微小RNA介导意大利蜜蜂工蜂对东方蜜蜂微孢子虫的跨界调控  

MicroRNA-Mediated Cross-Kingdom Regulation of Apis mellifera ligustica Worker to Nosema ceranae

作　　者：杜宇 范小雪 蒋海宾 王杰 冯睿蓉 张文德 余岢骏 隆琦 蔡宗兵 熊翠玲 郑燕珍 陈大福 付中民 徐国钧[1,2] 郭睿 DU Yu;FAN XiaoXue;JIANG HaiBin;WANG Jie;FENG RuiRong;ZHANG WenDe;YU KeJun;LONG Qi;CAI ZongBing;XIONG CuiLing;ZHENG YanZhen;CHEN DaFu;FU ZhongMin;XU GuoJun;GUO Rui(College of Animal Sciences(College of Bee Science),Fujian Agriculture and Forestry University,Fuzhou 350002;Apitherapy Research Institute,Fujian Agriculture and Forestry University,Fuzhou 350002)

机构地区：[1]福建农林大学动物科学学院(蜂学学院),福州350002 [2]福建农林大学蜂疗研究所,福州350002

出　　处：《中国农业科学》2021年第8期1805-1820,共16页Scientia Agricultura Sinica

基　　金：国家现代农业产业技术体系建设专项(CARS-44-KXJ7);福建农林大学杰出青年科研人才计划(xjq201814);福建农林大学科技创新专项(CXZX2017342,CXZX2017343);福建农林大学优秀硕士学位论文资助基金(杜宇);福建省大学生创新创业训练计划(202010389016,202010389158)。

摘　　要：【目的】东方蜜蜂微孢子虫(Nosema ceranae)感染意大利蜜蜂(Apis mellifera ligustica,简称意蜂)导致蜜蜂微孢子虫病。本研究结合前期已获得的miRNA和mRNA组学数据,通过生物信息学方法对意蜂工蜂中肠的差异表达miRNA(differentially expressed miRNA,DEmiRNA)靶向结合的东方蜜蜂微孢子虫的mRNA和差异表达mRNA(DEmRNA)进行预测、数据库注释和调控网络分析,以期在组学水平解析miRNA介导意蜂工蜂对东方蜜蜂微孢子虫的跨界调控机制。【方法】通过比较东方蜜蜂微孢子虫侵染7 d和10 d的意蜂工蜂中肠(AmT1、AmT2)和未受侵染的工蜂中肠(AmCK1、AmCK2)的miRNA组学数据筛选出宿主的显著性DEmiRNA,通过比较侵染意蜂工蜂中肠的东方蜜蜂微孢子虫(NcT1、NcT2)和东方蜜蜂微孢子虫纯净孢子(NcCK)的mRNA数据筛选出病原的DEmRNA。利用TargetFinder软件预测宿主显著性DEmiRNA靶向结合的病原mRNA和DEmRNA。利用相关生物信息学工具对上述靶DEmRNA进行GO和KEGG数据库注释。结合前期研究结果筛选出孢壁蛋白、极管蛋白、蓖麻毒素B凝集素、ABC转运蛋白、ATP/ADP移位酶和糖酵解/糖异生途径等毒力因子和能量代谢通路相关的病原DEmRNA及与其存在靶向结合关系的宿主显著性DEmiRNA,并构建和分析二者的调控网络。【结果】AmCK1 vs AmT1比较组中宿主的48条显著上调miRNA和36条显著下调miRNA分别靶向病原的1345和1046条mRNA;进一步分析发现,宿主的47条显著上调miRNA和34条显著下调miRNA可分别靶向NcCK vs NcT1比较组中病原的584条显著下调mRNA和265条显著上调mRNA,它们可分别注释到19和22个功能条目以及66和64条通路。AmCK2 vs AmT2比较组中宿主的56条显著上调miRNA和51条显著下调miRNA分别靶向病原的1260和1317条mRNA;进一步分析发现,宿主的52条显著上调miRNA和49条显著下调miRNA可分别靶向NcCK vs NcT2比较组中病原的587条显著下调mRNA和336条显著上调mRNA,�【Objective】Nosema ceranae infects Apis mellifera ligustica and causes microsporidiosis.In this study,to reveal the mechanism of miRNA-mediated cross-kingdom regulation of A.m.ligustica worker to N.ceranae,prediction,GO and KEGG database annotation as well as regulatory network analysis of N.ceranae mRNAs and differentially expressed mRNAs(DEmRNAs)targeted by differentially expressed miRNAs(DEmiRNAs)of A.m.ligustica workers’midguts were conducted by bioinformatic approaches based on previously gained miRNA and mRNA omics data.【Method】Significant host DEmiRNAs were screened out by comparison of miRNA omics data from A.m.ligustica workers’midguts at 7 d and 10 d post N.ceranae infection(AmT1,AmT2)and corresponding uninfected midguts(AmCK1,AmCK2).DEmRNAs of pathogen were screened out through comparison of mRNA omics data from N.ceranae infecting A.m.ligustica worker’s midgut(NcT1 and NcT2)and pure fungal spores(NcCK).mRNAs and DEmRNAs of N.ceranae targeted by significant host DEmiRNAs were predicted using TargetFinder software.GO and KEGG database annotations of aforementioned targets were conducted using related bioinformatics tools.On basis of our previous findings,pathogen DEmRNAs associated with spore wall protein,polar tube protein,ricin B lectin,ATP/ADP translocase,ABC transporters and glycolysis/gluconeogenesis,and their target significant DEmiRNAs of host were filtered out,followed by construction and investigation of regulatory network.【Result】In AmCK1 vs AmT1 comparison group,48 significantly up-regulated miRNAs and 36 significantly down-regulated miRNAs could respectively target 1345 and 1046 mRNAs of N.ceranae;additionally,47 significantly up-regulated miRNAs and 34 significantly down-regulated miRNAs of host could target 584 significantly down-regulated mRNAs and 265 significantly up-regulated mRNAs in NcCK vs NcT1;these targets were involved in 19 and 22 functional terms as well as 66 and 64 pathways.In AmCK2 vs AmT2 comparison group,56 significantly up-regulated miRNAs and 51 signific

关 键 词：意大利蜜蜂 东方蜜蜂微孢子虫 微小RNA 跨界调控 调控网络 免疫防御 

分 类 号：S895.236[农业科学—特种经济动物饲养]