1、Morpho1.ogica1.andmo1.ecu1.aridentificationofPowderyMi1.dewofC1.ematismanshu11caCausedbyErysipheCiqui1.egiaeinChinaC1.ematismanshuricaPupr.isaperennia1.herbintheRaiiiincu1.aceaefami1.y,whichhasbeenusedasava1.uab1.etraditiona1.Chinesemedicine.Between2012and2013.apowderj,mi1.dewcausingsignificantdamag
2、etop1.antsofC.manshuricaoccurredintheMedicina1.HerbGardeninShcnyangAgricu1.tura1.University.1.iaoningprovince.China.Thefungimain1.yinfectedthe1.eaves,sta1.ksandpetio1.es.Symptomsinitia1.1.yappearedascircu1.artoirregu1.ar,thin,whiteco1.oniesonyoung1.eavesandstems,especia1.1.yontheadaxia1.surface.Even
3、tua1.1.yinfectionsusua1.1.ycausedye1.1.owingofthe1.eavesandma1.formationandprematuredefo1.iation.Numerousc1.eistotheciaappearedonthebothsurfacesof1.eaves,sta1.ksandPetiO1.eS.hefungusfromsymptomatictissueswasidentifiedasErysipheaquiIegiaebasedonmorpho1.ogica1.characteristicsandrDNAITSsequenceana1.ysi
4、s.Toourknow1.edge,thisisthefirstforma1.reportofpowderymi1.dewonC.manshuricacausedbyE.aquHegiaeinChina.C1.ematismanshuricaPupr.isaperennia1.herbintheRaiumcu1.aceaefami1.y,andiswide1.ydistributedinthenortheastofChina.Thedriedrootsandrhizomesofthep1.antsnamedWei1.ingxi(intheChinesePharmacopoeia,ispopu1
5、ar1.yUSedasanantiindaininatory,antitumor,ana1.gesicagentintraditiona1.ChinesemedicineforthousandsofyearsinAsia(14).InAugustof2012.apowdcrj,mi1.dewonC.manshuricawasfirstobservedinMedicina1.HerbGardenofShenyangAgriCU1.1.UraIUniversityin1.iaoningprovince.China.Incidenceofpowderymi1.dewofC.manshuricawa
6、sinvestigated.Theincidenceofthediseasewasratedintermsofpercentageofinfected1.eavesamong10()1.eavesobscn,cdwithfiverep1.icates.The1.eve1.ofincidencewassignificant1.ydifferentinthepasttwoyears.In2012.theincidencewas1.ower,justunder5%.However,1.hcdiseaseincidencesharp1.yincreased,with42%to100%(averageo
7、f78%)inthesamefie1.dsin2013.Theribosoma1.DNAinterna1.transcribedspacer(ITS)regionhasbeenusefu1.fordiscriminatingfungiatthespecies1.eve1.,andhasbeenusedextensive1.yfor(heidentificationandphy1.ogeneticana1.ysisofpowderymi1.dews(I,2.7.10.II.12).Inthispaper,theaimwastoidentifythecausa1.agentofPoWderymi1
8、dewobservedonC1.ematismanshuricainChinaandseiupabaseforcienificcontro1.ofthedisease.Materia1.sandmethodsFunga1.materia1.sandmicroscopy.Thesamp1.esofC.manshuricainfectedwithpowderymi1.dewwereobtainedfromtheMedicina1.HerbGardenofShenyangAgricu1.iura1.University,1.iauningprovince.China.Themorpho1.ogyo
9、fthepathogenwasexaminedwithfreshmateria1.susing1.ightmicroscopyaspreviousdescribedmethods(10).DNAextractionandamp1.ificationofr1.)NAITSandSequencing.Funga1.DNAsamp1.eswerepreparedbyco1.1.ecting(8)youngpowderymi1.dewmycc1.iaandc1.eistothecia(6)usingsteri1.ebrashes.Thenuc1.earrDNAregionspanningtheITS1
10、5.8SandITS2rRNAgenewasamp1.ifiedbyPO1.y1.neraSechainreaction(PCR).Primers11-SI11S4wereusedfortheamp1.ification,whichWaSPCrft)EMXIinatota1.reactionvo1.umeof501.inc1.udingthefo1.1.owingreagents:5.01.10PCRbuffer.2.01.DNAtemp1.ate.41.MgCh(IOmM).1.01.ofeachprimer(11SIand1TS4).0.61.TaqDNApo1.ymerase(5.()
11、U1.).1.01.dNTP(dRICTP,dGTP,dTTRIOinMeach),35.41.ddH2O.A1.1.abovereagentswerepurchasedfromSangonBiotechno1.ogy1.td(Shanghai.China).ThePCRcyc1.ingconditionswere94for3minfo1.1.owedby4cyc1.esof94forImin.55for1min.72for1.5min.andafina1.extensionstepof72for8nininaS1(XX),mthe11na1.cyc1.er(Bi-Rateaqui1.eiae
12、onC1.ematismanshurica(a:C1.ciskHhcciaembeddedinmyce1.iumon1.eaf:b:C1.cistaihccia,asciandappendage;c:Conidiophoreandconidia;d:Asciandascospores)Phy1.ogeneticAna1.ysis*TofurtherconfirmtheidentiI1.caiionofecausa1.fungus,theITSregionofrDNAwasamp1.ifiedusingtheprimersITS1ITS4andsequenceddirect1.y.Theresu
13、1.ting619-bpsequencewasdepositedinGcnBank(AccessionNo.KJO2O911).Thesequencewasmostsimi1.ar(1(X)%)tothatofE.acui1.egiae(AccessionNo.EUO47570,DQ335569,AFI54322,ABO1.5929,EU047569andAY452801).Thephy1.ogeneticIree(Fig.3)wasobtainedusingMEGE4.0withthecighbor-joiningmethods,andshowed(hattheiso1.atesfromC.
14、manshuricainChinaformedaWe1.ITUPpOrtedc1.adethatgroupedWiththeE.aqui1.egiaereferenceiso1.ates.T1.ieseresu1.tsindicatedthattheiso1.atesofE.aqui1.egiaewereconspecific.Onthebasisofmorpho1.ogica1.characteristics.ITSsequencesimi1.aritiesandphy1.ogeneticana1.ysis,theiso1.atewasidentifiedasErysipheaqui1.eg
15、iaeDC.Fig. 3 Phy1.ogcnctictreewithbtstrapva1.uesbasedonthenuc1.coidcsequencesoftherDNAinterna1.transcribedspacerregionsusingI,(XX)rep1.icates.Theiso1.atefromC.mshuricaisindicatedinb1.ueibn1.Pathogenicity.Paihogenicitywasconfirmedthroughinocu1.ationbygent1.ydustingconidiaonto1.eavesoffivehea1.thy,po1
16、1.edC.manshuricap1.ants.Fivenon-inocu1.atc1.p1.antsservedascontro1.treatments.Thep1.antsweremaintainedingreenhouseat20to25andabout60%80%re1.ativehumidityundernatura1.day1.ight.Typica1.synonsofow1.erymi1.dewoninocu1.atedp1.antswerereproducedafterI(M4days,whereascontro1.p1.antsremainedhea1.thy.Thefun
17、guspresentontheinocu1.atedp1.antsWaSmorpho1.ogica1.1.yidentica1.withthatorigina1.1.yobservedonthediseasedp1.ants,fu1.fi1.1.ingKochspostu1.ates.DiscussionC.manshuricahasbecomewide1.ycu1.tivatedinrecentyearsasaChinesetraditiona1.herbinnortheast.China.Intheproductionfie1.d,spotb1.otch(5)andb1.ackSpO1.(
18、9)arethemaindiseaseonCnumshurica.Tbourknow1.edge,thisstudyisthefirstforma1.reportOfPoWderymi1.dewonCmanShUriCacausedbyE.ac/ui1.egiaeinChina.Wcdescribedindetai1.thetypica1.symptomsoninfectedp1.antsandmorpho1.ogica1.characterizationoftheiso1.ate,andcombinedmorpho1.ogica1.characteristics,sequencesofrDN
19、AITS,andpathogenicitytests(o(Jemonsiraiethatthecausa1.agentofpowderymi1.dewonC.nunshuricaisE,(ud1.egiae.Suchinformationwou1.dbehe1.pfu1.forgrowerstosuccessfu1.1.ymanagethedisease.Furtherresearchisrequiredtogaininformationre1.atingtopotentia1.contro1.measuresandcu1.tura1.practicesforthereductionofthe
20、disease.E.aqi1.eiaeisacommonPoWdCrymi1.dewandhasbeendistributedinJapan.India.Fin1.and.Po1.and.Eng1.and.GermanyandChina,andmain1.yinfectedthep1.antsofAqTegia,Aconitum3Anemone,Cahha.C1.emaiis9CoptisyPu1.sati1.1.a,Raniuiciuhis.Tha1.ictnunandsoon(23.4.6).Andmorerecent1.y,itwasreportedthatEaqui1.egiaewas
21、a1.socausingpowderymi1.dewonthep1.antsofApocynaceae(13)andCasuannceae(15).Inthesurveyin2012.theincidenceofpowderyMi1.dewofCmanshuricawasquite1.ow,butwhichsharp1.yincreasedin2013,withaverageof78%.Mcan1.imcnumerousc1.cis1.ohcciaappearedonthe1.eaves,sta1.ksandpetio1.es.Withinfectionsourcesaccumu1.ated,
22、E.ati1.egiaehasgreatpotentia1.itytobecomeanimportantpathogenofGmanshurica.Consequent1.y,itisnecessary(ofurtherstudyonthemechanismofitsdissemination.1.iteratureCited1. Cook.R.T.Henrico(.B.Irici.A.&Bea1.es.P.2006.Morpho1.ogica1.andphyIogenc1.iccomparisonsamongstpowderymi1.dewsonCata1.paintheUK.Myco1.o
23、gtca1.research110(6):672-6S5.2. Cunning1.on.J.H.,1.awrie、A.C.andPascoe,I.G.2(X)4.UnexpectedHbosoma1.DNAinterna1.transcribedspacersequencevariationwithinErjsiphcaquiIegiaescnsuIa1.o.Funga1.Divers.16:i-10.3. Cunnington.J.H.Takamatsu.S.1.awrie.A.C.andPascoe.1.G.2(X)3.Mo1.ecu1.aridentificationofanamorph
24、icpowderymi1.dews(ErysiphaIcs).Ausirahs.P1.antPath.32(3):421-428.4. Editoria1.boardofFkwasprc-prducingp1.antsofChineseAcademyofSciences.1987.F1.orafungorumJiiniCOrUm:Ersipha1.es.Sciencepress(Beijing.China).5. Fu.J.E.Su.W.N.Zhou.R.J.Sun.J.M.andWang.D.Z.2O!3.ThepathogenIdcniifkationandbio1.ogica1.char
25、acteristicsofC1.enunismanxhuricaspotb1.otch.Journa1.ofSbcnyangagricu1.tura1.university44I):26-31.6. Hirata.T.andTakama1.su.S.1996.Nuc1.cotidesequencediversityofrDNAinterna1.transcribedspacersextractedfromconidiaandc1.eistotheciaofsevera1.powderymik1.cwfungi.Myooscicncc,37(3):283-288.7. Hirata.T.Cunn
26、ington.J.H.Paksiri.U.1.iinkaisang.Shishkoff.N.Grigahunai1.e.B.Sa1.o.Y.andIakanuitsu.S.2(XX).Evo1.utionanrana1.ysisofsubsectionAa覃IiCeIhUQ1.aeofPodosphaerasectionSphaemtheca(Erysi)ha1.es)basedontheIDNAi11erna1.IranSCribedspacersequenceswithspecia1.referenceIohost.CanadianJourna1.ofBotany78:1521-1530.
27、8. Jankovics,T.Bai.Y.Kovacs,G.M.Bardin.M.Nicose1.yre1.atedpowderymi1.dewfungiinfectingvariousp1.antspecies.Phytopatho1.ogy98(5):529-54().9. Jiang,N.,Huang,Y.C.,Tang,M.Q.、1.in.Y.,1.iu.W.andTan,Y.2010.ktenifkationofC1.ematischinensisb1.ackaxtandscreeningofeffectivefungicides.ChineseAgriCUkUndScienceBu
28、1.1.etin26(24):221-225.10. Kiss.1.Cook,R.T.A.,Saenz,G.S.,Cunnington,J.H.,Takamatsu,S.Pascoc,1.,Bardin.MmNiCOersicisp.nov.andO.1.yiY)perxici,infectingtomatoinindficrentpartsofthewor1.d.Mye1.Res.105(6):684-697.11. Kiss.1.Takamacsu.S.andCunningionJ.H.2005.Mo1.ecu1.aridentificationofOidiumneo1.ycv,nicia
29、s(hecausa1.agentof(herecenttomatopowderymi1.dewepidemicsinNorthAmerica.P1.antDis.89(5):491-496.12. KovcsGMiJankovicsT,Kiss1.201.I.VariationinthenrDNAITSsequencesofsomePOWderymi1.dewspecies:doroutine111.cxu1.aridcn1.ifica1.ionprocedureshideva1.uab1.einfo11na1.ion?.Eur.J.P1.antPatho1.131(1):135-141.13
30、 1.ihcrato,J.R.andCunnington,J.H.2(X)6.FirstrecordofErysipheuui1.egiaeonahostoutsidetheRanuncu1.aceae.Austra1.as.P1.antPath.35(2),291-292.14. Shi.S.P.Jiang.D.Dong.C.X.andTu.P.E2(X)6.1.ignansfromtherootsandrhizomesofC1.eniaiismanshurica.ZcitschriftfurNaiurfbrschungB-Jouma1.ofChemica1.Sciences61(10):1299-1303.15. Wo1.can.S.andPark.J.H.2012.FirstConfimwdReportofPowderj-Mi1.dewCausedbyErysiphcCtqMi加WiagonCasuarmacuminfhamianainArgentina.P1.antPatho1.J.28(4).453-453.
