RNA干扰MDCht9基因的家蝇幼虫的转录组分析  

Transcriptome Analysis of Musca domestica Larvae with RNA Interference MDCht9 Gene

作　　者：李妍 国果 苏佩佩[1] 赵欣宇 杨隆兵 赵文静 修江帆 Li Yan;Guo Guo;Su Peipei;Zhao Xinyu;Yang Longbing;Zhao Wenjing;Xiu Jiangfan(The Key and Characteristic Laboratory of Modern Pathogen Biology,School of Basic Medical Sciences,Guizhou Medical University,Guiyang,550025;School of Food Science,Guizhou Medical University,Guiyang,550025)

机构地区：[1]贵州医科大学基础医学院,现代病原生物学重点特色实验室,贵阳550025 [2]贵州医科大学食品科学学院,贵阳550025

出　　处：《基因组学与应用生物学》2021年第2期569-577,共9页Genomics and Applied Biology

基　　金：国家自然科学基金项目(81560337,81760647)资助。

摘　　要：家蝇(Musca domestica)属于双翅目昆虫,广泛分布于世界各地,目前家蝇抗药现象较为严重。为了筛选及挖掘与防治有害医学昆虫相关的分子靶标,本研究以家蝇幼虫为实验对象,采用RNAi方法沉默家蝇几丁质酶MDCht9基因后,分析幼虫mRNA转录组表达变化,初步探讨MDCht9的功能。研究结果表明,在注射dsRNA 24 h后,幼虫MDCht9 mRNA的表达显著下降85%,提示干扰有一定效果。实验组与对照组相比共筛选出378个基因差异表达显著,其中上调基因162个,下调基因216个,占所有被检测到转录RNA基因的百分比为2.66%。通过GO和KEGG相关生物信息学分析,挖掘出涉及幼虫时期生长发育、蛋白质的消化与吸收、雄虫性成熟、物质代谢等相关通路基因。采用实时荧光定量PCR对差异基因进行验证,与转录组结果一致。本实验通过RNAi方法成功降低MDCht9基因mRNA表达,MDCht9基因参与幼虫时期生长发育、蛋白质的消化与吸收、雄虫性成熟、物质代谢等功能。本研究结果有助于后续生物信息学分析及为家蝇生长发育关键基因的挖掘提供基础数据,为害虫防治提供新的靶标途径。Musca domestica belongs to Diptera insects and is widely distributed throughout the world.At present,the resistance of M.domestica is serious.In order to screen and excavate the molecular targets related to the prevention and control of harmful medical insects,this study used M.domestica larvaes as the experimental object,and then used the RNAi method to silence MDCht9,and analyzed the transcriptome expression changes of the larvaes mRNA,and initially discussed the MDCht9 features.The results showed that the expression of MDCht9 mRNA in larvae decreased significantly by 85%after 24 h of dsRNA injection,suggesting that RNA interference has an effect.Compared with the control group,the experimental group screened 378 genes with significant differential expression,including 162 up-regulated genes and 216 down-regulated genes,accounting for 2.66%of all transcribed RNA genes detected.Through GO and KEGG related bioinformatics analysis,genes related to growth and development,protein digestion and absorption,male sexual maturity and material metabolism were discovered.The differential genes were verified by RT-qPCR,which was consistent with the transcriptome results.In this study,the mRNA expression of MDCht9 was successfully reduced by RNAi method.MDCht9 was involved in the growth and development of larvae,digestion and absorption of protein,male sexual maturity and material metabolism.These results contribute to the subsequent bioinformatics analysis and provide basic data for the excavation of key genes in the growth and development of M.domestica,providing a new target pathway for pest control.

关 键 词：MDCht9 RNA干扰 转录组 

分 类 号：R184.3[医药卫生—流行病学]