为了研究烟草脉斑驳病毒(TVMV)侵染本氏烟的抗病分子机制,本文建立了TVMV转录组数据库,挖掘抗病相关基因以及代谢和信号通路,采用Illumina Novaseq 6000高通量测序平台对侵染TVMV的本氏烟进行转录组测序,并进行生物信息学分析;利用R package:edge R等软件分析了有关抗病的差异表达基因,对差异表达基因进行基因本体论(GO)及京都基因和基因组百科全书(KEGG)基于途径的通路富集分析.基于TVMV侵染本氏烟转录组的测序结果中共获得4 593个差异表达基因,其中3 564个基因上调表达,1 029个基因下调表达.共筛选出10个抗病相关的差异表达显著基因,6个上调,4个下调.GO数据库中注释到生物学过程、细胞组分及分子功能等三大类共50个功能组.其中,在第三大类分子功能中,蛋白质结合功能以及ATP结合功能所涉及的差异表达基因最显著,分别为501个和453个差异基因.KEGG共富集20条通路,注释到核糖体途径的差异基因富集程度最高,基因最显著,差异表达基因数为307个.蛋白质结合、ATP结合、核糖体、真核生物核糖体的生物反应、DNA复制(DNA replication)等通路可能在调控本氏烟抗TVMV病毒中起着重要作用.通过对转录组中筛选的差异表达基因进行分析研究,为后续本氏烟抗TVMV病毒的深入研究奠定了重要的基础.
In order to study the molecular mechanism of tobacco vein mottling virus(TVMV)infection of Nicotiana ben-thamiana,this study established a TVMV transcriptome database,excavated disease-resistant genes,metabolic and signaling pathways,and used the Illumina Novaseq 6000 high-throughput sequencing platform to perform transcriptome sequencing and bioinformatics analysis Nicotiana benthamiana infected with TVMV.We used software such as R package:edge R to analyze differentially expressed genes related to disease resistance,and conducted gene ontology(GO)and Kyoto encyclopedia of genes and genomes(KEGG)pathway enrichment analysis on differentially expressed genes in the KEGG.A total of 4 593 differen-tially expressed genes were obtained based on the transcriptome sequencing results of TVMV infection,among which the 3 564 genes were up-regulated and 1 029 genes were down-regulated.A total of 10 differentially expressed genes related to disease resistance were screened,6 were up-regulated and 4 were down-regulated.A total of 50 functional groups have been annotated in the GO database in three categories:biological processes,cell components and molecular functions.Among them,in the third class of molecular functions,the differentially expressed genes involved in protein binding function and ATP binding function were most significant,there were 501 and 453 differential genes,respectively.KEGG was enriched in 20 pathways,annotation to ribosome pathway has the highest degree of differential gene enrichment.307 differentially expressed genes were more sig-nificant.Protein binding,ATP binding,ribosome,eukaryotic ribosome biological reaction,DNA replication and other pathways may play an important role in the regulation of TVMV.The analysis of differentially expressed genes screened in transcriptome has laid an important foundation for further research on the resistance of Nicotiana benthamiana to TVMV virus.