 was identified from A.ipsilon transcriptome dataset.The encoded protein exhibited characteristic thioredoxin features and demonstrated the closest resemblance to Spodoptera litura’s Trx2.Recombinant AiTrx2 protein was successfully expressed in E.coli and the protein displayed reductase activity.Moreover,overexpression of AiTrx2 enhanced the tolerance of E.coli to oxidative stress.The mRNA transcripts of AiTrx2 could be detected in different tissues and developmental stages,with the highest transcription levels in larval fat body and adult stages.The expression level of AiTrx2 was significantly upregulated in A.ipsilon larvae after treatment with a LC_(50)concentrations of chlorantraniliprole,lambda-cyhalothrin,and phoxim.In this study,the molecular characteristics and expression patterns of AiTrx2 are clarified.The transcription of AiTrx2 is dramatically enhanced following exposure to chlorantraniliprole,lambda-cyhalothrin,and phoxim,These results suggest that AiTrx2 plays an important role in defensing oxidative stress induced by insecticides.

关 键 词：小地老虎 硫氧还蛋白 杀虫剂 抗氧化活性 表达模式 

分 类 号：S433.82[农业科学—农业昆虫与害虫防治]