First Report of Nothophoma quercina Causing Leaf Spot Disease of Phellodendron amurense in China.docx
IFirstReportofNothop1.iomaquercinaCausing1.eafSpotDiseaseof2 Phe1.1.odendronamurenseinChina345 Wcn1.IJiao.RuJunZhou94JunFanFu.IIaiJiaoXu.Departincn1.ofP1.an1.Patho1.ogy.Co1.1.egeofP1.an1.6 Protection,ShcnyangAgricu1.tura1.University,Sbcnyang,1.iaoning,110866,China;NingHao,Departmentof7 ChineseHerba1.MedicineCu1.tivationandIdcntif1.cation,Co1.1.egeofHorticu1.ture,ShenyangAgricu1.tura1.8 University,Shenyang.1.iaoning.110866,China91()Phe1.1.odendmnamurenseRupr(Riuaceae)isaperennia1.herbdisiributedin1.iaoning,Ji1.in.11Hei1.ongjiang,andtheInnerMongo1.iaprovincesofChina,whichwasusedasatraditiona1.Chinese12 medicinetotreatjaundiceandgastrointestina1.diseases(Zhangcta1.2012).A1.theendofAugust13 2013.typica1.signsandsymptomsof1.eafspotdiseaseonP.amurensewereobservedatthe14 Medicina1.HerbGardeninShenyangAgricu1.1.ura1.University(SYAU),Shenyang,1.iaoning,China.15 Thediseaseincidencerangedfrom45%to87%ratedbythepercentageOfinfcctcd1.eavesamonga16 tota1.of5001.eaves(5rep1.icates,eachwith1001.eaves).Symptomsinitia1.1.yappearedassma1.1.17 ova1.-circu1.ar,ye1.1.ow-brown1.esionson1.ower1.eaves,subsequent1.yen1.argedandweresurroundedby18 afaded-greenmargin.1.aterthe1.eaveseventua1.1.ywitheredanddroppedoffthep1.antear1.y.19 Symptomatic1.eaveswerewashedwithsteri1.ewater,surface-steri1.izedin75%ethano1.for30s.1%20 sodiumhyx)ch1.oritefor3-5min.andthenwashed3timeswithsteri1.ewater.1.eaffragments21 (approximate1.y5×5nn)wereaseptica1.1.ycutfromthemarginsofthe1.esionsp1.acedonto<a(o22 dextroseagar(PDA),andincubatedat25.Theaeria1.myce1.iumwaswhite,andthemyce1.ium23 waso1.ivaceousneartheco1.onycenter.65minindiameterafter7donoatmea1.agar(O).Ma1.t24 extractagar(MEA)co1.onieshadregu1.armargins,andwhi1.e1.opa1.egreyaeria1.myce1.iumcovered25 theentireco1.ony.70mmindiameterafter7d.B1.ackpycnidiawerescat1.crox1.overthemediumat1526 d.hepycnidiawereg1.obose.g1.abrous,ostio1.ate.and85.7-130.5×97.4-210.8m.Conidiawere27 Subg1.obosetoova1.,(hin-va1.1.e<1.,smooth,andasep(ate,measuring5.2-8.5×3.1-5.2m,with228 minu(cguttu1.cs.Tbconfirmpathogenicity.IOhea1.thy1.eaveswerespray-inocu1.atedwithspore29 suspension(1('conidia/m1.).whi1.eIOcontroI1.eavesweretreatedwithsteri1.ewater.A1.1.1.eaveswere30 coveredwithpo1.yethy1.enebagsomaintainhumidconditions(>80%RH)andincubatedina31 incubator(25)undera12hphotopcritx1.Typica1.spotsappearedona1.1.theinocu1.ated1.eaves32 after14d.whereasthecontro1.p1.antsremainedasymptomatic.Thesamefunguswasre-iso1.ated33 fromtheinocu1.ated1.eaves,satisfyingKoc1.Vspostu1.ates.Formo1.ecu1.arana1.ysis,the1.argesubunit34 (1.SU)andinterna1.Iranscribcdspacers(ITS)oftheribosoma1.DNAwereamp1.ifiedwithprimers35 1.R0R1.R7and1TS4ITS5respective1.y,andtheamp1.iconsweresequencedanddepositedin36 GenBankundertheaccessionnumbersKY437156andKY435896.SequencequeriesontheB1.ASTsearchengineofNCBIGcnBank(WWW.ncbi.n1.m.nih.gov/griibank/confirmedthatthespeciesbe1.ongstothegenusNothophoma.Phy1.ogenetictreebasedonneighbor-joining(NJ)ana1.yseswiththecombineddatasetsofITSand1.SUusingMEGA4.0showedhighsimi1.aritywiththesequenceofNoihophomaquercina(CBS633.92).Therefore,onthebasisofmorpho1.ogica1.,cu1.tura1.characteristicsandphy1.ogeneticana1.ysis,thecausa1.agentof1.eafspoton1.UmMrenSewasidentifiedasNothophomaquercina(Aveskampea1.,2010;Cheneta1.,2015).Toourknow1.edge,thisisthefirstreportof1.eafspotonP,amurensecausedby/V.(uerci11ainChina.ReferencesZhangGY,DongRJ,1.ianI.,eta1.Journa1.ofShenyangPharmaceutica1.University.10:013,2012.AvcskampMM,DcGruytcrJ,WoudcnbergJHC,cta1.StudMyco1.65:I-60.2010.373839404142434445464748ChcnQ.JiangJRtZhangGZ.cta1.StudMyco1.82:137-217.2015.
