Identification and characterization of Phoma fungicola causing leaf spot disease of Phellodendron amurense Rupr in China.docx

IekntifkatiOnandcharacterizationofPhoinafungico1.acausing1.eafspotdiseaseofPhe1.1.adendnHiarnurenseRuprinChinaA1.ntractPhf1.1.otictuImfXamtrcnseRraperennia1.herbinC<>njx>sicacfami1.y,>chixIKCdforjaundiceandgastrointestina1.discaws.In2015.u1.eafspo<diseaseWaSQbSCrVCdonp1.antsofP.etnuremeinaproductionfie1.dofShenyang1.iaoningProvic.ChinaTbcdisschadaspeck1.edappea11i!xreiniIia1.1).Evemuu1.1.y.infec1.irusua1.1.yCaUMrdye1.1.owigofIiieka%,eundprvnu1.urudet<>1.iatin.Ihecau5a1.agentofinf&1.iunnh(swaxidenh11edasP/iojtmJint(ic(>1.abased»miHinrphoIogicM刖dCUkUra1.chaw1.cristies,pathogenicityCcMsarx1.phy1.ogenetica11A1.ysix.To<wrknow1.edge,thisisthefirstreportofakafspo<diseaseonP.iwwvrsecausedbyP.fwigko1.ainChina.!nt11iuc1.i<>nPhd1.micndrrffx(jjnu/rnxcRuprixaperennia1.medicina1.herbIU1.ivCtoChina,whereithasbeengrownfoemorethan2.0(X>years.Commercia1.p1.unutk>narcmuin1.>diribcdin1.iaoningPiovinccofChina.TbCdriedrccbarkofP.arnurensehasbeenusedforcrcatmeniofjaundiceaidgxUinteUina1.dsNiiw、inIradi1.iiiiu1.Chinesemedicine.Wi1.iifurthernreavhingini1.activeingredientaniextractionhasrecent1.ybeenQUdiCdfbriMa111.i(umnr.antimicrobia1.an<in1.herbio1.ogica1.activities.Insummerof2014.typica1.signsandsymptomsof1.eafspotdisease心UhmgmMSiOnSOahcIeavexofI*amunrnstfvrec»b>tfrveda!IheMedicina1.IktbGardeninShenyasigAgricu1.tura1.Univcrvi1.y(SYUXShenyanj.1.ianning,China.Sincethen,itsincidcrxx:inthesubsequentyear*Signifkunt1.yincrcnwdIn2015,thediseaseimposedUdirect(hrcuttoWMainabIcp<duion.Asthedisease<gfcsscd.IbC1.eafSPOgcncn11y0>a1.csccdtocauseb1.ighton1.eaves(issuesandhcfungu¼.PHtna.WaSiso1.atedfromIbeSC1.egions.thsuspectedha(WaSthecawa1.PaIbOgen“thediseaM!.PhonwisVrryimportantpytopathngcn(Avcxkiimpcta1.201Q)andthetypespeciesRherhnrnmWsctubtp(1857)wasOi1.aMishedbySaocurdo1.ISSO),MuchresearcheshnsbeenCaIr1.edoutta<consistsofa1.argenumber<cdophytcs.SaPfDbCSaidp1.antPaIhogCits.butIheCkUNiiiCakanofspecie%inPhtMm1.atx1.a1.1.iedgeneraissti1.1.c<Mi1.roveria1.Asubdivision<>1.theawxua1.gcnsPhomathatkcurrencywide1.yapp1.ieddividesthegcwsintoninew1.innx,inc1.udingthesectionsPjMinm、Hcenswra.Macnnra.Paraphww.PGrwtdktea、PhyU(KiiicU)IdestPiJWia.PIetKnhrmtsandSe1.enMne1.1.tiBocicnu1997).Confusingcharactersarereportedamongsevera1.PhomtiMXIiUIarx1.Ithtcdgtrtera>urhasAscwhyUhPhtNnopxix、Phy1.kziim.PkigphtNna.PyrtfntHrhatrRandSta)(NUMpt)n.KeuiI1.N<>JanextensiveMudyHe1.wtxnPkttmttandPhy1.kziictan1.ybased<)nIheh>Mp1.ain,invitroChaKICWrV卜UftberIIMire.i1.IabeenuggeaedIha1.thePhnmaC1.aviHca1.innyxtcmWOU1.dbeimpenvedbyadding11Nphy1.ogeneticdata,a11iUc1.inca1.in:morenatura1.gr<p.(Grondona以u1.1997;Torrcwia1.2005j.一Xfc阿JThediseasehda1.readybeendiscoveredandIUimedas1.eafspotm(年份).accordingtoIhCposi1.icmatk1.nup!x>k>>ca1.ikn1.1.ca1.in.Ihep1.hgewasdividedIIMDPhy1.kztida.Butt1.mg1.COCnbmXionofmoho1.ogica1.andmn1.ec1.irmethodsinchistudy,furtherMudiesshowedIha1.Ihcpathogenbe1.ongstoPhna.Materia1.sandmethodsObservationOfsymptom、andco1.1.ectiunofnmp1.e¼FO1.IoWinWinGc1.dob%cnuk>nnfdisc:Nedevck>p<ncn1.fivetnwendiyedp1.antsWCnrrandom1.ySC1.a1.Cdfrom11c1.dsintheShcnyangAgricu1.tura1.Univcn>ity.InnerSbcnyang.1.iaoningprovince.China.MftS±1.Ch4C1.rruMK<<PhomHjoNdcotoCWKit»wt"如ovWc>Q5IhO1.atiOntheCaUMdfed1.ttU“I咏rmctft20MBocrema,6.K.deGn>tecJ.N<wtoot,M.6.,&Kw,M.£.C.20049banqA*“*b<E<mu*dPtn*w*c*(ndr>M>raftMGCJtUrw.M1.InOOE:CABIPUdUNR,(Vpica1.1.yHymp1.oinatic1.eaveweresurtces1.eriIizedbydappedinto75%CU1.anD1.1.<w30s.rinsingthreetime*withsteri1.edisti1.1.edwater,andthenMKIkinginI%sodiumhypoch1.oriteso1.utionfor2minandrinsedinsteri1.edisti1.1.edwater.Sma1.1.piecesof(issues(approximate1.y5×5mm>WCrCaseptica1.1.ycutfrommarginsofthe1.esionsadp1.acedonpoutodextroseagar(PDA)(20Ogpotatoes.20gdextrose.20agnr/1.IJiMiI1.cdwa1.er"TbcwdhhcwereincubatedMHena1.u1111.ightantiobserveddai1.y.After5day、ofincuhatin11.thegrowingfungiWCTCiv>b1.cdantipurifiedonPDAusingthesing1.esporetechnique,transferredtoPDAs1.antsandkeptuc4Xforfurtherstudies.Thecu1.tureswereinvestigatedforco1.onydiameter.PyCnidiUinandCemidia1.InDrPiM1.k塔y.(Cu1.tura1.Charactcr1.Micsandmorpho1.ogica1.ThePhomaiso1.atewasfirM1.yexaminedmorpho1.ogica1.1.y.Theiso1.atewascu1.turedonPDA»<Mrtnca1.gar<OAj(15gofO1.iUnCa1.20gagan,1.Idisi1.1.edWdICfJundnw1.tagar(MEA;40gma1.1.extractOxid.15gOX(Iidagar.111.apwa1.er)incubated31.25'Cucoxitinuousi1.Iuinuia1.ion.atdexaminedco1.onydiiimctcOntheOAax!(bePnAafter7d.Twoweek、1.ater,thetn<>rphn1.c>gyofthePyCnid山andconidiaweirMitdicdfromtheOAandIhCMEAcu1.tures.Imagesoftheco1.onicsoftheiso1.atewerecapturedUstngacamcra.Theexperimentrepea1.edtwicewithRmrrep1.icates.DNCX1.ruc1.HMi.PC?RandNeqUenCinXDNAexIrac1.iun.(heMudiedu1.1.uxe%wetvgro11PD11edtumatroomIempcra1.urvuarMaryshakerfor7dayx.ECnCmiCDNAswereexIrxr1.aifrommyce1.iabygn«TIAMGENI>、AsecureP1.antKitDcconiingtothemanufacturer'sinanitions.1.a