梨小食心虫几丁质合成酶2基因的克隆与表达研究  

Molecular cloning, sequence analysis and expression of the chitin synthase 2 gene in the oriental fruit moth Grapholitha molesta(Busck)

作　　者：杨静[1] 庾琴[1] 高越[1] 刘中芳[1] 史高川[2] 张鹏九[1] 范仁俊[1] 

机构地区：[1]山西省农业科学院植物保护研究所农业有害生物综合治理山西省重点实验室,太原030031 [2]山西省农业科学院棉花研究所,运城044000

出　　处：《应用昆虫学报》2017年第3期407-416,共10页Chinese Journal of Applied Entomology

基　　金：山西省农业科学院攻关项目(YGG1433);山西省农业科学院博士研究基金(YBSJJ1513);山西省农业科技成果转化和推广示范工程(2015CGZH041);国家公益性行业专项(201103024)

摘　　要：【目的】研究梨小食心虫Grapholitha molesta(Busck)几丁质合成酶2(GmCHS2)基因序列和表达特性,为进一步研究其功能及新型杀虫剂的开发提供理论依据。【方法】利用转录组数据和RACE技术首次获得GmCHS2基因cDNA序列(命名为GmCHS2,GenBank登录号为KY242360),利用其他昆虫同源序列构建系统发育树,利用实时荧光定量PCR技术研究GmCHS2在不同发育阶段和不同组织的表达特性。【结果】GmCHS2基因序列全长4991bp,开放阅读框为4554bp,其中5′端非编码区长267bp,3′端非编码区长170bp,共编码1517个氨基酸,包含了14个跨膜螺旋。系统发育树结果表明该基因属于几丁质合成酶2类基因。GmCHS2基因在试虫不同发育阶段都有表达,预蛹期和成虫期表达量最高;不同组织中,前肠和中肠的表达量最高,其次是后肠,其余组织表达量很少或不表达。【结论】本文研究了GmCHS2基因在梨小食心虫不同发育阶段和组织部位的表达特性,该基因可能在梨小食心虫生长发育过程中具有重要作用。[Objectives] To analyze the mRNA expression characteristics of the oriental fruit moth, Grapholitha molesta （Busck） chitin synthase 2 gene （GmCHS2） and provide a theoretical basis for studying its physiological function and developing new pest control methods. [Methods] The cDNA of GmCHS2 （GenBank accession number： KY242360） was obtained based on the transcriptome dataset and rapid amplification cDNA end（RACE）technology. A phylogenetic tree was constructed based on the homologous sequences of other insects. Relative expression levels of the GmCHS2 gene were detected in different development stages and tissues using real-time quantitative PCR. [Results] GmCHS2 cDNA was 4991bp in length, including a 267 bp 5＇ terminal UTR and a 170 bp 3＇ terminal UTR, encodes 1517 amino acids and contains 14 transmembrane helices. A phylogenetic tree indicates that GmCHS2 belongs to the chitin synthase 2 gene family. Analysis of GmCHS2 expression in different development stages revealed that the highest expression levels occurred in prepupae and adults. Analysis of the expression of GmCHS2 in different tissues indicates that it was highly expressed in the foregut and midgut, followed by the hindgut, and only slightly expressed, or not expressed, in other tissues. [Conclusion] GmCHS2 may play an important role in the growth and development of G. molesta.

关 键 词：梨小食心虫 几丁质合成酶 表达谱 荧光定量 PCR 

分 类 号：S433[农业科学—农业昆虫与害虫防治]