烟粉虱MEAM1隐种磷脂氢谷胱甘肽过氧化物酶基因克隆与表达分析  

Molecular cloning and expression profiling of phospholipid hydroperoxide glutathione peroxidase genes in Bemisia tabaci MEAM1 (Hemiptera: Aleyrodidae)

作　　者：纠敏[1] 汪伦记[1,2] 任利娜 代威 李晶晶 赵君峰[1] 张敏[2] JIU Min;WANG Lun-Ji;REN Li-Na;DAI Wei;LI Jing-Jing;ZHAO Jun-Feng;ZHANG Min(College of Food and Bioengineering,Henan University of Science and Technology,Luoyang,Henan 471023,China;Key Laboratory of Microbial Resources Development and Utilization,College of Food and Bioengineering,Henan University of Science and Technology,Luoyang,Henan 471023,China)

机构地区：[1]河南科技大学食品与生物工程学院,河南洛阳471023 [2]河南科技大学食品与生物工程学院微生物资源开发与利用重点实验室,河南洛阳471023

出　　处：《昆虫学报》2019年第2期141-149,共9页Acta Entomologica Sinica

基　　金：国家自然科学基金项目(31672036;31101443)

摘　　要：【目的】本研究旨在从烟粉虱Bemisia tabaci中东-小亚细亚1隐种(Middle East-Asia Minor 1, MEAM1)中克隆磷脂氢谷胱甘肽过氧化物酶(phospholipid hydroperoxide glutathione peroxidase, PHGPX)基因,鉴定其在烟粉虱不同发育阶段及吡虫啉处理不同时间后雌成虫体内的表达情况,明确其在烟粉虱应对外界环境压力中的功能。【方法】利用3′RACE克隆和测定烟粉虱MEAM1隐种内PHGPX基因的cDNA全长序列,并对其编码的氨基酸序列进行生物信息学分析;利用定量RT-PCR技术对该基因在烟粉虱MEAM1隐种不同发育阶段及吡虫啉处理不同时间后雌成虫体内的表达量进行分析。【结果】获得了烟粉虱MEAM1隐种两个磷脂氢谷胱甘肽过氧化物酶基因的全长cDNA序列,分别命名为BtB-PHGPX1(GenBank登录号:KY312116)和BtB-PHGPX2(GenBank登录号:KY312117)。序列分析表明,BtB-PHGPX1基因开放阅读框全长732 bp,编码243个氨基酸;BtB-PHGPX2基因开放阅读框全长567 bp,编码188个氨基酸。序列比对结果表明两基因的编码蛋白内均具有谷胱甘肽过氧化物酶保守的半胱氨酸、谷氨酰胺和色氨酸残基位点。BtB-PHGPX1在烟粉虱MEAM1隐种卵内表达量显著高于其在若虫、伪蛹、雌成虫和雄成虫内的表达量,BtB-PHGPX2在烟粉虱MEAM1隐种卵内的表达量显著低于其在若虫、伪蛹和雌成虫内的表达量(P<0.05)。BtB-PHGPX1和BtB-PHGPX2在雌成虫内的表达量均显著高于雄成虫内。吡虫啉处理雌成虫2 h时两基因的表达量均较对照显著提高(P<0.05),处理后5, 10和24 h时其表达量均较对照显著下降(P<0.01)。【结论】本研究克隆了烟粉虱MEAM1隐种两个PHGPX基因的序列全长,明确了其在不同发育阶段及吡虫啉处理不同时间后雌成虫体内的差异表达,推测PHGPX在烟粉虱抵御环境压力及杀虫剂胁迫时可能发挥着重要的防御作用。【Aim】The objective of this study is to clone phospholipid hydroperoxide glutathione peroxidase(PHGPX)genes from Bemisia tabaci MEAM1 cryptic species,to identify their expression levels in whiteflies at different developmental stages and in female adults treated with imidacloprid for different time,and to elucidate their function in the whitefly in response to environmental pressure.【Methods】The cDNAs of PHGPX genes were cloned by 3′RACE from B.tabaci MEAM1 cryptic species,and the putative amino acid sequences were analyzed by bioinformatics methods.The expression levels of PHGPX genes in the whitefly at different developmental stages and in female adults treated with imidacloprid for different time were analyzed by quantitative reverse transcription-polymerase chain reaction(qRT-PCR).【Results】The full-length cDNA sequences of two PHGPX genes were obtained from B.tabaci MEAM1 cryptic species,and named as BtB-PHGPX 1(GenBank accession no.:KY312116)and BtB-PHGPX 2(GenBank accession no.:KY312117),respectively.Sequence analysis indicated that the open reading frame of BtB-PHGPX 1 is 732 bp in length encoding 243 amino acids,while that of BtB-PHGPX 2 is 567 bp in length encoding 188 amino acids.Sequence alignment result showed that the encoded proteins of both genes have conserved cysteine,glutamine and tryptophan residues of glutathione peroxidase.The mRNA level of BtB-PHGPX 1 in egg was significant higher than those in nymph,pseudopupa,female adult and male adult of the whitefly,while that of BtB-PHGPX 2 in egg was significant lower than those in nymph,pseudopupa and female adult(P<0.05).The mRNA levels of both BtB-PHGPX 1 and BtB-PHGPX 2 in female adults were significant higher than those in male adults(P<0.05).Furthermore,the expression levels of BtB-PHGPX 1 and BtB-PHGPX 2 significantly increased in female adults at 2 h after treatment with imidacloprid as compared with their respective control(P<0.05),but decreased significantly at 5,10 and 24 h after treatment(P<0.01).【Conclusion】In this study,

关 键 词：烟粉虱 MEAM1隐种 磷脂氢谷胱甘肽过氧化物酶 氧化应激 吡虫啉 

分 类 号：Q966[生物学—昆虫学]