First Report of Nothophoma quercina causing leaf spot disease of Phellodendron amurense in China(1).docx

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1、IFirstReportofNothophomaquercinacausing1.eafspotdiseaseofPhe1.1.odeiidron2 amurenseinChina345 Wcn1.IJiao.RuJunZhou94JunFanFu.IIaiJiaoXu.Departincn1.ofP1.an1.Patho1.ogy.Co1.1.egeofP1.an1.6 Protection,ShenyangAgricu1.tura1.University,Sbcnyang,1.iaoning,110866,China;NingHao,Departmentof7 ChineseHerba1.

2、MedicineCu1.tivationandIdcntif1.cation,Co1.1.egeofHorticu1.ture,ShenyangAgricu1.tura1.8 University,Shenyang.1.iaoning.110866,China91()P1.te1.1.odendmnanttreseRuprisaperennia1.herbinRutaceaefami1.ydistributedin1.iaoning.11Ji1.in,Hei1.ongjiangandInnerMongo1.iaprovinceofChina,whichusedto(rea1.jaundicea

3、nd12 gastrointestina1.diseasesas(hecommon1.ytraditiona1.Chinesemedicine(Zhang,c1.a1.2012).Inthe13 endofAugust2013.typica1.signsandsymptomsOf1.CafspotdiseaseonP.amurensewereobserved14 attheMedicina1.HerbGardeninShenyangAgricu1.tura1.University(SYAU),Shenyang,1.iaoning,15 China,withtheincidenceofdisea

4、sed1.eafrangingfrom45%to87%,Symptomsinitia1.1.yappeared16 assma1.1.,ova1.-circu1.ar,ye1.1.ow-brown1.esionson1.ower1.eaves,whichgradua1.1.yen1.arged,coa1.esced17 andsurroundedbyfaded-greenmargin,witheredanddefo1.iatedear1.ier.Symptomatic1.eaveswere18 washedwithwatersurfacc-s(cri1.izcdin75%ethano1.for

5、30sthen20%sodiumhypoch1.oritefor19 3-5min.and(henwashedthreetimeswithsteri1.izeddisti1.1.edwater.Thefragments(approximate1.y20 55mm)wereaseptica1.1.ycutfromthemarginsofthe1.esionsandthenp1.acedontopotatodextrose21 ager(PDA),andincubatedat25.Aeria1.myce1.iumwhi1.e,myce1.iumo1.ivaceousneartheco1.ony22

6、 centreand65mmdiamafter7donOA.MEAco1.oniesweremarginregu1.ar,aeria1.myce1.ium23 coveredthewho1.eco1.ony,whitetopa1.egrey.70mmdiamafter7dandb1.ackpycnidiaarescattered24 overthemediuminI5d.T1.iepycnidiawerea1.wayssuperficia1.onorimmersedintoagar,g1.obose25 andg1.abrous,measuring85.7-130.597.4-210.8m,w

7、ithstio1.atc.Conidiaweresubg1.oboseto26 ova1.,thin-wa1.1.ed,smoothandasetate,measuring5.28.53.15.2n,with2minuteguttu1.es.To27 confirmKoch,spostu1.ates,K)hea1.thy1.eaveswereinocu1.atedbysprayingwithconidia1.suspension28 (IO6COnidia/m1.)countedunderahacmocytomctcnIOcontro1.1.eavesweretreatedwithsteri1

8、e29 disti1.1.edwater,thena1.1.the1.eaveswerecoveredwithpo1.yethy1.enebagstomaintainhighmoist30conditions(8()%RH)andincubatedinthei1.1.uminationincubator(25QC)undera12hooperio1.31TheIypica1.spotsappearedona1.1.theinocu1.ated1.eavesaftertwoweeks,whereasthecontro1.p1.ants32remainedasymptomatic.Thesame

9、funguswasre-iso1.atedfromtheinocu1.ated1.eaves,satisfying33Koc1.fspostu1.ates.Formo1.ecu1.arana1.ysis,the1.argestsubunit1.argesubunitribosoma1.(1.SU)and34interna1.transcribedspacer(ITS)wasU1.nP1.ifiCdwithprimers1.R0R1.R7andITS4ITS5,35respective1.y,andsequenced.Phy1.ogcnctictreewasproducedusingMEGA4.

10、0.and36neighbor-joining(NJ)ana1.yseswereperformedusingthecombineddatasetsofthetworegions.he37 resu1.tsofphy1.ogeneticana1.ysisshowedhighsimi1.arwithsequenceofNothop1.iomaquerciu(CBS38 633.92).The1.SU(1.(X)3b)and1S(538b)oftherepresentativeiso1.ateweredepositedto39 GenBankasKY437I56andKY435896,respect

11、ive1.y.Therefore,onthebasisofmorpho1.ogica1.,40 cu1.tura1.ChUraCtCriSIiCSandphy1.ogeneticana1.ysis,thecausa1.agentof1.eafspotonP.amtrftsewas41 identifiedasNothophomuquercitu(veskampeta1.2010:Cheneta1.,2015).Toourknow1.edge.42 thisisthefirstreportof1.eafspotonP.amuren.secausedbyjV.quercinainChina.4344 Referencest45 ZhangGY.DongRJ.1.ian1.eta1.Journa1.ofShenyangPharmaceutica1.University.10:013,2012.46 AveskampMM,DeGruyterJ,WoudenbergJHC,eta1.StudMyco1.65：I-60,201().47 ChcnQ,JiangJR.ZhangGZ,C1.a1.StudMyco1.82:137-217,2015.