中华稻蝗几丁质合成酶1基因的mRNA表达及功能  

mRNA Expression and Physiological Function of Chitin Synthase1 from Oxya chinensis(Thunberg)

作　　者：余志涛[1] 刘晓健[1] 马恩波[1] 郭亚平[1] 张建珍[1] 

机构地区：[1]山西大学应用生物学研究所,太原030006

出　　处：《中国农业科学》2012年第5期877-884,共8页Scientia Agricultura Sinica

基　　金：国家自然科学基金项目(30970410;31071980);山西省回国留学人员科研项目(2009);山西省科技攻关项目(20110311010)

摘　　要：【目的】研究中华稻蝗(Oxya chinensis(Thunberg))几丁质合成酶1(CHS1)基因的表达特性及其在生长发育过程中的作用,从而为实现基于RNAi的蝗虫有效控制提供理论依据。【方法】根据已知中华稻蝗几丁质合成酶1基因(OcCHS1)保守区域部分cDNA序列(GenBank登录号:HM214491)设计特异性表达引物,运用RT-PCR和qPCR研究时空表达特性;采用RNA干扰技术研究其生物学功能。【结果】OcCHS1在中华稻蝗各龄期都有表达,其体表为高表达,其次是气管,其它组织部位表达量低。RNA干扰试验发现,4龄第4天若虫注射OcCHS1的双链RNA(dsRNA)后,与对照组相比,OcCHS1 mRNA表达量被沉默了70.8%,稻蝗出现蜕皮时间延迟、不能完成蜕皮或腹部皱缩死亡等现象,注射dsOcCHS1组死亡率为85.2%,与对照组(7.4%)相比差异显著。【结论】几丁质合成酶1对中华稻蝗的生长发育具有重要作用,其主要参与体表和气管几丁质的合成。采用RNA干扰技术使该基因沉默后可导致中华稻蝗死亡。【Objective】The objective of this study is to analyze the mRNA expression characteristics and physiological function of chitin synthase1 gene(OcCHS1) from the Chinese rice grasshopper,Oxya chinensis(Thunberg),and to provide a theoretical basis for effective pests control based on RNAi.【Method】Based on the known partial conservative cDNA fragment of OcCHS1(GenBank accession number: HM214491),primers were designed for mRNA expression analysis.The developmental and tissue-specific expression patterns of OcCHS1 were determined by RT-PCR and qPCR.RNAi experiment was performed to explore the physiological function.【Result】 OcCHS1 was expressed at all the developmental stages.Tissue-specific analysis showed that OcCHS1 was highly expressed in the integument,followed in the trachea,and lowest in other tissues.RNAi-based gene silencing experiment showed that the fourth day of the 4th instar nymphs injected by OcCHS1-speci?c dsRNA could repress the transcript levels of OcCHS1 by 70.8% compared to those in the control nymphs.Moreover,dsRNA injections signi?cantly increased abnormalities,such as stunted development,uncompleted molting,crimpy abdomen and eventually died.Consequentially,85.2% of mortalities were observed,while the mortality in the control nymphs was only 7.4%.【Conclusion】OcCHS1 is essential for development and mainly responsible for chitin synthesis in cuticle and trachea of O.chinensis.Moreover,silencing of chitin synthase 1 gene by RNAi is lethal to O.chinensis.

关 键 词：中华稻蝗 几丁质合成酶基因1 MRNA表达 RNA干扰 

分 类 号：S433.2[农业科学—农业昆虫与害虫防治]