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1、植物病理学报ACTAPHYTOPATHOLOGICASINICA37(4):368-376(2007)SequenceAnalysisoftheCoatProteinGeneofBeanyellowmosaicvirusIsolatesfromFababeaninYunnan,ChinaandSyria1,21*2QIPeng,WANGXiao-ming,HEYue-qiu(1TheNationalKeyFacilityforCropGeneResourcesandGeneticImprovemen,tNFCRI,Instit
2、uteofCropScience,ChineseAcademyofAgriculturalSciences,Beijing100081,China;2PlantProtectionCollege,YunnanAgricultureUniversity,Kunming650201,China)Abstract:ViraldiseasesareoneofthemajorfactorsforyieldlossoffababeaninYunnanprovince,China.Beanyellowmosaicvirus(BYMV)was
3、identifiedinfababeansamplesbyTissueBlotImmunoassay(TBIA).AccordingtoBYMVsequencesubmittedtoGenBank,apairofprimerwasdesignedandusedtoamplifythecoatprotein(CP)geneofBYMVinsixfababeanisolates,fivefromYunnanprovinceandonefromSyria,byreversetranscriptionPCR(RT-PCR).Theam
4、plifiedfragmentwasclonedandinwhich822nucleotideswasforCPgene.TheCPgenewasfoundtobe86.4%to100.0%and96.7%to100.0%ho-mologousamongsixBYMVisolatesatboththenucleotideandaminoacidleve.lTheCPgeneofsixse-quencedisolatesshowed79.1%to97.9%and83.5%to98.5%nucleotideandaminoacid
5、identitieswith34BYMVisolatesinGenBank.ThecomparisondataofCPgenesequencesshowedthatBYMV-ChinesefababeanisolateswerecloselyrelatedtotheBYMV-Japanesefababeanisolates.TheNAGmotif,abasicstructureforaphidtransmissionoftheviruslocatedattheN-terminusofCPgeneinBYMV,waspresen
6、tedinallofsixisolates.Keywords:Beanyellowmosaicvirus;fababeanisolate;coatprotein;sequenceanalysis1,212菜豆黄花叶病毒中国和叙利亚蚕豆分离物外壳蛋白的序列分析亓鹏,王晓鸣,何月秋12(中国农业科学院作物科学研究所国家农作物基因资源与基因改良重大科学工程,北京100081;云南农业大学植物保护学院,昆明650201)摘要:病毒病是影响云南省蚕豆生产的重要病害。对采集的蚕豆病毒病标样进行了组织印迹法检测,表明菜豆黄花叶病毒(BYMV
7、)是最主要的病原。据此,以BYMV基因的保守序列设计了一对特异性引物,用BYMV的5个中国云南蚕豆分离物和1个叙利亚蚕豆分离物侵染的蚕豆叶片总RNA为模板,RT-PCR扩增获得了长度为907bp的目标片段。序列分析显示,此片段中包含822bp的外壳蛋白序列。6个分离物间的外壳蛋白核苷酸和推导编码蛋白质的氨基酸序列的同源性分别为86.4%~100.0%和96.7%~100.0%。与GenBank登录的34个具有完整外壳蛋白序列的BYMV分离物进行同源性和系统进化树分析的结果表明,6个分离物在核苷酸和氨基酸水平上与其它分离物的同源
8、性分别为79.1%~97.9%和83.5%~98.5%,BYMV中国蚕豆分离物与日本蚕豆分离物同源性最高。外壳蛋白基因的序列特征揭示,在BYMV中Receiveddate:2006-02-25;Reviseddate:2006-08-14Foundationitem:ACIA