植物病原真菌致病機制2014年5月27日11、什么是真菌?2、哪些是植物病原真菌?3、植物病原真菌的本質(zhì)?

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、植物病原真菌致病機制研究手段7、禾谷鐮孢致病策略提綱2什么是真菌?3Fusariumverticillioides4FungiHeterotrophic,lackingchloroplastsPossessacellwallandvacuolesLifecycle,haploidanddiploidHyphalgrowth,tipgrowthVersatileinmetabolitesbiosynthesisSurfaceexpertDimorphicgrowthswitch(yeast,hyphae)Fungalcellwallcomposedofglucansandchitin,butnocellulosemodifiedfromMeyerowitz2002ScienceFungi0.1Gadicotmonocotsplit5animalplantfungi6ModifiedfromHedges,2002,NatRevGenet3:838-849;Schochetal.,2009,SystBiol58:224-239盤菌亞門(Pezizomycotina)散囊菌綱外囊菌亞門（Taphrinomycotina）酵母菌亞門Ascomycota子囊菌門Basidiomycota擔(dān)子菌門銹菌綱黑粉菌綱Aphylogenyoffungi7gAphylogenyofplants8Plant-FungalinteractionsPathogen(病原菌)Symbioticfungi(共生菌)Endophyte(內(nèi)生菌)Saproph(腐生菌)9PlantproducerFungusdegraderEcosystemAnimalconsumerFungiarethemostcommonparasitesofplants.Allhigherplantspeciesaresusceptibletooneormorefungaldiseases,andmostcropplantshavemany.PlantpathogensFungiBacterialVirusOomyceteNematode10哪些是植物病原真菌?1112Magnaportheoryzaeafilamentousascomycetefungus,isthecausalagentofriceblastdisease,themostdestructivediseaseofriceworldwide13由子囊類真菌引起的水稻病害稻瘟病稻曲病胡麻斑病惡苗病爛秧病稻瘟病菌稻綠核菌稻平臍蠕孢串珠鐮刀菌禾谷鐮刀菌尖孢鐮刀菌…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擔(dān)子菌子囊菌Ug99Sweet

wines(Sauternes,Tokaj),

upto€500/bottle.附著胞appressorium吸器haustorium麥類葉斑枯病Sclerotiniasclerotiorum核盤菌29黑麥麥角菌Clavicepspurpurea麥角生物堿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植物病原真菌的本質(zhì)?

fungithathavedevelopedstrategiestocolonizetheplantsandobtainnutrientsfromthem“Everythingshouldbemadeassimpleaspossible—butnosimpler!”AlbertEinstein31PlantpathogencategoriesbasedonnutritionmodeHemibiotrophyNecrotrophyMelampsoralarici-populina(青楊葉銹病菌)BasidiomycetePucciniagraminis(稈銹菌)BasidiomyceteUstilagomaydis

(玉米瘤黑粉菌)

BasidiomyceteBlumeriagraminis

(小麥白粉菌)ascomyceteBotrytiscinerea(灰霉菌)ascomyceteBiotrophyFusariumoxysporum(尖孢鐮孢）

ascomyceteFusariumgraminearum(禾谷鐮孢）

ascomyceteSclerotiniasclerotiorum

(核盤菌)ascomyceteColletotrichumgraminicola(玉米炭疽莖腐病菌)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?

生長獲取營養(yǎng)對抗植物免疫操控植物代謝與運輸效應(yīng)蛋白次生代謝物38FungalCellWallFungi:mastersoflignocellulosesaccharification394041植物病原真菌致病工具分子效應(yīng)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謝物/真菌毒素Metabolites/mycotoxin51草酸5253545556真菌活性物質(zhì)的種類LovastatinAflatoxinCyclosporinGibberellincarotenoidpenicillinPolyketides（聚酮類）fattyacid-derivedcompounds，iterativetypeIPKSsAflatoxin,LovastatinNon-ribosomalpeptides（非核糖體肽類）aminoacid-derivedcompounds，singlemultimodularenzymes(NRPSs)Cyclosporin，penicillinTerpenes（萜類）Composedofseveralisopreneunits,thekeyenzymeisterpenecyclase

Gibberellin,carotenoidAlkaloids（生物堿）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水稻惡苗病Fusariumsolani

CM3100:Fusarioses

ofhumanFusariumpseudograminearum:37Mb,12488genescrownrotdiseaseofwheatFusariumcircinatum:44Mb,15713genesPitchcankerofpineinAfrica松樹潰瘍病Eudicot,Monocotdiverge59DONisavirulencefactorforwheatheadblight60616263SignalTransductionandTranscriptionalRegulation64656667植物病原真菌致病機制研究手段68科赫法則（Kochpostulates），通常是用來確定侵染性病害病原物的操作程序。1在每一病例中都出現(xiàn)相同的微生物；2要從寄主分離出這樣的微生物并在培養(yǎng)基中得到純培養(yǎng)（pureculture）；3用這種微生物的純培養(yǎng)接種健康而敏感的寄主，同樣的疾病會重復(fù)發(fā)生；4從試驗發(fā)病的寄主中能再度分離培養(yǎng)出這種微生物來。如果進行了上述4個步驟，并得到確實的證明，就可以確認該生物即為該病害的病原物。德國細菌學(xué)家羅伯特·科赫（RobertKoch，1843～1910年）69柯赫氏法則已被移植并成為植物病理學(xué)中一項經(jīng)典法則1.共存性觀察：被疑為病原物的生物必須經(jīng)常被發(fā)現(xiàn)于病植物體上。2．分離：必須把該生物從病植物體分離出來，在培養(yǎng)基上養(yǎng)成純培養(yǎng)，純培養(yǎng)即只有該種生物而無其它生物的培養(yǎng)物。3．接種：用上述純培養(yǎng)接種于健康植物上，又引起與原標本相同的病害。4．再分離：從上述接種引起的病植物再度進行分離而得純培養(yǎng)，此純培養(yǎng)與接種所用純培養(yǎng)完全一致（第4點是在柯赫氏以后由斯密斯（E．F．Smith）補充的）。70WhatIcannotcreate,Idonotunderstand.RichardP.Feynman(1918–1988),anAmericanphysicist71TargetgenehphhphhphPrimerUPrimerDPrimerIFPrimerIRHYYG同源重組基因敲除及回復(fù)Youshouldnotbelievesomethingjustbecauseyoucanexplainit.-ArthurKornberg72Seeingisbelieving73

生化和代謝Trichothecenebiosynthesispathwaygenes(Tri5etc.)®ulatorygenes(Tri6etc)Tri5Tri4Tri101Tri11Tri3Tri13Tri7Tri1Tri8Tri1Tri874禾谷鐮孢致病策略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:fromourlabLys?e,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禾谷鐮孢侵染小麥幼苗策略隱蔽入侵全面占領(lǐng)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細胞表面蛋白CFEM1對于小麥幼苗致病和穗部致病都有貢獻野生菌株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線粒體檸檬酸轉(zhuǎn)運蛋白FgCIC1對于小麥幼苗致病和穗部致病都有貢獻Zhang,X.-W.,Jia,L.-J.,Zhang,Y.,Jiang,G.,Li,X.,Zhang,D.,andTang,W.-H.*(2012)Inplantastage-specificfungalgeneprofilingelucidatesthemolecularstrategiesofFusariumgraminearumgrowinginsidewheatcoleoptiles.PlantCell.24:5159-5176122非核糖體多肽類次生代謝產(chǎn)物合成基因簇FG3_54對于小麥幼苗致病和穗部致病都有貢獻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謝產(chǎn)物推測為非核糖體多肽類真菌毒素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