【word】 苦瓜青枯病病原菌的鉴定（英文）

【word】 苦瓜青枯病病原菌的鉴定（英文） 苦瓜青枯病病原菌的鉴定(英文) 植物病理 ACTAPHYTOPATHOLOGICASINICA40(3):315-316(2010) IdentificationofPathogenCausing BalsamPearBacteriaIWlIt YUANGao—qing,XIONGYing,YEYun—feng’,LIQi—qin,LINWei (CollegeofAgriculture,GuangxiUniversity,Nanning530004,China;Guang~BotanicalGardenofMedicinalPlant,Nanning530023,China) Abstract:BalsampearbacterialwiltwasobservedinNanning,Guangxi,China.Basedonbacteriological identification,pathogenicitytestsand16SrDNAsequencinganalysis,thepathogenwasidentifiedasRalstonia solanacearum(Smith)Yabuuchieta1.ThisisthefirstreportofR.solanacearumcausingbacterialwilton balsampearinChina. Keywords:balsampearbacterialwilt;Ralstoniasolanacearum;pathogenidentification 苦瓜青枯病病原菌的鉴定袁高庆,熊英,叶云峰一,黎起秦,林纬(广西 大学农学院,南宁 530004;.广西药用植物同,南宁530023) 文章编号:12-0914(2010)03-0315-02 Balsampear(MomordicacharantiaL.)isone ofthemostpopularvegetablesinChina.In2006,a newwiltdiseasewasobservedonbalsampearin Nanning,Guangxi,China.Diseaseincidence rangedfrom7%to15%,withthesymptomap— peared5-7weeksaftertransplanting.Inorderto controlbalsampearwilt,apreliminarystudyon identificationofthepathogenwascarriedout. 1MaterialsandMethods 1.1Pathogenstrains Pathogenstrainskgqk一1,kgqk-2andkgqk?3 wereisolatedfromdiseaseplantscollectedfromWu— tangtownofNanning,withNAmediumcontaining (g/L)peptone,5;yeastextract,l;beefextract, 3;glucose,10;agar,18.StrainLHGI(isolated fromSiraitiagrosvenoriiwilt)andTM一5(isolated fromtomatobacteria1wilt)wereusedascontro1for bacteriologicaldetermination. 1.2Bacteri0IOgicaIdetermination Bacteriologicalcharactersofstrainkgqk?1in— cludingculturalcharacteristics,morphology,stain— ingreaction,physiologicalandbiochemicalproper— ties.weredeterminedbyconventionalmethods{,. 1.3Pathogenicdetermination Pathogenstrainskgqk—l,qk一2andkgqk一3 wereinoculatedon20balsampear,spongegourd, waxgourd,pumpkin,cucumber,tomato,tobacco andkidneybeanplantsbythestemtechniquerespec— tively.Theconcentrationofbacteriumsuspension wasl0cfu/mL.Thecontrolplantswereinoculated inasimilarwaywithautoclavedwater. 1.416SrDNAsequencinganalysis TheDNAregioncoding16SribosomalRNAof strainkgqk一1wasamplifiedwiththeuniversal Receiveddate:2009-06一I9;Reviseddate:2010-02-22 Correspondingauthor:LIQi—qin,Professor,focusOnthebiologicalcontrol ofplantdiseases;E-mall:qqli5806@gxu.edu.cn 316植物病理40卷 primersf27andr1492(forward:5’-AGAGTTT— GATCCTGGCTCAG一3:Reverse:5一TACGGC— TACCTTGTTACGACTT-3,andthenthe16S rDNAsequencewascomparedwiththoseofrelated speciesinGenBankbyBLASTsoftware. 2Results 2.1Symptomsofbalsampearbacterialwilt infectednaturally Initially,theupperleavesorsideshootsofin— 4dayslater,allthe fectedplantsbecamewilted.3— leavesandsideshootsofdiseasedplantswiltedbut keepingleavesgreen,vascularbundlebecamebrown andoozedoutonthecutsurfaceasgreyishwhite masses.Thewholeplantswiltedanddied6—9days afterthefirstsymptomsappearance. 2.2BacteriolOgicaIcharacters ThesinglecolonyofallthreestrainsonNAme— diumwerenearlyround,swellup,smooth—edged, greyishwhite,notransparent,andtheircolourbe camelightpinkafterculturingfor3daysandbrown atlast.OnNAmediumcontaining0.1%2,3,5一 Triphenyltetrazoliumchloride(TrC),allsingleco? lonieswerenearlyround,swellup,greyishwhite withlightpinkcentre.Theseculturalcharacteristics werethesaffleasthoseofcontrolstrains.Thecells ofpathogenweregramnegative(G一)andappeared shortbacillaryinshapewith4polarflagella, (0.8—1.35)×0.37ixm一0.56ixminsize. Strainkgqk一1couldusemannitol,sorbitol,dul— citol,maltose,lactose,cellobiose,reducednitrate, saccharose,D—fructoseanddextroseascarbon sourceandtryptone,beefextract,yeastextract,po— tassiumchloride,potassiumnitrateandureaasnitro— gensourceforitsgrowth.D—xyloseandpectinose werenotsuitableforitsgrowth.Thisstraincould notgrowinthemediumcontainingammoniumox— alateandammoniumchloride.Thestraingrewatthe temperaturerangeof16—40?andpH5.5—9.5.with theoptimaltemperature28—31?andpH7.0. 2.3Pathogenicity Typicalsymptomswereseenoninoculatedbal- sampear,spongegourd,waxgourd,tomatoandto? baccoplants4daysafterinoculation.andin7一l0 daysallplantswiltedanddied.Bacteriawiththe samecolonymorphologyasthoseinoculatedwere reisolated.Nosymptomsdevelopedoncontrol plants.All3strainsinducedhypersensitivereactions ontobacco.However.thesestrainscouldnotin. fectpumpkin,cucumberandkidneybeanplants. 2.416SrDNAsequencinganalysis Endsof1428bpPCRproductofstrainkgqk一1 weresequenced(GenBankaccessionnumber FJ184057).Thesequenceobtainedwere99%iden— ticalwiththecorresponding16SrRNAgenese— quencesofmanyRalstoniasolanacearumstrainsin GenBank(GenBankaccessionnumberDQ924953, EF585154,AL646053,AB024607,FJ210681, U28220,AY464977andsoon). Basedonbacteriologicalidentification,patho— genicitytestsand16SrDNAsequencinganalysis.the pathogencausingbalsampearbacterialwiltwas identifiedasRalstoniasolanacearum(Smith) Yabuuchieta1..ThisisthefirstreportofR.so— lanacearumcausingbalsampearbacterialwiltin China. References DongXZ,CaiMY.Systematicdeterminativemanual ofgeneralbacteria(inChinese)[M].Beijing: SciencePress.(北京:科学出版社),2001.1—419. FangZD.Researchmethodofplantpathology(Third Edition)(inChinese)[M].Beijing:ChinaAgricul— turePress(北京:中国农业出版社),1998.156— 232. LaneDJ.16S/23SrRNAsequencing[A].Stacke— brandtE,GoodfellowM.NucleicAcidTechniquesin BacterialSystematics[M].Chichester,UK:John Wiley&Sons.1991.115—175. KlementZ,FarkasGL,LovrekovichL.Hypersensi— fivereactioninducedbyphytopathogenicbacteriainthe tobaccoleaf[J].Phytopathology,1964,54:474— 477. 责任编辑:于金枝 ]J]J]J]j-二