植物生理与分子生物学：植物病原真菌致病机制

植物病原真菌致病机制2014年5月27日11、什么是真菌?2、哪些是植物病原真菌?3、植物病原真菌的本质?

4、植物病原真菌致病策略?

5、植物病原真菌致病工具分子6、植物病原真菌致病机制研究手段7、禾谷镰孢致病策略提纲2什么是真菌?3Fusariumverticillioides4FungiHeterotrophic,lackingchloroplastsPossessacellwallandvacuolesLifecycle,haploidanddiploidHyphalgrowth,tipgrowthVersatileinmetabolitesbiosynthesisSurfaceexpertDimorphicgrowthswitch(yeast,hyphae)Fungalcellwallcomposedofglucansandchitin,butnocellulosemodifiedfromMeyerowitz2002ScienceFungi0.1Gadicotmonocotsplit5animalplantfungi6ModifiedfromHedges,2002,NatRevGenet3:838-849;Schochetal.,2009,SystBiol58:224-239盘菌亚门(Pezizomycotina)散囊菌纲外囊菌亚门（Taphrinomycotina）酵母菌亚门Ascomycota子囊菌门Basidiomycota担子菌门锈菌纲黑粉菌纲Aphylogenyoffungi7gAphylogenyofplants8Plant-FungalinteractionsPathogen(病原菌)Symbioticfungi(共生菌)Endophyte(内生菌)Saproph(腐生菌)9PlantproducerFungusdegraderEcosystemAnimalconsumerFungiarethemostcommonparasitesofplants.Allhigherplantspeciesaresusceptibletooneormorefungaldiseases,andmostcropplantshavemany.PlantpathogensFungiBacterialVirusOomyceteNematode10哪些是植物病原真菌?1112Magnaportheoryzaeafilamentousascomycetefungus,isthecausalagentofriceblastdisease,themostdestructivediseaseofriceworldwide13由子囊类真菌引起的水稻病害稻瘟病稻曲病胡麻斑病恶苗病烂秧病稻瘟病菌稻绿核菌稻平脐蠕孢串珠镰刀菌禾谷镰刀菌尖孢镰刀菌…142.BotrytiscinereaBotrytiscinereaPersoon:Fries[teleomorphBotryotiniafuckeliana(deBary)Whetzel],knownasgreymould,caninfectmorethan200plantspecies.Thefungusisconsideredasatypicalnecrotroph,whichco-optsprogrammedcelldeathpathwaysinthehosttoachieveinfection.Botrytiscinereaismostdestructiveonmatureorsenescenttissuesofdicotyledonoushosts.Underspecificclimaticconditions,B.cinereamaycausenoblerotingrapeberries,whichareusedtoproducesweetwines(Sauternes,Tokaj).ThemostprestigiousBotrytiswinesaresoldforpricesupto€500/bottle.153.Pucciniaspp.Threerustdiseasesoccuronwheat,namelystem(black)rust(causedbyPucciniagraminisf.sp.tritici)(Pgt)(Fig.5),stripe(yellow)rust(P.striiformisf.sp.tritici)(Pst)(Fig.6)andleaf(brown)rust(P.triticina)(Pt).Pgt,PstandPtareobligate,biotrophicbasidiomycetefungiwithmacrocyclic,heteroeciouslifecyclesTheoccurrenceofraceUg99ofPgtinEastAfricawithvirulenceforSr31(Pretoriusetal.,2000),acommonlyusedresistancegene,hasrenewedstemrustresearch(Singhetal.,2011).Historically,stemrusthasbeenmostnotoriousfordamagingwheatcrops.ThediseasewasfearedinancientRomewhererituals(‘Robigalia’)wereperformedtosavecropsfromrust(Zadoks,1985).16WheatstriperustPucciniastriiformisWestendf.sp.tritici(PST)17TheascomyceteFusariumgraminearum(teleomorphGibberellazeae),whichresidesintheorderHypocreales,isahighlydestructivepathogenofallcerealspecies().Locally,F.graminearumco-existsandco-infectswithotherFusariumspecies.Fusariumgraminearumproducesseveraltrichothecenemycotoxins,themostimportantofwhicharedeoxynivalenol(DON),acetylatedDONderivatives,nivalenolandthephytoestrogenzearalenone.DONbindstothepeptidyltransferaseproteinintheribosomeandinhibitsproteintranslation.Differentnaturalisolates(termedchemotypes)producedifferentmycotoxintypes(Alexanderetal.,2011).ControlofFusariumfloralinfectionsremainsproblematic.Inmostcerealspecies,theresistancesourcesidentifiedareonlypartiallyeffectiveandaremajorQTLbased(Buerstmayretal.,2009).Someazolefungicidesaremoderatelyeffective,butspraycoverageandthetimingofapplicationsremaindifficult.4.Fusariumgraminearum185.FusariumoxysporumFusariumoxysporumSchlecht.isaubiquitoussoil-bornepathogenthatcausesvascularwiltonawiderangeofplants.Characteristicdiseasesymptomsincludevascularbrowning,leafepinasty,stunting,progressivewilting,defoliationandplantdeath(Agrios,2005).196.BlumeriagraminisBlumeriagraminisisanascomycetebelongingtotheErysiphales.Itcausespowderymildewsofgrasses(Fig.10),includingwheatandbarley.207.MycosphaerellagraminicolaTheascomyceteMycosphaerellagraminicola(anamorphSeptoriatritici)isintheorderDothidealesandcausesSeptoriatriticiblotch(STB)diseaseofwheat.218.Colletotrichumspp.22Shenetal.,2001,Mycol.Res.23CornAnthracnoseStalkRotFromwebsiteofPlant&PestDiagnosticLaboratoryatPurdueUniv.(Author:PeggySellers,PhotobyGregShaner)24ColletotrichumgraminicolaThecausalagentofmaizeanthracnosestalkrotandleafblight.Symptomsofmaizeanthracnosestalkrot(left)andleafblight(right)----fromBergstromandNicholson1999PlantDisease83:596-608Fluorescent-taggedC.graminicolawithinamaizeleaf----fromCzymmeketal.2005MethodsinMicrobiology34:27-61(Fig2.6A)259.UstilagomaydisFarmersinMexicoinfectcornartificiallytoharvestinfectedcobsforthepreparationofHuitlacoche,atraditionaldishpopularinpre-Hispanictimes.amodelforbiotrophic,plant-pathogenicbasidiomycetes26CornSmut2710.MELAMPSORALINIIncontrastwithmostoftheotherpathogensonthis‘Top10’list,flaxrustismorefamousthaninfamous.Althoughitwasinitiallystudiedasapathogenhavinganimpactontheflaxandlinseedindustries,itsmainimpactonfoodandfibreproductioncame(andcomes)fromitsroleasamodelsystem,providinginsightsintothemolecularbasisofplantimmunity.28Magnaportheoryzae2.Botrytiscinerea3.Pucciniaspp.4.Fusariumgraminearum5.Fusariumoxysporum6.Blumeriagraminis7.Mycosphaerellagraminicola8.Colletotrichumspp.9.Ustilagomaydis10.MelampsoraLini担子菌子囊菌Ug99Sweet

wines(Sauternes,Tokaj),

upto€500/bottle.附着胞appressorium吸器haustorium麦类叶斑枯病Sclerotiniasclerotiorum核盘菌29黑麦麦角菌Clavicepspurpurea麦角生物碱ergotalkaloidTheancientGreeksofEleusis,acityabout30kmNorth-EastofAthens,heldaritualcalled‘Eleusinianmysteries’(15thcenturyBC-4thcenturyAD)inwhichtheycelebratedtheannualreturnofPersephone,daughterofDemeter(theGreekgoddessofagriculture);thatrepresentedthereturnofplantsandlifetotheEarthduringthespring.Duringthisritual,whichtookplaceafteraperiodoffasting,theywoulddrinkaspecialbeveragecalledkykeon,madeofbarleyandotherherbs,whichissuspectedtohavecontainedpsychoactivealkaloidsfromthemouldClavicepspurpureaorClavicepspaspaligrowingonthebarley,orPaspalumdistichumgrowingontheherbs.Thatwouldhaveproducedvisionsandhallucinationslinkedtorevealingmentalstateswithastrongspiritualandintellectualcontent.Therelationbetweenmycotoxinsandthe‘Eleusinianmysteries’wasbeautifullydescribedinthe1978classicalwork‘TheroadtoEleusis’(Wassonetal.,1978)whichwasrecentlyextendedandre-editedontheoccasionofitsthirtiethanniversary(Wassonetal.,2008).30植物病原真菌的本质?

fungithathavedevelopedstrategiestocolonizetheplantsandobtainnutrientsfromthem“Everythingshouldbemadeassimpleaspossible—butnosimpler!”AlbertEinstein31PlantpathogencategoriesbasedonnutritionmodeHemibiotrophyNecrotrophyMelampsoralarici-populina(青杨叶锈病菌)BasidiomycetePucciniagraminis(秆锈菌)BasidiomyceteUstilagomaydis

(玉米瘤黑粉菌)

BasidiomyceteBlumeriagraminis

(小麦白粉菌)ascomyceteBotrytiscinerea(灰霉菌)ascomyceteBiotrophyFusariumoxysporum(尖孢镰孢）

ascomyceteFusariumgraminearum(禾谷镰孢）

ascomyceteSclerotiniasclerotiorum

(核盘菌)ascomyceteColletotrichumgraminicola(玉米炭疽茎腐病菌)ascomyceteMagnaportheoryzae(稻瘟病菌)ascomyceteObligatebiotrophy3233Processofinfection34BiotrophicgrowthphaseandtheentrainmentofplantmetabolismOnceinsidethehost,andwithinternalstoreslikelyexhausted,thepathogenneedstoestablishitselfrapidlybymobilizingmechanismsthatwillensureadequatenutrientuptakefromthehost.Fortheobligatebiotrophs,thisphaseisinitiatedbytheformationofahaustorium.Forhemibiotrophs,deficientinsuchfeedingstructures,somerecentevidencesuggeststhattheyhavedevelopedmechanismsthatentrainhostmetabolismtoestablishnewsinktissuestosuittheneedsofthepathogen.Invertasesandhexosesugartransportersplayakeyroleindefiningaleafasa‘source’orasa‘sink’ofcarbohydrates.35NecrotrophicgrowthphaseandentrainmentofplantmetabolismShortlyafterpenetration,necrotrophicplantpathogenicfungi,aidedbyproductionoftoxins,hydrolyticenzymesandnecrosis-relatedproteins,causecelllysisanddeathofhostcells.Onemaysurmisethatthelysisofhostcellswouldleadtoanincreaseinaccessiblenutrients;However,asevidencedfromexaminationofauxotrophicmutants,certainaminoacidssuchasmethionineandhistidinearenotsuppliedinsufficientquantitybythehosttissueandtheirbiosynthesisiscarriedoutbythefungus.36SporulationSporulationmarksthecompletionofthepathogeniclifecycle.Truebiotrophicfungicompletetheirlifecyclebymaintainingabiotrophicstate,whereashemibiotrophsandnecrotrophscompletetheirlifecycleinanecrotrophicstate.Invitrostudieshaveshownthatsporulationcanbetriggeredbyexhaustionofnitrogenandcarbonsources.WheatBarley37植物病原真菌致病策略?

生长获取营养对抗植物免疫操控植物代谢与运输效应蛋白次生代谢物38FungalCellWallFungi:mastersoflignocellulosesaccharification394041植物病原真菌致病工具分子效应蛋白Effectorproteins

42EffectorsofbiotrophicfungalplantpathogensThedefiningfeatureofpathogeneffectorsistheiralterationofthestructureandfunctionofthehostcellduringtheinfectionprocess.43Pep1isanapoplasticeffector.Pep1isshowntointeractdirectlywithPOX12andtoinhibititsactivity.EffectorsofcornsmutfungusCmu1wasshowntobeachorismatemutase.Chorismateisthebranchingmetaboliteoftheshikimatepathway.rechannelingofthechorismateflow4445CornsmutfungusTin246474849theflaxrustfungusMelampsoralini:effectorAvrL567wiltpathogensFusariumoxysporumf.sp.lycopersici:theAvr2effector50植物病原真菌致病工具分子代谢物/真菌毒素Metabolites/mycotoxin51草酸5253545556真菌活性物质的种类LovastatinAflatoxinCyclosporinGibberellincarotenoidpenicillinPolyketides（聚酮类）fattyacid-derivedcompounds，iterativetypeIPKSsAflatoxin,LovastatinNon-ribosomalpeptides（非核糖体肽类）aminoacid-derivedcompounds，singlemultimodularenzymes(NRPSs)Cyclosporin，penicillinTerpenes（萜类）Composedofseveralisopreneunits,thekeyenzymeisterpenecyclase

Gibberellin,carotenoidAlkaloids（生物碱）NRPSsDerivedfromtryptophananddimethylallylpyrophosphateergopeptides5758Fusarium(Hypocreales,Nectriaceae)isoneofthemosteconomicallyimportantgroupsofmycotoxigenicphytopathogensandemergenthumanpathogens.Modified

fromGeiser,etal.2013.Onefungus,onename:definingthegenusFusariuminascientificallyrobustwaythatpreserveslongstandinguse.Phytopathology103:400–8;Maetal.,2013,Annu.Rev.Microbiol.67:399–416;Fusariumgraminearum:36Mb,13332genesFusariumheadblight(FHB)ofwheat,barleyGibberellastalkrotofmaizeFusariumtucumaniae,Fusariumviguliforme:SuddendeathsyndromeofsoybeansFusariumverticillioides:42Mb,14179genesEarrotandFusariumstalkrotofmaizeFusariumsolani:51Mb,15707genes

RootrotofpeaFusariumoxysporum:61Mb,17735genesvascularwiltonawiderangeofplantsFusariumfujikuroi:44Mb,14813genesbakanaediseaseofrice水稻恶苗病Fusariumsolani

CM3100:Fusarioses

ofhumanFusariumpseudograminearum:37Mb,12488genescrownrotdiseaseofwheatFusariumcircinatum:44Mb,15713genesPitchcankerofpineinAfrica松树溃疡病Eudicot,Monocotdiverge59DONisavirulencefactorforwheatheadblight60616263SignalTransductionandTranscriptionalRegulation64656667植物病原真菌致病机制研究手段68科赫法则（Kochpostulates），通常是用来确定侵染性病害病原物的操作程序。1在每一病例中都出现相同的微生物；2要从寄主分离出这样的微生物并在培养基中得到纯培养（pureculture）；3用这种微生物的纯培养接种健康而敏感的寄主，同样的疾病会重复发生；4从试验发病的寄主中能再度分离培养出这种微生物来。如果进行了上述4个步骤，并得到确实的证明，就可以确认该生物即为该病害的病原物。德国细菌学家罗伯特·科赫（RobertKoch，1843～1910年）69柯赫氏法则已被移植并成为植物病理学中一项经典法则1.共存性观察：被疑为病原物的生物必须经常被发现于病植物体上。2．分离：必须把该生物从病植物体分离出来，在培养基上养成纯培养，纯培养即只有该种生物而无其它生物的培养物。3．接种：用上述纯培养接种于健康植物上，又引起与原标本相同的病害。4．再分离：从上述接种引起的病植物再度进行分离而得纯培养，此纯培养与接种所用纯培养完全一致（第4点是在柯赫氏以后由斯密斯（E．F．Smith）补充的）。70WhatIcannotcreate,Idonotunderstand.RichardP.Feynman(1918–1988),anAmericanphysicist71TargetgenehphhphhphPrimerUPrimerDPrimerIFPrimerIRHYYG同源重组基因敲除及回复Youshouldnotbelievesomethingjustbecauseyoucanexplainit.-ArthurKornberg72Seeingisbelieving73

生化和代谢Trichothecenebiosynthesispathwaygenes(Tri5etc.)®ulatorygenes(Tri6etc)Tri5Tri4Tri101Tri11Tri3Tri13Tri7Tri1Tri8Tri1Tri874禾谷镰孢致病策略75HowdoesaFungusPersuadeHostPlanttoAllowitsGrowthInside?76-ElucidatingFusarium

graminearum

infection

strategiesbasedonfungalexpressionprofilingat4inplanta

stagesFusariumgraminearum:oneofthemostdestructiveplantpathogensworldwideTrichotheceneMycotoxin77Fusariumgraminearumgenome78Cuomo,C.A.,etal.(2007).TheFusariumgraminearumgenomerevealsalinkbetweenlocalizedpolymorphismandpathogenspecialization.Science317:1400-1402.Ma,L.J.,etal.(2010).ComparativegenomicsrevealsmobilepathogenicitychromosomesinFusarium.Nature464:367-373.Wong,P.,etal.(2010).FGDB:revisitingthegenomeannotationoftheplantpathogenFusariumgraminearum.NucleicAcidsRes.34:D637–D639KnownpathogenicityorvirulentgenesMoresignalingcomponentsheterotrimericGprotein,RasGTPase,histinekinase,proteinphosphataseDevelopmentalregulators(FTL1,FgStuAp,etc)Cellcycleregulators(CID1)Growthregulators(GzSNF1,FgPac1,etc)Plantcellwalldegradingenzymes(FgPG1,FgPG2,etc)Fungalgrowthprimarymetabolicpathways(methioninesynthesis)Otherlyticenzymes(FGL1,encodesanextracellular

lipase)ROSproducing(NOS1)Steroltrafficking(NPC1)Ironuptake(SID1,NPS6)Othertranscriptionfactors(ZIF1etc.)Nitratetransport(CrnA)Trichothecenebiosynthesispathwaygenes(Tri5etc.)®ulatorygenes(Tri6etc)MAPKcascadegenes7942proteinkinasegenes;62transcriptionfactorgenes80What’sknown?Kazan,K.,Gardiner,D.M.,andManners,J.M.(2011).Onthetrailofacerealkiller:recentadvancesinFusariumgraminearumpathogenomicsandhostresistance.Mol.Plant.Pathol.13:399-413CurrentUnderstandingonF.graminearumPathogenesisWhat’sstillunclearThetimingandsequentialrelationshipofproducingplantcellwalldegradationenzymes.About70plantcellwall-degradingenzymes(CWDE)havebeenidentifiedinFusariumspecies,althoughnonehaveyetbeenshowntoberequiredforpathogenicity.Thespecificroleofreactiveoxygenspecies(ROS).Howdoesthepathogenadjustprimarymetabolismtoachievegrowthinplanta?SecondarymetabolitesotherthanDON.81Invitrostagespecificgeneexpressionprofiling82Seong,K.Y.,Zhao,X.,Xu,J.R.,Güldener,U.,andKistler,H.C.(2008).ConidialgerminationinthefilamentousfungusFusariumgraminearum.FungalGenet.Biol.45:389-399.Differencebetweenpreviousinplantaandinvitroprofiling83ExpressedF.graminearumprobesetsduringinvitrogrowthSeong,K.Y.,Zhao,X.,Xu,J.R.,Güldener,U.,andKistler,H.C.(2008).ConidialgerminationinthefilamentousfungusFusariumgraminearum.FungalGenet.Biol.45:389-399.72h:fromourlabLysøe,E.,Seong,K.Y.,andKistler,H.C.(2011).ThetranscriptomeofFusariumgraminearumduringtheinfectionofwheat.Mol.PlantMicrobe.Interact.24:995-1000.1mm^3tissuecontains125,000cells.Infectedhostcells:Un-infectedhostcells=1:1250.20umInanearlyinfectionstage,thefungalhyphaemayonlyinvade~100cells.Thechallenge:profilingfungalgeneexpressioninaplantSignalNoise1:1dilution0.0011SignalNoise1:1250dilution11.2584Stage-SpecificInPlantaFungalGeneExpressionProfilingMolecularStrategiesofthePathogenGrowinginsideHostPlantsPlant–FungalInteractionGFP-taggedFusariumgraminearumPH-185Stage-specificfungalgeneprofilingelucidatesthemolecularstrategiesofFusariumgraminearumgrowinginsidewheatcoleoptilesWheatcoleoptiles86Structureofcoleoptile87LeafepidermissubepidermisColeoptile88F.graminearumcanenterwheatcoleoptileswithoutwoundingInoculationprocedure89BeforecuttingAftercuttingAfterinoculationF.graminearuminvasivegrowthinsidecoleoptiles90Lasercapturemicrodissectionofinplantagrowingfungiforgeneprofiling9116hpiLasercapturemicrodissectionofinplantagrowingfungiforgeneprofiling92BeforeLCMAfterLCMLCMcaptured40hpiMicroarraydatawithquality40hpi64hpi16hpi240hpiconidiaHyphaeinmedia72hUnsupervisedclustertree93Inplantaandinvitrogrowingtranscriptomesaredistinct940hpi0h2h8h24h72h240hpi16hpi64hpi40hpiPC1(32.6%ofvariance)0-0.2-0.4-0.60.20.40.60.80-0.2-0.40.80.60.40.2PC2(20.6%ofvariance)0h2h8h24h0hpi16hpi40hpi64hpi72h240hpiDatafromSeongetal.,2008ThisreportInvitroInplantaEscape/suppresshostdefense

Grab/stealnutritionAltermetabolismFungusPlanthostSecretedagentsLyticenzymes

ROSToxinsSecondarymetabolites"I'mafirmbelieverthatwithoutspeculationthereisnogoodandoriginalobservation"C.DarwintoA.R.Wallace,185795QuestionstobeaddressedThetimingandsequentialrelationshipofproducingplantcellwalldegradationenzymes.About70plantcellwall-degradingenzymes(CWDE)havebeenidentifiedinFusariumspecies,althoughnonehaveyetbeenshowntoberequiredforpathogenicity.Thespecificroleofreactiveoxygenspecies(ROS).Howdoesthepathogenadjustprimarymetabolismtoachievegrowthinplanta?SecondarymetabolitetoxinsotherthanDONorZearalenone.96Putativesecretedandcell-surfaceproteingenes97Secretedproteins(1355)RelativeexpressionlevelSecretedproteases(134)Plantcellwalldegradingenzymes(133)101214GPCR(81)CPGRP(42)CFEM-motifcontaining(21)1214161810Wheatcoleoptilecellwallstructure98CelluloseFerulicacidXylanPectinMLGXyloglucanMiddlelamellaPrimarycellwallPlasmamembraneVogel,J.(2008).Uniqueaspectsofthegrasscellwall.Curr.Opin.PlantBiol.11:301-307.Pectinrelevantenzymesexpression99CelluloseandHemicellulosesrelated100PlantCellWallDegradingEnzymes10116hpi40hpi64dpi240hpiconidiaF.graminearumgrowinginsideofwheatInvitroTwooftheinplanta-preferentially-expressedCWDEgenesarerequiredforfullvirulenceonwheatcoleoptiles102103SupplementalFigure5.(continued).(F)LivecellimagesofF.graminearummutantsonwheatcoleoptiles.Arrowspointtotheinvadedhyphae.Scalebar=50μm.16hpi40hpi64hpi96hpi144hpiPH-1∆eng1∆cbhC1040080012001600016406496144WT∆eng1Inplanta(hpi)Distancefromcuttingedgetoinfectionfront(μm)040080012001600016406496144WT∆cbhC1Inplanta(hpi)Distancefromcuttingedgetoinfectionfront(μm)Twooftheinplanta-preferentially-expressedCWDEgenesarerequiredforfullvirulenceonwheatcoleoptilesQuestionstobeaddressedThetimingandsequentialrelationshipofproducingplantcellwalldegradationenzymes.About70plantcellwall-degradingenzymes(CWDE)havebeenidentifiedinFusariumspecies,althoughnonehaveyetbeenshowntoberequiredforpathogenicity.Thespecificroleofreactiveoxygenspecies(ROS).Howdoesthepathogenadjustprimarymetabolismtoachievegrowthinplanta?SecondarymetabolitetoxinsotherthanDONorZearalenone.104ROSrelevanttoF.graminearum-coleoptileinteraction105Mock,woundedF.graminearuminoculated,woundedMock,not-woundedF.graminearuminoculatednot-woundedIndexofDABstain00.20.40.6woundednot-woundedMockF.graminearuminoculated106Extracellularreactiveoxygenspeciesscavengecomesaheadofproduction8101214ROSproducingenzymes(cytosolic)(14)ROSproducingenzymes(secreted)(9)8101214Relativeexpressionlevel6H2O2scavengingenzymes(cytosolic)(14)H2O2scavengingenzymes(secreted)(6)81012O2-scavengingenzymes(cytosolic)(4)O2-scavengingenzymes(secreted)(2)C02824720164064240Invitro(h)InPlanta(hpi)Extracellularreactiveoxygenspeciesscavengecomesaheadofproduction10726912Log2transformedtranscriptlevelExtracellularROSproducingenzymesExtracellularH2O2scavengingenzymesExtracellularO2-scavengingenzymes02824720164064240Invitro(h)Inplanta(hpi)QuestionstobeaddressedThetimingandsequentialrelationshipofproducingplantcellwalldegradationenzymes.About70plantcellwall-degradingenzymes(CWDE)havebeenidentifiedinFusariumspecies,althoughnonehaveyetbeenshowntoberequiredforpathogenicity.Thespecificroleofreactiveoxygenspecies(ROS).Howdoesthepathogenadjustprimarymetabolismtoachievegrowthinplanta?Secondarymetabolitetoxins(DONandbeyond).108Shiftinprimarymetabolismduringearlystageofcoleoptileinfection109Oxidativephosphorylation(60)TCAcycle(28)Glyoxylatecycle(17)Fattyacidoxidation(25)Glycolysis/Gluconeogenesis(27)

FGSG_01421putativemitochondrialcitratecarrier121416181086Relativeexpressionlevel02824720164064240Invitro(h)InPlanta(hpi)GlycolysisandGluoconeogenesis110Glycolysis164064240inplanta(hpi)0121416722824invitro(h)0Gluconeogenesis68101214164064240inplanta(hpi)0722824invitro(h)0Log2transformedtranscriptlevelinvitro(h)2824720164064240inplanta(hpi)025913Shiftinprimarymetabolismduringearlystageofcoleoptileinfection111Shiftinprimarymetabolismduringearlystageofcoleoptileinfection112040080012001600016406496144WTΔcic1Inplanta(hpi)Distancefromcuttingedgetoinfectionfront(μm)QuestionstobeaddressedThetimingandsequentialrelationshipofproducingplantcellwalldegradationenzymes.About70plantcellwall-degradingenzymes(CWDE)havebeenidentifiedinFusariumspecies,althoughnonehaveyetbeenshowntoberequiredforpathogenicity.Thespecificroleofreactiveoxygenspecies(ROS).Howdoesthepathogenadjustprimarymetabolismtoachievegrowthinplanta?Secondarymetabolitetoxins(DONandbeyond).113TrichothecenebiosyntheticpathwayareprobablyturnedoffGenesnotincludedinthepathwayAlexanderetal.,200902004006008001000TRI5114Specificsecondarymetaboliteclusterswereinducedat64hpiduringinfection115aurofusarinFG3_54clusterisrequiredforvirulenceoncoleoptile11616hpi40hpi64dpi240hpiconidiaCovertPenetrationOvertDestructionRapidproliferationReproductionF.graminearumgrowinginsideofwheatPlantcellwalldegradationSucroseimportLactoseimportExtracellularO2-scavengeExtracellularROSproducingExtracellularH2O2scavengePutativetoxinproteinproducingSecondarymetabolitesproducing117stage-dependentdeploymentofplantcellwall-degradingenzymesearlymitigationandlaterproductionofROSashiftinenergymetabolismtowardsglyoxalatecyclelateronsetofpotentiallyphytotoxicsecondarymetaboliteproductionInfectionstrategies禾谷镰孢侵染小麦幼苗策略隐蔽入侵全面占领Zhang,X.-W.,Jia,L.-J.,Zhang,Y.,Jiang,G.,Li,X.,Zhang,D.,andTang,W.-H.*(2012)Inplantastage-specificfungalgeneprofilingelucidatesthemolecularstrategiesofFusariumgraminearumgrowinginsidewheatcoleoptiles.PlantCell.24:5159-5176118Stage-SpecificInPlantaFungalGeneExpressionProfilingMolecularStrategiesofthePathogenGrowinginsideHostPlantsPlant–FungalInteraction119Youshouldnotbelievesomethingjustbecauseyoucanexplainit.-ArthurKornbergArthurKornberg(March3,1918–October26,2007)wasanAmerican

biochemistwhowontheNobelPrizeinPhysiologyorMedicine1959forhisdiscoveryof"themechanismsinthebiologicalsynthesisofdeoxyribonucleicacid(DNA)"togetherwithDr.SeveroOchoaofNewYorkUniversity.120细胞表面蛋白CFEM1对于小麦幼苗致病和穗部致病都有贡献野生菌株ΔFgcfem1野生菌株ΔFgcfem1(FGSG_02077)M3M2M1Zhang,X.-W.,Jia,L.-J.,Zhang,Y.,Jiang,G.,Li,X.,Zhang,D.,andTang,W.-H.*(2012)Inplantastage-specificfungalgeneprofilingelucidatesthemolecularstrategiesofFusariumgraminearumgrowinginsidewheatcoleoptiles.PlantCell.24:5159-5176121线粒体柠檬酸转运蛋白FgCIC1对于小麦幼苗致病和穗部致病都有贡献Zhang,X.-W.,Jia,L.-J.,Zhang,Y.,Jiang,G.,Li,X.,Zhang,D.,andTang,W.-H.*(2012)Inplantastage-specificfungalgeneprofilingelucidatesthemolecularstrategiesofFusariumgraminearumgrowinginsidewheatcoleoptiles.PlantCell.24:5159-5176122非核糖体多肽类次生代谢产物合成基因簇FG3_54对于小麦幼苗致病和穗部致病都有贡献Zhang,X.-W.,Jia,L.-J.,Zhang,Y.,Jiang,G.,Li,X.,Zhang,D.,andTang,W.-H.*(2012)Inplantastage-specificfungalgeneprofilingelucidatesthemolecularstrategiesofFusariumgraminearumgrowinginsidewheatcoleoptiles.PlantCell.24:5159-5176FG3_54基因簇合成的次生代谢产物推测为非核糖体多肽类真菌毒素123AcknowledgementsFundedby:ChineseAcademySciences,973,863,theMinistryofAgricultureXiao-WeiZhangDongZhangYanZhangTing-LuYuanLei-JieJiaGangJiangJuanHe124Shen-HuaYaoShi-LanZhaoZhi-YongZhangThanksto:JonDuvick,Rong-XiangFang,Jin-RongXu,Zong-HuaWang,Zu-HuaHe,You-LiangPeng,Cheng-ShuWang,Li-HuangZhu,Zhi-HuaZhou,Lai-GenLi,ChenYang,SheilaMcCormick,XuanLi,YongWang,Ren-TaoSong,Xiao-YanGaoPenicillin:thefirstmiracledrugManyofyouarehereonlybecausepenicillinsavedyourlife,orthelifeofoneofyourparentsorgrandparents.EarlypenicillinculturefacilityattheSirWilliamDunnSchoolofPathology,Oxford,England.Chainsofconidia(spores)producedbyhyphalbranchfrommycelium

SirAlexanderFleming,1952Thankyou!125126Around100,000speciesoffungihavebeenformallydescribedbytaxonomists,buttheglob