BOOK OF ABSTRACTS

OPENINGSESSIONOpeninglecture1Globalchangeisachallengingparameterforplantpestriskassessment.CharlesManceau1

1Anses – French agency for food, environmental and occupational health & safety, 7 rue JeanDixméras,F-49044AngersCedex01,France

Globalchangereferstoplanetary-scalechanges intheEarthsystemwhich includeshumansociety.Globalchangeencompassesclimate,landuse,biodiversity,tradeofplantandplantpartsandhasastrongandwell defined impactonplanthealthdespite the short time frameunder consideration.Emergingplantdiseasespresentmanyseriousissuesforhumanwell-being,whetherinagricultural,forestry, environmental or regulatory arenas. The temporal and spatial scales of plant diseaseemergence are defining features related to local, national and global drivers. These include theincreases inglobal trade,ofcourse,butalso the introductionofnovelcrop,changes inproductionsystems,interactionsoccurringatthelandscapelevelandtheimpactofclimatechange.AgroecologyisaoneofthedriversthatraisequestionsaboutthepredictabilityofemergingplantdiseasesPestriskanalysis(PRA)istheprocessusedbyNationalPlantProtectionOrganizations(NPPOs)asthetechnical justification for phytosanitary measures. PRA is defined by the International PlantProtectionConvention(IPPC)as“theprocessofevaluatingbiologicalorotherscientificandeconomicevidence to determinewhether a pest should be regulated and the strength of any phytosanitarymeasurestobetakenagainstit.”Theprocessrequiresariskassessmenttocharacterizetheriskandriskmanagementtodetermineappropriatemeasures.PRAsaremainlyperformedinthecontextoftheinternationaltrade.Anintroducedpathogeninoneregionthatleadstoanemergingdiseasemayhavebeenendemic,widespreadandsometimescrypticinanother.Besides,itmaybere-emergenceofadiseasewhichhaddisappearedbecauseofmodificationsinproductionsystems.Geneticchangesthroughhybridizationcanleadtohostshiftsandadaptations.Lessoften,theemergingdiseasemaybecausedbyapathogenthatishithertonewtoscience.Keywords:emergingdiseases,Pestriskanalysis,IPPC

Openinglecture2

ValuingBiodiversityandBioticInteractionsforCropprotectioninAgroecologyPhilippeLemanceau1,FrançoiseLescourret2,SandrinePetit11Agroécologie,AgroSupDijon,INRA,Univ.BourgogneFranche-Comté,F-21000Dijon,France2UnitéderecherchePlantesetSystèmesdeCultureHorticoles,INRA,DomaineSaint-Paul,F-84914Avignon,France

Theintensiveagricultureinitiatedafterthesecondworldwarisnowconsideredasbeingunsustainablebecauseoftheerosionofresources(e.g.soils,biodiversity,fossilenergy)andthehighuseofinputs(pesticides,fertilizers,water)withpossibledeleteriouseffectsbothontheenvironmentandonthequalityofagriculturalproducts.Accordingly,anincreasinginterestisgiventoagroecologywiththedevelopmentofcroppingsystemswhichvaluebiodiversityandbioticinteractionsinagroecosystems.Variousillustrationsandprospectswillbegivenontheroleofbiodiversityandbioticinteractionsin(i)soilbornediseasesuppression,(ii)thebalancebetweenthedisservicesandservicesprovidedbyweeds,(iii)keepingthelevelofpestsbelowaharmfulnessthreshold.Thispresentationwillencompasstheevaluationof(i)thebeneficialeffectsoncropprotectionofbiodiversity(levelandcomposition)andbioticinteractions(includingtrophicnetworks),andof(ii)theenhancementoftheseeffectsbyappropriatecropmanagement(includingthechoiceofplantgenotypes)attheplotandlandscapelevels,allowingthedecreaseofpesticideusewhilekeepingtheproductivityhighenough.

Session1.FROMPLANT-MICROBEINTERACTIONSTOINTERACTIONSWITHINPHYTOBIOMES(SalleJeanBart)

Keynotelecture1

Phytobiomeecology:developingnovelapproachesforsustainablediseasemanagementLindaL.Kinkel,DepartmentofPlantPathology,UniversityofMinnesota,St.Paul,MN55108

Microbesandplantsexistwithincomplexnetworksofinteractingplantandmicrobialspecies,thephytobiome.Ourworkexplorestheinteractingrolesofplantcommunitydiversity,planthost,andmicrobialspeciesinteractionsindeterminingthepathogen-suppressivepotentialandcompositionofsoilmicrobiomes,andtheconsequencesforplantproductivity.Usingculture-basedapproaches,wefoundthatrhizosphereStreptomycesassociatedwiththesameplanthostweresignificantlymorepathogen-suppressivewhenthehostgrewinmonoculturevs.withinahigh-diversityplantcommunity.Incontrast,populationsofStreptomycesintherhizosphereofplanthostsgrowinginhigh-diversitycommunitiesweremoreniche-differentiatedthanpopulationsassociatedwiththesamehostinmonoculture.Thesedatasuggestthatplantcommunitydiversityplaysacriticalroleindeterminingthelikelihoodofantagonisticarmsracecoevolutionvs.nichedifferentiationamongsympatricsoilpopulations,withsignificantimplicationsforplantdiseasesuppression.Ampliconsequencingofrhizospherecommunitiesassociatedwithdifferentplanthostsprovideinsightsintonon-culturedtaxaassociatedwithdiseasesuppression.Intotal,ourworkillustrateshowdiffusenetworksofspeciesinteractionsoverdiversespatialscalescontributetodeterminingthepathogen-suppressiveandplantgrowth-promotingpotentialofindigenoussoilmicrobes,andsuggestsspecificcropmanagementapproachestargetingspeciesinteractionsthatofferpotentialforsustainablediseasecontrol.

Keywords:phytobiome,diseasesuppression,coevolution,suppressivesoil,antagonist,Streptomyces

Keynotelecture2Individual-basedecologyofthephyllomicrobiomeJohanLeveau1

1UniversityofCalifornia,DepartmentofPlantPathology,Davis,CA95616,USAThe interactions of plant-associated microorganisms among themselves and with their host arecomplexandtakeplaceacrossawiderangeofspatialscales.Thereisalimitedunderstandingofthetypes, mechanisms, and outcomes of these interactions at the lowest scale, i.e. at the level ofindividualmicrobes, and how these interactions link to observations at higher-level spatial scales.Here,Iwillexplorerecentprogressthathasbeenmadeinthefieldofindividual-basedecologyofthephyllomicrobiome,i.e.theplantleafsurfaceasahabitatformicroorganisms.Iwillpresentexamplesfromourownworktohighlightthevalueoftheindividual-basedapproachaswellastheuseofnewresearchtoolsthatallowthesingle-cell interrogationofamicrobe’sexperienceof itsenvironment,thedeconstructionofleafsurfacecomplexitytounderstandtheimpactofmicroscalefactorssuchasleafsurfacetopography,andtheinsilicosimulationandpredictionofindividual-basedinteractionsinaheterogeneousenvironment.Keywords:phyllosphere,individual-basedecology

Session1Oral1

Plant-microbeinteractionsinstrawberrygrowninsustainableanddiseasesuppressivesubstratesJaneDebodea,CarolineDeTendera,AnaSheinLeeDiaza,b,TinaKyndtb,BartVandecasteelea,HildeMuylleaandMartineMaesa

aInstituteforAgriculturalandFisheriesResearch(ILVO),PlantSciencesUnit,Merelbeke-Melle,Belgium.bGhentUniversity,DepartmentMolecularBiotechnology,ResearchgroupEpigenetics&defence,Gent,Belgium.

Peat based substrates are not sustainable and often fail to support biological control, evenwheninoculatedwithbiocontrolagents.Locallyproducedplantfibresmayallowforpartiallyreplacingpeatinsubstrates.Moreover, theymayeasilybecolonizedbymicroorganismsand in thiswaypromotetheinstallationofbiologicalcontrolagentsresultingindiseasesuppressiveness.Strawberrypottrialswereconductedusingmiscanthusstrawasa20%v/vamendmentinpeat.Themiscanthus straw was extruded and pre-colonized with the biocontrol fungus Trichodermaharzianum (MSEXTRI treatment). Strawberry plants grown in this treatmentwere significantly lesssusceptibletoBotrytiscinereainoculatedonthefruitsascomparedto100%peatandpeatwith20%v/v extruded miscanthus straw (MSEX treatment). Both bacterial and fungal rhizospherecommunitieswerestudiedusing16SrDNAV3-V4andITS2metabarcoding,respectively.Thespecificpresence ofTrichoderma in the rhizospherewas studied using platings on semi-selectivemedium.Thestrawberrydefenceresponsewasstudiedthroughgeneexpressionanalysis.Adding extruded miscanthus straw significantly reduced the fungal diversity in the strawberryrhizosphere.Surprisingly, intheMSEXTRItreatment,therelativeabundanceofthefungalHumicolaspp.washighlyincreasedascomparedtothe100%peatandMSEXtreatment,whereastherelativeabundance of theTrichoderma spp.was low in all treatments.Humicola spp. are known for theirpotential in lignocellulose degradation and induced resistance. Plating on semi-selective mediumshowed that only the strawberry roots of theMSEXTRI treatment were consistently colonised byTrichoderma. Bacterial communities maintained their diversity and did not exhibit significantchanges, although some genera shifted in relative abundance between the treatments. Thestrawberry plants grown in the MSEXTRI treatment showed an up-regulation of defence genesFachi2-1,Fachi2-2andFaPALascomparedtotheothertreatments.Thismightbecorrelatedwiththeobservedchangesintherhizospheremicrobiomeanddiseasesuppressiveness.

Keywords:bacteria,fungi,miscanthus,metabarcoding,geneexpression,Trichoderma

Session1Oral2OxylipincommunicationbetweenthefungalendophyteParaconiothyriumvariabileandthephytopathogenFusariumoxysporum.

MargotBärenstrauch1,StéphaneMann1,DidierBuisson1,SoizicPrado1,CarolineKunz1,2

1SorbonneUniversités,MuséumNationald’HistoireNaturelle,CNRS,LaboratoireMoléculesdeCommunicationetAdaptationdesMicroorganismes(MCAM),UMR7245CNRS-MNHN,Paris,France2SorbonneUniversités,UPMCParis06,UFRdeBiologie,Pariscedex05,France

It ismoreandmoreevident that inorder to studyplantpathogen infectionprocesses,onehas totakeintoaccountthepresenceofthirdprotagonists,theendophyticandepiphyticmicrobiota.Howfungalendophytescommunicatewithinvadingplantpathogens,themicrobiotaandthehostplantisthetopicofourresearchproject.

WehaveatourdispositionarangeofisolatedfungalendophytesfromtheconifertreeCephalotaxusharringtoni1and inparticular,a foliar species,Paraconiothyriumvariabile (Dothidiomycetes). Initialresearch on this endophyte indicates antagonistic activity towards phytopathogens, inhibition ofmycotoxinproductioninF.oxysporum2andplantmetabolitebiotransformationtotheadvantageoftheendophyte3.During the competitionbetweenP. variabile andF.oxysporum, twooxylipins,13-keto-9,11-octadecadienoic acid (13-KODE) and 13-hydroperoxy-9,11-octadecadienoic acid (13-HpODE)areoverproduced,whichisaccompaniedbyadecreaseinbeauvericinsecretion,oneofthemostpotentmycotoxinsofFusariumspeciesandavirulencefactoroninfectedplants.

To elucidate the role of the twooxylipins in the fungal antagonistic interaction,we identified andcloned two lipoxygenase genes (Pvlox1 and Pvlox2) of P. variabile potentially involved in thebiosynthesisof13-KODEand13-HpODE.Weexpressed the twoPvlox genes inEscherichia coli andcurrently study the biochemical activity of the PvLOX1 and PvLOX2 enzymes. In a complementaryapproach, we study the expression patterns of the two Pvlox genes in P. variabile during itsantagonisticinteractionwithF.oxysporum.Toexpandtheresearchonan inplantatripartitemodelsystem,we started inoculation tests onArabidopsis thaliana plants to followP. variabile infectionmicroscopicallyandtestitsinfluenceontheplantfitness.

1. Langenfeld,A.etal.(2013),FungalBiol.117,124–1362. Combès,A.etal.(2012),PLoSONE7,e473133. Tian,Y.etal.(2014),Phytochemistry108,95–101

Keywords:Fungalendophyte,oxylipinsignalling,lipoxygenase,mycotoxin,Fusariumoxysporum,Arabidopsisthaliana

Session1Keynotelecture3VirusesinthePhytobiome:AbundanceandEcologicalRoles.MarilynJ.Roossinck

1CenterforInfectiousDiseaseDynamics,PennStateUniv.,UniversityPark,PAWehaveanalyzedplantvirusesinabiodiversityhotspotinnorthwesternCostaRicawithabout10,000speciesofnativeplants,andalowdiversitynaturalareaintheUnitedStates:theTallgrassPrairiePreserve.Wefindmanyvirusesthataredistantlyrelatedtoknownviruses,anequalorgreaternumberofcompletelyunknownviruses,andafewknownviruses.Persistentvirusesarethemostcommon,andappeartohaveverylongrelationshipswiththeirhostswithnearly100%ratesofverticaltransmission.Acutevirusesarefoundineachstudysite,somewithwide-spreadinfections,butknownarelinkedtoanysymptoms.Mixedinfectionsarealsocommon.

MembersofthefamilyPartitiviridaearethemostcommonvirusesfoundinbothstudiesofwildplants,andarealsoverycommonincrops.WeareexploringtheimpactsofthesevirusesoncropplantsusingJapapeñapeppers,whicharealwaysinfectedwithPeppercrypticvirus1.Beneficialeffectsofvirusinfectionwillbediscussed,includingtheimplicationsforagriculture.

Session1Oral3LipopeptidesinvolvedintherhizospherefitnessofB.amyloliquefaciensbiocontrolstrainsMarcOngena

University of Liège, Gembloux Agro-Bio Tech Faculty, Microbial Processes and InteractionsLaboratory,Passagedesdéportés2,B-5030Gembloux,Belgium

Bacilli are among the genera most commonly identified in the bacterial rhizosphere

communityassociatedwithplantroots.Interestingly,someisolatesfromparticularspeciessuchasB.amyloliquefaciensclearlyrepresentbeneficialpartnersforthehostplantbasedontheirpotentialtocombat phytopathogens and/or to directly promote growth and are promising candidates for thedevelopment of microbial alternatives to chemical pesticides or fertilizers. Such plant protectioneffect is associated with their capacity to form a wide array of biocontrol secondary metabolites(BSM)retainingantimicrobialandhostimmunityelicitingactivities.HoweveritalsoprimarlyreliesontheirpotentialtoestablishandpersistintherhizosphereatthresholdpopulationsnecessarytoformrelevantamountsoftheseBSM.

We will provide an overview of bacterial traits involved in such rhizosphere fitness andpresentourrecentworksshowinghowsomeunsuspectedplantmolecularpatternsmayalsoimpactthephenomenon.WecombinedimagingandotherMStechniqueswithgeneticapproachestoshowthat B. amyloliquefaciens can boost the synthesis of a particular lipopeptide-type antibiotic(surfactin)uponperceptionofsomepolysaccharidespresentintherootcellwall.Afastaccumulationofthismultifunctionalantibioticintherootenvironmentisactuallyofclearbenefitforbothpartners.Ononeside,itfavorstheecologicalfitnessofthebacteriumbyimprovingmotility,biofilmformationandearly root tissue colonizationandon theother side, the lipopeptide rapidly reaches thresholdconcentrationsnecessaryfortriggeringsomeimmune-relatedmolecularresponsesinthehostplantleading to a significantly lower susceptibility to phytopathogen ingress. These data provide newinsights into the chemical dialoguebetween the twoorganisms andmorebroadly, into the subtlemolecular mechanisms driving the multitrophic plant-microbe interactions in the rhizosphereenvironment. It also opens the door to further investigations for better understanding howbacilliandotherrhizobacteriacansensetheirenvironmentandnaturalplanthosts.Keywords:cellwallpolymers,rootexudates,lipopeptides,surfactin,plantimmunity

Session1Oral4Agrobacteriumopines:fromtumormarkerstomicrobialecologytoolsSamuelMondy1,2,AuroreLenglet1,3,CélestinLibanga1,PascalRatet1,4,DenisFaure1andYvesDessaux11Instituteforintegrativebiologyofthecell,CEA,CNRS,UniversitéParisSud-UniversitéParis-Saclay,Avenuedelaterrasse,91198GifsurYvette,France2Genosol,UMRAgroécologie,Inra,17RueSully,21000Dijon,France3Departmentofplantmolecularbiology,UniversityofLausanne,Routede laSorge,Biophore,1005Lausanne,Switzerland4Institute of Plant Sciences Paris-Saclay – Université Paris-Saclay Building 630, rue de Noetzlin,PlateauduMoulon,91190Gif-sur-Yvette,France

Opinesarelowmolecularweightmoleculesthataretypicallyfoundincrowngalltumorsand

hairy root hyperplasia incited by members of the Agrobacterium and Rhizobium genera. Opinepresence in the transformed cells creates an environment favorable to the pathogen, promotingbothitsgrowthandthedisseminationofthepathogenicity.

AsidefromtheirroleinAgrobacteriumecology,opineshavebeenusedforsome20yearstoinvestigatehowplantexudationshapesthediversityoftheassociatedmicrobiota.Transgenicplantsproducing opines have been produced and installed in nonsterile “real” soil. Generally speaking,while the carrying capacity of the plants and the concentration of members of several microbialpopulations remained unaffected, a drastic stimulation of the opine-degrading community wasobserved,whatevertheopineproduced,theplantgenusandthesoilcharacteristics.However,thetaxonomicpositionofthemembersoftheopine-degradingcommunitiesvariedasafunctionoftheopinesproducedbytheplant.Thistypeofsimpleexperimentallowedustoverifythevalidityofthe“rhizosphere effect” and to demonstrate that the composition of the associated microbiota maydependonplantexudatesthatmaynotbeproducedanymoreatthetimeoftheanalysis.

More recent experiments based on a similar experimental patternwere set up to analyzemore finely the changes that affected the plant microbiome, using 16rRNA analyses and nextgenerationsequencingtechnologies(NGS).Usingbothtransgenicplantsthatproducedtwodifferentconcentrations of opine and an artificial exudation device, we demonstrated that the amount ofcarbonaffectedbythemodificationoftheexudationisacriticalparameterinvolvedinthereshapingofmicrobialpopulations.Wealsodemonstratedthatroot-associatedbacteriamayhavedevelopedtwodifferentstrategiestocolonizetheplantsurface,i.e.copiotrophy(theabilitytouseawiderangeof substrates) and oligotrophy (the ability to utilize very efficiently only a limited number ofsubstrates).

Keywords:Agrobacterium,opines,NGS,rootcolonization,rhizosphere,microbiota

Session1Oral5Effectofplantmycorrhizationon the rhizosphericmicrobiome structure ina contextofpollutedsoilsHacèneMeglouli1,JoëlFontaine1,FrédériqueChangey1,MarylineMagnin-Robert1,BenoitTisserant1,FrédéricLaruelle1,ThomasLerch2,MohamedHijri3,AnissaLounès-HadjSahraoui1

1. Université du Littoral Côte d’Opale, EA 4492 – UCEIV – Unité de Chimie Environnementale et

InteractionssurleVivant,SFRCondorcetFRCNRS3417,F-62228Calaiscedex,France.2. UMR 7518 « IEES, Institut d'Ecologie et des Sciences de l'Environnement» (CNRS-UPEC-

AgroParisTech-ENS-UPMC-IRD), Université Paris-Est Créteil, 60, avenue du Général de Gaulle,94010CréteilCedex.

3. Institut de recherche en biologie végétale, Université de Montréal and Jardin botanique deMontréal,4101RueSherbrookeEst,Montréal,QCH1X2B2,Canada.

Polychlorinated dibenzo-p-dioxins (PCDD) and polychlorinated dibenzofurans (PCDF), collectivelytermed dioxins, are the most toxic group of persistent organic pollutants (POP). Due to theirstructuralstability,derivedfromtwoaromaticringscontainingonetoeightchlorineatoms,PCDD/Fare widespread in the environment and are considered to be harmful both for environment andhuman health (immunotoxicity, neurotoxicity, carcinogenicity). In the current work, ArbuscularMycorrhizal Fungi (AMF) –assisted phytormediation is proposed to restore aged PCDD/F pollutedsoil. This technique is environmental friendly, potentially cost effective and not altering the soilmatrix.Moreover,AMF improvesthephytotechnologyeffectivenessbyprovidingprotectiontotheplants against the pollutant toxicity and by enhancing pollutant dissipation. The contribution ofmycorrhizal inoculumadditionontheabilityoffourplantspecies(Clover,Alfalfa,RyegrassandTallfescue) to grow and to dissipate PCDD/Fwas assessed after 20weeks ofmicrocosm culture. Ourfindings showed that the four plant species were able to grow on the contaminated soil and theadditionofarbuscularmycorrhizalinoculumprovidesahighcolonizationratedespitethepresenceofPCDD/Finthesoilattheconcentrationof200ng/kg.ThevegetationledtoPCDD/Freductionof38.6ng/kgwiththeTallfescue.Thisdissipationwasfoundtobeduetothestimulationofthemicrofloraevaluated through microbial enzymatic activities (hydrolases and dehydrogenases) and specificbacteriallipidmarkers(PhospholipidFattyAcids).Inparallel,theevolutionofstructureanddiversityof microbiomes refers to the whole community of bacteria, archaea and fungi afterphytoremediationofsterilizedornon-sterilizedsoilwereexamined.Itwasshowedthatbacterialandfungal community structures were significantly affected by arbuscular mycorrhizal inoculation,vegetationandtheinitialmicrobialstateofthesoil.

Keywords:dioxins,phytoremediation,arbuscularmycorrhizalfungi,sterilization,vegetation,rhizospheremicrobiome.

Session 2. SPACE-TIME AND MULTI-SCALES APPROACHES: DIAGNOSTIC, EPIDEMIOLOGYANDECOLOGYINTHEFIELDSKeynotelecture4

Non-invasivesensorsanddigitaltechnologiesforcharacterizingplant-pathogeninteractions-applicationsforprecisioncropprotectionandplantphenotyping

Anne-KatrinMahlein,MatheusKuska,StefanThomas,DavidBohnenkamp,AnnaBrugger,EliasAlisaac,MirwaesWahabzada,JanBehmann,UweRascher,KristianKersting

Thedetectionandidentificationofplantdiseasesisafundamentaltaskinsustainablecropproduction.Anaccurateestimateofdiseaseincidence,diseaseseverityandnegativeeffectsonyieldqualityandquantityisimportantforcropproduction,horticulture,plantbreedingorfungicidescreeningaswellasinbasicandappliedplantresearch.Remoteandproximalsensingtechniqueshavedemonstratedahighpotentialindetectingdiseaseandinmonitoringcropstandsforinfectedplantareas[1].Mostpromisingsensortypesarethermography,chlorophyllfluorescenceandhyperspectralsensors.Thisvarietyofsensorsystemsavailableprovideshighresolutiondataofagriculturalcropstandsorsingleplantorgansandcanconstitutethebasisforanearlydetectionandidentificationofplantdiseases.Particularlyhyperspectralimagingofdiseasedplantsoffersinsightsintoprocessesduringpathogenesis.Fungalleafpathogensinfluenceboth,thestructuralcharacteristicsandtheleafchemistrywhichinturnisreflectedintheopticalpropertiesofplants.Byhyperspectralimagingandsubsequentdataanalysisroutinesitwaspossibletorealizeanearlydetection,identificationandquantificationofdifferentrelevantplantdiseases[2].However,thecomplexweboffactors,influencingopticalcharacteristicsofdiseasedplantsremainpoorlyunderstood.Additionallyscientistworkingwiththesetechnologiesregularlyconfronttheproblemofdealingwithmassive,high-dimensional,andtemporalobservations,posingachallengeinscalability.Toutilizethefullpotentialofthesehighlysophisticated,innovativetechnologiesand,complexdataforprecisioncropprotectionorplantphenotyping,amulti-disciplinaryapproach—includingplantpathology,engineering,andinformatics—isrequired[3].

[1]Mahlein,A.-K2016.Plantdiseasedetectionbyimagingsensors-parallelsandspecificdemandsforprecisionagricultureandplantphenotyping.PlantDisease100(2),241-251

[2]Rumpf,T.,Mahlein,A.K.,Steiner,U.,Oerke,E.C.,Dehne,H.W.,Plümer,L.(2010).Earlydetectionandclassificationofplantdiseaseswithsupportvectormachinesbasedonhyperspectralreflectance.ComputersandElectronicsinAgriculture,74(1),91-99.

[3]Wahabzada,M.,Mahlein,A-K.,Bauckhage,C.,Steiner,U.,Oerke,E-C.,Kersting,K.(2015):Metromapsofplantdiseasedynamics-automatedminingofdifferencesusinghyperspectralimages,PLOSOne10(1):e0116902

Session2Oral6LocalandlandscapeeffectsonEuropeangrapevinemothandGrapepowderymildewinvineyardsDuduChakuki,1,2andLiorBlank11DepartmentofPlantPathologyandWeedResearch,AgriculturalResearchOrganization,VolcaniCenter,BetDagan50250,Israel2DepartmentofGeographyandHumanEnvironment,TelAvivUniversity,RamatAviv,TelAviv,69978,IsraelAgriculturalareasarecharacterizedbyaheterogeneouslandscapewhichincludesavarietyofagriculturalcropsalongsidenaturalareas,forests,builtareasetc.Thelandscapeandtheenvironmenthaveanimpactonthedynamicsofpestsandpathogensinspaceandtime.Untilrecently,studieswerebasedonsmallnumberofobservationscollectedinthefieldorfromthelaboratory.Duetothecomplexityofthebiologicalsystem,itisdifficulttodefinetheimportanceofthevariablesincontrolledtrialsorinsmallextentstudiesandtoconcludereliablerelationships.Agriculturalresearchisincreasinglybecomingadata-intensivescience,relayingonmassiveamountofdatacollectedinthefield.Inthisstudyweexaminedpredictorsofthedistributionoftwoimportantpestsinvineyards-Europeangrapevinemoth(LobesiaBotrana)andGrapepowderymildew(Uncinulanecator)atbothlocalandlandscapescales.Weusedmonitoringdatacollectedin220commercialvineyards.Inthelocalscalewefoundthatthemostinfluentialvariables,forbothpests,werethegrowerandthecultivar.Forthelandscapescale,wecharacterizedthelandscapecontextaroundtheplotsin10nestedcirclesrangingfrom100to1500minradius.WeobtainedsignificantpositivecorrelationsbetweenEuropeangrapevinemothincidenceandthetotalareaofvineyards(peakingat100m),orchards(200m),forestareas(1500m)andbuiltareas(1500m).Incontrast,Grapepowderymildewwasnotcorrelatedtolandscapecontext.Theseresultsdemonstratethatlandscapestructurehavedirecteffectsonpests.Theseeffectsdependonboththespeciesandthescale.Understandingtheeffectsoflandscapecontextcanimproveourunderstandingregardingpestbiologyandofferspowerfulinformationforfutureresearchaboutdispersal-relatedcharacteristicsofpestsandcanimprovepestcontrolmanagement.Keywords:land-use,LobesiaBotrana,Uncinulanecator

Session2Oral7Theroleofinfectedflowerpetalsandleaves,andprecipitationinpredictingSclerotiniastemrotofoilseedrapeAndreaFicke1,ChloéGrieu1,MayBenteBrurberg1,GuroBrodal1,1NorwegianInstituteofBioeconomyResearch(NIBIO),Sclerotiniastemrot(SSR)isanincreasinglyimportantdiseaseofoilseedrapeinmanypartsoftheworld.Sclerotiniasclerotiorum,knownasthecausalagentofSSR,hasanextremelywidehostrangeincludingmanyvegetablecropsandweeds.Thefungusproducessclerotia,whichcansurvivedormantinthesoilformanyyears.Sclerotiagerminateunderwetconditionsanddischargeascosporesfromapothecia.Thesporesarethoughttoinfecttheirrapeseedhostviaflowerpetalscaughtinleafaxils.Availabilityofresistantvarietiesislimited,hencerotationsaswellasfungicidesarethemostcommonwaystocontrolthedisease.Aligningfungicideapplicationswiththeactualneedforcontrolhasledtothedevelopmentofseveraldiseaseriskpredictionmodels.However,theaccuracyofpredictionformostofthesemodelsisnotsatisfactory.Theobjectiveofourworkwastoinvestigatesomeofthekeyfactorsinthepathogen’slifecycle:theimportanceofprecipitationandtheroleofflowerpetals.Wegrewspringoilseedrape(Brassicanapus)cultivar‘Mosaik’inthegreenhouseuntilfloweringandplacedtheminthefield.Plantswereexchangedeveryseventotendaysduringthegrowingseasonwithhealthyfloweringplants.Afterbeingexposedtofieldconditions,theplantswerecoveredwithplasticbags,broughtinsideandincubatedfortwotothreeweeksbeforevisualassessmentforSSRinfection.WerepeatedthisexperimentoverfouryearsandatthreedifferentlocationsinSoutheastNorway.Localweatherstationsprovideddataonprecipitation,temperatureandrelativeairhumidity.Neitherprecipitationduringexposureinthefields,norprecipitationoneandtwoweeksbeforeexposuretime,correlatedsignificantlywithSSRinfectionoftheplants.FlowerpetalsandgreenleavesofrapeseedplantswerecollectedfromninedifferentfieldtrialsovertwoyearsinSoutheasternNorwayandtestedwithqPCRforpresenceofS.Sclerotiorum.SeverityofSSRinfectioninthefieldtrialswereassessedatharvestandcorrelatedwithpercentageofpositiveleafand/orpetalsamplestested.TherewasnocorrelationbetweenSSRseverityinthefieldandpercentageofpositiveleaforpetalsamples.SeveralSSRriskmodelsuseprecipitationandfloweringasdeterminingfactorsforSSRinfection.Ourstudiesindicatethatprecipitationisnotaspredictiveforinfectionasgenerallythought.SoilhumidityorleafwetnessmightbemoreimportantfactorsinassessingtheriskforSSRinfectionandspread.FlowerpetalsplayanimportantroleintheSSRepidemic,buttherelationshipbetweenpetalinfectionandSSRinfectionappearsnottobeasdirectaspreviouslyindicated.Keywords: Sclerotinia sclerotiorum, sclerotinia stem rot, rapeseed, canola, oilseed rape,epidemiology,infectionperiods.

Session2Oral8Whenaplantpathogenrunsdownariver:populationgeneticsofthepoplarrustepidemicsintheDuranceRivervalley.Pascal Frey1, Constance Xhaard1,2, Michaël Pernaci1,3, Ronan Becheler1, Katherine J. Hayden1,4,StéphaneDeMita1,FabienHalkett1.1UMRIAM,INRA,UniversitédeLorraine,54000Nancy,France2Presentaddress:Unité1018INSERM,InstitutGustaveRoussy,94800Villejuif,France3Presentaddress:PalmElitSAS,ParcAgropolis,34980Montferriez-sur-Lez,France4Presentaddress:RoyalBotanicGardenEdinburgh,Edinburgh,EH35LR,UKTracking pathogen movement, identifying source populations and understanding environmentalfactors, including human activities, that influence pathogen spread are central tasks in diseaseecology. Herewe focus on the spread of a plant pathogen in a wild pathosystem. Every year wemonitoredan annual epidemicof the Europeanpoplar rust fungus,Melampsora larici-populina, intheDuranceRiver valley, in the FrenchAlps. This valley is particularlywell suited for the study ofrecurrent biological invasions: the need of an alternate host plant (larch) to perform its sexualreproduction restricts the resident pathogen population upstream the river, in a poplar-larchsympatry area. Then, a clonal epidemic phase spreads downstream the valley during fivemonthsalongaca.200kmnaturalriparianstandofblackpoplar,Populusnigra.Thislandscapealsoincludesa few cultivated stands with poplars carrying qualitative resistances, thus exerting a peculiarselection pressure on pathogen populations. In this study we used epidemiology and populationgeneticstoolsto(i)sortM.larici-populinaindividualsaccordingtotheirwildorcultivatedorigin,(ii)todescribethespreadoftheepidemiconthewildpoplarstands,(iii)toassesstheevolutionofthegenetic composition of the pathogen populations along the epidemicwave, and (iv) to assess theevolutionoflifehistorytraitsduringtheepidemic.Theseresultsarediscussedinthelightofrecentstudies focusing on the relative effects of demographic and selection events on the evolutionarychangesaccompanyingbiologicalinvasions.Keywords:disease ecology, landscape epidemiology, colonization, dispersal, range expansion,wildpathosystem

Session2Oral9

Spatiotemporal patterns of Phytophthora megakarya infections in newly established cacaoplantationsinCameroonNdoungué Djeumekop M.M1,2, Ngo Bieng M.A4, Nembot C.G5, Petchayo S1,5, Neema C2, and TenHoopenG.M3

1IRADBP2123Yaoundé,Cameroun2Montpellier SupAgro,UMRBGPI,Campus InternationaldeBaillarguet,34398MontpellierCedex5France3CIRADUPRBioagresseurs,CampusInternationaldeBaillarguet,34398MontpellierCedex5France4CIRADUMRSystem:TropicalandMediterraneanCroppingSystemFunctioningandManagement2PlaceViala,Bât.27,34060Montpellier,France5UNIVERSITEDEYAOUNDE1,FacultédesSciences,BP812Yaoundé,CamerounStudying spatial and temporal plant disease dynamics helps to understand pathogen dispersalprocesses, a necessary undertaking in epidemiology in order to improve disease controlrecommendations.Inthisstudy,plantationsdevoidofprimaryinoculumofPhytophthoramegakarya(causalagentofcacaoblackpoddisease),uponestablishmentin2006,weremonitoredforpresenceof disease on a weekly basis from 2009 to 2016. Isolates of P. megakarya collected in theseplantations were genotyped with 14 SSR markers. Ripley’s K functions were used to characterizespatialdiseasedynamics.TheunivariateK-functionwasusedtodescribespatialdiseasepatternsandthe bivariate K12-function was used to describe the relation between healthy and diseased cacaotrees. Disease distribution maps show aggregated disease patterns in all plots. The K-functionconfirmed these results although it was not significant in all patterns, probably due to a limitednumber of diseased cocoa trees. Healthy and diseased cacao trees were mostly negativelycorrelated, indicating that cacao black pod disease dispersal is a clustered process preferentiallyaffecting neighbors of already infected trees. Basedonobservations it appears that occurrenceofblackpoddisease isnotacompleterandomizedprocess.Theneighboringenvironmentcangreatlyinfluencediseasedispersalprocesses.For instance, closeness toalready infectedcacaoplantationscan favor dispersal of disease propagules while the presence of a river can increase the diseaseincidenceandpathogendiversity.According,totheresults,blackpoddiseaseismainlyspreadoverrelatively short distances. Isolation of newly established cacao plantations from infected onesappearsthereforetobeaneffectiveapproachtocontrolofblackpod.Keywords:Phytophthoramegakarya,firstinfections,spatialpattern,temporalevolution,cacao

Session2Oral10DisseminationofXanthomonasfragariaeinastrawberryfieldcrops JanvanderWolf1,PieterKastelein1,BertEvenhuis1&ArnoldMoene2

1WageningenPlantResearch,P.O.Box16,6700AAWageningen,EJan.vanderWolf@wur.nl2WageningenUniversity,DepartmentofEnvironmentalSciences,MeteorologyandAirquality.P.O.Box47,6700AAWageningen

Xanthomonasfragariae (Xfr) is thecausativeagentofangular leafspot instrawberry,aquarantineorganisminEurope(EPPOA2).Thedisseminationofthepathogeninsymptomaticstrawberryfieldsbymachineries,splashwaterandaerosolswasstudied.A rotarymowerwas used to clip leaves of symptomatic plants. The cutter blades of themachinebecame heavily contaminated. Use of these contaminated machine resulted in (symptomless)infectionsofplantsinitiallyfreeofXfr,evenat10mdistancefromthesymptomaticplants.Cleaningofthemowerwithawaterhosewasnotsufficienttoremovethepathogen.SpreadofXfrfromsymptomaticplantsbysplashwaterduringoverheadirrigationatmoderatewind,resultedinaspreadoveradistanceofatleastfourmeter.Atwindlessweatherconditionsandlightrainasimilarminimumdistanceofspreadwasfoundbutbacterialdensitiesrecoveredwerelower.AnexponentialdecreaseofXfrdensitywasfoundwithdistancefromtheinoculumsource.Inartificiallycreatedaerosols,Xfrcouldbecollectedbyanairsampleruptoadistanceof100mfromthe sourceand those createdbymowinga symptomatic cropup toadistanceofminimally25m.Aerosols releasedby a sprayer, carrying in total ca. 1011 cfuofXfrwas able to cause infectionsofstrawberryplantsuptodistanceofatleast10mofthesprayer.ItisconcludedthatthereisaconsiderableriskofspreadofXfrfromsymptomaticplantsbymachinesand wind-driven aerosols within and between fields. Splash dispersal during rain or overheadirrigationcanresultofspreadXfrwithinthefield.Keywords:Angularleafspot,machines,aerosols,airsampler,disinfection,splashingwater

Session2Oral11(invitedlecture)SeasonalascosporereleasebythewheateyespotpathogensOculimaculayallundaeandO.acuformisintheNorthwestUSATimothyD.Murray

DepartmentofPlantPathology,WashingtonStateUniversity,Pullman,WA99164-6430USA

EyespotdiseaseofwheatiscausedbyOculimaculayallundae(OY)andO.acuformis(OA).ApotheciaofOAandOYwerefirstreportedincommercialwheatfieldsintheU.S.PacificNorthwest(PNW)in2003and2014,respectively,buttheirroleineyespotepidemiologyremainsunknown.PopulationgeneticstudiesofOAandOYinthePNWconcludedthatbothspecieswereundergoingsexualreproduction,butitsfrequencyandroleinthediseasecyclearenotknown.Apreviousascosporetrappingstudyconductedininoculatedfieldplotsdemonstratedthatascosporereleaseoccurredinspringandautumn,withmoresporestrappedinspring.Inthisstudy,ascosporetrappingwasconductednearnoninoculatedwheatfieldstodeterminewhetherandwhenascosporeswerepresent.AscosporeswerecollectedwithBurkardvolumetricsporesamplersduringthe2014-15and2015-16cropseasonsatfourlocationsnearPullman,WA(S15,S16,PP15,andPC16).Stickytapeswerecollectedweekly,examinedmicroscopically,andthenDNAwasextractedandRT-PCRusedtoquantifyDNA,whichwasconvertedtonumberofascospores.

AscosporesofOAandOYwerepresentinbothcropseasons,withmoresporescollectedin2015-16than2014-15atalllocations.Thegreatestreleaseofascosporesoccurredinautumnatthreelocations(S15,PP15,andPC16)andwasequalinautumnandspringattheotherlocation(S16).MoreascosporesofOYthanOAwerecollectedinbothyearsatalllocations;basedonDNAcontent,OYwas2.3to4.8timesmorefrequentthanOA.Itisnotclearwhymoreascosporeswerepresentin2015-16,butmayberelatedtoaboveaveragetemperaturesduringspring2016.Itisclearfromthesedatathatascosporesarepresentduringtheautumn,whichistheprimaryinfectionperiod,andmayhelpexplainthehighgenotypicdiversityobservedinpreviousstudies.

Keywords:wheat,eyespot,epidemiology

Session2Oral12Apossibleevolutionaryresponseofleaffungipathogensofcerealstochangesinfertilizationlevel.Pierre-AntoinePrécigout1,2DavidClaessen2CorinneRobert11:UMRINRA/AgroParisTechEcoSysInstitutNationaldelaRechercheAgronomiqueRoutedelaFermeF-78850Thiverval-Grignon2:UMRMathématiquesEco-EvolutivesInstitutdeBiologiedel'EcoleNormaleSupérieure(IBENS)46rued'Ulm75005ParisWeuseamodelof lifehistoryevolutionofcrop fungalpathogensaiming tounderstandhowtheyadapttofertilization.Wefocusonasinglepathogenlifehistorytrait,thelatentperiod,importantforpathogen resource allocation strategies and canopy colonization.We implement three fertilizationscenarios corresponding to major effects of nitrogen fertilization on cereals: increase in (i)metabolitesinleaves,(ii)leaflifespanand(iii)canopyleavesdensity.Wefindthatsporesproductionincreases with higher fertilization.We use two fitness measures to identify putative evolutionaryresponses of latent period to changes in fertilization. When fitness is defined as annual sporesproduction,wepredictapositivecorrelationbetweentheoptimallatentperiodandfertilization.Bycontrast, the model predicts a negative relationship between the optimal latent period andfertilizationwhen fitness isdefinedas thewithin-seasonexponentialgrowth rateof thepathogen.Wehypothesizethat if thecriticalstep intheco-dynamicsofmultiplestrains is thecolonizationofnewfieldsthenthe limitingstepwill likelybetheamountofspores innextyear’s inoculum,whichwillinfluencetheearlystepsofepidemics.Inthatcase,thestrainthatproducesmostsporesoveraseasonislikelytowinthecompetition,andevolutionshouldmaximizeannualsporesproduction.If,on the other hand, the competition between strains is mainly experienced by within-seasoncompetitionforhealthycroptissue,thentherateofspreadthroughthecanopywillbecritical.Wepredict that reducing fertilization will change the aggressiveness profiles of different biotrophicfungiusspeciesorstrainsinthefield.Keywords:Epidemiology,Fertilization,LatentPeriod,LifeHistoryTheory,Resource-basedModel,Structured-populationsModel

Session2Oral13Genetic diversity and population structure of Zymoseptoria tritici in France at different spatialscalesAliSiah1,BenoitTisserant2,MyriamBomble,PatriceHalama1andPhilippeReignault21ISA,EA7394-ICV-InstitutCharlesViollette,F-59000Lille,France2 Unité de Chimie Environnementale et Interactions sur le Vivant (EA 4492), Université du LittoralCôted’Opale,CS80699,F-62228,Calaiscedex,FranceZymoseptoriatriticiisthemostfrequentlyoccurringanddamagingfoliarpathogenonwheatcropsinEurope,especiallyinFrance,whereenvironmentalconditionsaresuitablefordiseasedevelopment.Morethan1500isolatesofthisfungus,isolatedinFranceoveraten-yearperiod(2005-2015)andatdifferentspatialscales(country,Hauts-de-Franceregion,field,30cm2square,plant,leafandlesion),werecharacterizedusingSSRmarkersandmatingtypeidiomorphsinordertogaininsightintofungalpopulationgenetic features.Results revealedoverallhighandsimilar levelsofgenicandgenotypicdiversitiesatthecountry,regionandfieldscales,butdiversityvaluesgraduallydecreasedfromthefield to finer scales, such as 30 cm2 square, plant, leaf and lesion. Bayesian and non-Bayesianstatistical analyses revealed significant structuration of the global French population into threegeneticclusters,andtheHauts-de-Francepopulationintothreesub-clusters,alldistributedaccordingto their geographical origin. However, no genetic differentiation was found at finer scales. Bothmatingtypeswerefoundatequalfrequenciesatthecountry,regionandfieldscalesandfoundtoco-occur together at all other investigated scales. This study revealed a high potential for sexualrecombination and genetic diversification for Z. tritici in France, as well as a structuration of itspopulations at largest scales, in accordance with local adaptation to agro-climatic conditionsfacilitatedbyhighfitnessdegreeofthepathogen.Keywords:Zymoseptoriatritici,geneticdiversity,populationstructure,SSRs,matingtypes.

Session2Oral14

LeafblotchondurumwheatinFrance:populationgenetics,hostspecializationandfungicideresistanceR.VALADE1, T. CAPARROY1, C. VITRY1,O. BEAUDOUIN1,M.MASSOT1, T. LEVEFRE2, J-P. LEGOFF2, F.LACOUDRE3, C. ANDRE4, M. COCHARD5, J. CONFAIS6, G. GAZEAU6, V. LAVAL 6, A-S WALKER6, T.MARCEL6,L.GOUT6,7

1ARVALIS Institut du Végétal Avenue Lucien Brétignières, Bâtiment INRA Bioger, 78850 Thiverval-Grignon,France.2RAGT2n,RueEmileSingla,SitedeBourran,BP3336,12033RODEZCedex9,France3LIMAGRAINFermedel’Etang77390Verneuill’Etang,France4SYNGENTA,12chemindel’Hobit,31790Saint-Sauveur,France5FLORIMONDDESPREZVeuve&Fils,3rueFlorimondDesprez,59242Cappelle-en-Pévèle,France6INRA,UMR1290BIOGER,UniversitéParis-Saclay,CampusAgroParisTech,78850Thiverval-Grignon,France7AGROPARISTECH, UMR1290 BIOGER, Université Paris-Saclay, Campus AgroParisTech, 78850Thiverval-Grignon,France

Leaf blotch, caused by Zymoseptoria tritici and Parastagonospora nodorum, is a major diseasecomplex ofwheatworldwide. In France, these diseases have been reported to cause severe yieldlossesinbreadwheat,mostofwhichwereduetoZ.tritici.Incontrast,ourknowledgeofthefungalspeciescausingleafblotchondurumwheatislimitedwhereastheoccurrenceofepidemicshasbeenincreasing in recentyears.Werecentlyconducteda3years surveyof leafblotchondurumwheatand breadwheat in fourmain French growing regions.More than 1100 isolates of Z. tritici weresampledfromnaturallyinfectedfields(treatedanduntreatedfields)ofbreadanddurumwheatandgenotypedusing 12microsatellitemarkers. The frequencies of occurrence among sites of the twofungal species in leaves were also evaluated using microbiological isolation methods and qPCRdetection.Lastly,thelevelofhostspecializationandaggressivenessofasubsetofP.nodorumandZ.triticiisolateswerealsodeterminedbycrossinoculationexperimentsonapanelofbreadanddurumwheat varieties. Based onmolecular test results and isolation assays, Z. tritici was detected in allFrench regions on durumwheat and breadwheat. P. nodorum was detected on durumwheat inmostFrenchregionsbutwasquantifiedonlyat low levelsbyqPCRonbreadwheat.Frenchdurumwheat cultivars were highly susceptible to the French P. nodorum isolates tested. The geneticdiversity of Z. tritici was structured by host and location of sampling. Inoculation experimentspointed out specialist, generalist and maladapted strains of Z. tritici in relation with populationgenetic groups. No differences between Z. tritici bread wheat populations and durum wheatpopulationswereobservedforfungicideresistance.Theseresultsshowthatcontrolofleafblotchondurum wheat, in France, needs to take into account the presence of both species and growingregions.Theseresultsmayhelptoadaptsustainablediseasemanagementstrategies.

Keywords:Z.tritici,P.nodorum,durumwheat,resistance,populationgenetics

Session2Oral15

Spatio-temporal diversity of thermal responses in populations of the wheat pathogenZymoseptoriatriticiAnne-LiseBoixel1,MichaëlChelle2andFrédéricSuffert1

1UMRBIOGER,INRA,AgroParisTech,UniversitéParis-Saclay,78850Thiverval-Grignon,France.2UMRECOSYS,INRA,AgroParisTech,UniversitéParis-Saclay,78850Thiverval-Grignon,France.

Amongtheenvironmental factorsaffectingthedevelopmentofplantpathogens,temperature isofcriticalimportanceasithasimpactsonallstagesoftheirinfectiouscycle.Pathogenpopulationsareconstantlyexposedtothermal fluctuationsatvariousspatio-temporalscales.Tobetterunderstandhowtheycanrespondtothesechanges,theiradaptivepotentialshouldbetakenintoaccount.Thischallengeisbecomingincreasinglyessentialwiththedevelopmentofagro-ecologyinthecontextofclimatechange,whichwillimpactthethermalenvironmentoffoliarpathogens.Theobjectiveofthisstudywastoassessthediversityofthermalresponsesbothbetweenandwithinnaturalpopulationsof thewheat pathogenZymoseptoria tritici. For this purpose, ninepopulationsof 30 isolates eachwerecollectedbyconsideringboth: (i)atemporalscalewithtwopopulationssampled inthesamefieldatthebeginningandattheendofanannualepidemic(seasonaleffect);(ii)aspatialscalewithsevenpopulationscollectedalongtwothermalgradientsattheEuropeancontinentalscale(climaticzone effect). The response to temperature of each of these isolateswas characterized in vitro bymonitoring their multiplication rate in liquid culture media over four days and under 12temperaturesrangingfrom6to34°C,then insilicobyfittingathermalperformancecurveforeachisolate. Key parameters were analyzed to compare the intra- and inter-population diversity ofthermalresponses.Thus,thisstudycontributestobetterapprehendthediversityofresponsesandthe existence of local adaptation patterns to temperature ofZ. tritici at a population level. Thesefindings will be used to infer the potential response of given populations to changes in thermalconditions and could be taken into consideration to improve prediction model of septoria triticiblotchepidemics.

Keywords:temperature,thermalperformancecurve,adaptivepotential,spatio-temporalgradients,Zymoseptoriatriticipopulations

Session2Oral16PopulationbiologyofPucciniacoronata–implicationsfortheepidemiologyofcrownrustonoatsAnnaBerlin1,JonasTörngren1,Ann-CharlotteWallenhammar2,andBjörnAndersson1

1SwedishUniversityofAgriculturalSciences,Dept.ForestMycologyandPlantPathology,Box7026,S-75007Uppsala,Sweden2RuralEconomyandAgriculturalSociety,POBox271,S-70145Örebro,Sweden

ThefungusPucciniacoronataisthecausalagentofthecrownrustonoats(Avenasativa)andgrasses. Thedisease is amajorproblem inoatgrowing regionsand itmay causedevastatingyieldlosses.Westudied thepopulationbiologyofP. coronata inoat fieldsandon thealternatehost indifferent regions to get a deeper understanding of the importance of the alternate hosts in theepidemiology of the disease. Samples were collected during a seven-year period, and collectionsweremadefrombothoatsandthealternatehostbuckthorn(Rhamnuscatharitcia).

To explore if the role of the alternate host, on which the sexual cycle of the life cycle iscompleted, sampleswere collected inmultiple years at the same farms. Inaddition, samplesbothfrom the alternate host, R. catharitcia, and oats in adjacent fields to the alternate host werecollectedinoneyearwithinaspecifiedregion.

We found that the genotypic variation of P. coronata within oat fields is high, as well asbetweenoatfieldswithinandbetweenregions.Thesamplescollectedonthealternatehostdidnotbelongtothesamepopulationasthesamplescollected intheadjacentfields; suggestingthatthepresenceofthealternatehostgreatlyincreasestheoverallgeneticdiversityofthefungus,however,onlyafewgenotypesoriginatingfromR.catharticawillsuccessfully infectoats.The lackofgeneticstructure within and between samples collected from different fields in the different regions,suggests thatwind-spreadspores fromother R.catharticahostswithina regionwill contribute tothegeneticdiversitywithinoatfields.ThisimpliesthatthealternatehostmustbeeradicatedwithinalargerareatodecreasethegeneticdiversitywithinP.coronataandlimitthespreadofinoculumtosusceptibleoatfields.

Keywords:P.coronataf.sp.avenae,populationbiology,epidemiology

Session2Oral17Assessment of Verticillium flax inoculum in agroecosystem soils using real-timePCR assay: fromdiagnosistoevaluationofcropsystem.MélanieBressan,ManonPeyré,AdrienBlum,LisaCastel,JéromeAilhas,IsabelleTrinsoutrot-Gattin,KarineLaval,ChristopheGangneux.

Verticillium wilt, due to the soilborne fungus Verticillium dahliae, is a persistent diseaseaffectingflaxcropsinNormandy.Thispathologyhasincreasedsincethelastdecade,leadingtoyieldlossesforflaxproducers.InpartduetothelongsurvivalofV.dahliaeinsoilandthedifficultyofearlydiagnosis, Verticillium flax wilt management remains problematic. Pathogen avoidance and thereductionofsoil inoculumthroughadaptedculturalpracticesarethetwobestalternativestofightagainstVerticilliumwilt.

Here,theobjectivesweretodeveloparapidandspecificassaytomeasureV.dahliaedensityin soil. A real-time PCR assay was optimized and validated to provide a sensitive and reliablequantification ofV. dahliae in a range of artificially inoculated soilswith known inoculumdensity.Thisassaywasthensuccessfullyappliedtostudythe insiturelationshipbetweenpathogendensityanddiseasedevelopment in flax fields. Somestudieshavealready reported thedirect relationshipbetween Verticillium densities in soil and disease severity on several important crops (potato,cauliflower…).Butthislinkappearedtobemorecomplexforflax.Theimportanceofsoilreceptivity,linkedtochemicalandmicrobialcharacteristics,havetobeparticularlyconsider.

Finally, the real-time PCR assaywas also used to evaluate and compare pathogen load infieldsfromdiversemanagementsystems:conventional,integratedandorganic.Verticilliumdensitiesappeared to be impacted by agricultural practices, and particularly tillage. The influence of thepreviouscroponpathogenloadhadalsotobeconsideredwithattentiontomanageefficientlythisdisease throughcrop rotation.MeasuredV.dahliaedensities inall fieldspresentedan intra-parcelheterogeneity, emphasizing the importance of an adapted sampling strategy to assess pathogendensity. Such knowledge of pathogen density in soils could provide critical information forstakeholderstoidentifyinfestedfieldsandpredictdiseasedevelopment.Keywords:Real-timePCRassay,Verticilliumdahliae,Flax,soil,cropfields

Session2Oral18IncidenceofrootpathogensassociatedtocloverrootrotinSwedenCharlottaAlmquist1,2,EvaStoltz3andAnn-CharlotteWallenhammar3,4

1DepartmentofPlantBiology,UppsalaBiocenter,SwedishUniversityofAgriculturalSciences,POBox7080,SE-75007Uppsala,Sweden2LantmännenReppeAB,Jungmansgatan8,SE-53140Lidköping,Sweden3RuralEconomyandAgriculturalSociety,HSKonsultAB,POBox271,SE-70145,Örebro,Sweden4PrecisionAgricultureandPedometrics,DepartmentofSoilandEnvironment,SwedishUniversityofAgriculturalSciences,POBox234,SE-52323Skara,SwedenRedclover(Trifoliumpratense)isanimportantforagelegume,however,thepersistenceinSwedishleysislimitedbyrootrotassociatedwithseveralfungalpathogens.Foridentificationofsoil-bornepathogenscausingdiseasesymptomsinrootsfastandspecificdiagnosticmethodsweredeveloped.Theobjectivesweretoidentifythecomplexofsoil-bornepathogensassociatedtorootrotandtomonitordiseasedevelopmentbyqPCRinasetofredclovercultivarsinfield-infectedredcloverroots.Afieldexperimentwassetupwithelevenfutureandmarketcultivars.Pathogenidentificationandquantificationbyreal-timePCRwasperformedinrootsofthreeselectedcultivarssampledatdifferenttimepointstheseedingyearandonceduringthefirstandsecondharvestyear.DNAfromFusariumavenaceum,Phomaspp.andCylindrocarpondestructanswereidentifiedonallsamplingoccasions,whileF.culmorumoccurredsporadically.SignificantdifferencesintheamountofdetectedpathogenDNAwerefoundwithinthefirstsamplingyearandbetweenyearsforeachofthepathogens,whilenodifferencewasshownbetweencultivars.WehavedemonstratedthatqPCRprovidesausefultoolforevaluatingthesusceptibilityofredclovercultivarsmonitoringthepathogenscausingdiseasesymptoms,andconstitutesatoolforthefirststeptowardsadiseaseriskassessmentofcloverrootrot.

Roundtable1

Oral19

TheuseofTaqManPCRandNextGenerationSequenceTechnolgytoevaluatethepresenceofplantpathogensintomatoseeds:What’sinaPipeline?

PeterBonants,ElsVerstappen,IlseHouwers,AnnetteDullemansandTheovanderLee

WageningenPlantResearch,Droevendaalsesteeg1,6708PBWageningen,TheNetherlands

Objectives:EvaluateNGStechnologyfortheuseofdetectionofplantpathogensintomatoseedscomparedto

real-timeTaqManPCRMethodsused:DNAandRNAextractionfromspikedandunspikedtomatoseeds,TaqManPCR,IlluminaHiSeqsequence,pipelineanalysisofsequencedataResults:Seedsaretransportedallovertheworld.Manydifferentpathogensoftomatocanbetransmittedbyseedandthereforemonitoringseedhealthisessentialtoreducecontaminationrisks.Forthedetectionoftheseseedtransmittedpathogensmanymethodsareavailable,e.g.ELISA,IF,dilutionplating,PCRandreal-timePCR.WeevaluatedTaqManPCRmethodsoftomatoseedtransmittedpathogensandtheuseofNextGenerationSequenceTechnologywithafocuson:Clavibactermichiganensissubsp.michiganensis,Xanthomonaseuvesicatoria,X.perforans,X.vesicatoriaandX.gardneri,Pseudomonassyringaepv.tomato,PepinoMosaicVirusandPotatoSpindleTuberViroid.TheTaqManPCRassayswereshowntobespeciesspecificwhentestedonaseriesofpathogensandrelatedspecies.DifferentDNAandRNAextractionkitswereevaluatedincombinationwiththeKingFisherplatform.Validationofthecompletemethodwasperformedforthedifferentperformancecharacteristics:analyticalsensitivity,analyticalspecificity,repeatability,reproducibility,selectivityandrobustnessusingseedbagscontaininghealthyseedlotsspikedwiththedifferenttomatopathogens.ObtainedDNA-andRNAextractswereevaluatedwithIlluminaHiSeqanalysisforthepathogensincludedinthestudy.Conclusions:AnefficientDNA/RNAextractionprocedurewasdevelopedfortomatoseeds.TaqManPCRswerevalidatedonasetoftomatoseedtransmittedpathogens.TheseDNAextractswerealsousedforIlluminaHiSeqanalysis.WefoundagoodcorrelationbetweenresultsobtainedbytheTaqManassaysandtheNGSanalysistodemonstratethepresenceofthepathogens.

6keywords:DNA/RNAextraction,tomatoseeds,TaqManPCR,NGS

Roundtable1Oral20Theapplicationofnext-generationsequencing(NGS)offersgreatopportunitiesforthedetectionanddiagnosisofvirusesandviroidsinplantsKrisDeJonghe,YoikaFoucart,AnneliesHaegemanandMartineMaesFlandersresearchinstituteforagriculture,fisheriesandfood(ILVO),PlantSciencesUnit,Burg.VanGansberghelaan96,9820Merelbeke,BelgiumThecurrentwidelyusedmolecularmethodsfordetectionandidentificationofvirusesandviroidsinplants,mostlyRT-(q)PCRandSangersequencing,havetheirlimitations.Firstofall,theneedtomaketherightchoiceofthevirus-specificmoleculartesthasanimportantimpactonthereliabilityofthetestresult.Secondly, incaseofmultipleinfections,thecorrectconclusionisnotalwaysdrawn,andthirdly, unexpected presence of viruses or viroids may easily remain unresolved and relatedsymptomsmayleadtothefalseconclusionofa“nonparasiticcause”.Nextgenerationsequencing(NGS) or deep sequencing follows a massively parallel sequencing (MPS) approach, a powerfultechnologyofferingahugepotentialforholisticvirus/viroididentificationinallkindsof(suspected)planttissuesandthiswithoutprecedingknowledgeonthetargetsequences.Withintheframeworkof the transnational EUPHRESCOproject “NGSDetect”, a groupof 15partners isworking towardsdevelopingandadaptinga standardizedNGS technology for virusesandviroids inplants. Ina firstseries of experiments, ILVO has assessed several key aspects, starting from the subsamplingprocedure,uptothecomparisonofdataanalysisstrategies,followingNGSonanIlluminaplatform.Severalhostplantswereused(e.g.apple,pear,potato,cherry),bothwithknownandunknownvirusinfections.Initially,partialvirusenrichment(purificationbyultracentrifugation)wascomparedwithadirectRNAextractionontheplanttissue.However,theviruspurificationproceduredidnotleadtosufficientRNAofyieldandintegritytopassthequalitycontrol(QC)andwasabandoned.Further,theeffectofthreetotalRNAextractionmethods(Trizoland2kitextractionmethods,RNeasy(Qiagen)andmirVana(Ambion))ontheNGSresultsweremutuallycompared.TheresultsofthesemethodsbasedontotalRNAextractionwerealsocomparedtoaprotocolstartingfromsmallRNAsextracteddirectly from theplant tissue (mirVana (Ambion) siRNAextraction). Inaddition, theeffectof rRNAdepletion was evaluated. Library preparation and NGS were outsourced (Admera Health, USA).Finally, thedatawereanalysedusing in-housescriptsand throughanautomatedpipeline forvirusdetection intheNGSdata(VirusDetectpipeline;Zhengetal.2017).Currently,acomparisonwithasecond automated pipeline (Virtool;©2016 Canadian Food Inspection Agency) is being evaluated,both for thedetectionofknown,and“new”viruses.AllobtainedNGS resultswereconfirmedandevaluatedby specific detectionwith the available classicmolecular tools. The current evolution incontinuouscostreductionfortheroutineuseofNGSalsoletspresumethatintheverynearfuture,onceanoptimised strategywill be available, the technologymaybewidely implementedbyplantvirusresearchanddiagnosticlabs.

Keywords:nextgenerationsequencing,NGS,deepsequencing,bioinformatics,siRNA

RoundTable1Oral21RapidandaccurateidentificationofRalstoniasolanacearum“speciescomplex”byMALDI-TOFMS.J.L.J.vandeBilt1,M.Wolsink1andM.Bergsma-Vlami11DutchNationalPlantProtectionOrganization(NPPO-NL),P.O.Box.9102,6700HCWageningen,theNetherlandsRalstonia solanacearum “species complex” (RSSC) is a soil-borne plant pathogenic bacterium,consistingofdiverseandwidespreadstrainsthatcausebacterialwiltonawiderangeofhostplants,amongthemmanyornamentalplants.Recently,wereportedauniquefindingofRSSC,PhylotypeI,resulting in stunted,yellowingandwiltedornamentalRosa sp.plants cultivatedundergreenhouseconditions. IdentificationofRSSC isolates fromRosa sp.was, amongother tests,performedat theproteomiclevelusingMatrix-AssistedLaserDesorptionIonizationTime-Of-FlightMassSpectrometry(MALDI-TOFMS).Prior to the routine identificationofRSSC isolates fromornamentalRosa sp.andassociated water samples, we created spectral database entries (Mass Spectra Profiles, MSP’s)specific for theRSSC, that represented all the four phylotypes. All spectrawereobtained in linearpositive-ionmodewithanm/zrangeof2000–20,000Da.ValidationdatademonstratedthatMSP’screated were specific for the identification of the representatives inside the RSSC, clearlydiscriminatingthemfromotherrelatedRalstoniasp.,suchastheR.picketii.Additionally,theproteinprofilesweacquiredinthepresentstudyaccuratelyidentifiedRSSCatthespecieslevel,includingR.pseudosolanacearum (phylotypes I and III), the“real”R. solanacearum (phylotype II) andR. syzygii(phylotypeIV),describingthediversityoftheR.solanacearum“speciescomplex”.Forroutineuse,theMALDI-TOFMSwasadditionallyshowntoberobust,quickandreproducible.Keywords:MALDI-TOF,RSSC,phylotypes,Ralstoniasolanacearum,Ralstoniapseudosolanacearum,Rosasp.

Roundtable1Oral22ComparisonofMultispectralimagingandNearinfraredReflectanceSpectroscopymethodsforphenotypingresistanceofcerealstoFusariumHeadBlight(FHB)andDeoxynivalenol(DON)ValérieCadot1,PatriciaLEM2,MarlèneFaure1,ClémenceGalon1,ThomasBaldwin1,PhilippeduCheyron3,NellyDEHAIS2,Jean-PhilippeMaigniel41.GEVESBeaucouzé–25rueGeorgesMorel,49071Beaucouzé-France2.BioGEVES,LeMagneraud–SaintPierred’Amilly–CS4005217700Surgères–France3Arvalis–RoutedeChâteaufortZAdesgraviers91190–VilliersleBacle4GEVESDomainedel’Anjouère-LaPouëze–49370Erdre-en-Anjou-FranceToencouragedevelopmentofcerealvarietiesresistanttoFusariumHeadBlightandmycotoxinsfortheNationalList,twonewmethodsofphenotypingweredevelopedbyGEVESonwinterwheat.Theyare more reliable and faster than in-field visual assessment and cheaper than LCMS-MS formeasuringDONcontent.Multispectralimagingisbeingdevelopedtopredict/quantifydamagefromdiseasesonseveral species.Near-InfraredSpectroscopy isanefficientandnon-destructivemethodforseedqualityanalysis.Theaimwas toassess thepotentialof these technologies forquantifyingFusariumgraminearumdamage,comparedtovisualassessmentandDONcontentinkernels.i)WithMultispectral imaging (VideometerR), the algorithm “Fusa-spectrale wheat”, was developed toevaluate the percentage of FusariumDamaged Kernels (FDK) atmaturity for varietal classificationusing Canonical Discriminant Analysis between kernels infected/not infected, on three years atseveral sites. FDK is strongly correlated with percentage of scabbed spikelets assessed by visualscorings (R>0.87) and with DON content (R>0.88). Correlation between FDK and Fusariumgraminearum biomass assessed by real-time PCRwas stronger in trials with only F. graminearum(R=0.97)thantrialswithacomplexFusarium-Microdochium.Fordurumwheatandtriticale,studiesare ongoing. ii) NIR calibration was developed from a sample set of varieties produced in twolocations over two crop years. Calibration models (visual scoring, DON content) were developedusing Partial Least Square Regression (PLS). Eachmodel was checked by internal cross validation.Their performance was assessed by the coefficient of determination in calibration (R2cal) andstandarderrorincrossvalidation.TheR2calobservedwas0.90for“DONcontent”modeland0.79for“visualscoring”model.These two technologies show potential to evaluate FHB andDON resistance on cereals. TheNIRSmodels require additional data to include geographical, genotyping and years effects. Withmultipectralandhyperspectralimaging,researchisongoingtoquantifyMicrodochiumspp.Keywords:Fusariumgraminearum,Phenotyping,Deoxynivalenol,Near-infraredspectroscopy,Multispectralimaging,Cereals.

RoundTable1Oral23StableinsertionandtransientexpressionofgreenfluorescentproteininAphanomyceseuteiches,thecausalagentofpearootrotdiseaseYohanaLaloum1,2,3,IsabelleTrinsoutrot-Gattin1,3,AzeddineDriouich2,3,Marie-LaureFollet-Gueye2,3,KarineLaval1,3andChristopheGangneux1,31UniteAgri'Terr,UniLaSalle,3rueduTronquet,76134Mont-Saint-Aignan,France.2LaboratoireGlycobiologie&MatriceExtracellulaireVegetaleUPRES-EA4358,UniversitedeRouen,76821MontSaintAignan,France3GRR-Vegetal,Agronomie,Sol,etInnovation(VASI),RegionNormandie,FrancePea(Pisumsativum)rootrotdiseaseiscausedbytheoomycetepathogenAphanomyceseuteiches.A.euteiches is one of themost destructive soilborne pathogens of pea crop. Since its discovery, noefficient control methods have been reported. Currently, the details of A. euteiches infectionmechanismsarestillnotwellknown.InordertostudytheinteractionbetweenA.euteichesandpearoot,anA.euteichesstrainexpressing the reportergenegfp (GreenFluorescentProtein)hasbeenobtained.A largequantityofprotoplasts fromayoungA.euteichesmyceliumwasproducedusinglysing enzymes from Trichoderma harzianum and 10% could regenerate on PDA mediumsupplementedwithMannitol.TheprotoplastsweretransfectedbyPEG/CaCl2-mediatedmethodwithAequorea victoria gfp gene under the control of a strong constitutive promoter from the BremialactucaeHam34geneandfusedtotheterminatorHam34gene.ThedatashowforthefirsttimethattransientexpressionofgfpgenecanbeobtainedinA.euteichesmycelium.Thisresultconfirmsthatapromoter from a Peronosporale oomycete can be expressed in a Saprolegniale oomycete. qPCRanalysisandconfocalmicroscopyobservationconfirmedboth insertionandexpressionofgfp inA.euteichesmycelium. However, despite the multiple insertion of the gfp gene in the A. euteichesgenome, only a transient cytoplasmic expression of GFP could be observed. The loss of the GFPfluorescent signal may result from a gene silencing process as previously demonstrated for thePhytophthorainfestanselicitininf1gene(vanWestetal.,1999).ReferencesP.vanWestetal.,1999.Mol.Cell3,339–348Keywords:Aphanomyceseuteiches,oomycetes,soilbornepathogen,transfection,GFPAcknowledgmentsThisresearchissupportedbytheRegionalFederationforResearch“Nomandie-Vegetal”.GRR-VASIisacknowledgedforfinancialsupport.

Session3.EMERGINGDISEASESANDNEWINSIGHTSINTAXONOMYANDPHYLOGENYOFPLANTPATHOGENSINEVOLVINGGLOBALCONDITIONSKeynotelecture5Keepingupwiththeplantdestroyersinthepost-genomicseraSophienKamounThe Sainsbury Laboratory, Norwich Research Park, Norwich, United Kingdom.http://www.KamounLab.netInfectiousplantdiseasescausehavoctoworldagricultureandthreatentoslowlaudableeffortstolaunchasecondgreenrevolutiontomeetthefoodsecurityneedsofaboomingworldpopulation.Pathogenssuchasthericeblastfungus,wheatstripeandstemrust,theIrishpotatofaminepathogen,andmanyotherscontinuetotriggerrecurrentepidemicswithfarreachingconsequences.Whenfacedwithopponentslikethese,weneedtoknowouradversary.Thegenomesequenceofaplantpathogenisadeeplookintoitssoul.FromimportantandoftenunexpectedinsightsintothebiologyofthepathogentothetoolsneededtodevelopsurveillanceanddiagnosticDNAmarkers,thegenomeisaninvaluableresourcethatacceleratesresearchandoutputdelivery.WiththecostofgenomesequencingdecreasingevenfasterthanMoore’slaw,thecost-benefitcalculationisevident.Forinstance,countlesstimeandmoneyarespentindevelopingDNAmarkers,investigatingpopulationstructures,debatingthepathogenorigin,etc.–activitiesthatcanbegreatlyhastenedbythegenomesequence.Inthistalk,Iwilldiscusssomeofwaysinwhichgenomebiologyimpactsplantpathology.Inparticular,Iwilladdresshowpathogengenomicscandrivebasicandappliedplantpathology,andhowstateoftheartfindingsonpathogenbiologycanbeexploitedtodrivethedevelopmentofnewapproachestobreedingdiseaseresistantcrops.Detailedknowledgeofthepathogengenomecoupledwithnovelmethodsofplantgenomeeditingisusheringtheeraofnext-generationdiseaseresistancebreedinginplants.Kamoun, S. 2012.Genomics of emerging plant pathogens: too little, too late.Microbiology Today,39:140.Kamoun, S. 2014. Keeping up with the plant destroyers. PlantVillagehttps://medium.com/@plantvillage/keeping-up-with-the-plant-destroyers-9c0047899683Cooke,D.E.L., Cano, L.M., Raffaele, S., Bain, R.A., Cooke, L.R., Etherington,G.J.,Deahl, K.L., Farrer,R.A.,Gilroy,E.M.,Goss,E.M.,Grunwald,N.J.,Hein,I.,MacLean,D.,McNicol,J.W.,Randall,E.,Oliva,R.F.,Pel,M.A.,Shaw,D.S.,Squires,J.N.,Taylor,M.C.,Vleeshouwers,V.G.A.A.,Birch,P.R.J.,Lees,A.K.,and Kamoun, S. 2012.GenomeAnalyses of anAggressive and Invasive Lineage of the Irish PotatoFaminePathogen.PLoSPathogens,8:e1002940.Islam, T., Croll, D.,Gladieux, P., Soanes,D., Persoons, A., Bhattacharjee, P., Hossain, S., Gupta,D.,Rahman,Md.M.,Mahboob,M.G.,Cook,N.,Salam,M.,Surovy,M.Z.,BuenoSancho,V.,Maciel,J.N.,Nani, A., Castroagudin, V., de Assis Reges, J.T., Ceresini, P., Ravel, S., Kellner, R., Fournier, E.,Tharreau,D., Lebrun,M.-H.,McDonald,B., Stitt,T., Swan,D.,Talbot,N., Saunders,D.,Win, J.,andKamoun,S.2016.EmergenceofwheatblastinBangladeshwascausedbyaSouthAmericanlineageofMagnaportheoryzae.BMCBiology,14:84.

Session3Oral26Genomic study shows endemic and pandemic rice blast lineages and lineages between rice andsetariainfectinggenepoolsThierry1M,Rieux2A,Ravel1S,Cros-Arteil3S,Adreit1H,Milazzo1J,Terauchi4R,Ioos5R,Tharreau1D,Fournier2E,Gladieux3P.1CIRAD,UMRBGPI,Montpellier,France;2CIRAD,UMRPVBMT,StPierredelaReunion,France;3INRA,UMR BGPI, Montpellier, France; 4Iwate Biotechnology Research Center, Kitakami, Iwate, Japan;5ANSES,Malzéville,France. The rice blast fungus Magnaporthe oryzae is the most damaging rice pathogen, and atextbookexampleofwidelydistributed,rapidlyadaptingpathogen,despitelimitedgeneticdiversity.The aim of our study was to elucidate the factors and evolutionary changes underlying theemergence,diversificationandspreadofM.oryzaeonrice.Analysesofpopulationstructurebasedon Infinium-genotyping of 5300 SNPs for 970 isolates collected on rice on the five continentsidentified 13 lineageswithinM. oryzae. Three lineageswere pandemic inmultiple continents andone of them was the only lineage detected in Europe. Multiple lineages with more restricteddistributions insub-SaharanAfricaandAsiawerealso identified.Wholegenomeresequencingofasubset of 80 rice-infecting isolates combinedwith 12 isolates collected on Setariamillet revealedseveral lineages with intermediate positions between the previously identified rice- and Setaria-infecting genepools, questioning theSetaria originof rice-infectingM.oryzae and suggesting thattheemergenceof riceblastmaybemore recent thanpreviously thought. Because the sequencedisolateswerecollectedbetween1973and2009andrecombinationislimited,wewillusedatedtipstocalibratetreenodeswithinaphylogenetic frameworktoelucidatethetimingofemergenceandglobal dispersal of M. oryzae. Most lineages were highly clonal, but we found evidence forrecombinationinawidelydistributedlineageinfectinguplandriceinYunnan,LaosandThailand.Wewill use genome scans for genetic exchanges to test thehypothesis that recombining lineages aremore likely to receive geneticmaterial from other lineages. Ourwork provides a population-levelgenomic framework for defining molecular markers to assist in the control of rice blast and forinvestigatingthemolecularunderpinningsofphenotypicandfitnessdifferencesbetweendivergentlineages.Keywords:Magnaporthe,Setaria,Rice,Populationgenomics,Recombination,Geneflow

Session3Oral27AssessingthephenotypicandgenotypicdiversityofSclerotiniasclerotioruminFranceChristelLeyronas1,MagaliDuffaud1,ClaireTroulet1,FrançoisVilleneuve2,MarcBardin1andPhilippeNicot1.1PathologieVégétale,INRA,84140,Montfavet,France2Ctifl,CentredeLanxade,24130,Prigonrieux,FranceSclerotinia sclerotiorum is a highly polyphagous ascomycete that can attack more than 400 plantspecies among which vegetables and arable crops. In order to improve the efficiency of plantprotectionagainstwhitemould,knowledgeontheS.sclerotiorumpopulationshastobeacquired.Inthe framework of national project “Scleroleg”, S. sclerotiorum isolates were collected onsymptomaticplantsinseveralFrenchregions.Theirgeneticandphenotypicdiversitywasassessed.The aggressiveness of 103 isolates collected frombean, canola, carrot, lettuce,melon andwitloofchicorywascomparedonmelon,tomatoandlettuceincontrolledconditions.Allisolateswereableto cause symptoms on detached leaves of the three plant species. The relative level ofaggressiveness of an isolate, compared to theothers, tended to be similar regardless of theplantspeciesonwhichitwasassessed.Onaverage,theisolatescollectedfromlettucewereslightlymoreaggressivethantheothersonallthreehostplantstested.Fortherestoftheisolates,therewasnolinkbetweenhostoforiginandaggressiveness,suggestingthatthereisnomarkedhostspecializationofthefungus.Thesedatawerepartlysupportedbytheresultsofgeneticcharacterizationof200S.sclerotiorum isolates with 16 microsatellites markers. While there was no significant geographicdifferentiation among isolates (some haplotypes were shared by isolates collected on differentspeciesandseparatedbyupto600km), the lettuce isolatesshowedstrongandsignificantgeneticdifferentiationwithallothers.Thereasonsforthesedifferencesremaintobeelucidated.However,ourstudyclearlysuggeststhatsclerotiaofS.sclerotiorumgeneratedononehostcropwill likelybecapableofproviding inoculumforthenextsusceptiblecropgrowninthesameplotandpossiblyforcropsinneighbouringfields,viatheproductionofairborneascospores.Keywords:whitemould,epidemiology,aggressiveness,geneticdifferentiation

Session3Oral28ModellingthedispersalofMonochamusgalloprovincialis,thevectorofthepinewoodnematode,andassessingtheeffectivenessofclear-cuttingmeasuresRobinetC.,Castagnone-SerenoP.,MotaM.,RouxG.,SarniguetC.,TassusX.andJactelH.

Working Group on Bursaphelenchus xylophilus, French Agency for Food, Environmental andOccupationalHealth&Safety(Anses),14ruePierreetMarieCurie94701MaisonsAlfort

Following the detection of a pinewood nematode outbreak, Implementing Decision 2012/535/EUimposes eradicationmeasures for at least four years,mainly consisting of the establishment of: i)clear-cutswitha radiusof500maround the infestedplants and ii) intensive surveillancewithinaradius varying from 6 to 20 km around the infested zone. To estimate the effectiveness of thesemeasures,asimulationmodelwasdevelopedandcalibratedusingexperimentaldataspecifyingthedispersalcapabilitiesoftheinsectvectorMonochamusgalloprovincialis.Thesedatawereobtainedinthelaboratory(onaflightmill)andinnatura(mark-and-recaptureexperiments)inthecontextoftheLandesdeGascogneforest(David,2014andDavidetal.,2014),oneoftheforestareasmostexposedto the risk of introduction of the pinewood nematode in France. The simulations thatwere thencarried out under different scenarios (preventive or curative) showed that the clear-cuts currentlyrequestedbyEuropeanandFrenchregulationswouldnotbeeffectiveinalandscapeconfigurationofcontinuousplantations ofmaritimepine. Indeed,with the recommended radius of 500m, at best11%ofnematodetransmissionsbytheinsectvectorwouldbeavoided.Moreover,toobtainapinewoodnematodetransmissionrate lowerthan0.1%, itwouldbenecessaryto implementclear-cutswith a radius of 15 - 38 km (Anses, 2015). These simulations, carried out in the context of acontinuousforest,shouldhoweverbesupplementedbyascenarioinvolvinghighlyfragmentedpineforests. In this regard, it would be interesting to include biological data from Spanish forestmanagers,astheyaredirectlyinvolvedinimplementingnematodeeradicationmeasuresinthistypeoflandscape.AlternativemanagementmeasurestocontrolthespreadofthepinewoodnematodewerealsoassessedbytheWorkingGroup.

Keywords: Pinewood nematode, Monochamus galoprovinicalis, Dispersion, Clear cuts, Riskassessment

ANSES. (2015). Request for an opinion on "the control strategy imposed by Implementing Decision2012/535/EUof26September2012onemergencymeasurestopreventthespreadwithintheEuropeanUnionofBursaphelenchusxylophilus"50p,https://www.anses.fr/fr/system/files/SVEG2014SA0103RaEN.pdf

DavidG.(2014).ÉtudedescapacitésdedispersiondeMonochamusgalloprovincialisvecteurdunématodedupinBursaphelenchusxylophilus.Thèse,UniversitédeBordeaux,173pp.

David G., Giffard B., Piou D. and Jactel H. (2014). Dispersal capacity ofMonochamus galloprovincialis, theEuropeanvectorofthepinewoodnematode,onflightmills.JournalofAppliedEntomology,138:566-576.

Commission ImplementingDecision2012/535/EUof26September2012onemergencymeasures topreventthespreadwithintheEuropeanUnionofBursaphelenchusxylophilus(SteinerandBuhrer)Nickleetal.(thepinewood nematode) (notified under document C(2012) 6543) (2012/535/EU).Official Journal of the EuropeanUnionL266of02.10.2012,pages42-52.

Session3Keynotelecture6RiskassessmentandmitigationoftheintroductionandspreadofnewplantpathogensinachangingworldArienaH.C.vanBruggenEmergingPathogensInstituteandPlantPathologyDepartment,UniversityofFlorida,Gainesville,FL32611,USA.

Anoverviewwillbegivenofmajorglobaltrends,includingincreasinglyunevenresourcedistribution,increasedwarandmigration,globalizationoffoodproduction,tradeandconsumption,increasinginequalitiesandfoodinsecurity,environmentaldegradation,globalclimatechange,emergingpathogens,andincreasingineffectivenessoffungicidesandantibiotics.ThenIwillzoominonemergingplantpathogensassociatedwithsomeoftheseglobalchanges.Iwilldiscusscurrenttoolsforriskassessmentofthepotentialintroductionandspreadofnewplantpathogens,withafocusoncorrelativepredictionmodelscombinedwithGISandmicrobialriskassessmentmodelsandtheassociateduncertainties.IwilluseBananaXanthomonaswilt,CitrusHuanglongbingandBlueberrytwigblightasexamples.Mitigationoptionswillfocusonqualityandsustainabilitycertification,quarantinemeasures,earlydetectionusinginternet-basedelectronictools,removalofinfectedplants(whenandwhereappropriate),developmentanduseofplantresistance,andpreventionofspreadbyenhancingagroecosystemresiliencebyincreasedplantandmicrobialdiversity.Thepresentationwillendwithcallingonplantpathologiststocontributetosolvingglobalproblems,inparticularenvironmentaldegradationandfoodinsecurity.

Session3Oral29BiOR²:adatabase/softwareprocessdedicatedtoplantpestsandpathogensrankingSession6:Emergingtoolsforthemanagementofplantdiseases:agroecologyanddiseasemanagementChristineTayeh1,DavidGuinehut1,AmandineLêVan1,XavierTassus1,EmmanuelGachet1andCharlesManceau11ExpertiseandBiologicalRisksUnit,PlantHealthLaboratory,FrenchAgencyforFood,EnvironmentalandOccupationalHealthandSafety(ANSES),7rueJeanDixméras,49044,AngersCédex01,FranceInthecontextofglobalizationofplantstrade,modelsforrankinginvasivepestsaccordingtotherisksforcropsandenvironmentareimportanttoriskmanagersastheyhavetoadaptquicklymanagementstrategiesaccordingtoriskypathways.Tothisend,anewmethodologywasdeveloped:BiOR²,whichstandsfor“BiologicalOrganismsdataRetrievalandRankingsystem”.BiOR²combinesadatabasetographicinterfacesandmulti-criteriadecisionaiding(MCDA)method.First,thedatabaserunsunderthedatabasemanagementsystem(DBMS)PostgreSQL.Itconsistsof55tablescontaininginformationrelativetoplants’trade,landuse,legislation,pestinterceptionsandclimateclassification.Thedatabaseislinkedtoagraphicuserinterface(BiOR²Form)thatallowsfillingindataonhostplants,pathogensandpest-plantinteractionsthroughaquestionnaire.Next,anothergraphicuserinterface(BiOR²Statistics)constructsamulti-criteriamatrixbyrelyingon2800linesofcodes,enablethelinkbetweentheavailabledataaccordingtospecificcriteriaandwhoseuncertaintycanbealsoquantifiedbyBiOR².Abaseof24independentcriteria,definedaccordingtoFAOISPMN°11document,coversriskofentry,establishment,spreadaswellaseconomic,socialandenvironmentalimpacts.TherankingisfinallyachievedthroughVisualPROMETHEE(MCDAmethod)withthepossibilityofconductingseveralscenariosaccordingtotheoccurrenceofthepestintheareaofinterestorthroughanadaptablecriteriaweightingsystem.TheprocesswasappliedtometropolitanFranceandtheFrenchOverseaDepartmentsonrespectively278and110plantpestsandpathogens,whichdemonstratesitsapplicabilitytoseveralregions.ThecontributionofBiOR²topolicyrecommendationsuchaspestprioritizationwasalsoinvestigated.Overall,BiOR²isintendedtobeobjective,generic,basedonsoundscience,highlyadaptabletomeetvariouskindsofquestionsaboutthemanagementandimpactofpestsandpathogens.

Keywords: Plantpestsandpathogens,planthealth, ranking, riskassessment,database,multi-criteriaanalysis

Session3Oral30LargehostjumpsinsomepowderymildewspeciesrevealedbyphylogenyandcrossinoculationsMarie-Laure Desprez-Loustau1, Marie Massot1, Nicolas Feau2, Tania Fort1, Antonio de Vicente3,JuanAntonioTorés4,DoloresFernándezOrtuño3,4

1.UMR1202BIOGECO,INRA,UnivBordeaux,69routed'Arcachon,33610Cestas,France2. Department of Forest and Conservation Sciences, British Columbia, The University of BritishColumbia,Vancouver,BCV6T1Z4,Canada3.InstitutodeHorticulturaSubtropicalyMediterráneaLaMayora(IHSM-UMA-CSIC),DepartamentodeMicrobiología,FacultaddeCiencias,29071,Málaga,Spain,4. Instituto de Horticultura Subtropical y Mediterránea La Mayora (IHSM-UMA-CSIC), Algarrobo-Costa,29750,Málaga,SpainPowderymildew fungihave longbeenconsideredashighly specificpathogens, i.e. agiven speciescaninfectonlyanarrowrangeofhostplants,usuallyinthesamegenus.Theavailabilityofmolecularmarkers has dramatically improved and changed our vision on these fungi, especially that hostspecificityisnotassimpleandnarrowaspreviouslybelieved.Forexample,recentphylogenieshavestronglysuggestedthatspeciescausingoakandmangopowderymildewarecloselyrelated.Hereweanalysed mango powdery mildew samples from southern Spain (6 different orchards in Malagaregion)bycombiningmulti-genephylogeny(ITS+4single-copycodinggenes)andcrossinoculationsbetweenoakandmango.Basedongeneticanalyses,twoErysiphespecieswereidentifiedinmangosamples, which are the species usually associated with oak powdery mildew in Europe. Erysiphequercicolawasdominantwith97%positivesamples,whereasE.alphitoideswasfoundinonly11%ofall samples, 3% alone and 8% inmixturewith E. quercicola. Cross inoculations between oak andmango, which led to typical symptoms, further supported the conspecificity of oak and mangopowdery mildew species. These results constitute the first report of mango powdery mildew inmainland Spain and mainland Europe, caused by E. quercicola and E. alphitoides. Our studyconfirmedthebroadhostrangeand/orhostshiftingabilityofbothE.quercicolaandE.alphitoides,which may explain their invasive success. Furthermore, it opens interesting prospects to theelucidation ofmolecularmechanisms underlying host range since the two closely relatedErysiphespeciesbelongtoasmallcladewithbothgeneralistandspecialistpowderymildews.Keywords:powderymildew,hostrange,hostshift,biologicalinvasion

Session3Oral31IdentificationandcharacterizationofPhytophthorahybridsusinggenotyping-by-sequencingKrisVanPoucke1,ThomasGoedefroit1,AnneliesHaegeman1,TomRuttink2,KurtHeungens1

1InstituteforAgriculturalandFisheriesResearch,PlantSciencesUnit,CropProtection,Burg.VanGansberghelaan96,9820Merelbeke,Belgium2InstituteforAgriculturalandFisheriesResearch,PlantSciencesUnit,GrowthandDevelopment,Caritasstraat39,9090Melle,BelgiumSincethemid-1990’s,severalPhytophthorahybridshavebeendescribedinmultiplephylogeneticclades.Asaresultofintensifiedinternationalplanttrade,whichmightbringpreviouslyseparatedspeciesincontactwitheachother,anincreaseinthenumberofhybridizationeventsissuspected.Thesehybridsmightbemoreaggressiveorhaveanexpandedhostrangeincomparisontotheparentalspecies.Earlydetectionofnewhybridspeciesisthuswarranted,butisnotalwayssuccessfulviatraditionalPCRampliconsequencingofindividualloci.Wehaveusedtwo-enzymegenotyping-by-sequencing(GBS)toreliablyidentifyandcharacterizehybridsinaEuropeancollectionofseveralhundredsofPhytophthoraisolates.DNAwasextracted,digestedandGBS-adapterswereligated.AfterPCRamplification,GBSlibrarieswerequantified,pooledandsequencedusingtheIlluminaHiSeq3000technology.ReaddatawaspreprocessedusingcustomscriptsandsubsequentlysubjectedtotheGibPSspipelinetoidentifyaglobalsetofGBStagsacrossallsamples.Subsequently,persampletheabsence/presenceprofileswerescoredtogetherwithSNPprofilesforeachGBStag.Themethodwashighlyreproducibleandgeneratedabout15000to30000tagsforanon-hybridspecies.WhenisolatessharealargenumberofGBStagswithatleasttwoPhytophthoraspecies,thisisindicativeofahybridizationevent.Usingthisapproach,wedetectedapreviouslyunidentifiedhybridofP.chlamydosporaandP.lacustris.Evenwhentheparentalspeciesarenotknown,alargenumberofGBStagsinasamplecanpointtoahybridizationevent,ashybridscontaintagsfromeachoftheparentalspecies.Wethusidentifiedanumberofpotentialnovelhybridspecies,which impliesthathistorichybridizationeventsmightbemore common in Phytophthora than previously assumed. Genotyping-by-sequencing can thus beusedtoreliablyidentifyandcharacterizePhytophthorahybrids

Session4.MICROBIALDETERMINANTSOFPATHOGENANDSYMBIOTICINTERACTIONSKeynotelecture7UnderstandingpathogenadaptationtotheplanthostAntonioDiPietro1

1Departamento de Genética, Universidad de Córdoba, Campus de Rabanales C5, 14071 Córdoba,Spain

Filamentousplantpathogensposeaseverethreattoglobal foodsecurity.Theseorganisms

often show exquisite host adaptation, but also undergo rapid evolution leading to shifts orexpansions in thehost range.Thegeneticmechanismsofpathogen-hostadaptation remainpoorlyunderstood. Inthesoil-inhabitingvascularwilt fungusFusariumoxysporum, individual isolatestendtoexhibithighspecificitytowardsagivenplanthost,whilethespeciescomplexcollectivelyattacksmorethanahundreddifferentcrops.Inaddition,F.oxysporumisalsoanemerginghumanpathogenthat provokes lethal systemic infections in immunocompromised individuals. Remarkably, a singlefieldisolateofthisfunguscankilltomatoplants,immunodepressedmiceandinsects.Byfollowingacombination of reverse genetics and experimental evolution approaches, we found that F.oxysporum uses multiple strategies to adapt to different host environments. These includerecruitmentofconserved fungalsignalingpathwaysorhijackingofhost regulatorymechanisms fornewvirulence-relatedfunctions.Strikingly,fungalpopulationsevolvedafterserialpassagingthroughdifferent environments displayed large-scale chromosomal reorganizations in transposon-richaccessory regions of the genome, suggesting that chromosome plasticity could act as a majorevolutionary driver in F. oxysporum. Understanding the geneticmechanisms that govern virulenceevolution and host adaptation may reveal new ways to control diseases caused by filamentouspathogensandimproveplanthealth.Keywords:evolution,fungus,Fusarium,host,pathogen,virulence

Session4Oral32Identification of atypical chitin synthase genes horizontally transferred in microbial plantpathogensIsabelle R. Gonçalves1,2, Sophie Brouillet3, Marie-Christine Soulié4, Simonetta Gribaldo5, CatherineSirven2,NoémieCharron2,MartineBoccara3andMathiasChoquer1,21Univ Lyon, Université Claude Bernard Lyon 1, CNRS UMR5240, Microbiologie Adaptation etPathogénie,BâtimentAndréLwoff,10rueRaphaëlDubois,F-69622Villeurbanne,France2BAYERS.A.S.,CentredeRecherchedelaDargoire,F-69263Lyon,France3Sorbonne Universités, UPMC Univ Paris 06, UMR 7205 (MNHN, UPMC, CNRS, EPHE), Atelier deBioinformatique,F-75231Paris,Cedex05,France4Sorbonne Universités, UPMC Univ Paris 06, INRA-AgroParisTech UMR1318, F-78026 Versailles,France5Institut Pasteur, Unité Biologie Moléculaire du Gène chez les Extrêmophiles, Département deMicrobiologie,25rueduDocteurRoux,F-75015Paris,France

Chitin, the secondmostabundantbiopolymeronearthafter cellulose, is found inprobably allfungi,manyanimals(mainlyinvertebrates),severalprotistsandafewalgae,playinganessentialrolein the development of many of them. It is produced by type 2 glycosyltransferases, called chitinsynthases(CHS).Inphytopathogenicfungi,severalCHSisoenzymesarefoundandtheyarenotonlythoughttobeessentialfortheirgrowthbuttheyarealsoconsideredtoparticipateasdeterminantsoftheirpathogenicity.

Weperformedagenome-wideanalysisoftheCHSmultigenicfamilyandcreatedadatabankofputative CHS. Phylogenetic analyses first allowed to propose a robust and unifying fungal CHSclassification that is easily accessible through a dedicated website(http://wwwabi.snv.jussieu.fr/public/CHSdb). These analyses also permitted to study theevolutionaryhistoryofCHS.Thisfamilyhasmainlyevolvedviaduplicationsandlosses.However,itislikely that severalhorizontalgene transfers (HGT)alsooccurred ineukaryoticmicroorganismsand,evenmoresurprisingly,inthegenomesofbacteria.Moreover,somefungalCHSarehighlysimilartoCHS of giant algae-infecting viruses. Interestingly, many of these atypical CHS are found in plantpathogens, bacteria or fungi. Different characteristics of these CHS genes might be associated tovirulencefactorsimpliedinplantinteractions.Keywords:Chitinsynthase,Evolution,Bacteria,Viruses,Fungi,Horizontalgenetransfer.Gonçalvesetal.BMCEvolutionaryBiology(2016)16:252.

Session4Oral33Colonizationofplantsbypectobacteria:morethanjusta“bruteforce”Vladimir Gorshkov, Amina Daminova, Rim Gubaev, Bakhtiyar Islamov, Olga Petrova, NataliaGogoleva,NadezhdaTarasova,YuriGogolev

KazanInstituteofBiochemistryandBiophysics,KazanScienceCentre,RussianAcademyofSciences,Lobachevskystr.,2/31,Kazan,Russia,420111

Pectobacteriaareconsideredas“tough”phytopathogens thatcolonize theirhostsapplying

“bruteforce”–extracellularenzymesdamagingplanttissues.However,someobservationspointtothe significance of pectobacteria “stealth” behavior in planta. An important property ofmicrobialpopulationsisrelatedtotheirheterogeneity.Herewith,differentmorho-physiologicalcelltypesarefunctionalpartsofthepopulation.Theresearchoftheformationofdifferentiatedmicrobialcellsisinitsinfancy,especiallywithinhost/pathogensystems.

We tested if the colonization of plants is coupled with dissociation of pectobacteriapopulationandtriedtoobtaininformationoncausesandconsequencesoffunctionalspecializationofpectobacteriacellsinplanta.

Wehave“lookedinside”infectedplantsusingavarietyofcomplementarymethods:varioustypes of microscopy and chromatography, gene cloning and their heterologous expression,mutagenesis,geneexpressionanalyses(qPCR,RNA-Seq).

Ourresultsshowthatdifferentplantcompartmentsformspecificsignalingbackgroundsthatdrivethebehaviorofmicrobesresulting in inplantadissociationofbacterialpopulation.Herewith,various sub-populations display functional specialization (blocking the vessels, stress adaptation,tissue maceration, cell migration, etc.). Novel biofilm-like structures – bacterial emboli weredescribed and discussed to play significant roles in the formation of pathosystem. In plantadissociation of bacterial population is coupled with plant susceptible response that providesconditioningofthecolonizedcompartment.Ourobservationsarestrengthenedanddeepenedbythewhole-transcriptomeanalysis of bothplant andpathogenduring their interactions. This studywassupportedbyRSF(15-14-10022).

Keywords:Softrot,dissociationofpopulation,biofilms,bacterialemboli,plantsusceptibleresponse

Session4Oral34FungaleffectorsandplantregulatorsofthebarleysusceptibilityfactorRACBRalphHückelhoven1,BjörnScheler1,VeraSchnepf1,CaroHöfle1,TinaReiner1,MathiasNottensteiner1

1Technical University of Munich; Chair of Phytopathology, Emil-Ramann Straße 2, 85354 Freising-Weihenstephan

ThegrasspowderymildewfungusBlumeriagraminis,asahaustorium-formingectoparasite,

has to overcome pre- and post-invasive host immunity. Additionally, as an obligate biotroph, B.graminishastoreprogramthehostforchangesinhostcellarchitectureandphysiology,butlittleisknownaboutvirulenceeffectorfunctionsthatsupportthis.

The signalling G-protein RACB is a barley susceptibility factor and a regulator of hostcytoskeletondynamics.ActivatedRACB supports fungal invasion intoepidermal cellsofbarleyandhaustorium expansion. RACB´s function as a susceptibility factor is tightly controlled in the host.MICROTUBULE-ASSOCIATED ROP GTPASE ACTIVATING PROTEIN1 controls RACB at the level ofsignalling activity whereas cytoplasmic ROP-BINDING KINASE1 controls RACB protein abundance.However,thephysiologicalfunctionofRACBisnotfullyunderstood.

RecentresultsshowthatRACBdoesnotfunctionasanegativeregulatorofPAMP-triggeredimmune responses inbarley. Instead,RACB is required forpolardevelopmentofbarleyepidermalcells.Strikingly,wefoundthatB.graminisexpressesaneffectorwecalledROP-interactivepeptide1(ROPIP1) from a non-autonomous retroelement. ROPIP1 interacts with RACB in planta, istranslocated into the host cytoplasm and destabilizes host microtubules. ROPIP1 has virulencesupporting functionswhenover-expressed in thehost, andhost-induced gene silencingof ROPIP1limitsfungalpenetrationsuccess.

ApparentlyB.graminisevolvedanoveltypeofvirulenceeffectorbyneo-functionalizationofarepetitiveelement.ThiseffectortargetsRACB,whichotherwisefunctionsinpolarcellgrowthandmicrotubule organization. This suggests that RACB´s function is co-opted by B. graminis forreprogrammingofhostcellcytoskeletonanddevelopmentduringingrowthofthehaustorium.

Keywords:Blumeria graminis, barley, small GTPase;microtubules, effector-triggered susceptibility

Session4Oral35ThebiologicalandevolutionarybasisofsystemicplantpathogenesisinXanthomonasJonathanM.Jacobs1,2,4,5*,CélinePesce1,2,3,TacaVancheva1,2,JillianM.Lang4,AlvaroPerez-Quintero1,Caitilyn Allen5, Jan E. Leach4, Lionel Gagnevin1, Boris Szurek1, Sébastien Cunnac1, Claude Bragard2,RalfKoebnik11IRD-CIRAD,UMRInteractionsPlantesMicroorganismesEnvironnement,Montpellier,France2Earth&LifeInstitute,UniversitéCatholiqueLouvain-la-Neuve,Louvain-la-Neuve,Belgium3DepartmentofMicrobiology,UniversityofNewHampshire,Durham,NH,USA4Department of Bioagricultural Sciences and Pest Management, Colorado State University, FortCollins,CO,USA5DepartmentofPlantPathology,UniversityofWisconsin—Madison,Madison,WI,USA

Pathogenicmicrobescausesystemicandnon-systemicdiseasesofplantandanimalhosts.Systemicdiseases are particularly destructive because the pathogen moves through the host vasculaturecausing widespread infection; meanwhile non-systemic pathogens remain restricted to the non-vascular tissue near the site of infection. The basis of systemic and non-systemic pathogenesis isunclear.HerewedescribetheroleofcellwalldegradationintheevolutionandbiologyintheGram-negative phytobacterial genus Xanthomonas. Xanthomonas comprises a diverse group of vascularand non-vascular pathogens of over 200 plant species. We demonstrate that a single, vascularpathogen-unique cell wall degrading enzyme called CelA contributes to systemic pathogenesis inmultiplepathogeniclineagesinthisdiversegenus.WedeterminedthatCelA1wasconservedonlyinsystemicpathogenicbacteria in thegeneraXanthomonas,Xylella andRalstoniabutabsent innon-systemicGram-negativeplantpathogenicbacteria.Mostnotablyadditionofthiscellwalldegradingenzyme to two distinct non-systemic pathogen species, barley-infecting Xanthomonas translucensand rice-infecting Xanthomonas oryzae, permitted systemic pathogenesis of their respective hostplants. Further genomic analysis of non-systemic Xanthomonas pathogens appear to haveinactivated this trait suggesting that they arose from related vascular subgroupsuponadapting tothenon-vascularplantenvironment.Overall thisworkprovidesa framework todescribepathogenemergencebasedonsymptomdevelopmentandtissue-specificityinanimportantpathogengenus.

Keywords:Evolution,Xanthomonas,Systemicpathogenesis

Session4Oral36AnewproteinaceousPAMPfromAscomycetesinducescelldeathinSolanaceaeBarbaraFrancoOrozco1,AdokiyeBerepiki1,PaulBirch2,KostyaKanyuka3andAnnaAvrova11JamesHuttonInstitute,CellandMolecularSciences,InvergowrieDundee,DD25DA,UK2UniversityofDundee(atJHI),DivisionofPlantSciences,InvergowrieDundee,DD25DA,UK3RothamstedResearch,PlantBiologyandCropScienceDepartment,Harpenden,AL52JQ,UK

Recognition of conserved pathogen- ormicrobe-associatedmolecular patterns (P/MAMPs)

byplants initiatesPAMP-triggered immunity (PTI). SomePAMPselicitdefence responses inawiderange of plant species, while response to others is restricted to certain plant species, probablyreflecting the presence of a particular cell surface localized pattern recognition receptor (PRR).AlthoughanumberofmicrobialPAMPshavebeenidentifiedthefullrepertoireremainsunknown.

The causal agent of scald, Rhynchosporium commune, is one of the most destructive andeconomically important pathogens of barley. It is a hemibiotrophwith an extended asymptomaticphase.FollowingconidiagerminationandcuticlepenetrationR.communehyphaespreadbetweenbarleyepidermalcellswithoutdirectlypenetratingthem.

Sequencing of the R. commune transcriptome from an early time point during barleycolonisation revealed a highly abundant transcript encoding a small secreted fungal protein ofunknown functionwith four cysteine residues,whichwe called RcCDI1 (Cell Death Inducing). It ismosthighlyabundantattheonsetofbarleyinfectionwithR.commune.Pichiapastoriswasusedtoproduce RcCDI1 and its homologues from different fungal species, including Zymoseptoria tritici,MagnaportheoryzaeandNeurosporacrassa.AlloftheseproteinsexhibitedPAMPactivity inducingcelldeathinSolanaceaebutnotinotherfamiliesofdicotsormonocots.Virus-inducedgenesilencing(VIGS)ofknowncomponentsofPTIinNicotianabenthamianashowedRcCDI1-triggeredcelldeathtobe BAK1, SOBIR1 and SGT1 dependent. However, the cell death was not suppressed by thePhytophthorainfestanseffectorsPiAVR3aorPexRD2,suggestingthat itdoesnotrequireNbCMPG1or NbMAPKKKe. Identification of the plant receptor involved in RcCDI1 recognition in N.benthamianawillprovideavaluableresourceforengineeringnon-hostresistanceinmonocots.Keywords:celldeath,fungal,PAMP,pathogen,Rhynchosporiumcommune,Ascomycete

Session4Oral37UnusualevolutionarymechanismstoescapeEffector-Triggered-ImmunityinthefungalphytopathogenLeptosphaeriamaculansPlissonneauClémence1,2,BlaiseFrançoise1,OllivierBénédicte1,RouxelThierry1andBalesdentMarie-Hélène11INRA,UMRINRA-AgroParisTech1290BIOGER,78850Thiverval-Grignon,France.2PlantPathology,InstituteofIntegrativeBiology,ETHZurich,Zürich,Switzerland

Leptosphaeriamaculansisthefungusresponsibleforthestemcankerdiseaseofoilseedrape

(Brassica napus). AvrLm3 and AvrLm4-7, two avirulence genes of L. maculans, are involved in anunusual relationship: AvrLm4-7 suppresses the Rlm3-mediated resistance. Surveys of populationssubmittedtotheRlm7selectionpressurerecentlyshowedthatthelossoftheAvrLm7specificitywasaccompaniedbythegainofavirulencetowardRlm3.

Inordertobetterunderstandthemolecularbasesofthisunusualrelationship,andfollowingthecloningofAvrLm7andAvrLm3,weassessedthemolecularpolymorphismofAvrLm3inaworld-wide collection of 235 L. maculans isolates. No field isolates exhibited deletion or inactivatingmutationsinAvrLm3,asobservedforotherL.maculansavirulencegenes.Ahighlevelofnucleotidicpolymorphismwasalsofound,andelevenisoformsoftheAvrLm3proteinwerefound.SignaturesofpositiveselectionwereidentifiedinAvrLm3.

In isolates virulent towards both Rlm3 and Rlm7 (a3a7), the loss of the Rlm3-mediatedresponse was due to two distinct mechanisms. When AvrLm4-7 was inactivated (deletion orinactivating mutations), amino acid substitutions in AvrLm3 generated virulent isoforms of theprotein,responsibleforvirulencetowardsRlm3.However,halfofthea3a7isolatesstillcontainedanavirulentalleleofAvrLm3,alongwithpointmutationsinAvrLm4-7.Directedmutagenesisconfirmedthat some point mutations in AvrLm4-7 were sufficient for the fungus to escape Rlm7-mediatedresistancewhilemaintaining the suppressionof theAvrLm3phenotype. The complex evolutionarymechanisms enabling L. maculans to escape Rlm3-mediated resistance while preserving AvrLm3integrity, alongwithobserved reducedaggressivenessof isolates silenced forAvrLm3, suggest thiseffectorplaysamajorroleinpathogenicitytowardsB.napus.Thisexamplecontributestocomplexifythe gene-for-gene concept of plant-pathogen evolution with a “camouflaged” model allowingretentionofnon-dispensableavirulenceeffectors.

Fundings:CTPS-ONEMA“ICOSCOP”,INRAmetaprogrammeSMaCH(K-masstecproject).ANR(LabExBASC;ANR-11-LABX-0034).Keywords:Leptosphaeriamaculans,evolution,Rlm7,Rlm3,avirulencegenes,Brassicanapus.

Session4Oral38Clathrin, a key role in the delivery of virulence factors in the phytopathogenic fungus BotrytiscinereaEytham SOUIBGUI1-2, Cindy DIERYCKX1, Christophe BRUEL1, Marie-Pascale LATORSE2 and NathaliePOUSSEREAU1(1) University Claude Bernard Lyon 1 CNRS-UMR5240 Microbiologie Adaptation Pathogénie, (2)BAYERSAS:14impassePierreBaizet,BP99163,69263LyonCedex09,FranceFungiarethemostimportantplantpathogensonagriculturalandhorticulturalcrops.Studyofthesepathogens remains essential to understand pathogenic process and control plant diseases. Theseorganisms evolved a repertoire of secreted enzymes involved in plant decomposition to colonizetheirhosts.SecretoryproteinsaretransportedfromEndoplasmicReticulumandGolgiapparatustoextracellular space through intracellular vesicles. In filamentous fungi, intracellular vesicles wereobservedusingelectronmicroscopybuttheirbiogenesisprocessisstillunknown.Theelucidationofthis process and the identification of proteins involved in secretory vesicles biogenesis remains achallenge to understand virulence factors delivery. A nonpathogenic mutant altered in theexpression of a gene encoding clathrin heavy chain was selected in a random mutant librarygeneratedinthenecrotrophicpathogenBotrytiscinerea.Thisgeneisessentialinmanyorganisms.Aclathrin dominant negativemutantwas generated inB. cinerea and confirmed the nonpathogenicphenotype observed on several host plants. In eukaryotic cells, clathrin heavy chain is mainlydescribed in the endocytic pathway, but it is also essential for high-density secretory vesiclesformationinS.cerevisiae(Gurunathanetal.,2002).CharacterizationofthetwomutantsofB.cinerearevealed a secretion defect of numerous proteins including known virulence factors, as Plant CellWallDegradingEnzymesandelicitors.Aclathrin-coatedvesicleenrichedfractionwas isolatedfromthe wild type and mutants strains. Using a proteomic approach, we analyzed and compared theproteomiccompositionoftheseclathrin-coatedvesiclesandpresentheretheresults.Thisstudydemonstratedforthefirsttimetheessentialroleofclathrinintheinfectiousprocessofafungalpathogenanditsimportanceinvirulencefactorssecretion.Keywords:clathrin,vesicles,secretion,LC-MSanalysis,virulence

Session5.FROMPLANTIMMUNITYTOINNOVATIVEPLANTBREEDINGKeynotelecture8BiotechnologyforPlantDiseaseControl-NoveltoolsandperspectivesDavidB.Collinge1

1DepartmentofPlantandEnvironmentalSciencesandCopenhagenPlantScienceCentre,UniversityofCopenhagen,Thorvaldsensvej40,DK-1871FrederiksbergC,DenmarkFaced with the multiple challenges of (1) predicted world population growth, (2) urbanisationreducing available area of agricultural land, (3) an increased demand for meat-based diets andtherefore fodder and (4) climate change and catastrophicweather events, global agriculturemustadopt more dynamic, efficient and sustainable production methods to increase food and fodderproductioninordertosupportagrowingpopulation(FAO).Biotechnology platforms already play an integral role in improving agricultural yields and foodquality.Withtheuseofmolecularbreedingandmutationaltool,farmersareproducingrecordyieldsthan ever previously. By providing both innovative tools in plant breeding and uncoveringfundamental knowledge of plant-microbes interactions, these new approaches also pave the wayforwardinadvanceddiseasemanagement.Iwillprovideaninsightintotechniquesincludingandbeyondtransgenicapproachesinplantbiotechapplications. CRISPR-Cas9will be highlighted as a key biotechnological but non-transgenic tool forthedevelopmentofdiseaseresistantplants.Innovationsinthisfieldstandtoimproveplantdiseasecontrolmethodsforfuturefarmingsystems.CollingeDBBiotechnologyforPlantDiseaseControl,NewYorkandLondon:Wiley(2016)440pages.ISBN978-1-118-86776-1–URLforBook:http://eu.wiley.com/WileyCDA/WileyTitle/productCd-1118867769.html

Session5Oral39Uncovering the priming potential of the Green Leaf Volatile Z-3-HAC in wheat, ametabolomicsapproachMaartenAmeye1,2,LievenvanMeulebroek3,LynnVanhaecke3,GuySmagghe1,GeertHaesaert2,KrisAudenaert2

1LaboratoryofAgrozoology,DepartmentofCropProtection,FacultyofBioscienceEngineering,GhentUniversity,CoupureLinks653,B-9000Ghent,Belgium2DepartmentofAppliedBiosciences,FacultyofBioscienceEngineering,GhentUniversity,ValentinVaerwyckweg1,B-9000Ghent,Belgium3LaboratoryofChemicalAnalysis,DepartmentofVeterinaryPublicHealthandFoodSafety,FacultyofVeterinaryMedicine,Salisburylaan133,B-9820Merelbeke,BelgiumBecause of growing environmental concerns and following legislative pressure (2009/128/EC), theneed for innovative cropprotection toolshasgrownduring the lastdecade.Within the contextofintegratedpestmanagement(IPM),greenleafvolatiles(GLVs)maybeofparticularinterest.GLVsareubiquitouslyproducedthroughoutthewholeplantkingdomandhavebeenattributedanimportantroleindirectandindirectdefenseresponsesandintheprimingofplantdefense.Primingreferstoamechanismwherebyplantsaresensitizedtorespondfasterand/ormorestronglytofuturepathogenattack.Inpreviousresearch,wedemonstratedthatpre-exposuretothegreenleafvolatile(GLV)Z-3-hexenylacetate (Z-3-HAC) primed wheat (Triticum aestivum L.) for enhanced defense against subsequentinfection with the hemibiotrophic fungus Fusarium graminearum. GLVs may thus constitute apromisingagronomictool.However,notmuchisknownabouttheunderlyingprimingmechanisms.Building further on these findings, we attempt to unravel themechanism of priming by Z-3-HAC.Here, we used an untargetedmetabolomics approach, to identify changes in themetabolome ofwheat after exposure to Z-3-HAC and a subsequent infection with F. graminearum. Our analysisrevealedalargeupregulationoftheproductionofglycosylatedcompoundsuponZ-3-HACtreatmentandadownregulationoftheproductionofthebenzoxazinoidDIMBOA.Inadditiontoanuntargetedmetabolomics,wewereinterestedintheresponseofaselectionofseveralplantdefensehormonesand metabolites of the glutamate biosynthesis. We observed a large induction of salicylic acidbiosynthesis,whereasotherplanthormoneswerenotaffectedbyZ-3-HAC.Concurrently,wefoundalargedecreaseincomponentsinvolvedinglutamatemetabolism.Together, these findings illustrate that exposure to Z-3-HAC has both a large effect on primary Nmetabolism and induces glycosylation of metabolites, which may contribute to the increaseddefense.Keywords:Volatile,priming,innovativecropprotection,Fusarium,metabolomics,wheat

Session5Oral40Discovering coherency of plant reflectance, gene expressions and enzyme activities duringdifferentbarley–powderymildewinteractionsMatheus Thomas Kuska1, Jan Behmann1, Dominik KilianGrosskinsky2, ThomasGeorg Roitsch2, andAnne-KatrinMahlein1

1UniversityofBonn,InstituteofCropScienceandResourceConservation(INRES),Phytomedicine-PlantDiseaseandPlantProtection,Nussallee9,53115Bonn,Germany2UniversityofCopenhagen,SectionforCropSciences,DepartmentofPlantandEnvironmentalSciences,HoejbakkegaardAllé13,2630Taastrup,DenmarkThe Mildew locus o (mlo) gene based resistance of barley is successful against infestation byBlumeria graminis f.sp.hordei (Bgh), the causal agent of barley powderymildew, by fast cellwallappositions (papilla). Modified epidermal cells remain intact. Contrary, barley Mla gene basedresistance leads to a single cell or tissue hypersensitive reaction. These resistance responses arebasedonacascadeofgen-proteinsignaling.Theobjectivesofthisstudyaretocharacterizeresistancereactionsagainstpowderymildewusingahyperspectral-imaging microscope and to uncover relevant wavebands for the transcriptionalpathway.ExperimentswereconductedwithH.vulgarenearisogeniclinescv.Ingridwildtype(WT),amlogenebased resistant genotype and cv. Pallas Mla gene based resistant genotype. Hyperspectralreflectancewasmeasured every 3 hours until 48 hours after inoculation (hai) withBgh and dailymeasurementsuntil5daysafterinoculationtoassessearlyinteractionsites.Parallelgeneexpressionandenzymeactivityprofilingwereusedtodeterminebarleypathogenesisandresistancestages.Differencesintherelevanceamongsinglewavebandsforhyperspectralcharacterizationofbarley–powderymildewinteractiontypeswereassessedusingtheReliefalgorithm.Significant changes of relevance occur at specific key moments of barley – powdery mildewinteraction 12, 24, 48 and 72 hai. Further data driven analysis indicated simultaneous processesbetween changes in hyperspectral reflection and observed gene expressions and enzyme activityovertime,respectively.These results enable an exhaustive interpretation of plant hyperspectral reflectance in compatibleandincompatibleplant-pathogensystemsfornon-invasiveplantphenotyping.Keywords:Cropresistance,phenotyping,genotyping,hyperspectralimaging,Blumeriagraminis

Session5Oral41ControlofImmunityduringlegumeRhizobiumsymbiosisFathi Berrabah1,2,3, Jeoffrey George1,2, El Hosseyn Aït-Salem1,2, Sophie Massot1,2, Thierry Baillau4,MichelZivy4,MarieGarmier1,2,BenjaminGourion1,2,5,PascalRatet1,21Institute of Plant Sciences Paris Saclay IPS2, CNRS, INRA, Université Paris-Sud, Université Evry,UniversitéParis-Saclay,Batiment630,91405Orsay,France2Institute of Plant Sciences Paris-Saclay IPS2, Paris Diderot, Sorbonne Paris-Cité, Bâtiment 630,91405,Orsay,France3DepartmentofBiology,FacultyofNatureandLifeSciences,ZianeAchourUniversity,17000Djelfa,Algeria.4Plateforme d’Analyse Protéomique de Paris Sud-Ouest, Unité Mixte Génétique Quantitative etEvolution-LeMoulon,F-91190,Gif/Yvette,Paris,France5Laboratoire des Interactions Plantes Microorganismes, Université de Toulouse, INRA, CNRS,CastanetTolosan,FranceRhizobiumandLegumesestablishasymbioticinteractionsresultingintheformationofrootnodulesinwhichrhizobiafixnitrogen.Itisgenerallypresumedthatrhizobia/legumessymbiosisevolvedfrompathogenicinteractions.However,evenwhenrhizobialpopulationsreachveryhighdensitiesinthenodules,theydonotelicitvisibledefensesreactions.Withtheaimtobetterunderstandthemolecularmechanism(s)thatprevent(s)thedevelopmentofdefensesduringrhizobia/legumesinteractions,westudiedthesymCRKmutantofMedicagotruncatulawhichdevelopsdefensereactionsinnodules.Proteomicsanalysiswasperformedonfractionatednodulesandtheproteomesofbothpartnerswascharacterized.Theresultssuggestapotentialinvolvementofethyleneinthedefensesdevelopedbythismutant.InagreementwithapotentialroleforethyleneinthesymCRKphenotype:i)ACCoxidaseactivity,whichisresponsibleforethylenesynthesis,ishigherinthesymCRKnodulesthanintheWTonesii)ethylenetreatmentsonmaturenodulesmimicthesymCRKmutationandiii)theinhibitionofethylenesynthesisreducestheintensityofdefensesdevelopedinthemutantnodules.TogetherourresultssuggestthattheethylenesignalingpathwayhastobeinhibitedinmaturenodulesinordertopreventthedevelopmentofdefensereactionsandthatSymCRKplaysaroleinthisinhibition.Wearenowfurthertestingthishypothesisortheactionofotherdefensehormonesincontrolofthesymbioticimmunity.

Keywords:Symbiosis,Rhizobium,legumeplants,immunity,ethylene.

Session5Oral42CharacterizationofIndividualReceptor-LikeKinaseDomainsinIOS1fromArabidopsisLaïlaGiordano*,ValérieAllasia,AndriaPietri,ElodieNaessens,SophieHok,andHaraldKellerUMR1355-7254InstitutSophiaAgrobiotech,06900SophiaAntipolis,France

Oomycetes are devastating filamentous pathogens that impact ecosystems and agriculture. Ourresearchaimsatcharacterizingthemolecularmechanismsthatgoverntheestablishmentofdiseasein host plants. To sense the environment, plant cells possess more than 200 plasma membrane-spanning receptors, which are composed of extracellular leucine-rich repeats (LRRs) and anintracellularkinasedomain.WepreviouslyidentifiedtheArabidopsisreceptor-likekinase"ImpairedOomyceteSusceptibility1"(IOS1),whichdownregulatesabscisicacid(ABA)signalingandcontributesto the infection successof filamentous,biotrophicpathogens suchasoomycetesand thepowderymildewfungus (Hoketal.,2011;Hoketal.,2014).Theextracellular regionof IOS1 iscomposedofLRRs and a domain, which shares similarities with malectin from animals. Animal malectins bindcarbohydratesandparticipateinmonitoringtheglycosylationstateofproteinsduringtheirtransitinthe endoplasmic reticulum (ER). We observed retention of IOS1 in the ER, which appears to bemediated through the malectin-like (ML) domain. Yeast two-hybrid screens with the extracellularIOS1 domain identified several proteins, among which some localize to the ER. Results from thescreenandsubsequentstudieswillbepresentedanddiscussed.Hoketal.(2014).PlantPhysiol.166,1506-1518.Hoketal.(2011).PlantCellEnviron.34,1944-1957.*L. Gioradano is supported by the Labex “Signalife” [#ANR-11-LABX-0028-01] and the INRA SPEdepartment.Keywords:Arabidopsis,oomycete,receptor,malectin,endoplasmicreticulum,abscisicacid

Session5Oral43ARPP8-likeRgenecontrolsPotatovirusY-inducedveinalnecrosisintobaccoV.Michel1,E. Julio²,T.Candresse1, J.Cotucheau²,C.Decorps²,R.Volpatti², F.DorlhacdeBorne2,V.

Decroocq1,andS.GermanRetana1.1UMRBFP1332(BiologieduFruitetPathologie),EquipedeVirologie,CentreINRAdeBordeaux,CS20032,33882Villenaved’OrnonCedex2ImperialTobaccoBergerac(ITB),LaTour,24100Bergerac,FrancePotatovirusY isoneofthemostdamagingvirusontobaccoworldwide.Aggressivenecroticstrainscan lead to considerable yield losses. The va locus in tobacco conferring resistance to PVY wascharacterized[1].However,PVYisolatesovercomingthisresistanceandinducingnecroticsymptomswere observed in several countries. In this context, it is important to find new va-independentresistancemechanisms.Therefore, phenotyping testswere conducted on 163 varieties of the ITB collection. Resistance toPVYwasestimatedbysymptomsurvey,andthescreeningwasperformedwithdifferentPVYnecroticstrains,includingoneabletoovercomeva-resistance.Ten cultivars that do not carry the va locus failed to show the typical vein necrosis infectionphenotype.PositiveELISAtestsshowedaviralaccumulationintheseplants,despitetheabsenceofnecrotic symptoms. A differentmechanism, involving tolerance to the necrosis induced by PVY isthereforeinvolved.FinemappingonaF2segregatingpopulationshowedthatthistolerancetraitisinheritedasasinglerecessive gene and allelism tests showed that eight out of ten varieties carry the same gene.Anchoring the linkage map to the tobacco genome physical map allowed the identification of acandidategene(RPP8-likeRgene)withaSNPpolymorphism inthose8cultivars.FunctionalassaysusingTILLINGmutantsintheRPP8geneconfirmedtheroleofthiscandidateintolerancetonecrosisinduced by PVY. The vein necrosis symptoms may be due to a RPP8-mediated HypersensitiveResponse (HR) inducedbyPVY,although inefficient to impedeviralpropagation.SuchsystemicHRwould not occur when RPP8 is mutated in the tolerant varieties. Experiments are in progress toconfirmthishypothesis.This gene could be used in future tobacco breeding programs to minimize the impact of varesistance-breakingstrainsandtolimitcroplosses.Keywords:potyvirus,tolerance,tobacco,vein-necrosis,R-gene[1]E. Julio, J.Cotucheau,C.Decorps,R.Volpatti,C.Sentenac,T.Candresse&F.DorlhacdeBorne.(2014). A Eukaryotic Translation Initiation Factor 4E (eIF4E) is responsible for the “va” Tobaccorecessiveresistancetopotyviruses.PlantMolBiolRep.DOI10.1007/s11105-014-0775-4.

Session5Oral44

Diversity in wheat Composite Cross Populations provides protection from the new yellow rust(Pucciniastriiformis)racesOdetteWeedon1,SarahBrumlop1,SvenHeinrich1andMariaRFinckh1

1UniversityofKassel,DepartmentofEcologicalPlantProtection,Nordbahnhofstr.1a,Witzenhausen.37213,Germany

CompositeCrossPopulations(CCPs)displayintra-specificdiversityandallowforevolutionarybreeding.Theirgeneticdiversityallowsthemtoadapttotheenvironmentalconditionsinwhichtheyaregrown.AnimportantquestioniswhetherCCPscanalsodealwithevolvingpathogenssuchasthenewyellowrust(Pucciniastriiformis)racesthathavedominatedmostEuropeancountriesafter2011.Three winter wheat CCPs, composed of 9, 12, or 20 parental varieties have been grown at theUniversity of Kassel in both a conventional and organic system,without conscious selection since2005 (F5). For comparison, several current commercial varieties were grown every year in bothsystems.Diseasepressurewasstill lowin2012(meanAUDPC=116)whenthenewraceswerenotyetevident locally.Yellowrustdominated in2014,2015,and2016 (meanAUDPC=345,438,232,respectively). In these three years,mean AUDPCs in the CCPswere 332 and 334 in the referencevarieties. AUDPCs varied by a mean of 54% among CCPs, while among reference varieties thevariationwas130%.Themoderncommercialvarieties, incomparison,wereeitherveryresistantorconsiderably more susceptible than the CCPs, most likely due to major genes and their geneticuniformity. These results indicate that the CCPs show low to moderate susceptibility to the newyellowrustracesandthatdiseaseincidencewasmorepredictablethanamongthemodernvarieties.This isdespite the fact thatmanyof theparentalvarieties thatweregrown insmallplots in2015,tendedtobehighlysusceptible,havingbeenbredbetweenthe1930sand2000,beforethesenewraces were present. The results illustrate the buffering capacity and potential of the geneticallydiverseCCPstoadapttothechangingbioticpressureoftheyellowrustpathogen.

Keywords:CompositeCrossPopulations(CCPs),EvolutionaryBreeding,Yellowrust,adaptation

Session5Oral45Developmentalprocessesalteredduringgallformationbyroot-knotnematodesRocíoOlmo1*,JavierCabrera1*,FernandoE.Díaz-Manzano1,MartaBarcala1,MariaFeAndrés2,AnaCláudiaSilva1,IsabelMartínez1,VirginiaRuíz1,CarmenFenoll1andCarolinaEscobar1

1 Facultad de Ciencias Ambientales y Bioquímica, Universidad de Castilla-La Mancha, Avenida deCarlos III s/n, 45071, Toledo, Spain 2 Departamento Protección Vegetal, Instituto Ciencias AgrariasCSIC,Serrano115,28006,Madrid,Spain.*Bothauthorscontributedequallytothiswork.

Plantendoparasiticsedentarynematodescauseseverelossesindifferentcrops.Amongthemost

harmfularetheroot-knotnematodes(RKNs;Meloidogynespp.).RKNsenterbytherootelongationzone andmigrate intercellularly. They stablish within the vascular cylinder and induce specializedfeeding cells, giant cells (GCs) from vascular cell precursors, still not well known, after repeatedmitosiswithpartialcytokinesis.ThecellsaroundtheGCsproliferateandthecortexcellshypertrophy,developingagall,anewlyformedpseudo-organ(Escobaretal.,2015).

Weobtained the transcriptomesof early-developingGCs (at 3 dayspost inoculation) and gallsinduced byM. javanica in Arabidopsis using laser capture microdissection (Barcala et al., 2010).Interestingly,GCsandgallstranscriptomessharedsimilaritiestothosefromthequiescentcentreandlateral root initial cells (Cabrera et al., 2014). Hence,we checkedmarker andmutant lines of keygenesforbothcelltypes,amongothers,lateralorganboundaries-domaintranscriptionfactors,LBDs,crucial during lateral root development, or HSFB4 (SCHIZORRIZA), a ground tissue and quiescentcentremarker.

Our results confirm that RKNs partially ‘hijack’ plant transduction pathways leading to theformation of lateral roots with common transducers as LBD16 or mRNA390, expressed in theproliferativecellswithinthegallsandintheGCs(Cabreraetal.,2014;2016),similarlytoHSFB4,andtootherkeymarkersforroottipmeristem.Moreover,themassivegenerepressionobservedintheGCs, ispartiallyregulatedthroughmiRNAsasmiR172,crucialduringflowering.Thisbringstogetherapparently unrelated and distant processes such as flowering in the aerial parts, lateral rootformation,ortherootmeristemmaintenanceinrootsandmorphogeneticresponsestopathogensinroots. All these suggests that common regulatory molecular partners at the cellular level mightcontrol different organogenetic processes triggered either by internal developmental cues or bybioticinteractions.

References

Barcala et al., 2010. Early transcriptomic events in microdissected Arabidopsis nematode-inducedgiantcells.PlantJ61(4):698-712.

Cabrera et al., 2014. A role for LATERAL ORGAN BOUNDARIES-DOMAIN 16 during the interactionArabidopsis-Meloidogyne spp.providesamolecular linkbetween lateralrootandroot-knotnematodefeedingsitedevelopment.NewPhytol203(2):632-645.

Cabreraetal.,2016.DifferentiallyexpressedsmallRNAsinArabidopsisgallsformedbyMeloidogynejavanica: a functional role for miR390 and its TAS3-derived tasiRNAs.New Phytol 209(4):1625-1640.

Escobaretal.,2015.OverviewofRoot-KnotNematodesandGiantCells.In:CarolinaE,CarmenFeds.AdvancesinBotanicalResearch:AcademicPress,1-32.

Keywords:Rootknotnematodes,lateralroot,giantcells,galls,rootmeristem

Session6.EMERGINGTOOLSFORTHEMANAGEMENTOFPLANTDISEASES:AGROECOLOGYANDDISEASEMANAGEMENT– partIKeynotelecture9European Platform of research and development of tools for biocontrol of phytopathogens:SMARTBIOCONTROLPhilippe Jacques1, Marc Ongena1, Essaid Ait Barka2, François Krier3, Jenny Neukermans4, SylvainDesprez5andallmembersofSMARTBIOCONTROLProject1 Terra Teaching and Research Centre, Microbial Processes and Interactions (MiPI), University ofLiège-GemblouxAgro-BioTech,B-5030Gembloux,Belgium2 Laboratory of plant stress, defence and reproduction, URVVC-EA 4707, University of ReimsChampagne-Ardenne,F-51687Reimscedex2,France3Université Lille1, InstitutCharlesViollette,Polytech’Lille, avenuePaul Langevin,CitéScientifique,59655Villeneuved’Ascq,France4 PCG - ProvinciaalProefcentrumvoor de Groenteteelt Oost-Vlaanderen vzw, Karreweg 6, 9770,Kruishoutem,Belgium5Materianova,ParcInitialis,AvenueN.Copernic1,7000,Mons,Belgium WithintheInterregFWVLPHYTOBIOproject(2010-2014)andmorerecentlythenewprojectportfolio SMARTBIOCONTROL (2016-2020), 26 partners from university laboratories, researchcentersoragricultural technical institutesandcompanydecidedtogathertheirskills todiscovernew biomolecules (mainly lipopeptides) retaining high potential to control efficiently, and in asustainable manner, a large panel of crop diseases. In the PHYTOBIO project, a dozen ofamphiphilicmetabolitesproducedbybacteria,mainlybelonging to theBacillusgenus, displayedpromisingactivitiesasbiocontrolagents,bothingreenhouseandfieldconditions. The new project portfolio SMARTBIOCONTROL is composed of 4 main scientific projectsworkingondifferentfieldsofbiocontrolinthecross-borderregionofFrance-Flanders-Wallonia: - the BioScreen project aims at discovering new multifunctional molecules of biologicaloriginsabletoeitherdirectlyinhibitmicrobialpathogensand/orinducesystemicresistanceinthecropsofinterest. - theBioProd projectwill optimize the production and purification of thesemolecules atpilot scale. In addition, the biodegradability and the toxicity of the new molecules will bemonitoredandmanyformulationswillbeassayedinordertogetthesemoleculescommercializedanddeliveredtofarmersinthemoststable,activeandeasy-to-handleforms. -theBioProtectprojectwillworktoimprovetheuseofbiologicalcropprotectionproductsto combat phytopathogens in a sustainableway and to reduce the amount of chemical in cropprotection. The project will also focus on the optimization of the efficacy of biological cropprotectionproductsin(field)trials. - the BioSens project will develop a new generation of biochips allowing to track thesebiocontrolagents (microorganismsand/orbiocidalmolecules) toevaluate theirpersistenceafterapplicationinthefield. Inthiskeynotelecture,wewillpresenti)themainresultsofthePhytobioprojectrelatedtothescreeningofthebioactivemolecules,theirmodeofaction,theirproductionprocessandtheirefficiency in greenhouses and fields trials and ii) the concept and strategy of theSMARTBIOCONTROLportfolio.

Keywords:SMARTBIOCONTROL,BIOSCREEN,BIOPROD,BIOPROTECT,BIOSENS,INTERREGVFWVL

Session6Oral46DevelopmentofanewbiologicalcontrolproductforpowderymildewcontrolincerealsJürgenKöhl1,CarinLombaers-vanderPlas1,TrudyvandenBosch1,HelenGoossen-vandeGeijn1,LiaGroenenboom-deHaas1,HuubSchepers2,UlrikeHilscher3,MariannWikström4

1 Wageningen University & Research, Business Unit Biointeractions and Plant Health,Droevendaalsesteeg1,6708Wageningen,TheNetherlands2WageningenUniversity&Research,BusinessUnitArableFarming,FieldproductionofVegetablesandMultifunctionalAgriculture,Edelhertweg1,8219PHLelystad,TheNetherlands3BayerCropScienceBiologics,Metkenberg6,23970Wismar,Germany4AgroPlantarum,Kärrarpsvägen410,26590Åstorp,Sweden

Objectives

The objective of our research as part of the EUproject BIOCOMES is to select new fungalantagonistsforcontrolofBlumeriagraminisf.sp.triticiinwheat.

Methodsused

More than 1200 fungal isolates were obtained from powdery mildew pustules on leavescollected inTheNetherlands,GermanyandSweden.Potentialcandidateantagonistswereselectedby high-throughput screening methods followed by efficacy tests under controlled and fieldconditions.Results

The pre-screening criteriawere fulfilled by 748 isolates. These criteriawere: no growth at36°C(safetyforhumans),germinationandgrowthat5°C(coldtolerance),germinationandgrowthat-7MPa (drought tolerance) and growth after exposure toUVb (UVb tolerance). In a next step 27isolateswereexcluded,becausetheybelongedtospeciesconsideredasnotsafeforhumans,animalsorplants.185representativeisolateswithhighsporeproductionwereselectedandtheirefficacytoreducepowderymildewwasassessedinbioassaysonwheatseedlings(cv.Julius)conductedat15°Candhighhumidity.NumbersofproducedB.graminis conidiaandthe leafcoverageswithpowderymildewpustuleswerequantified.The10 isolateswithhighestefficacyandmost consistent resultswerethentestedonpottedspringwheatplants(cv.Calixo)intheopenfieldfollowedbyanefficacytrialinaspringwheatcrop(cv.Heron).Thebestisolatessignificantlyreducedtheleafcoveragewithpowderymildewandtheincreaseofthepowderymildewepidemicintime.Conclusions

New antagonistic isolates were selected for the development of economically feasiblebiological control agents. Next research steps will focus on mass production and formulation,additionalfieldtestingandmode-of-action.Acknowledgement

ThisprojecthasreceivedfundingfromtheEuropeanUnion’sSeventhframeworkProgrammeforresearch,technologicaldevelopmentanddemonstrationundergrantagreementno612713.

Session6Oral47MultitrophiccontrolofFusariumHeadBlight:newacquisitionSabrina Sarrocco1, Riccardo Baroncelli2, SaraManfredini1, Rodolfo Bernardi1,MiriamHaidukowski3,AntonioMoretti3andGiovanniVannacci1

1UniversitàdegliStudidiPisa–DipartimentodiScienzeAgrarie,AlimentarieAgro-Ambientali,PlantPathology&Mycologylab-ViadelBorghetto80–56124Pisa,Italy.2 Université de Brest, EA 3882, Laboratoire Universitaire de Biodiversité et EcologieMicrobienne,IBSAM,ESIAB,TechnopôleBrest-Iroise,29280Plouzané,France.3ISPA-CNR,ViaAmendola,122/O-70126Bari,Italy

FusariumHeadBlight(FHB)isoneofthemostimportantdiseasesofwheat,causinglossesinyield and quality, this last due to mycotoxins production by the main causal agent Fusariumgraminearum (FG). The early detection and control of trichothecene-producing Fusarium spp. iscrucialtopreventtoxinsenteringinthefoodchain.BiocontrolofFHBisavalidstrategyandentailsthe treatment of crop residues and/or of spikes during anthesis with antagonists. The aim is topreventthepathogeninfectingplantsandcompetitionseemstobeoneofthemainmechanismsofaction.

Two beneficial fungi, T. gamsii 6085 (TG) - able to control F. graminearum growth andmycotoxinaccumulation,andtoreduceFHB infield -andFusariumoxysporum7121(FO)–agoodsaprotrophiccompetitor forcropresidues–havebeenused inamultitrophicapproach inordertocontrol F. graminearum growth (evaluated by Real-time PCR) and to reduce trichothecenesproduction(measuredbyHPLC)onnaturalsubstrates.Inaddition,NicheOverlappingamongthetwoantagonistsandthepathogenwasfurtherinvestigatedbyusingtheBiologmicroarraysystem.

The two beneficial fungi are able, to a different extent, to reduce pathogen growth andmycotoxinsaccumulationinnaturalsubstrates.Noantagonisticeffectshasbeenregisteredbetweenthetwoantagonistswhenusedtogether, leavingopenthepossibilitytousetheminamultitrophicapproachthatwillincreasetheprobabilityofsuccessofthediseasecontrol.BiologresultsshowthatFGandFOoccupythesamenicheandcompetefornutrients.FGdominatesonTGsuggestingthatnutrientcompetitionisnotthemainmechanismofactionforTG.

Finally, thanks to the availability of the genomes of both isolates, and the genome-editing(CRISPR-Cas) protocols,we are actually collecting new information and attempting to improve theefficacyofthetwoantagoniststowardsFHB.

Keywords:FusariumHeadBlight,Fusariumgraminearum,Trichodermagamsii,Fusariumoxysporum,mycotoxins,multitropicapproach,genomeediting

Session6Oral48Cellwallglycomoleculesandbordercells:sentinelsforrootdefenseagainstsoil-bornepathogens.VicréMaïté*,CastilleuxRomain,PlancotBarbara,ManasfiYoussef,PlassonCarole,BoulogneIsabelle,Follet-GueyeMarieLaure,DriouichAzeddineLaboratoireGlycobiologieetMatriceExtracellulaireVégétale(EA4358),FédérationdeRecherche«NormandieVégétal»etPlate-Formed’ImagerieCellulaire(PRIMACEN)deHaute-Normandie.NormandieUniversité,UniversitédeRouen,76821MontSaintAignanCedex,France.

Mostplantspeciesreleasethousandsoflivingrootbordercellsintherhizosphere(Hawesetal.,2000).Duetotheirveryspecialpositionattheinterfacebetweenrootsandsoil,bordercellsarekeyelementsinvolvedinrootprotectionagainstsoil-bornepathogens.Inpea(Pisumsativum),rootbordercellsarespecializedintheproductionofanti-microbialcompoundsincludingpisatin,arabino-galactanproteins(AGP)andextracellularDNA(Cannesanetal.,2011;2012;Driouichetal.,2013).Rootbordercellsandtheirassociatedmucilagewereshowntobeessentialinshapingsoil-bornemicrobialcommunitiessurroundingtheroottip(NguemaOnaetal.,2013).WehaveassessedtheimpactofAGPproducedbypearootcapandbordercellsonthedevelopmentcycleofAphanomyceseuteiches,apathogencausingrootrotdisease.InvitroassayshaverevealedthatAGPattractthezoospores,inducetheirencystmentandpreventtheirsubsequentgermination.ThesedataprovideevidenceforaroleofAGPinroot-microbeinteraction(Cannesanetal.,2012).Wehavealsoshownthatrootbordercellsfromdifferentplantspeciessecreteextensin,anotherhydroxyprolin-enrichedglycoproteinbelongingtothesamefamilyasAGP(Plancotetal.,2013;Koroneyetal.,2016).Inaddition,extensinsecretionseemstobeenhancedinresponsetoelicitors(Plancotetal.,2013,Castilleuxetal,unpublished).Weproposedamodeltermed“RootExtracellularTrap”orRETinwhichwepostulatedthatrootmucilagetogetherwithrootbordercellscouldfunctioninrootdefenseinawaysimilartothatofneutrophilextracellulartraps(NET)inmammaliancells(Driouichetal,2013).Cellwallglycomoleculesincludingcallose,AGPandextensinappeartoplayacriticalroleinrootimmunityandplanthealth.Inthiscommunication,wewillpresentanddiscussthefunctionalorganizationofrootbordercellsandtheirsecretionsinplantdefense.Acknowledgments:ThisworkwassupportedbyLeGrandRéseaudeRechercheVASI«Végétal-Agronomie-SolsetInnovations».Keywords:arabinogalactan-proteins,cellwall,extensin,oomycetes,rootbordercellsReferencesCannesanMA,GangneuxC,LanouxA,GironD,LavalK,HawesM,DriouichA,Vicré-GibouinM.(2011)AssociationbetweenbordercellresponseandlocalizedrootinfectionbyAphanomyceseutiches.AnnalsofBotany108:459-469CannesanMA,DurandC,BurelC,GangneuxC,LerougeP,IshiiT,LavalK,Follet-GueyeML,DriouichA,Vicré-GibouinM(2012)EffectofarabinogalactanproteinsfromtherootcapsofPisumsativumandBrassicanapusonAphanomyceseuteicheszoosporechemotaxisandgermination.PlantPhysiology159:1658-1670DriouichA,Follet-GueyeML,Vicré-GibouinM,HawesM(2013)Rootbordercellsandsecretionsascriticalelementsinplanthostdefense.CurrentOpinioninPlantBiology16:489-495

HawesMC,GunawardenaU,MiyasakaS,ZhaoX(2000)Theroleofrootbordercellsinplantdefense.TrendsinPlantScience5:128-133.KoroneyAS,PlassonC,PawlakB,SidikouR,MenuBouaouicheL,DriouichAandVicré-GibouinM(2016)RootmucilageofSolanumtuberosumisenrichedingalactose-containingmoleculesandimpactsthegrowthofPectobacteriumatrosepticum.AnnalsofBotany118(4):797-808Nguema-OnaE,Vicré-GibouinM,CannesanMA,DriouichA(2013)Arabinogalactanproteinsinroot–microbeinteractions.TrendsinPlantScience18:440-449PlancotB,SantaellaC,JaberR,Kiefer-MeyerMC,Follet-GueyeML,LeprinceJ,GattinI,SoucC,DriouichAandVicré-GibouinM(2013)Decipheringtheresponsesofrootborder-likecellsofArabidopsisthalianaandLinumusitatissimumtopathogen-derivedelicitors.PlantPhysiol.,163,1584-1597.

Session6Oral49ResistanceinducedbyulvansagainstplantpathogenicfungiMarcielJ.Stadnik1,MateusB.deFreitas1,LeonardoAraújo2,LuizAugustoM.Peruch2,JoãoAméricoWordell2,RobertaPaulert3andMiguelD.Noseda31FederalUniversityofSantaCatarina,UFSC,88.034-001Florianópolis-SC,Brazil2AgriculturalResearchCompanyofSantaCatarinaState,Epagri,Florianópolis,SC,Brazil3FederalUniversityofParaná,UFPR,Curitiba,PR,Brazil

Ulvans are water-soluble sulfated heteropolysaccharides extracted from the cell walls ofgreenmarinemacroalgaeUlvaspp.andareabletoinducesystemicresistanceinplants.Theaimofthis report is to summarize themain results of our research group, fromdiscovery to attempts inelucidating themechanismsof action.Abioprospecting study carriedoutbetween2002and2004identified Ulva fasciata Delile as a potential source of active compounds for controlling plantdiseases.Ingreenhouse,sprayofUlvaextractsreduced(about80%)thenumberofpowderymildewcoloniesonpre-treatedleaves.Whenappliedunderfieldconditions,spraysofthealgalextractswerealso able to control anthracnose by 50% in a highly susceptible bean cultivar. Follow-up studiesrevealedthatulvanswerethemainelicitormoleculesintheextractsofUlva.Ingreenhouse,ulvansdemonstratedtoprotectseveralcropplantssuchasbeans,apple,grapevineandcereals(e.g.wheatandbarley)againstabroadrangeoffoliarphytopathogens,includingbiotrophic,hemibiotrophicandnecrotrophic fungi. Pretreatment of barley leaveswith ulvans significantly reduced the severity ofBlumeria graminis by 80%, and in cereals, priming effectwas detected in assayswith suspension-culturedcells.Resistanceinducedbyulvansreducedthebeananthracnoseseverityby50%,lastingatleastuptoninedaysaftertreatment.Protectionlevelsof65%wererecordedinulvans-treatedappleplantsagainsttheGlomerellaleafspot,buttheeffectwasweakerinnot-directlytreateddistalparts.Jasmonic acid pathway as well the enzyme NADPH oxidase were triggered during ulvan-inducedresistance in Arabidopsis thaliana. These findings may open new horizons for using the algalpolysaccharidesasresistanceinducers.Keywords:seaweeds,anthracnose,powderymildew,ulvans,inducedresistance.

Session6Oral50ACombineduseofBiostimulantsandResistanceInducerstoimprovewheatprotectionAliceFourquez1,MarylineMagnin-Robert1,BéatriceRandoux1,PhilippeReignault1,AliSiah2,PatriceHalama2,MarieGacoin3,NathalieGaveau3,YukoKryzaniak4,SophieTrouvelot4,Marie-ClaireHeloir4,MarielleAdrian4,EstelleMoreau5,andothermembersoftheIris+consortium.1Univ.LittoralCôted’Opale,UCEIV-EAn°4492,UnitédeChimieEnvironnementaleetInteractionssurleVivant,SFRCondorcetFRCNRS3417,CS80699–F-62228Calaiscedex,France2EquipeBiotechnologieetGestiondesAgentsPathogènesenagriculture,LaboratoireCharlesViollette,SFRCondorcetFRCNRS3417,InstitutSupérieurd’Agriculture,Univ.LilleNorddeFrance,48BdVauban,–F-59046Lillecedex,France3UnitédeRechercheVigneetlesVinsdeChampagne(URVVC),LaboratoiredeStress,DéfensesetReproductiondesPlantes(SDRP),SFRCondorcetFRCNRS3417,Bâtiment18,MoulindelaHousse-BP1039-51687ReimsCedex2,France4Agroécologie,AgrosupDijon,CNRS,INRA,Univ.BourgogneFranche-Comté,17rueSully–F-21065Dijon,France5ParcTechnopolitainAtalante,CS4190835435Saint-Malo,France

InthecontextoftheEcophytoFrenchNationalActionPlan,Franceiscommittedtoreducegraduallytheuseofpesticidesonagriculturalcropswhilemaintainingahighproductionrateandgoodfoodquality.Asanalternativetopesticides,knowntoactdirectlyonpathogens,newelicitors,orResistanceInducers(RIs),arebeingdevelopedtoprotectplantsviathestimulationofplantdefenses.GenerallytheefficacyofRIstoprotectplantsisnowwellestablishedincontrolledandgreenhouseconditions,butmaysubstantiallydecreasewhenappliedtocropsinfieldconditions.WeaimhereatcombiningRIswithbiostimulants(BSs),inordertoincreasetheirprotectiveefficacybyoptimizingthephysiologyoftreatedplants.Despitetheirgrowinginterestinagriculture,themodesofactionofBSsonplantshavebeenpoorlycharacterizedsofar.IntheIRIS+project,wefocusononemajorcropofgreateconomicinterest:breadwheat(TriticumaestivumL.)to(i)evaluatetheimpactofthreenewmoleculesconsideredasBSsonplantgrowthandphysiologyand(ii)screenasetoffivenewmoleculesconsideredaspotentialsRIsagainsttwofoliarcryptogamicdiseases:powderymildew(PM,causedbyBlumeriagraminisfsp.tritici)andSeptoriatriticiblotch(STB,causedbyZymospetoriatritici),(iii)selectatleastoneefficientBS/RIcombinationinordertoimprovethelevelofwheatprotectionagainstthetwodiseases,comparedtoatreatmentwithaRIalone.AmongthefiveRIstestedtoprotectwheatagainstPMandSTB,SDN2,SDN3andSDN5wereefficientagainstPM,withprotectiveeffectsof20-30%,20-40%and65%respectively.Moreover,SDN3andSDN5werealsoefficientagainstSTB,theydecreasednecroticleafareaby50%andalmost80%respectively,andalsoreducedsporulationby30-70%and40%respectively.ThethreeBShavebeentestedindifferentconditions(methodsofapplications,substrate,nutrition)fortheireffectonplantgrowthparametersandleafpigmentcontent,butnoglobalpositiveeffectcouldbedetected.ThedirectactivityofRIsandBSsonthetwofungusstrainshavealsobeentestedinvitro:SDN2weaklyinhibitedB.graminissporesgermination,SDN3totallyinhibitedZ.triticimyceliumgrowthandSDN5totallyinhibitedB.graminissporegerminationandstronglydelayedZ.triticimyceliumgrowth.WestillhavetodeterminewhethertheprotectiveeffectsofRIsobtainedinplantaareduetothestimulationofwheatdefenses,directactivityonB.graminisandZ.tritici,oracombinationofbothmodesofaction.ConcerningBSs,wehavetooptimizeamethodofapplicationrelevanttoanevaluationofphysiologicalandgrowthparametersbeforeconsideringacombineduseofRIsandBSs.Keywords:Breadwheat,Powderymildew,Septoriatriticiblotch,Resistanceinducer,Biostimulant

Session6Oral51Burkholderia genomemining forNRPS reveals a great potential for lipopeptide productionwithbiocontrolapplicationsQassimEsmaeel1,2,MaudePupin3,4,FrançoisKrier1,PhilippeJacques5andValérieLeclère1

1: Univ. Lille, INRA, ISA, Univ. Artois, Univ. Littoral Côte d’Opale, EA 7394-ICV- Institut CharlesViollette,F-59000Lille,France2:LaboratoiredeStress,DéfensesetReproductiondesPlantesURVVC-EA4707,UFRSciencesExactesetNaturelles,UniversityofReims-Champagne-Ardenne,Reims,France3.Univ.Lille,CNRS,CentraleLille,UMR9189-CRIStAL-CentredeRechercheenInformatiqueSignaletAutomatiquedeLille,F-59000Lille,France4:Inria-LilleNordEurope,Bonsaiteam,F-59655Villeneuved’AscqCedex,France5: TERRA Research Centre, Microbial Processes and Interactions (MiPI), Gembloux Agro-Bio TechUniversityofLiege,B-5030Gembloux,Belgium

BacteriabelongingtothegenusBurkholderia live invariousecologicalnichesandpresentasignificant role in theenvironments through theexcretionof awide varietyofmodular secondarymetabolites including non ribosomal peptides (NRPs) and polyketides (PKs). These secondarymetabolites representan important sourceofnaturalproducts that canbeused topromoteplantgrowthorreplacechemicalpesticidesastheyhaveantifungalactivitiestogetherwithbiosurfactantproperties.

A genomeminingwas performed on 48 fully sequenced genomes ofBurkholderia speciespubliclyavailable.Thisinsilicoscreeningfornewsecondarymetabolites,usingspecificbioinformaticstools, revealed a total of 161 clusters containing nonribosomal peptide synthetases (NRPSs) andpredictedthesynthesisofatleast11novelproducts.Mostofthemaresiderophoresorlipopeptides,two classes of compounds with potential application in biocontrol. A dozen of synthetasesharbouringsignatures forcyclic lipopeptideproduction (presenceofCstarter,anddualC/Edomainsforepimerisationofcorrespondingmonomers)werepredictedfromgenomesequencesofthesinglestrain, displaying theability todecrease surface tension to27mNm-1. Consequently, this strain isconsidered as a very good candidate for biocontrol agent development. Other lipopeptides likeburkhomycin and burkholdin, attractive for its antifungal activity, were shown to be produced byvariousstrains.Keywords:Burkholderia,Biocontrol,lipopeptides,NRPS,genomemining

Session6.EMERGINGTOOLSFORTHEMANAGEMENTOFPLANTDISEASES:AGROECOLOGYANDDISEASEMANAGEMENT– partIIKeynotelecture10RNA-basedcontroloffungaldiseases:applicationandmechanismsKarl-HeinzKogel,JafargholiImani,AlineKoch

Justus Liebig University Giessen, Department of Phytopathology, Heinrich-Buff-Ring 26, D-35392Giessen,Germany

RNA interference has emerged as a powerful genetic tool for scientific research. The

demonstration that agricultural pests, such as insects and nematodes, are killed by exogenouslysupplied RNA targeting their essential genes has raised the possibility that plant diseases can becontrolled by lethal RNA signals.We have shown that transgenic expressing1 (Host-Induced GeneSilencing) or spraying2 (Spray-Induced Gene Silencing) a 791 nt long double-stranded (ds)RNA(termedCYP3-dsRNA)thattargetsthethreeessentialfungalergosterolbiosynthesisgenes(CYP51A,CYP51B, CYP51C), efficiently inhibited the necrotrophic fungus Fusarium graminearum on barley.Strong inhibition of fungal growth required an operational fungal RNA interferencemechanism asdemonstrated by the fact that a Fusarium DICER-LIKE-1 mutant was insensitive to CYP3-dsRNA.FurtheranalysisshowedthatCYP3-dsRNAwassystemicallytranslocatedintheplantviathephloemandefficientlytakenupbythefungusindistal,non-sprayedleafareas.Consistentwiththisfinding,itwas recently shown that spray applications of RNA to plants, including Arabidopsis, tomato andtobacco, israthereffective incontrollingplantdiseasescausedbymicrobialpathogens.3Moreover,optimizationofRNAdeliverybycertainnanostructure-basedformulationshasbeenshown.4I will discuss recent developments in the use of RNA as a new chemical for the control of plantdiseases.

1. Kochetal.(2013)PNAS110:19324-193292. Kochetal.(2016)PLOSPath.DOI:10.1371/journal.ppat.1005901:1be8a6b457c73d5c3. Wangetal.(2016)NaturePlants,doi:10.1038/nplants.2016.154. Mitteretal.(2016)DOI:10.1038/nplants.2016.207

Keywords:Fusarium,smallRNA,DICER,HIGS,SIGS

Session6Oral52

TowardsagroecologicalmanagementofriceblastdiseaseinMadagascarHighlandsSesterM.1,RamanantsoanirinaA.2,RaboinLM1,DusserreJ.1,RavelosonH.2,TharreauD.3

1:CIRAD.UPRAIDA.TAB-115/02.34398Montpelliercedex5.France2:FOFIFA.SRRAntsirabe.BP230.Antsirabe110.Madagascar3:CIRAD.UMRBGPI.TAA-54/K.34398Montpelliercedex5.FranceRiceisthestaplecropandfoodinMadagascar.Sincethe1980s,ajointbreedingprogramconductedbyCIRADandFofifa,hasledtothecreationofnewvarietiesadaptedtothehighaltitudeandatthesametimetorainfedcroppingenvironment(Raboinetal.,2014).Butthefirstreleasedvarietiesrapidlybecamesusceptibletoriceblast,afungaldiseasecausedbyMagnaportheoryzae,andhadtobeabandoned.Inthe2000and2010s,newvarietiestoleranttoblastwereproposedtofarmers.Uplandricecroppingsystemhasbecomeanimportantcomplementtoirrigatedrice,andawaytoimproveself-sufficiencyinriceandfoodsecurity.Riceblast,afungaldiseasecausedbyMagnaportheoryzae,isamajorconstraintforriceparticularlyinuplandcroppingconditions.IndevelopingcountrieslikeinMadagascar,nochemicalsolutioncanbeconsidered,sothereisacrucialimportancetofindoutagroecologicalsolutionstolimittheriskofepidemics.TheinteractionsbetweenuplandriceandblastepidemicshavethenbeenstudiedinamultidisciplinaryteaminthehighlandsofMadagascarformorethan10years.Conservationagriculturecroppingsystemswerestudied(Sesteretal.,2014,Dusserreetal.,inpress)andshowedaninterestingimpactonricesusceptibilitytoblast.Complementaryfieldexperimentsshowedthatricesusceptibilitywasaffectedbysoiloriginandcropdensityandthatinfestedcropresiduesappearedassourcesofprimaryinoculum(Ravelosonetal.,2013).Experimentsoncultivarmixturesshowedpromisingresultstocontrolblastepidemics(Raboinetal.,2012).Thesedataweremeasuredinfieldexperimentsandintegratedtogetherinasimulationmodelatthelandscapescale(Sesteretal.,2016).

Keywords:blast,Rice,AgronomicManagement,Foodsecurity

Session6Oral53:canceledandconvertedintoPoster168MosaicsofplantdiseaseresistancegenesareamoreversatilemeansofachievingdiseasecontrolthanpyramidsinmostagriculturallandscapesRamsesDjidjou-Demasse1,BenoîtMoury2andFrédéricFabre11IRDINRA,UMR1065,F-33882Villenaved'Ornon,France2INRA,UR407,PathologieVégétale,F-84140Montfavet,France

Thebreakdownofplantvirusresistancegenesisamajorissueinagriculture.Weinvestigated

whether a set of resistance genes would last longer when stacked into a single plant cultivar(pyramiding)orwhendeployedindividuallyinregionalmosaics(mosaicstrategy).

Wemodeledthegeneticandepidemiologicalprocessesshapingthedemo-geneticdynamicsof viruses under a multi-locus gene-for-gene system, from the plant to landscape scales. Thelandscape consisted ofmany fields, was subject to seasonality, and of a reservoir hosting virusesyear-round.

Strategy performance depended principally on the fitness costs of adaptive mutations,epidemic intensitybeforeresistancedeploymentandlandscapeconnectivity.Mosaicswereat leastas good as pyramiding strategies in most production situations tested. Pyramiding strategiesperformed better only with slowly changing virus reservoir dynamics. Mosaics are more versatilethanpyramidingstrategies,andwefoundthatdeployingamosaicofthreetofiveresistancegenesgenerallyprovidedeffectivediseasecontrol,unlesstheepidemicsweredrivenmostlybywithin-fieldinfections.

We considered the epidemiological and evolutionary mechanisms underlying the greaterversatilityofmosaicsinourcasestudy,withaviewtoprovidingbreedersandgrowerswithguidanceastothemostappropriatedeploymentstrategy.

Keywords: Durable disease resistance, Pyramids of plant resistance, Mosaic of plant resistance,Landscapeepidemiology,Regionaldeploymentstrategy

Session6Oral54RiskassessmentandmanagementofBananastreakvirusesinGuadeloupeGuillaumeFort1,KaïssaPlaisirPineau1,ChantalGuiougou1,GersendePressat1,FrédéricLambert1,MarieUmber2,CécileDubois3,FrédéricSalmon1,Pierre-YvesTeycheney1

1CIRAD,UMRAGAP,F-97130CapesterreBelle-Eau,Guadeloupe,France2INRA,URASTRO,DomainedeDuclos,F-97170Petit-Bourg,Guadeloupe,France3CIRAD,UMRAGAP,F-34380Montpellier,Franceteycheney@cirad.fr

SeveralspeciesofBananastreakvirus(BSV)occurinbanana.Theycauseawiderangeofsymptomson leaves, pseudostem and fruits, although the impact of infections on yield and fruit quality hasneverbeenproperlyassessed.BSVsarenaturallytransmittedbymealybugs.However,spontaneousinfections occur in interspecific varieties such as plantains, following stress-induced activation ofinfectiousendogenousBSVsequences(eBSVs)integratedinB(Musabalbisiana)genomes.

ThekineticsofactivationofinfectiouseBSVswasmonitoredinGuadeloupeinanexperimentalplot,using a random block design. It showed that infectious eBSVs display differential activationpotentials in plantain varieties French Clair and Pelipita, pointing to a role of plant geneticbackground in the activation process. It also showed that the multiplication mode of plantingmaterial influences activation levels monitored under field conditions and that infection had nosignificantimpactonplantgrowthandfruitproductionofbothvarieties.A wide range prevalence study of BSVs undertaken throughout Guadeloupe‘s plantations, Creolegardens,abandonedfieldsandwildareasamongvarieitesrepresentativeofthemaindessertbananaandplantain types grown inGuadeloupe showed that overall BSVprevalencewere low in dessertbanana and cooking banana. Comparedwith a similar survey carried out in 2006, prevalencewasverysimilarfordessertbananabutsignificantlylowerforplantains,whichcarryeBSVsthatinterferewithmoleculardiagnosticandcausefrequentfalsepositives.ItislikelythattherecentoptimizationofBSVmoleculardiagnosticincreasedtheaccuracyofdetection.Overall,theseresultssuggestthatBSVshavealowprevalenceandunmeasurableimpactondessertbananaandplantain inGuadeloupe,owing to lowvector-borne transmissionand lowactivationofinfectious eBSVs. These results also lead to recommendation regarding the management of BSVsthroughsafemultiplicationmodesofplantainplantingmaterial.Keywords:Bananastreakviruses;endogenousviralelements;activation;prevalence;riskassessment

Session6Oral55INATREQ™ACTIVE–anewfungicideforcontrolofSeptoriatriticiblotch(Zymoseptoriatritici)LiseNistrupJørgensen1,ClaudeMaumene2&AndrewLeader3

1 Department of Agroecology, Aarhus University, Flakkebjerg, 4200 Slagelse, Denmark; 2Arvalis Institut du Végétal,Station Expérimentale, 91720 Boigneville, FRANCE, 3 Dow AgroSciences,CPC2CapitalPark,Cambridge,UnitedKingdom.

Septoriatriticiblotch(STB)(Zymoseptoriatritici)isaseriousandverycommonpathogeninwheatcrops.Inoculumispresentinallintensivewheatproducingcountriesandthelevelofseveritydependsonhumidityandraineventsduringthegrowingseason.Between1and4fungicidetreatmentsarecommonlytargetingcontrolofSTBperseasoninWesternEuropedepending on the actual risk. In recent years the control of STB is challenged followingresistance development to both strobilurins and triazoles. More recently isolates withreduced sensitivity to SDHI’s in UK and Ireland have also been detected. Of additionalconcern in the coming years is the possibility that somemolecules that are widely usedtodaywillbeunderregulatorypressureandmayexitthemarket.

DuetothehighincidenceoffungicideresistanceinZ.tritici,itisseenasverypositive,thatanewfungicide– Inatreq™Active(fenpicoxamid)withauniquebiochemicalmodeofactionatanewtarget site in this segmenthasbeendiscoveredbyDowAgroSciencesandshouldbeavailabletocerealgrowersin2019/2020.Thenewactiveisderivedfromanaturalcompoundproduced by fermentation of anActinomycete (Streptomyces spp.)which thenundergoesaminoralterationtostabilizetheproduct. Inatreqshowsnocross-resistancetoexistingcerealfungicides,includingtriazoles,strobilurinsandSDHIs.However,astheactivein Inatreq is a target site inhibitor, the product should only be used in combinationwithotheractivestominimizetheriskofresistancedevelopment.

Inatreqhas been tested in early development trials inDenmark and France. Thesetrials have consistently confirmed excellent control of STB and yield benefits under bothpreventive and curative conditions when applied as a T1 or T2 spray. Inatreq’s strongresidualeffectandcurativeactivityonSTBhavebeenshowntoofferflexibilityindoseandapplicationtiming.InDenmark,strategieswithInatreqhavegivensignificantlybettercontrolofSTBaswellassignificantlybetteryieldscomparedtoallexistingstrategies. Keywords: disease management, resistance management, wheat, Septoria tritici blotch,Inatreq

Session6Oral56

Managementofhairyrootdisease(crazyroots)intomatoStefanVanKerckhove1,AnneleenPaeleman1,ChristineVos1,LienBosmans2,HansRediers2,BartLievens2,BartVanCalenberge3,LieveWittemans3,WendyVanlommel4,RobMoerkens4

1ScientiaTerraevzw,B-2860Sint-Katelijne-Waver,Belgium2Laboratory for Process Microbial Ecology and Bioinspirational Management (PME&BIM),DepartmentofMicrobialandMolecularSystems(M2S),KULeuven,CampusDeNayer,B-2860Sint-Katelijne-Waver,Belgium3ResearchStationforVegetableProductionvzw,B-2860Sint-Katelijne-Waver,Belgium4ResearchCentreHoogstratenvzw,B-2328Meerle,Belgium

The hairy root disease, often termed ‘crazy roots’, has in recent years become a serious

problem in greenhousehydroponic cultures of tomato, eggplant and cucumber inmany Europeancountries. The typical symptoms include excessive development of roots, which leads to morepronouncedvegetativegrowthof theplantsandsignificant lossesofmarketableyield.Theprojectconsortiumworkingonthecrazyrootstopicaimstofindintegratedmanagementsolutionstotacklethisproblem.Anoverviewoftheirresultssofarwillbepresented.

In a first step, isolatesof rhizogenicAgrobacteriumwere collected fromdifferent growers,establishing a collection of over 100 isolates. The isolates were subsequently genotypically andphenotypically characterized, which indicated the highly complex nature of this pathogen.Subsequently,theeffectofvariousculturalmethodswastested,suchastheuseofdisinfectantsandchoice of substrate. In addition it was observed that the persistent nature of the Agrobacteriumstrainsisinmanycasesduetotheirabilitytoformbiofilmsintheirrigationsystem,addinganotherlayer of complexity to their control. Finally, a screening was also performed to identify possiblebiologicalcontrolorganismsagainstrhizogenicAgrobacterium.Theknowledgedevelopedwithintheconsortiumhasbeenappliedintopracticeatdifferentgrowerswithpromisingresults.

Keywords:rhizogenicAgrobacterium,crazyroots,tomato

SESSION1.FROMPLANT-MICROBEINTERACTIONSTOINTERACTIONSWITHINPHYTOBIOMESSession1Poster1BestPass:boostingplant-endophytestability,compatibilityandperformanceacrossscalesDavidB.Collinge1andtheBestPassconsortium1DepartmentofPlantandEnvironmentalSciencesandCopenhagenPlantScienceCentre,UniversityofCopenhagen,Thorvaldsensvej40,1871FrederiksbergC,DenmarkFaced with multiple challenges, global agriculture must adopt more dynamic, efficient andsustainable production methods to increase food and fodder production to feed a growingpopulationwithfewerresources(FAO).TheBestPassproject is aMarieCurie “Early trainingnetwork”with15PhD studentshostedby12beneficiariesa plus 7 partner organisationsb. The 15 PhD projects concern endophytes in plants.Model plants include tomato, Salicornia and grasses (Festuca and Lolium) and both bacterial andfungalendophytes includingbothnewlydiscoveredandmodelorganisms.Theprojects range fromfundamental studies of the biology of endophyte-host interactions to understanding the factorsunderlyingthedevelopmentofstablemicrobialproductsthatcanboostplantperformance.a Beneficiaries:ABiTEP,ACIB,Austrian Instituteof Technology,AarhusUniv, BOKU,CSIC,DLF, IGZ,INOQ,NCU,TUGraz,UnivAmsterdamaswellasUCPHbPartnerOrganisations:Universities:Cologne,Massey,MichiganStateandWageningen,aswellasSMEs:Biotenzz,Biofungitek,RoombioticTheprojecthasreceivedfundingfromtheEuropeanUnion’sHorizon2020researchandinnovationprogrammeundertheMarieSklodowska-CuriegrantagreementNo676480.

Session1Poster2MicrobiomediversityincropdebrisandpotentialinteractionswiththefungalpathogensZymoseptoriatriticiandLeptosphaeriamaculansLydieKerdraon1*,Marie-HélèneBalesdent1,MatthieuBarret2,ValérieLaval1,FrédéricSuffert1

1UMRBIOGER,INRA,AgroParisTech,UniversitéParis-Saclay,78850Thiverval-Grignon,France2UMRIRHS,INRA,AgroCampusOuest,Universitéd'Angers,49071Beaucouzé,FranceDespitetherecognizedcontributionofcropdebrisasinoculumsourceofmanyplantpathogens,littleisknownaboutthestructureandfunctionofmicrobialentitiesassociatedtothisplanthabitat,andtheirinteractionswithresidue-borneplantpathogens.Theobjectiveof thiswork is to characterizebacterial and fungal assemblagesassociated towheatandoilseedrapedebrisandtoassesstheirrelative impactsonthebuild-upofprimary inoculumofZymoseptoria tritici (septoria leaf blotch of wheat) and Leptosphaeria maculans (stem canker ofoilseedrape)duringtheinter-croppingperiod.Thistwo-yearsstudyisbasedonafielddesigncomposedofthreeplots(wheatmonoculture,oilseedrape-wheat rotations)established in theexperimentaldomainofGrignon (Parisbasin).Wheatandoilseedrapecropdebrisweresampledfourtimesayearduringtheascosporereleaseperiod(fromOctobertoMay).Thestructureofdebris-associatedmicrobialassemblageswasassessedthroughacombinationofclassicalmicrobiologicalapproachesandMiSeqsequencingoftwomolecularmarkers(16SrRNAgeneandITS1regionofthefungalinternaltranscribedspacer).Communityprofilingapproachwasperformedonapproximately100samplescollectedonthe firstyear of the experiment. After suppressing rare OTUs (Operational Taxonomic Units), around 110bacterialOTUsand70fungalOTUswereidentifiedpersample.Accordingtobetadiversityanalysis,thecrop(wheat,oilseedrape)wasthemain,significantfactorstructuringdebris-associatedmicrobialassemblages.Thesamplingdatewasalsoshapingamicrobialcommunitycomposition.These first results,whichwill be combined to thoseof the second yearof sampling, reinforceourview on the community composition, and give an opportunity to identify microbial speciespotentiallyactingonprimaryinoculumofZ.triticiandL.maculans.Keywords: Zymoseptoria tritici, Leptosphaeria maculans, microbial diversity, plant debris, primaryinoculum,metabarcoding

Session1Poster3ScreeningofwheatendophytesasbiologicalcontrolagentsagainstFusariumheadblightMorganeComby1,2,3,MarieGacoin2,MathildeRobineau2,FanjaRabenoelina3,SebastienPtas2,JoëlleDupont1,CamilleProfizi2,FabienneBaillieul3

1InstitutdeSystématique,EvolutionetBiodiversité,ISYEB–UMR7205–CNRS,MNHN,UPMC,EPHE,MuseumNationald’HistoireNaturelle,SorbonneUniversités,57rueCuvier,CP39,75005Paris,France2SouffletBiotechnologies,QuaiSarrail,10400NogentsurSeine,France3LaboratoiredeStress,DéfensesetReproductiondesPlantes–EA4707,FacultédesSciences,MoulindelaHousse,BP1039,51687Reims,FranceFusarium head blight (FHB), caused by phytopathogenic fungi such asFusarium graminearum andFusariumculmorum, isoneof themost importantdiseasesonwheatcropsworldwide.Theaimofthepresent studywas to find endophytes ofwheat candidates as biological control agents (BCAs)againstFHB,usingtwodifferent invitro tests.Thecommon invitro testbydualcultureassayswasusedtoconductalarge-scalescreeningof86strains,isolatedfromwheatplants,towardsFusariumspp.Inaddition,aninvitroscreeningtestondetachedwheatspikelets,easytohandlebutclosertoreallife,hasbeendevelopedonasubselectionof22strains.Bothinvitrotestsidentified13strainspromisingforthecontrolofFHB,ofwhich10belongedtothreespeciesnotreportedbeforefortheirantagonistic capacities against Fusarium spp. However, the efficacy of some strains turned outdifferent between both in vitro tests, raising the importance of finding the most appropriatescreening approach for the search of BCAs. This study pointed out the interest of the test ondetached wheat spikelets that provided information about a potential pathogenicity, the growthcapacityandefficacyoftheendophytestrainsonthetargetedplant,atearlyscreeningstep.Keywords:Fusarium,biocontrol,endophytes,screening,real-timePCR

Session1Poster4Fungal endophyte diversity in chestnut galls and surrounding tissues in plotswith different treecompositionPilarFernandez-Conradi1,ThomasFort1,XavierCapdevielle1,BastienCastagneyrol1,HervéJactel1andCécileRobin1

1Biogeco,INRA,Univ.Bordeaux,F-33610,CestasFungalendophytesarepotentialbio-controlagentsforpestinsects.Yet,theirdiversityandfunctionsneedfurtherunderstanding.Weexploredfungalendophytediversityingallsinducedbyaninvasiveinsect,Dryocosmus kuriphilus, and in surrounding chestnut leaf tissues, sampled inmature forestplotswhere chestnutswhere growing aloneormixedwith pines, oaks or ashes.Wehypothesizedthatendophytescommunitiesingallsdifferswithinplotcompositionandthatgallstissuesshelteramorediverseandrichendophytecommunitythanleaftissues.Weselected28chestnutplotsconsistingin10chestnutmonoculturesand18two-speciesmixtures.Ineachsite,wesampledgallsandleaftissues(3samplespertreeand3treespersiteofeachtype).Wedisinfected leafandgallsamplesandextractedDNA.FungalendophyteswerecharacterizedbyIlluminasequencingoftheInternalTranscribedSpacer1(ITS1)region.Wefoundatotalof1,378differentOTUs inoursamples.ThemostcommonOTUcorrespondedtoGnomoniopsis sp. which may be associated to D. kuriphilus gall necrosis. Endophyte communityrichness and diversity on chestnut leaves and galls were independent of plot composition.Endophytesrichnessingalltissueswasreducedcomparedtosurroundingleaftissues.These results suggest that plot composition do not impact endophyte communities from chestnutgallsandthat,interestingly,galls,commonlythoughtasasourceofnutrimentwhichcandirectlyorindirectlyattractfungalendophytesandactasafilterforfungalendophytesfromchestnutleaves.Abetter understanding of their functioning is important to improve biocontrol agents for gallinginsects.Keywords:endophytes,forestdiversity,plant-fungus-insectinteractions,chestnut

SESSION2.SPACE-TIMEANDMULTI-SCALESAPPROACHES:DIAGNOSTIC,EPIDEMIOLOGYANDECOLOGYINTHEFIELDSession2Poster5AvrLm7 and AvrLm3 frequency evolution in French Leptosphaeriamaculans populations: a 20year’ssurveyMarie-HélèneBalesdent1,ClémencePlissonneau1,LaurentCoudard1,LoïcLeMeur2,MartineLeflon3,XavierPinochet3,MagaliErmel4,RégineDelourme4,ThierryRouxel11INRA,UMRINRA-AgroParisTech1290BIOGER,78850Thiverval-Grignon,France2Agrosolutions(groupeInVivo),83avenuedelaGrandeArmée,75782ParisCedex16,France3TerresInovia,78850Thiverval-Grignon,France4INRA,UMR1349IGEPP,DomainedelaMotte-BP35327,35653LeRheuCedexPhomastemcanker,causedbythefungusLeptosphaeriamaculans, ismainlymanagedthroughthedeploymentofresistantvarieties.Specificresistancegenes(Rlm)arepresentincommercialvarietiesandtheireffectiveness isafunctionofthefrequencyofthecorrespondingavirulenceallele infieldpopulationsof thepathogen.ThedurabilityofagivenRlmgenemayvary,dependingontheplantgenetic background (Brun et al. 2009), the fitness cost linked to the loss of the avirulence gene(Huangetal.2010)andagronomicpractices(Daverdinetal.2012).AftertheveryrapidbreakdownofRlm1inthe90’sinFrance,itwasquestionedwhetherallreleasedRlmgenescouldbeovercameatthesamespeed.Rlm7 was introduced in commercial hybrids in France when most of the isolates possessed theavirulentalleleAvrLm7(Balesdentetal.2006).ThefrequencyofvirulentisolateswasmonitoredinL.maculanspopulationsofatanationalscale(8to20sitesperyear)from2000to2015.While only one virulent isolate towardRlm7 (“avrLm7”)was found in 2000, theirmean frequencyreached4%in2010,20%in2013and41%in2015.Regionalvariationswereobserved.Forinstancein 2013, avrLm7 frequency varied from 0% (Britany) to 45% (Center region). Due to the negativeinteractionbetweenAvrLm3andAvrLm7 (Plissonneauetal.2016),nearlyallavrLm7 isolateswereavirulenttowardRlm3.Only1.8%ofthecurrentFrenchL.maculanspopulationcaninfectbothRlm3andRlm7varieties.ComparedtotheRlm1breakdownthathappened inonly3growingseasons inFrance,10yearsofintenseuseofRlm7wereneededtoreachameanfrequencyofvirulentisolatesofonly20%atthenationallevel.ThissurveyalsoconfirmsthenegativeinteractionbetweenAvrLm3andAvrLm7,whichoffersgreatperspectivesfordurablemanagementofRlmgenesinoilseedrape.Keywords: Leptosphaeriamaculans, resistance breakdown,Rlm7,Rlm3, avirulence genes,Brassicanapus.

Session2Poster6Quantitative analysis of distributionofFusariumgraminearum andMicrodochium spp. inwinterwheatTatianaGagkaeva1,XeniaCheredova1,AlexandraOrina1andOlgaGavrilova11All-RussianInstituteofPlantProtection(VIZR),St.-Petersburg–Pushkin,Podbelskogosh.,3,196608,RussiaBecausesimilarsymptomsofplantdiseasecanbeproducedbyFusariumandMicrodochiumcausalagents, the use of symptoms alone is an inadequate method for disease identification in fields.Extensivecolonizationofleaftissueandheadsofwinterwheatwasoccurredin2016inSouthregionofRussia.Thenecrotic lesionsofflag leafandpinkcoloredsporulationoffungionheadhavebeenobservedinmanyfields.The quantitative PCRwas used for analysis of fungal DNA content in different tissue (head, grain,peduncle, flag leaf, and stem) of wheat. Deoxynivalenol (DON) amounts in plant tissues wereanalyzedbyELISA.F.graminearumwaspresentedinallpartsofthewheatplantsoftheZadoksstage85(thecontentoffungalDNAvariedinlimits1.4-74.0×10-7ng/ngoftotalDNA).ThelargeamountsofF.graminearumDNAcontentswerefoundinheads.Asignificantpositivecorrelationbetweentheamountsof fungalDNAandDON inplant tissuewasfound (r=+0.96,p<0.001).The stemsandpeduncleofall analyzedwheat cultivarshad lowerDONcontentincomparewiththeotherplanttissues.IntheallanalyzedplanttissuestheDNAofbothM.nivaleandM.majushavebeenfound.Totally,contentofM.majusDNAwassignificantlyhigher(from8.5×10-4to1.2×10-1ng/ngoftotalDNA),thanM. nivale (from 8.1×10-5 to 3.6×10-3 ng/ng of total DNA) in all parts of vegetative plants. The flagleaves contained the maximum of DNA of both Microdochium species. A significant positivecorrelationbetweenthecontentsofDNAofM.nivaleandM.majuswas found (r=+0.57,p<0.05).Thespeciesdiversity,resistanceofwheatcultivarsandstepstominimizefungalinfectioninfieldarediscussed.TheinvestigationwassupportedbytheRussianScienceFoundation(No.14-26-00067).Keywords:fungi,Fusarium,Microdochium,winterwheat,organs,real-timePCR

Session2Poster7ModelingimpactsoffertilizationonepidemicscausedbyahemibiotrophicfungalpathogenChristopheGigot1,2,DavidClaessen2,Pierre-AntoinePrécigout1,2,CorinneRobert1,21UMRÉcoSys,INRA-AgroParisTech,Routedelaferme,78850Thiverval-Grignon,France2Institut de Biologie de l'École Normale Supérieure, CNRS-ENS-INSERM UMR8197, École NormaleSupérieure,46rued'Ulm,75005Paris,FranceA modeling framework focused on life history of foliar plant pathogens is being developed tosimulatewithinandbetweenseasoninteractionsbetweencropgrowthanddiseasedevelopment.Itisaresource-basedmodelrelyingonaphysiologicallystructuredpopulationapproach.Thesimulatedpathosystem is, in particular, able to respond to both plant architecture and fertilization. Theimplementationtakesplaceatthetwo-complementarypatchandcanopyspatiallevels.Apatchisasmall infectableunitof leaf tissue (about the sizeofamature infectionor “lesion”).Productionofnew patches over time is a function of the available resource produced by the other patchesdepending of parameters such as photosynthetical activity and disease intensity. The pathogenpropagulescancontaminatenearbyhealthypatches.Thedefinitionofpatchproximitydependsonthe dispersal ability of the pathogen which is determined by parameters such as dispersal mode(splashorwind)andcanopydensity.This framework isused to studyspatiotemporal changesofapathosystem with a hemibiotrophic fungal pathogen, namely Zymoseptoria tritici which causesSeptoriatriticiblotchonwheat.Thisdiseaseischaracterizedbyacomplexinfectioncycleconsistingof a long symptomless incubation period after the infection, followed by chlorosis and necrosissymptoms before fungal sporulation. A sensitivity analysis over fertilization parameters (i.e. patchresource carrying capacity, patch green lifespan, and maximum patch creation rate) is beingperformed in order to quantify the effects of crop resource level on plant physiology and diseasedynamics. Such a resource-based modeling framework using an ecological approach studying thestructure, function and dynamics of a system to address epidemiological questions is currentlymissinginthephytopathologicalcommunity.Thiskindoftoolsmaybeveryvaluabletodesignmoreresilientagroecosystems,predictingpathogendynamicsunderdifferentfertilizationconditions.Keywords: Septoria tritici blotch, fertilization, epidemiology, life history theory, resource-basedmodel,structuredpopulationmodel

Session2Poster8BiomarkersforRamularialeafspotdiseaseinbarleyManojKamble1,MikkelThygesen2,AndreaLorentzen3,PiaJensen3,PeterKristensen4,ReneLemcke5,Elisabet Sjokvist6, Jean-Baptiste Lopez1, Neil Havis6, Peter Roepstorff3, Michael Foged Lyngkjær5,SimonaRadutoiu1

1DepartmentofMolecularBiologyandGenetics,AarhusUniversity,Denmark2DepartmentofChemistry,CopenhagenUniversity,Denmark.3DepartmentofBiochemistryandMolecularBiology,SouthernUniversityofDenmark,Denmark.4DepartmentofEngineering,AarhusUniversity,Denmark.5DepartmentofPlantandEnvironmentalSciences,CopenhagenUniversity,Denmark.6DepartmentofPlantProtection,Scotland’sRuralCollege,Scotland.Ramulariacollocygni (Rcc),ahemibiotrophandcloserelativeofZ.tritici isapathogenofbarley inEurope and South America. It causes the disease, Ramularia leaf spot (RLS), and involves thesecretionofaphotoactive toxin, rubellin, in theplantapoplast leading tonecrotic leaf lesionsandeventuallyup to35%yield losses.R.collo-cygni,a seed-born fungusgrowsasanendophyte in theearlydevelopmentalstages, thereforenocompleteresistance isavailable inbarleygermplasmandfungicides are proven to be inefficient due to the symptomless infection. Molecular mechanismsunderlyingthediseaseestablishmentandtheswitchbetweenendophyticandpathogenicstatusarecurrentlyunknown.My PhD project focuses on the identification of key gene-protein families and their function inbarley-Ramularia collo-cygni interplay using “omics” approaches. I am deploying the promisingcandidates for disease tolerance to develop biomarkers based on promoter/gene sequences.Alternatively, I am using rubellin toxin to design an antibody to develop a diagnostic approach todetectthefungusinthebarleycultivarsatdifferentdevelopmentalstages.Transcriptomicsandproteomicsanalysesoftwocontrasting(tolerantvssusceptible)barleycultivarswereperformed to identify the active genes/proteins andpathways duringdifferent stages of thedisease establishment. Preliminary results from the transcriptome analyses allowedus to pinpointspecific gene families, biological and molecular functions (receptors, transporters, transcriptionfactors)aredifferentially regulatedbetweencultivars, suggesting theirmajor role inRLSdisease inbarley. Candidates identified and confirmed from omics data analyses will be used as potentialcandidates for genetic marker development in the plant breeding programs. We also found thatdifferent barley cultivars respond differently to rubellin treatment indicating the involvement ofcultivarspecificbarleygene(s).Togetheralltheseresultswithwillbepresentedattheconference.Keywords:Ramulariacollo-cygni,Ramularialeafspotdisease,rubellin,omics,biomarker.

Session2Poster9MolecularvariabiltyandgeneticstructureofIYMVinBurkinaFasoM. Koala1,2, D. Sérémé1,2, F. Vignols3, E. Lacombe3, M. Bantgratz2,3, B.J. Neya1,2, C. Brugidou2,3 N.Barro4,andO.Traoré1,2Laboratoire de Virologie et de Biotechnologie Végétale (LVBV), Burkina Faso, Institut del’EnvironnementetdeRecherchesAgricoles(INERA)01BP476Ouagadougou01,BurkinaFaso.Laboratoire Mixte Internationale (LMI) Patho-Bios, Institut de l’Environnement et de RecherchesAgricoles(INERA)01BP476Ouagadougou01,BurkinaFasoUMR IPME (UM, IRD, CIRAD), Institut de Recherche pour le Développement, IRD, 64501, 34394Montpelliercedex5,FranceLaboratoired’EpidémiologieetdeSurveillancedesbactériesetvirusTransmissiblepar lesAlimentsetl’eau.LabESTA/UFR/SVT,UniversitéOuagaIProfesseurJosephKi-Zerbo,01BP7023Ouagadougou01,BurkinaFasoImperatayellowmottlevirus(IYMV,Sobemovirus)wasfirstdescribedin2008inthesouth-westernregionofBurkinaFaso(WestAfrica).ThegeneticdiversityofIYMVwasnotdocumenteduptoday.Inthisstudy,thevariabilityofCPofIYMVwasevaluatedthroughthemolecularcharacterizationof39isolatescollectedinthewesternpartofBurkinaFaso.ComparisonofsequencesofthesenewisolatesandoneIYMVsequenceavailableinGenBankrevealedthattheaveragenucleotidediversitywaslowforaplantvirus.Theratioofnonsynonymousoversynonymousnucleotidesubstitutionspersitewaslow, indicating a CP diversification under strong purifying selection. In spite of the low nucleotidediversity,phylogeneticanalysisrevealedsegregationofIYMVisolatesintosixmajorclade.Studyofspatial distributon and genetic structure revealed no correlation between genetic variation andgeographicaloriginofIYMVisolates.ThisisthefirststudyofthegeneticdiversityofIYMV.Keywords: Imperata yellow mottle virus (IYMV)-Coat protein(CP)-genetic variability-Geneticdifferentiation

Session2Poster10CoexistenceofLeptosphaeriamaculansandL.biglobosaonoilseedrapecropsGeorgiaK.Mitrousia1,MarkWinter2,3,MalgorzataJedryczka4,LakshmiH.Gajula1,JoannaKaczmarek4,SitiN.M.Sidique1,YongjuHuang1,BruceD.L.Fitt1

1SchoolofLifeandMedicalSciences,UniversityofHertfordshire,Hatfield,AL109AB,UK2Plant Pathology and Crop Protection Division, Department of Crop Sciences, Georg-August-UniversityGöttingen,37077,Germany3DepartmentofPlantPathology,UniversityofMinnesota,StPaul,MN55108,USA4InstituteofPlantGenetics,PolishAcademyofSciences,Strzeszynska34,60-479Poznań,PolandLeptosphaeriamaculansandL.biglobosaareassociatedwithphomastemcankerofoilseedrape.Inmanycountries,L.maculansisconsideredtobemoredamagingthanL.biglobosa,whichisgenerallyassociatedwithupperstemlesions.However,L.biglobosaisanimportantpathogeninPolandandinChina the disease is associated only with L. biglobosa. This work studied the coexistence ofLeptosphaeriaspp.onwinteroilseedrapecropsoverthreetofivecroppingseasonsinGermany,theUK and Poland. The relative contribution of the two Leptosphaeria spp. to phoma leaf spot andphoma stem canker severity was examined on cultivars differing in their resistance against L.maculans, includingcultivarswiththeRlm7resistancegene.TherewasextensivecolonisationbyL.biglobosa on cultivars with the Rlm7 gene. Effective control of L. maculans, by using cultivarsresistant against this pathogen, may increase the possibility of severe epidemics caused by L.biglobosainthefuture.CombineddatafortheabundanceofthetwoLeptosphaeriaspp.inair-borneinoculum and their contribution to disease severity showed that L. biglobosa has an increasinglyimportantroleindevelopmentofdiseaseepidemics.Keywords:phomastemcanker,Leptosphaeriamaculans,Leptosphaeriabiglobosa,oilseedrape

Session2Poster11Deathoftheassumptionthat‘latentperiod’isfixedoverthecourseofaplantdiseaseepidemicFrédéricSuffert11UMRBIOGER,INRA,AgroParisTech,UniversitéParis-Saclay,78850Thiverval-Grignon,France

Thelatentperiodisdefinedasthetimebetweeninfectionandtheonsetofsporulationdueto that infection. It is a crucial life history trait, particularly for polycyclic diseases, because itdetermineshowmanycompleteinfectiouscyclescanoccurduringanepidemicanditsfinalintensity.Manystudieshavefocusedonthevariationoflatentperiodwithpathogenorhostgenotype,oritsplasticity inresponsetoenvironmentalfactors.Thefocusontheseaspectsisunsurprising,asthesefactors form the apices of the epidemiological triangle (host, pathogen, and environment).Experimentsincontrolledconditionsaregenerallyusedtoassesspathogenpathogenicity(virulenceandaggressiveness)andhostsusceptibility.Onceestimatedforoneorseveralpairsofhost-pathogengenotypes,thevalueoftheparameter‘latentperiod’isimplicitlyconsideredtobefixedandisusedin epidemiological models. Paradoxically, most epidemiological studies do not consider the latentperiodofapathogenpopulationtobeintrinsicallyvariable.Mythesishereisthatthelatentperioddisplaysnon-negligiblevariationoverthecourseofadiseaseepidemic,andthatthisvariabilityhasmultiplesources,someofwhichhaveantagonisticimpacts.Idevelopargumentsforfoursourcesofvariability challenging the implicit assumption that the latent period remains constant: dailyfluctuations in leaf temperature, thenatureof the inoculum,hoststageor theageofhost tissues,intra-population competition and selection for aggressiveness traits. I focus on experiences withSeptoriatriticiblotch(Zymoseptoriatritici),makinguseofempiricaldatasetscollectedduringvariousresearchprojects.Keywords: generation time, latent period, plant disease epidemiology, polycyclic disease, wheat,Zymoseptoriatritici

Session2Poster12Characterization of German Plasmodiophora brassicae populations and possible strategies tosuppresstheclubrootdiseaseinoilseedrapefieldsNazaninZamani-Noor11JuliusKühn-Institut,InstituteforPlantProtectioninFieldCropsandGrassland,Messeweg11-12,D-38104Braunschweig,GermanyClubrootcausedbyPlasmodiophorabrassicae,isoneofthemosteconomicallyimportantdiseasesofoilseedrape(OSR)andotherbrassicavegetablesworldwide.Diseaseoutbreakshavecausedseriousproblems in recent years inGermany. Therefore,monitoringand classificationofP.brassicae-fieldpathotypesinOSRcultivationareasisofprimeimportance.Between 2012 and 2016, 100 new clubroot-infested fields were identified in 12 German federalstates.Incidenceofclubrootvariedwithinthesefieldsfrom22%to90%.AmongallP.brassicae-fieldpopulations, fourdistinctvirulencepatterns (P1,P2,P3andP5)were identifiedaccording toSomé(1996).28isolateswereabletoovercometheresistanceofclubroot-resistantOSRcv.Mendel.TheinvestigationonsoilsamplesconfirmedthatclubrootappearsoverawiderangeofpH,from5.1to8.3, but that a significant negative correlation occurs between disease incidence and soil pH.Furthermore,morecasesofdiseaseandsevereincidenceswereobservedinsandyloamandloamysandascomparedwithothersoiltypes.In parallel field studies, resistant cultivars and soil amendmentwith calcium cyanamide andburntlime alone or in different combinations applied before or after sowing were examined for thepotentialstrategiesforsuppressingtheclubrootdisease.Soilmoisture,soiltemperatureandsoilpHattwodifferentdepths(15and30cm)weremeasuredatregularintervalsoverthegrowingseason.Clubroot disease incidence and severity were assessed visually for the development of root galls.Field results showed clear differences between the treatments. Resistant cultivars remained themore effective management strategy providing 60-80% disease control. Soil amendments gavevariable control between three field locations and years. Changing the time of application had asignificanteffectonthe final severityof thedisease. Incomparisonwithcalciumcyanamide,burntlimeapplicationhasasmallereffect.Keywords: Brassica napus, pathotype, disease severity index, soil amendment, soil pH, calciumcyanamide

Session2Poster13Phenotypic variability between some isolates of Botrytis cinerea and Botrytis pseudocinereacollectedintheregionofBejaia(NorthernAlgeria)AhmedAdjebli1andKamelAissat1

1Laboratoire d’EcologieMicrobienne, Faculté des Sciences de laNature et de la Vie,Université deBejaia,06000Bejaia,AlgérieBotrytiscinerea isanubiquitouspathogenwhichcausessevere losses inmany fruit,vegetableandornamentalcrops.Thepathogeninfectsleaves,stems,flowersandfruits.Inthepresentstudy,eightsingle-spore strains of B. cinerea isolated from tomato greenhouses located in Bejaia regions(Northern Algeria). Isolates weremolecularly characterized by ninemicrosatellitemakers. IsolateswereassignedasB.cinereaandB.pseudocinereawithfour isolatesofeachspecies.Morphologicalcharacterizationwasestablishedusingtwoculturesmedia(PDAandMEA).AllisolatesinoculatedonPDAmediumwereexclusivelySclerotial,incontrastallisolateswereMycelialonMEAmedium.Thestrainsofthetwospeciesweresimilarinaggressivenessonbothhostspeciestested(tomatoleavesand apple fruits). Moreover B. cinerea isolates were more aggressive than B. pseudocinerea onlettuceleaves.Bothplants(tomatoleavesandlettuceleaves)weresignificantlymoresusceptibletoB. cinerea andB. pseudocinerea than those of apple fruits. A significant negative correlation wasestablished between aggressiveness and morphological type. The epidemiological consequencesconcerningpopulationsofB.cinereaandB.pseudocinereaintomatogreenhousesarediscussed.Keywords:Botrytispseudocinerea,Botrytiscinerea,Morphology,Agressiveness

Session2Poster14DetectionofSclerotiniasclerotioruminoilseedrapeusingreal-timePCRMariaAlgerin1,DianaTilevik1andAnn-CharlotteWallenhammar2

1UniversityofSkövde,SchoolofBioscience,POBox408,SE-54128Skövde,Sweden2R&D,RuralEconomyandAgriculturalSociety,HSKonsultAB,POBox271,SE-70145Örebro,SwedenSclerotiniasclerotiorumisaplantpathogencausingSclerotiniastemrot,whichisamajordiseaseinoilseedrapeinSweden.Thepathogencansurviveinthesoilassclerotiaforatleasttenyears.FourfieldtrialswereperformedinÖrebroCounty,Sweden,duringtheyears2012and2014-2016inorderto study the occurrence of the pathogen and the disease. Real-time PCR (qPCR) was used toquantitativelydetectS.sclerotioruminoilseedrapeleavesaswellasinairsamples.TheleavesoftheoilseedrapewerecollectedatBCCH63andairsampleswerecollectedusingaBurkardsporetrap.Thefieldexperimentsconducted2012and2014wereperformedinspringoilseedrapeandshowedalowtohighstemrotincidenceforyear2012(maximum24%)andverylowincidencefor2014(0-2%).TheresultsfromtheqPCRshowedavariationinincidenceofS.sclerotioumDNAonthespringoilseedrapeleavesfrom10to100%.During2015and2016,thefieldexperimentswereperformedinwinter oilseed rape and stem rotwas detected in all fields. In five out of ten andnine out of tenwinter oilseed rape fields the incidence of stem rotwas equal to or above 15%, respectively. Thehighestinfectionlevel,62%,wasassessedinafieldin2016.Leavesfromwinteroilseedrapearestillbeinganalyzed.TheresultsfromthesporetrapshowedthatS.sclerotioumDNAcanbedetectedinthe air from the beginning of May until the beginning of August. The obtained qPCR data incombinationwithstemrotincidenceandclimatedataarebeingemployedinthedevelopmentofacomputationalpredictionmodelthatcouldpotentiallybeusedtoimprovediseaseriskassessment.Keywords:Sclerotiniasclerotiorum,real-timePCR,oilseedrape,sporetrap,predictionmodel

Session2Poster15The EF1α region as a target to assess Fusarium diversity on cereals using a high-throughputsequencingtechnologyRyanBasler1,Anne-LaureBoutigny2,AngéliqueGautier1,FlorianDauthieux1,StephenLeite1,RomainValade3,JaimeAguayo2,RenaudIoos2,ValérieLaval11INRA,UMR1290BIOGER_CPP,Bat13,avenueL.Bretignière,78850Thiverval-Grignon,France2ANSES Laboratoirede la santédes végétaux,UnitédeMycologie,DomainedePixérécourt,54220Malzéville,France3ArvalisInstitutduvégétal,DomaineAgro-Paris-Tech,BâtimentBiogerCPP,78850Thiverval-Grignon,FranceFusarium head blight is a major cereal disease caused by a complex of Fusarium species. Thesespeciesvaryinimportancedependingonclimaticconditions,agronomicfactorsorhostgenotype.Inaddition,most of the Fusarium species are able to produce one ormoremycotoxinswith varyingdegreesof toxicity in thegrain. Thesemycotoxins constitute an important food safety concern, astheyhavehealthimplicationstobothhumansandanimals.FusariumspeciesinvolvedinFHBdifferintheirpathogenicity,abilitytoproducemycotoxinsandfungicidesensitivity.AccurateandexhaustiveidentificationofFusariumspeciesinplantaisthereforeofgreatimportance.Inthisstudy,anewsetofprimerstargetinganinformativeregionoftheEF1αgenewasdeveloped.TheprimerswereevaluatedfortheirspecificityonapanelofDNAextractsrepresenting30Fusariumspeciesand23nonFusariumspecies.HighthroughputsequencingprotocolusingIlluminatechnologyandbioinformaticsanalysesstepswereoptimizedandvalidatedonmockcommunitiesandinfectedgrain communities. Fusarium species could be retrieved from the mock and infected graincommunities and good reproducibility could be observed according to the different parameterstested (DNA extraction, wheat cultivar, PCR cycle number, sequencing runs). One infected wheatgraincouldberetrievedin10000uninfectedgrainsusingthistechnology.Thetoolwasfurthertestedon65fieldsamplesofdurumwheat,softwheatandbarleycollectedatvarious locations inFranceandupto17Fusariumspeciescouldbedetected.This new set of primers allows evaluating diversity of theFusarium complexon cereals usinghighthroughputsequencing. ItprovidesamoreexhaustivepictureoftheFusariumcommunitythanthecurrently used techniques based on isolation or species-specific PCR detection. This newexperimentalapproachmaybeusedtoshowchangesinthecompositionoftheFusariumcomplexordetecttheemergenceofnewFusariumspeciesinresponsetovariousenvironmentalfactorswhichisofgreatconcernformanagingthediseaseandpredictingmycotoxincontaminationrisks.Keywords:Fusarium,highthroughputsequencing,metabarcoding,wheat

Session2Poster16Barleysoilbornemosaicviruses:Identificationofpredominantvirusesaffectingyieldandmaltingquality,inordertoorientatebreedingtowardsasustainableresistanceValérie Cadot1, Julie Villemot1, Thierry Candresse2, Armelle Marais2, Michel Bonnefoy3, MarcSchmitt4, Christian Leveau5, Luc Didierjean6, Dominique Romelot9, Jacques Soyer1, Louis-MarinBossuet1, Olivier Herbert1, Romain Valade3, Jean François Herbommez7, Charles Snijders7, AmélieGenty7,EdouardTatara7,PhilippeLerebour7,ClairePerrot7,NathalieLeroy7,FrankRabenstein8,DenisMauny10,LucPelce3,EdouardBaranger3,AméliePetit11,FrançoisLevêque12,MathiasSexe13,GregoryCousin14,DamienRousseaux15,RégisFournier4,MathieuRolland11GEVES,RueGeorgesMorel,49071Beaucouzé&Domainedel’Anjouère-LaPouëze–49370Erdre-en-Anjou. 2INRA UMR1332 BFP, INRA, Université de Bordeaux, 33882 Villenave d’Ornon, France.3ARVALIS–3rueJosephetMarieHackin75016PARIS.4IFBM:7rueduBois-de-la-Champelle54500Vandœuvre-lès-Nancy&RoutedeSamadet-F-64410ArzacqArraziguet.5MalteursdeFrance:66ruedelaBoétie75008Paris.6BrasseursdeFrance,9BoulevardMalesherbes,75008Paris.7UFS:17,rueduLouvre,75001Paris.8JuliusKühnInstitut:FederalResearchCentreforCultivatedPlants-Instituteof Epidemiology and Pathogen Diagnostics -Erwin-Baur-Str. 27-06484 Quedlinburg – Germany.9Axereal: 36, rue de laManufacture CS 40639 45166OLIVET cedex. 10Soufflet Agriculture: Quai duGénéral Sarrail – BP 12 -10402Nogent sur Seine.

11SeineYonne :49, routed’Auxerre89470Moneteau. 12DijonCéréales,4boulevarddeBeauregard -BP407521604Longvic. 13EMC2:BP30045–55101Verduncedex.14Lorca:RoutedeMetz,57580Lemud.15Vivescia:2rueClémentAderBP101751685ReimsCedex2.Geneticresistanceistheonlywaytocontrolmosaicdamageinwinterbarley.Atleast18resistancegenesareknown,butuntil recently, rym4was themostusedbybreeders tocontrolBarleyyellowmosaicvirus-1(BaYMV-1),predominantinFranceandBarleymildmosaicvirus(BaMMV).However,since2009,new significantdamagehasbeenobserved inmaltingbarley in awideningproductionarea,indicatingpossibleovercomingofresistance,withthedevelopmentofBaYMV-2.Theobjectivesof theMosa-hordeumprojectwere to: identifynewvirusesorpathotypes;developnewdetectiontools; confirm cultivar resistanceobserved in field usingMB tools; confirmefficiency of resistancegenes cited in literary reviews; quantify impact on yield and malting quality. A complete viralinventory was carried out 2013- 2016 on affected barley crops in 108 sites using: real-time PCR;Sangersequencing;Next-GenerationSequencing;andserologicaltests.Anewtool,basedonderivedCleaved Amplified Polymorphic Sequences (dCAPS), developed to investigate BaYMV-1 and 2distributions,demonstratedthatBaYMV-2ispredominant(>95%)indiseasedsamples.BaYMV-1andBaMMVwere also identified in co-infectionwithBaYMV-2on susceptible cultivars. Comparisonofyieldcomponents,maltingandbeerqualityobtainedonhealthy/contaminatedplotswithcouplesofcultivarsrevealedvariationinyieldlosses,upto3t/ha,reductioninnumberofspikesandkernels/m²,smallerkernels,slightincreaseinproteincontentanddecreaseofmaltextract.Phylogeneticanalysesindicated that the rym4 resistance-breaking ability of BaYMV-2 independently evolvedonmultipleoccasions. In limitednumberof trials, rym5 resistancewasovercomebya variantofBaMMV.Theimplantationofdifferentialsin21contaminatedtrialsconfirmedtheefficiencyof11resistancegenesagainsttheBaYMV-2/BaMMVcomplex.TomonitorBaYMVandBaMMVresistanceforregistrationintheNationalListreal-timePCRappearsmoreefficientthandCAPS.Thesefindingswillhelpbreedersachievesustainableresistance.Keywords:BaYMV1&2detection,resistancegenes,yield,maltingquality,barley

Session2Poster17SpatiotemporaldynamicsofinoculumsofSigatokadiseaseofbananaatplotandplantscalesYolandeChilin-Charles1,HélèneBardou1,MaddlyMontoban1,CatherineAbadie11CIRAD–Centredecoopération InternationalenRechercheAgronomiquepour leDéveloppement,UMRBGPI"BiologieetGénétiquedesInteractionsPlante-Parasite",StationdeNeufchateau,SainteMarie97130CapesterreBelle-Eau,FranceSigatokadisease(SD),causedbythefungalpathogenMycosphaerellamusicola,isaverydestructivefoliar disease of bananas spread by conidia and ascospores. A good understanding of epidemicsprogress at different spatial scales is essential to improve present control practices.Ouraim is todescribeand toquantify the spatiotemporaldynamicsof theprimaryand secondaryinoculumsatplotandplantscalesonthreebananacycles.InGuadeloupe,800vitroplantswereplantedandobservedafternaturalinfectionsduring30months.Thesporedispersalwasdescribedbyfollowingtheincidenceatplotscaleandbyquantifyinglesionsatplantscale.Theprimaryandsecondaryinoculumsweremonitoredmensuallybyusingvolumetricsporetraps,attwoheights(aboveandunderplantapproximatingthebothinoculumcapture).Thesecondary inoculum was also quantified at plant scale by numerating and sporulating lesions ondifferentleaves.Results showed that the airborne inoculum is spatially randomly dispersed at plot scale, for theprimaryandsecondaryinoculumandfortwosuccessivebananacycles.Atplantscale,lesionshadthesamespatialdistributiononthedifferentcompartmentsoftheleaf,andtheyweremorenumerousonthetopcompartmentsthanonthebottom.Theconidiaconcentrationintheairwasalwaysunder60 conidia/m3. This conidia concentration was twice higher in the secondary than in the primaryinoculum.Therewasnotrelationshipbetweenthedynamicsoftheconidiaintheairandtherainfall.The ascospores concentration could not be achieved. At plant scale, the secondary inoculumquantifiedthroughtheaveragelesionnumberonthetopcompartmentvariedfrom3to225/25cm2.Thedynamicsoftheappearanceoflesionswasdifferentbetweenbananacycle.Theseresultsgavenewknowledgeonspatialdispersalatplotandplantscaleandonrelativepartofprimaryandsecondaryinoculums.Keywords:Sigatokadisease,spatiotemporaldynamics,sporetraps,lesionscounting

Session2Poster18Invitroexpressionofhumanphage-displayedscFvtodetectpotatovirusYSang-Ho Cho1, Eui-Joon Kil1, Sungrae Cho1, Haneul Seo1, Dong-Hoon Park1, Young Gyu Lee2 andSukchanLee11DepartmentofGeneticEngineering,SungkyunkwanUniversity,Suwon16419,Korea2Highland Agriculture Research Institute, National Institute of Crop Science, Rural DevelopmentAdministration,Pyeongchang25342,KoreaPotato virus Y (PVY), themost notorious plant virus, causes a very serious problem in potato. Toprevent the spread of plant viruses, there are so many diagnosis and detection techniques asserological procedures like enzyme-linked immunosorbent assay (ELISA). Serologicalmethods havebeenusedwidelywithwholeantibodyorrecombinantantibody likesingle-chainvariable-fragment(scFv).Screening fromrecombinantantibody (scFv) librarywithphagedisplay technologyhasbeenusedformoreconvenientantibodyproduction.However,therearesomedifficultiesinobtainingorexpressingspecificantigensusedforanimalimmunizationorscreening.Toovercomethislimitation,recombinantcoatprotein(CP)ofPVYdisplayedonyeastsurfacewasusedasantigen inthisstudy.Bio-panning for selection of anti-PVY scFv was performed with human scFv library againstrecombinantyeastantigen.Theantibody-antigenbindingactivityofscFvcandidateswasmeasuredby enzyme-linked immunosorbent assay (ELISA) test. Selected scFvwas expressed as both phage-displayedandsolublescFv.ThescFv-displayedphageswereprecipitatedbyusingPEG/NaClsolutionandused for expeditedevelopmentof virusdetectionmethod. For bacterial expressionof solublescFv,theselectedscFvgenesweresubclonedintopET26b(+)andproteinswerepurifiedusingaffinitychromatography.PVY-infectedsampleswasdetectedbyELISAand lateral flow immunoassay(LFIA)using the anti-PVY scFv. The purified scFv proteins showed better antigen-specific binding activityand lowercrossactivity thancommerciallyavailableantibodies.Thehorse radishperoxidase (HRP)wasclonedandexpressedwith scFvas fusionprotein to reduce reaction stepsand times.Purifiedenzyme-linked scFv showed chromogenic activity with 3,3 ,5,5 -tetramethylbenzidine (TMB)substrateformingbluecolor.TheseresultsdemonstratethatPVY-specificscFvproteinsisolatedfromhumanscFv libraryusingyeastsurfacedisplayandphagedisplaytechnologiesprovidenew invitroantibodyproductionsystemforplantvirusdiagnosis.Keywords:ELISA,scFv,Phagedisplay,PotatovirusY,Virusdetection,Yeastsurfacedisplay

Session2Poster19DevelopmentofarapidmethodtoidentifypectinolyticbacteriaisolatedfromblacklegsymptomsonpotatofieldJérémyCigna1,2,PaulineDewaegeneire2,AmélieBeury-Cirou2,VirginieGobert2andDenisFaure1

1CNRS-I2BC,ANR-15-CE21-0003,1avenuedelaterrasse,91198Gif-sur-Yvette,France2SemencesInnovationProtectionRechercheetEnvironnement(SIPRE),ruedeschampsPotez,62217Achicourt,FranceSoft rot pectinolytic bacteria induce blackleg disease on potato crop. Symptoms are caused byPectobacteriumandDickeyageneraandcompriseseveralspeciesorsubspecies.Rapidandaccurateidentificationisacrucialissueforpotatoindustry.Inthiswork,wedevelopageneticcharacterizationtool focus on fifty-threegapA genes of genomes publicly available. After alignment of sequences,thirty-fivesignaturenucleotidesspecificofagenus,speciesorsubspecieswerefound.Then,aPCRassay based on design of primers ongapA sequencewas effected on twenty-two strains isolatedfrom blackleg symptoms of French potato fields. Amplicons were sequenced and signaturenucleotides were analyzed. A phylogeny reconstruction has confirmed the identification of eachstrainbycomparisonwithgapAsequencesofreferentgenomesandvalidatetheapproach.Finally,themethod is useful for a rapid and accurate characterization of all soft rot pectinolytic bacteriaisolated in this study: P.atrosepticum, P.parmentieri, P.carotovorum subsp. brasiliense G1,P.carotovorumsubsp.carotovorumG2andG3,P.carotovorumsubsp.odoriferumG4,D.dianthicolaandD.solaniwereallidentifiedwiththedevelopedtool.Moreover,sequencescanbestoredandre-usedforfurtheranalyses.Keywords:identification,softrotpectinolyticbacteria,blacklegdisease,potatocrop,phylogeny

Session2Poster20LettuceinBelgiumhasfungalandnematodeproblemsJolien Claerbout1, Anneleen Volckaert2, Jenny Neukermans3, An Decombel4, Peter Bleyaert5, SofieVenneman6,IsabelVandevelde7,NicoleViaene8andMonicaHöfte91Ghent University, Coupure Links 653, 9000 Ghent, Belgium., ILVO (Institute for Agricultural andFisheries Research), Burgemeester Van Gansberghelaan 96, 9000 Merelbeke, Belgium. 2PCG(Provinciaal Proefcentrum voor de Groenteteelt Oost-Vlaanderen vzw), Karreweg 6, 9770Kruishoutem, Belgium. 3mans@pcgroenteteelt.be, PCG (Provinciaal Proefcentrum voor deGroenteteelt Oost-Vlaanderen vzw), Karreweg 6, 9770 Kruishoutem, Belgium. 4Inagro vzw,Ieperseweg 87, 8800 Rumbeke-BeitemSint-KatelijneWaver, Belgium. 7PSKW (Proefstation voor de,Belgium.5Inagrovzw,Ieperseweg87,8800Rumbeke-Beitem,Belgium.6PSKW(Proefstationvoordegroenteteelt), Duffelsesteenweg 101, 2860 groenteteelt), Duffelsesteenweg 101, 2860 Sint-KatelijneWaver,Belgium.8ILVO(Flandersresearchinstituteforagriculture,fisheriesandfood),Burg.Van Gansberghelaan 96, 9820 Merelbeke, Belgium); Faculty of Sciences, Ghent University, K. L.Ledeganckstraat35,9000Ghent,Belgium9GhentUniversity,CoupureLinks653,9000Gent,BelgiumLettuce is a very important crop in Belgium and is mainly produced in glasshouses with up to 5rotationsperyear.In2015theproductionoflettucegeneratedarevenueofalmost25millioneuro,withaproductionof42800tonneson200ha.Asaresultofthisveryintensiveproduction,thecropsuffersfromsoil-bornediseases,causedbyvariouspathogensincludingRhizoctoniasolani,Pythiumspp.,BotrytiscinereaandSclerotiniaspp.Thesepathogenscausesymptomscommonlydescribedasbasalrot.Inaddition,nematodessuchasParatylenchusspp.andPratylenchuspenetranscauserootdamage resulting in reduced growth. Since the complete ban ofmethyl bromide in 2006, controlstrategiesrelyonintensiveuseoffungicidesandnematicides,resultinginunwantedresiduesinthesoiland theendproduct.Furthermore, theBelgian lettuceproduction is since2015 threatenedbylettucewiltcausedbyanewraceofFusariumoxysporumf.sp.lactucae,forwhichthereiscurrentlyno control method available. To make disease control in intensive lettuce production more inaccordancetothecurrentIPMguidelines,wearecollaboringinaprojectcalledFUNSLAwiththeaimtodevelopadecision support system for soil-bornepathogens.Weare studying theepidemiologyandactivityofthesesoilbornepathogensindetail,sothattherightchemicalsareusedonlywhennecessary.Inaddition,thepotentialofbiocontrolagentsthatreduceinoculumbuild-upinthesoilisinvestigated.Damagethresholdsfornematodeshavebeenestablishedtoavoidunnecessaryuseofnematicides. To determine the activity of the different pathogens causing basal rot in differentseasons,threeglasshousesarecontinuallyreplantedwithlettuceandmonitoredwithoutanydiseasecontrol. Depending on the season, several different R. solani anastomosis groups could bedistinguished inasameglasshouse. Inwinter,wemainly isolatedR.solaniAGBI fromlettucecropswith rotting symptoms, while in other seasons R. solani AG1-1B, AG2-1 and AG4 appear to bedominant.Wearecurrentlystudyingthepathogenicityandtemperaturerangeoftheseanastomosisgroupsinmoredetail.PostersessionKeywords:Rhizoctoniasolani,Fusariumoxysporum,nematodes,basalrot,seasonality

Session2Poster21Virulence characterization of Podosphaera xanthii populations in main Mediterranean melonproductionareasIsabelle Justafré1, Carole Constant2, Mireille Buisson3, Sophie Perrot4, Valérie Grimault4, SylvieBaudracco-Arnas5, François Bertrand6, Denis Losdat7, Robert Legnani8, Michel Pitrat9 and ThierryJaunet10

1HM.Clause,LeMasSaintPierre,LaGaline,13210SaintRémydeProvence,France2SakataVegetablesEurope,DomainedeSablas,Ruedumoulin,30620Uchaud,France3GautierSemences,Routed’Avignon,13630Eyragues,France4GEVES,RueGeorgesMorel,49071Beaucouzé,France5LaboratoiresASL,755chemindeMeinajaries,84140Montfavet,France6Monsanto,,ChemindeRoquemartine,13670Saint-Andiol,France7Rijk-ZwaanFrance,LaVernède,30390Aramon,France8TakiiFrance,chemindelaCrau,13630Eyragues,France9INRA,DomaineSaint-Maurice,67alléedeschênes,84143Montfavet,France10HM.Clause,1cheminduMoulindesRonzières,49800LaBohalle,FranceMelonresistancetopowderymildew,causedbyPodosphaeraxanthii,isakeycomponenttocontrolthismajorworldwidemelonproductionconstraint.FiveracesbasedonfourmelonlineshavebeendescribedinP.xanthii inEurope.However,therelevanceofthisracesystemcanbequestionedbythe fast race evolution in the last decade and the limited knowledge of the virulence diversity inpopulations present today in European melon production areas, impeding resistant varietydevelopmentanddeploymentadaptedtotargetmarkets.TobetterunderstandP.xanthiivirulencedistribution inEuropeanddevelopsolidbasistoredefineracenomenclature,aFrenchAgricultureMinistryfundedprojecthasbeeninitiatedin2012.Throughthisproject,acollectionofmorethan200P.xanthiiisolateshasbeenbuiltfromsamplesofmelonandothercucurbitscollectedin2013and2014inmajormelonproductioncountries(France,Spain, Italy, Morocco, and Turkey). Within this collection, virulence of 125 isolates has beencharacterizedonapanelof30melonlines, includingthefour linesdefiningP.xanthii racesand26linesrepresentingmajorpowderymildewresistancesources.Virulencewasdeterminedbydetachedleafdiscassayaccordingtothevarietyregistrationprotocol.Based on the existing race nomenclature, melon isolate virulence appeared homogeneous in allregions,with75%isolatesbelongingtotherace3.5.However, ifconsideringthewholepanelof30lines, a very complex pattern of virulence profiles has been pointed out with more than 75% ofmelonisolatesvirulentonacombinationof15ormorelines.Inordertoreducethiscomplexity,lineandisolatesub-panelshavebeenselectedbystatisticalanalysisandcharacterizedondetachedleafdiscandwholeplant.Results fromthisanalysiswillbeusedtoredefinearacenomenclaturemoreadaptedtotheexistingP.xanthiivirulencesituation.Keywords:powderymildew,melon,virulence,racenomenclature

Session2Poster22A new diagnostic multiplex polymerase chain reaction scheme for three species of Pantoeathreateningriceproductioninsub-SaharanAfricaK.Kini1,2,R.Agnimonhan1,O.Afolabi1,R.Dossa1,D.Silué1,andR.Koebnik2

AfricaRice, Plant Pathology,Africa Rice Centre,01 B.P 2031,Cotonou, Cotonou, Atlantique,Benin,Institut de la Recherche pour le Développement (IRD), Montpellier, FranceAfrica Rice Centre, Plant Pathology,Africa Rice Centre,01 B.P 2031, Cotonou, Atlantique, Benin,AfricaRice, Plant Pathology, Cotonou, Atlantique, Benin, Institut de la Recherche pour leDéveloppement(IRD),Montpellier,FranceMembersofthegenusPantoeaareresponsibleformanydiseasesofeconomically importantcropsworldwide.EmergingdiseasesofriceduetoinfectionbyPantoeacausesignificantdamageinmostrice-growingareasinsub-SaharanAfrica.Theaimofthisstudywastodevelopadiagnosticmultiplexpolymerasechainreaction(PCR)assayforrapid,sensitiveandsimultaneousdetectionofthePantoeaspp.belongingtothreemajorspeciesofPantoea,Pantoeaananatis,PantoeastewartiiandPantoeaagglomerans.Genus-andspecies-specificprimerstargetingfourhousekeepinggenesofPantoeasppwere designed through a bioinformatics pipeline using multiple genome sequences. Sensitivity ofdetection was monitored on isolated DNA, on in vitro-grown bacterial cells, on artificiallycontaminatedriceseeds,onartificiallyinoculatedriceleavesandonsymptomaticandasymptomaticleaves collected fromaffected rice fields. The reaction parameterswere optimized for amultiplexPCRschemeandappliedonatotalof183Pantoeasppstrains.ThemultiplexPCRschemeaccuratelyrevealed the presence of pathogens on rice seeds and leaves. This is the first report of amethodallowing simultaneous detection of three important Pantoea spp., which will be useful inepidemiologicalsurveillanceprograms.Keywords:MultiplexPolymeraseChainReaction,Housekeepinggene,Pantoeaspp,rice

Session2Poster23Epidemiologyandcontrolofbeetrust(Uromycesbeticola)onsugarbeetRoseKristoffersen1,ThiesMartenHeick1,AnnemarieFejer Justesen1,AnneLisbetHansen2andLiseNistrupJørgensen1

1AarhusUniversity,InstituteforAgroecology,Forsøgsvej1,4200Slagelse,Denmark2NBRNordicBeetResearch,Sofiehøj,Højbygaardvej14,4960Holeby,DenmarkIncreasinglyhighdiseasepressurefrombeetrust(Uromycesbeticola)hasbeenregisteredinrecentyearsinDenmark.Thischallengethecommonbeliefthatbeetrustisaminordiseaseonsugarbeetswhich seldom requires control. Thus, there is aneed togain knowledgeof thepathogen’sbiologyand epidemiology including factors contributing to spread of the disease and the onset of anepidemic.AspartofaprojectinvestigatingIPMsolutionsinsugarbeet,theoccurrenceofU.beticolaspores in the air above sugarbeet fieldswasmonitoredusingBurkard7day recording volumetricspore traps and qPCR in four consecutive field seasons. Spores were detected throughout thesamplingperiodandwereconstantlypresentwhenfirstdiseasesymptomswereseeninthefield.Adiseasecyclewithallmajorstepswasestablished includingmeansofoverwinteringandspread. Incoastalregions,seabeetscanactasagreenbridgeandprovidetheprimaryinoculumtothesugarbeet crops. Locations with early onset of beet rust were investigated and indicated a connectionbetween early beet rust occurrence and the presence of local wild sea beets. It has beendemonstratedthatattacksofbeetrustcausesyieldloss,buthighdiseasepressuredidnottranslateunambiguously into decreased yield or economic return in all trials. In high risk areas diseasepressure can be reduced by growing less susceptible cultivars and by application of strobilurin ortriazolefungicides.Keywords:Betavulgaris,sporetrapping,fungicide,diseasecycle

Session2Poster24PredictingabundanceofBotrytiscinereaairborneinoculumtoforecastgreymouldepidemicsChristelLeyronas1,OlivierMartin2,PhilippeNicot1,SamuelSoubeyrand2

1PathologieVégétale,INRA,84140,Montfavet,France2BioSP,INRA,84914,Avignon,FranceBotrytiscinerea isanairbornedisseminatedascomycetethatcausesgreymouldonmorethan200species, among which some are cultivated worldwide and have a great economic importance(grapevine, lettuce, tomato). Predictive tools could help growers to rationalize their crop healthmanagementpractices,particularlythetimingforchemicalcontrolandthus lowerthecostofcropprotectionandpreservetheenvironmentwhilemaintainingtheefficacyofdiseasecontrol.Thereisaneed of a forecasting system that can predict abundance of viable inoculum in the air in Frenchagriculturalareas.Weconductedastudyovera3-yearperiodtocharacterizethetemporalevolutionoftheabundanceof B. cinerea inoculum on a non-cultivated site (region of Avignon) and to determine if climaticparameters can serve as predictive parameters in a future grey-mould risk forecast model. Localmeteorologicaldatawereacquiredcontinuouslyonaclimaticplatform.Backwardtrajectoriesofairmasses arriving in Avignon on the sampling days and climatic parameters these air massesencounteredalongtheirwaywerecomputedusingthesoftwareHYSPLIT.GeneralizedlinearmodelswithPoissondistributionwerethenevaluatedtomodelB.cinereaabundance.Modelsconditionaloneither local climatic variables or climatic variables frombackward trajectories or on both types ofvariableswereconsidered.Crossvalidationswereperformedtoidentifythebestmodel.Viableinoculumwasobservedfor96%ofthesamplingdays.Theabundanceofthisviableinoculumwas positively correlated with average daily relative humidity and negatively correlated with airtemperature and solar radiation. The analysis of backward trajectories suggested that air massesoriginating from theNorthor theSouthbroughtmoreviable inoculum than those from theWest.Moreover, the model providing the best prediction criterions was obtained when local climaticparametersbutalso,climaticparametersalongtheairmasstrajectoriesandtheiroriginweretakenintoaccount.Keywords:aerobiology,airmasstrajectory,climaticparameter,statisticalmodel

Session2Poster25NGS and virus diagnostic: does sequence analysis strategies really matter? Results of aninternationalproficiencytestingonsiRNASebastienMassart,KrisDeJonghe1,IanAdams,AnnalisaGiampetruzzi,IgorKoloniuk,PetrKominek,JanKreuze,DenisKutnjak, Leonidas Lotos,Hano J.Maree,ThibautOlivier,MikhailPooggin,AnaB.Ruiz-García,DanaSafarova,P.H.H.Schneeberger,NoaSela,EvaVarallyay,EevaVainio,EricVerdin,MarcelWestenberg,YvesBrostauxandThierryCandressea

1Flandersresearchinstituteforagriculture,fisheriesandfood(ILVO),PlantSciencesUnit,Burg.VanGansberghelaan96,9820Merelbeke,Belgiuma:thecontributorsofthiscommunicationarelistedbyalphabeticalorder,exceptS.M.,Y.B.andT.C.TheyaremembersoftheCoreGroupoftheCOSTActionandtheiraddressesarelistedintheCOST.Therecentdevelopmentsofhigh-throughputsequencing(alsocalledNextGenerationSequencing-NGS)technologiesandbioinformaticshavedrasticallychangedtheresearchonviralpathogensandisnow raisinga growing interest for virusdiagnostics. However, anydiagnostic techniquehas tobeincluded in standardizedprotocols.Currently,ahugediversityofbioinformaticsprotocols forvirusdiscovery has been reported in the scientific literature but, to date, without addressing theirreliability for diagnostic purpose. The objective of this work was therefore to compare theperformanceofexistingbioinformaticspipelinesandof theresult interpretationthroughadouble-blindedlargescaleproficiencytestingbasedonasetoftenfastqfilesandinvolving21laboratoriesfrom16countries.Thefastqfilescontained50,000(3),250,000(4)and2.5M(3)sequencesof21-24nt coming from 3 samples. The false positive rate was only 0.5% and mainly related to theidentificationof integrated sequencesormisinterpretationof the results. Theoverall sensitivityofdetectionwas57%andrangedbetween35and100%betweenlaboratorieswithamarkedeffectofrarefactionforsomelaboratories.Aprincipalcomponentanalysisandcorrelationstudiesunderlinedthe most important parameters for appropriate diagnostic. The repeatability of detectioncorresponded to 73%. This work also underlined (i) the complexity of discovering new viruses byNGS, (ii) the difficulty to detect viral pathogens with low number of siRNA reads, (iii) theinconsistenciesofdatabasesanditsimpactonresults.Overall,thisworkbringskeyinsightsintothereliability of bioinformatics pipelines and underlines some key parameters for achieving a reliabledetectionofvirusesinadiagnosticsettingusingsiRNAsequencing.Keywords:deepsequencing,massivelyparallelsequencing,bioinformatics

Session2Poster26Distribution and change in populations of Leptosphaeria species associated with phoma stemcankerintheCzechRepublicJanaMazáková1,MiloslavZouhar1,PavelRyšánek11Czech University of Life Sciences Prague, Faculty of Agriculture, Food and Natural Resources,DepartmentofPlantProtection,Kamýcká129,16500Prague,CzechRepublicIntheCzechRepublic,the increase inareasownwithoilseedrapeduringthe lasttwodecadeshasbeencontributingtotheincreasedappearanceofphomastemcanker.Thediseaseiscausedbytwoclosely related fungal pathogens, Leptosphaeria maculans and L. biglobosa. The objective of thisstudywas to assess the countrywide distribution of these two pathogens and their occurrence inoilseed rape tissues. In 2007/11, 1454 leaf spotswere visually identified based on symptoms andthenanalysedusing species-specificPCR.Outof these,39and15%weredetectedasL.maculans-and L. biglobosa-infected, respectively, in case of single species-infected samples, while 26%corresponded to theco-infectionbyboth species. Furthermore, some isolates, thatwerecollectedfrom selected leaf spots and maintained in pure cultures, were identified based on pigmentproductionduringculturingonsolidandinliquidmediaandPCRassay.Inthiscase,theco-infectionbyL.maculansandL.biglobosainasingleleafspotappearedaswell.Inyears2007/12,708basesofoilseedrapeplantsdividedintoupperstem,basestem,rootcollarandtaprootpartswereanalysedusing symptom identification and PCR. The proportion of plants in which L. biglobosa DNA wasamplifiedwasgreaterthanthatofplantswithL.maculansDNAand40%oftestedplantswerefoundtobeco-infectedbybothL.maculansandL.biglobosa.According toour results, itappears thatL.maculansisthepredominantspeciesinautumn,whileL.biglobosaismoresuccessfulspeciesthanL.maculansincolonizationofoilseedrapetissuesinlatergrowthstagesofaplantinconditionsoftheCzech Republic. Thisworkwas financially supported byMACRNAAR, projectsNo.QJ1310227 andQK1710397.Keywords:phomastemcanker,oilseedrape,Leptosphaeriamaculans,L.biglobosa,symptoms,PCR

Session2Poster27TheabilityoffungiisolatedfromcankersofpomefruittreestocausefruitrotsinthestorageIngaMoročko-Bičevska1,OlgaSokolova1,KristīneVēvere1,MārisJundzis11InstituteofHorticulture,LatviaUniversityofAgriculture,Graudustr.1,Dobele,LV-3701,LatviaApples andpears occupy an important niche in the fruit growing industry in Latvia. Emergence offruit rot diseases and increasing damages in the orchards and storages, lack of the data on theircauses and control possibilities are among themain concerns of the pome fruit growers. Severalpathogenicfungiareknowntocausecankersonthetreesinorchardsandalsofruitrotsinorchardsand storages (e.g.Neofabraea spp.,Monilinia spp.). The knowledge on these diseases is still notsufficient, and their significance inmanyareas is not known. Theaimof thepresent studywas toelucidateabilityofvariousfungiisolatedfromtreecankersofappleandpeartocausefruitrotsinthestorage. Four apple and four pear cultivars differing in tolerance to fruit rots were used for thestudies. Pathogenicity on fruits was characterized for 20 fungal isolates belonging toNeofabraeaspp.,Fusarium spp.,Diaporthe spp.,andValsa spp. in twostorageexperiments.Thedifferences inabilitytocausefruitrotwereobservedamongisolatesbelongingtothesamespeciesandamongthespecies.Elevenofthirteentestedisolatesoriginatingfromtreecankerswerealsoabletocausefruitrot.NeofabraeastrainsandoneFusariumsp.isolateweremostaggressive,whichcausedsignificantdamagesonmostofthetestedcultivars.Themoreaggressiveandvirulent(degreeofdamagesandabilityto infectmorecultivars)wereNeofabraeaspeciesand isolatesoriginatedfromtreecankers.Tests on plants are in progress to characterize aggressiveness and virulence of these isolates onplantsofthesameappleandpearcultivarsusedinstorageexperiments.Keywords:apple,pear,fruitrot,canker,Neofabraea

Session2Poster28FungiandmycotoxinsinannualandperennialgrassesofLeguminosaeAleksandraOrina1,OlgaGavrilova1,TatianaGagkaeva1,AlexeyBurkin2andGalinaKononenko2

1All-RussianInstituteofPlantProtection(VIZR),St.-Petersburg–Pushkin,Podbelskogosh.,3,196608,Russia2All-RussianInstituteforVeterinarySanitation,HygieneandEcology,Moscow,Zvenigorodskoesh.5,123022,RussiaThe aim of study was to evaluate the contamination of growing forage plants by fungi andmycotoxins. Seventy seven samples (13 plant species of Galega, Lathyrus,Medicago,Melilotus,Trifolium,andViciagenera)belongingtoannualandperennialgrassesofLeguminosae,collectedinthedifferent regionsof theEuropeanpartofRussiawereanalized.Comparativeevaluationof theinfection by Alternaria, Cladosporium, and trichotecenes producing Fusarium fungi in the plantssampleswascarriedoutbyusingreal-timeTaqManPCR.Theamountsof16mycotoxinsintheplantssampleswereanalyzedbyELISA.The Lathyrus spp. in comparison with the other members of the family, characterized by highcontents of the DNA of Cladosporium (8.7×10-4 ng/ng of total DNA) and Fusarium fungi (7.5×10-5ng/ngof totalDNA). The large amounts of theAlternaria DNA (7.8×10-5 ng/ngof totalDNA)werefound inplantsofgenusMelilotus.Theperennialplants (Galegaorientalis,Lathyrus spp.,Trifoliumspp.)containedmoreDNAofFusariumfungiincomparedtoannualcrops.Taking into consideration their frequency and concentration, themost importantmycotoxinswerealternariol and cyclopiazonic acid, their highest amounts were detected in plants from genusTrifolium. Mycotoxins produced by Fusarium fungi occurred with less frequency that othermycotoxins.ThegreatestvarietyoffungalmetaboliteswasdetectedinLathyrusspp.Thestatisticalanalysisshowedthesignificantimpact(p<0.05)ofplantspeciesonquantityofDNAofallgroupsoffungiandthemostmycotoxins.ApositivecorrelationbetweenthecontentoftheDNAofAlternariaandCladosporiumfungi(r=+0.30,p<0.05)wasrevealedinallsamplesoflegumes.InsomecasestheconnectionbetweenamountsofthefungalDNA,mycotoxincontent,theoriginofsamples,andthetimeoftheircollectionwereconfirmedstatistically.Keywords:legumes,fungi,DNA,real-timePCR,mycotoxins,ELISA

Session2Poster29Use of RAPD and ISSRmarkers in detection of genetic variation amongColletotrichum falcatumWentisolatesfromSouthGujaratIndiaPritteshKPatel1,RushabhShah1,RKrishnamurthy1,VkulbhushanBhaskar2

1UkaTarsadiaUniversity,CGBhaktaInstituteofBiotechnology,Bardoli,394350-Gujarat,India2JTES’SArtsCommerce,ScienceCollege,Jamner,Dist:Jalgaon,424206–Maharastra,India

Thepresentresearchworkaimsatfindinggeneticdifferences inthegenomesofsugarcaneredrotisolates Colletotrichum falcatum Went using Random Amplified Polymorphic DNA (RAPD) andinterspersed simple sequence repeat (ISSR) molecular markers. Ten isolates of C. falcatum fromdifferentredrotinfectedsugarcanecultivarsstalkwereusedinpresentstudy.Theamplifiedbandswerescoredacrossthelanesobtainedin15RAPDprimesand21ISSRprimessuccessfully.ThedatawereanalyzedusingNTSYSpc2.2software.Theresultsshowed80.6%and68.07%polymorphisminRPADandISSRanalysisrespectively.BasedontheRAPDanalysis,tengenotypesweregroupedintotwomajorclustersatacut-offvalueof0.75.GeographicallydistantC.falcatumisolatecfGANfromsouthGujarathadalevelofsimilaritywithCoimbatoreisolatecf8436presentedonseparatecladeofbootstrappeddendrograms.Firstandsecondclusterconsistedoffiveisolates(cfNAV,cfPAR,cfTIM,cfMAR, and cfVES) and three isolates (cfKAM, cfCHA and cfMAD) respectively, indicating the closerelation among them. Twentyone ISSRprimers produced119distinct and scorable loci in that 38were monomorphic. The number of scorable loci for each primer varied from 2 (ISSR822) to 8(ISSR807,ISSR823andISSR15)withanaverageof5.66lociperprimer.PrimerISSR835amplifiedthehighestnumberofbands(57),whileonly16bandswereobtainedbyprimersISSR822.FourprimersnamelyISSR830,ISSR845,ISSR4andISSR15showedthehighestvalueofpercentageofpolymorphism(100%). The results indicated that both of themarker systems RAPD and ISSR, individually can beeffectively used in determination of genetic relationship among C. falcatum accessions collectedfromdifferentpartsofsouthGujarat.Keywords:Colletotrichumfalcatum,ISSR,RAPD,RedRot,sugarcane

Session2Poster30EpidemiologyofPasmoandSeptorialinicolaresistanceinFrenchflaxcultivarsA.Penaud1,D.Paumier2,B.Bamme1,A.Petiteau2,E.Heritier3,F.Suffert4,R.Valade21TerresInovia,CentredeGrignon,78850Thiverval-Grignon,France2ARVALIS Institut du Végétal, Avenue Lucien Brétignières, Bâtiment INRA Bioger, 78850 Thiverval-Grignon,France3ARVALISInstitutduVégétal,Stationexpérimentale,91720Boigneville,France4INRA,UMR1290BIOGER,UniversitéParis-Saclay,CampusAgroParisTech,78850Thiverval-Grignon,FranceSeptoriaLinicolaisthecausalagentofPasmo,aworldwidediseaseofLinumusitatissimum(flax,i.e.‘linen’and‘linseeds’usedforfibersandfoodoil,respectively).Whilesevereoutbreakshaveoccurredin France in last years, biotic and abiotic factors favoring epidemics are little known and hostresistance level is still undetermined. A 3-year survey of Pasmo was performed in order tocharacterizethesourcesofprimaryinoculum,theearlystagesofepidemic,andhostresistancelevelforsomeofthemostdeployedFrenchcultivars.Cohorts of S. Linicola fruiting bodies were observed and accurately described (type, number,maturity) on linseed stubbles during fall and winter and disease was assessed in flax fields.Pseudothecia, asci and ascospores ofMycosphaerella linicola, the sexual form of S. linicola, wereidentifiedforthefirsttimeinFranceonlinseedstubbles.Pseudotheciaappearedlatesummer,withapeak in October. The temporal dynamic was notably similar for the three years, with however asignificant shift depending on the climatic conditions of early autumn (dryness). Ascospores areprobablyresponsibleforprimaryinfectiononwinterlinseed,whilepycnidiosporescontributemainlytotheepidemicdevelopmentonspringlinenandspringlinseed.Finally,weconcludethatbothtypesofsporesareinvolvedinFrenchconditions;theirrelativeimportanceprobablydependsonthetypeofcrop,theepidemicstage,andtheclimaticconditions.Undercontrolledconditions,22varieties(15linenand7linseed)wereexaminedforPasmoattacksoncotyledons,leavesandstemsusing6S.Linicolaisolates.Therangeofsusceptibilitywasrelativelywide and no significant, high level of resistance was detected. Nevertheless, linen appearedsignificantlymoresusceptiblethanlinseed.TheseresultshelptounderstandhowPasmodevelopsandtoidentifymanagementoptionsadaptedtothediversityofflaxcroppingpracticesinFrance.Keywords:flax,hostresistance,linen,linseed,Linumusitatissimum,Mycosphaerellalinicola,Pasmo,primaryinoculum,Septorialinicola,sexualreproduction

Session2Poster31Detection,identificationandquantitationofoomycetesinimportednurseryplantsAlexandraPuertolas1,StephenWoodward1,EricBoa1andPeterBonants21UniversityofAberdeen,InstituteofBiologicalandEnvironmentalSciences,DepartmentofPlantandSoilScience,Aberdeen,AB243UU,Scotland,UK2PlantResearchInternational,WageningenUniversity,Wageningen,6708PB,theNetherlandsInternational trade in hardy ornamental nursery stock, including Internet sales, has increased thedispersal of many plant pathogens, including highly damaging oomycetes. Potted plants with soilsubstrates are now recognised as themajor pathway of introduction of newpests and pathogensintoEurope.Wescreenedthediversityandpathogenloadsinsoilfrompottedwoodyplants,inrootsand in water using baiting assays. Classical isolation techniques were used as well as molecularassays,includingTaqManPCRchemistryandNextGenerationSequencingapproachesusingamulti-locusIlluminaMiSeq.Approximately90%oftestedplantscontainedatleastonespeciesofoomycete(Phytophthora, Pythium, Phytopythium), while 86% of asymptomatic plants tested positive foroomycetesinthegrowthsubstrate.TenPhytophthoraspecies,17Pythiumspp.and5Phytopythiumspp.Wereisolatedusingclassicaltechniques.TaqManassaysforthreedifferentloci(ITS,trnM-trnPtrnM and atp9-nad9), two of them Phytophthora spp. specific, revealed higher pathogen averagedensities in soils in comparison with the root and filter samples. The multi-locus Illumina assaydemonstrated the high diversity of oomycetes present in all samples analysed, includingasymptomaticplants,revealinghighabundanceofpotentiallyharmfulspeciessuchasPhytophthoraramorum and P. alni. These results provide worrying evidence of plant pathogens being moved‘silently’ between countries in nursery plants, and highlight the need for stronger regulation toreduceplantbiosecurityrisksinEurope.Keywords:oomycetes,multi-locus,TaqMan, IlluminaMiSeq,ornamentalplants, internationalplanttrade,Internetsales

Session2Poster32OccurrenceofPlasmodiophorabrassicaeWor.andvirusdiseasesofoilseedrape (Brassicanapussubs.napus)intheCzechRepublicVeronikaŘičařová1,JanKazda1,PetrBaranyk2,3,LenkaGrimová1andPavelRyšánek11DepartmentofPlantProtection,CzechUniversityofLifeSciencesPrague(CULS),Kamýcká129,16500Praha6–Suchdol,CzechRepublic2UnionofOilseedGrowersandProcessors (SPZO),NaFabiánce146,Praha8–Březiněves,18200,CzechRepublic3DepartmentofCropProduction,CzechUniversityofLifeSciencesPrague(CULS),Kamýcká129,16500Praha6–Suchdol,CzechRepublicClubroot, caused byPlasmodiophora brassicae (Wor.), has been spreading onwinter oilseed rape(Brassica napus L.) in the Czech Republic over the past six years. Clubroot infestation and spreadweremonitored over five years andmaps of infestationwere created. Experimentswith clubrootresistant cultivars of winter oilseed rape were carried out in the field and greenhouse. In thegreenhouse, six resistant cultivarswere grown in infested soil collected from various fields in theCzechRepublic,andassessedfordiseaseseverity.Thesoilsampleswerealsotestedforthepresenceand amount of P. brassicae inoculum by conventional and quantitative PCR analysis. In the fieldexperiment,sevenresistantcultivarsweregrownanddiseasedevelopmentwasmonitoredmonthly.Yieldsweremeasuredattheendofthecroppingseason.Finally,asetof17P.brassicaefieldisolatesfromacrosstheCzechRepublicwereassessedforpathotypedesignationonthedifferentialhostsofWilliams,Soméetal.,andtheEuropeanClubrootDifferentialset.The informationobtainedontheeffectivenessofhostresistanceandpathogenicdiversityofP.brassicaepopulationsfromtheCzechRepublicmayhelptomoreeffectivelymanageclubrootinthiscountry.Inautumn2016theunusuallyhighabundanceofgreenpeachaphid (Myzuspersicae)occurredonoilseedrapefieldsacrosstheCzechRepublic.Thisspecies isavectorofTurnipyellowsvirus (TuYV)and Turnip mosaic virus (TuMV), which are commonly found on oilseed rape. The nationwidemonitoring of these two viruses wasmade using detection by Triple Antibody Sandwich Enzyme-LinkedImmunoSorbentAssay(TAS-ELISA)andDoubleAntibodySandwichELISA(DAS-ELISA).ThetestrevealedhighoccurrenceofTuYV-93.7%oftestedsampleswerepositive.Ontheotherhand,theoccurrenceofTuMVwasvery low– just0,2%of sampleswerepositive.The springmonitoringofvirusoccurrenceisplanedaswellastestingofoilseedrapecultivars,whicharedeclaredasresistant.Keywords:clubroot-resistantcultivars,pathotypes,qPCR,Myzuspersicae,Turnipyellowvirus(TuYV),Turnipmosaicvirus(TuMV)

Session2Poster33ErucasativanaturallyinfectedbyTurnipmosaicvirusinBrazilMarcos Roberto Ribeiro Junior¹, Lívia Filomena Santos Baldini¹, Denise Nakada Nozaki¹, GiovanaCarolinaDouradoCruciol¹,KéssiadeFátimadaCunhaPantoja¹,BrunoRossittodeMarchi¹,MarceloAgenorPavan¹andRenateKrauseSakate¹¹DepartmentofPlantProtection,SãoPauloStateUniversity(UNESP),Botucatu,BrazilLeavesof rocket salad (ErucasativaMiller)withmosaic symptomswereobserved inacommercialcrop production and investigated for the presence of viruses. Associated to these plants we alsofound raphanus (Raphanus raphanistrumL.)withmosaic symptoms.Filamentousparticles typicallyofpotyviruswereobservedbyElectronMicroscopyandtheplantswerepositiveforELISATestusingthepotyvirusantiserum(Agdia,Inc).TotalRNAwasextractedbyBertheauetal.,(1998)followedbyRT-PCRusingtheuniversalprimersforpotyvirus(W-CIENandPV-1,describedbyGibbs&Mackenzie,1997). The amplified fragment showed 99% identity with Turnip mosaic virus (TuMV, accessionnumber EU734433.1). A couple of primerswere synthesized to amplify the complete coat proteinregion(forward:5’ACAGATGAGCAGAAACAGGC3’andreverse:5’AATCAAATGCGTACCGAGC3’)andidentity of 97%wasobservedwith TuMV (accessionnumberAB701725.1). Bayesian analysis usingdifferentreferencesequencesofTuMVisolateswasperformedandthis isolatewasgroupedintheBrassica-Raphanus (BR) clade according toNguyenet al., (2013). The viruswas sap transmitted toChenopodiumamaranticolor,Chenopodiumquinoathatdevelopedlocal lesions,Betavulgaris(locallesionsandsystemicnecrosis),Nicotianabenthamiana (mosaicsymptoms), torocketandraphanusplants thatdeveloped local lesions followedbymosaicandnecrosisof leaves.The isolateofTuMVwasalsoefficientlytransmittedtotworocketgenotypes(astroandrococó)andraphanusplantsbyAphisgossypii.Asfarasweknown,thisisthefirstreportofrocketsaladinfectedwithTuMVinBrazil.Keywords:Potyvirus,rocketsalad

Session2Poster34CharacterizationofgeneticdiversityofVenturiainaequalispopulationinLatviausingmicrosatellitesOlgaSokolova1andIngaMoročko-Bičevska11InstituteofHorticulture,LatviaUniversityofAgriculture,Graudustr.1,Dobele,LV-3701,LatviaApple scab, causedby theheterothallic ascomyceteVenturia inaequalis (Cooke)G.Wint. isoneofthemostimportantdiseasesofappleworldwide.DuetothelifecycleofV.inaequalis,ineachspringapple trees are infected with newly released ascospores representing new genotypes of thepathogen.Thisphenomenonhasbeenreportedtohaveoneof themajor impactsondiversityandformationofmoreaggressiveracesofV. inaequalis.Setof99singlespore isolatesoriginatedfromdifferent regions,orchardsandcultivarsweregenotypedwith14polymorphicmicrosatellites (SSR)markerstocharacterizegeneticdiversityofV.inaequalispopulationinLatvia.ThesetofisolatesalsoincludedseveralstrainsfromBelgium,France,Poland,Germany,Netherlandsobtainedfromculturecollections for comparisons. Resulting binary data set was analysed by cluster analyses UPGMA,NeighbourJoiningandprincipalcomponentanalysis(PCA).InUPGMAanalysesLatvianoriginisolatesclusteredaccording to samples, trees andpartly also toorchards. InPCAanalysis several separategroupswere identifiedamongLatvianorigin isolatesaccordingtotheregion,orchard,andcultivar.Some of the Latvian V. inaequalis isolates showed genetic similarity to isolates virulent on scabresistant cultivars from other European countries indicating the potential for further virulencedevelopment.Keywords:Applescab,SSRmarkers,fungaldiseases,Malusdomestica

Session2Poster35Comparison of crown rust resistance reaction of oat differential lines obtained from differentsourcesSylwiaSowa1,EdytaPaczos-Grzęda1,AnetaKoroluk11UniversityofLifeSciencesinLublin,InstituteofPlantGenetics,BreedingandBiotechnology,Akademicka15,20-950Lublin,Poland

Rustsareknownasthemostwidespread,destructiveandeconomicallyimportantdiseasesofcerealcrops worldwide. Necessary information about fluctuations in the racial composition, raceprevalence, arising of new races and effectiveness of resistance genes can be obtained by rustpopulationchangesmonitoring.Informationprovidedareespeciallyimportanttobreedingprogramsinordertodecideon,whatbreedingstrategyforresistanceshouldbetaken,whichgenotypesshouldbe cultivated and which genes should be combined in crossing. At the heart of reliablemeasurementsarereferencematerials.Theygiveabenchmarkforlaboratoriesthroughouttheworldto deliver accurate and comparable results of virulence surveys andmolecular characterization ofmajorcerealrusts.Thisstudycomparescrownrustresistancereactionofoatdifferential linescontaininggenes:Pc39,Pc40,Pc46Pc48,Pc50,Pc51,Pc58,Pc59,Pc94andPc96obtainedfromIowaStateUniversityinUSAandCerealResearchCentreAAFCWinnipeginCanadabymeansofhost-pathogentest.Assessmentwasconductedonten-days-old3-cm-longleavesfragmentsinthelaboratorytestsundercontrolledtemperature,humidityandlightingconditions.Forinoculation378P.coronataf.sp.avenaeisolatescollectedinPolandduringtheyears2013-2016fromrandomfarmfieldsandfieldplotswereused.The results indicate that majority of the tested lines showed a homogeneous reaction with theexceptionoflinespossessingPc50andPc51genes.IntheisolinePc50onthebasisofsegregationinresistantandsusceptibleplantstwolines,eachpossessingadifferentmajorgenePc50-2andPc50-4were already distinguished by Šebesta (1983). Both Pc51 lines in Polish conditions showed highefficiency, however when resistance was overcome, plant reaction differed. These results mayindicate thatA. sterilis Wahl No. 8, the donor of Pc51 line resistance could also have twomajorresistancegenes,whichwasrandomlyreselectedinlaterisolinegenerations.Duetopathogenevolutionconstantrustresearchshouldbeconductedinordertosupportdiseaseresistancebreedingaswellasstudycoevolutionofnaturalhostpathogensystems.Itisessentialforresearchers around the world to work with a uniform material to obtain accurately andunambiguouslycomparableresults.Keywords:crownrust,Pucciniacoronata,oat,differentiallines

Session2Poster36TheinfluenceofsoiltillageandcropmanagementintheagroecosystemsonsoilfungistasisagainstFusariumgraminearum.SkaidreSuproniene1,GrazinaKadziene2,AgneVersuliene2,DonatasSneideris3,Algirdas Ivanauskas3,JurgitaKelpsiene4,NeringaRasiukeviciute41Lithuanian Research Centre for Agriculture and Forestry, Institute of Agriculture, Department ofPlantPathologyandProtection,Institutoal.1,58344,Akademija,Kedainiaidistr.,Lithuania2Lithuanian Research Centre for Agriculture and Forestry, Institute of Agriculture, Department ofCropandSoilManagement,Institutoal.1,58344,Akademija,Kedainiaidistr.,Lithuania3NatureResearchCentre, InstituteofBotany,LaboratoryofPhytoviruses,Akademijosstr.2,08412,Vilnius,Lithuania4Lithuanian Research Centre for Agriculture and Forestry, Institute of Agriculture, Laboratory ofMicrobiology,Studentustr.15A,53362,Akademija,Kaunasdistr.,Lithuania

Recently a rapid spreadofFusariumgraminearum, themain causal agentof Fusariumheadblight(FHB) of cereals, has been observed inNorthern Europe. The pathogen drift to theNorth ismostlikelypromotedbychangesinclimateandfarmingpractices.Increaseofminimumtillage,continuouscropping of cereals or extremely poor rotations in many regions, along with warming climate,enables themanifestation of FHB on epidemic scale. The primary source of FHB infection is hostplant residues remaining in soil. Therefore the establishment of F. graminearum in differentagroecosystemsmay strongly dependon the soil capacity to suppress pathogendevelopment andsurvival. The aim of this study was to evaluate the influence of different soil tillage and cropmanagementsystemsonsoilfungistasisagainstF.graminearum.Soil sampleswerecollected three timesduring theplantgrowth season, fromsix long-termcerealbasedcroprotation field trialsand froma long-termexperimentsetupunderconventional tillage,reduced tillageandno-tillagemanagement. Soil fungistasiswasevaluated in termsof reductionofradialgrowthofF.graminearumininvitroassayandexpressedinpercent.Differences in soil fungistasis, among treatments investigated, in most cases were statisticallyinsignificant. F. graminearum growth on unfumigated soil was reduced by 80% compared tochloroform fumigated soil in the tillage management experiment and by 70% - in the fields ofdifferentcroprotations.Keywords:croprotation,fungistasis,soilsuppresion,tillageminimisation

Session2Poster37ReactionofstembasediseasescausativeagentstofungicidesandimpactofagronomicpracticestoeyespotincidenceoncerealsIvanaSvačinová1,PavelMatušinsky1,LudvíkTvarůžek11AgrotestFyto,Ltd.,Havlíčkova2787/121,76701Kroměříž,CzechRepublicThe control practice of foot-rot diseases is mostly based on the use of fungicides. In 18-year´smonitoringweanalysedthemainfactors influencingfoot-rotdiseasesoccurrence.ThehighestrateofM.nivaleinfectionwasfoundin2006(37.0%)andthelowestonein2014(13.0%).Oculimaculaspp.occurrencewasvariablebetweenyearswithmeanlevelaround6.0%,maximum(7.6%)in2008andminimumwhichdidnotexceed1.0%in2012.Earlydateofsowingandwheatasprecedingcropwerethemainfactorsincreasingeyespotinfection.This research was supported by Ministry of Agriculture of the Czech Republic, projects No.QJ1530373,QJ1310091andRO0211.Theresponsesofbothpathogenstoprochlorazwereassessedinperiod2012–2016.BothM.nivaleandOculimaculaspp.populationsweresensitivetoprochlorazwithmeanlevelsbetween0.01-0.06μgml-1and≤0.1μgml-1respectively.Keywords:cereals,fungicideresistance,agronomicpractices,precedingcrop,sowingterm

Session2Poster38PlasmodiophorabrassicaepropagatesatdifferentrateinWinterOilseedRapecroprotationsAnn-CharlotteWallenhammar1,2,LenaEngström2,JohanRoland2andAndersJonsson2,3

1RuralEconomyandAgriculturalSociety,HSKonsultAB,POBox271,SE-70145,Örebro,Sweden2PrecisionAgricultureandPedometrics,DepartmentofSoilandEnvironment,SwedishUniversityofAgriculturalSciences,POBox234,SE-52323Skara,Sweden3ResearchInstitutesofSweden,RISE,AgrifoodandBioscience,POBox63,53321Skara,SwedenClubroot caused by the soil-borne obligate endoparasite Plasmodiophora brassicae is nowconsidered as themost serious disease inOilseedRape production in several European countries,associatedwithappreciableyield losses.Outbreaksofclubrootarecontinuouslyreported inwinteroilseed rape (WOSR) in Swedenwith a sharp increase in 2016. Crop rotation is one of themostvaluabletoolsfordiseasemanagement.TheobjectivesweretostudytheinfluenceonyieldofWOSRincroprotationswhenWOSRwasgrownatdifferenttimegaps.WOSRwasgrownevery(a)second,(b)thirdand(c)sixthyearrespectivelyinthree7-yearrotationsalternatingwithwinterwheat.TwofieldtrialswereestablishedatexperimentalfarmsinsouthwestSwedenandWOSRwasgrownyear1and7.AtoneexperimentalsiteWOSRwasreplacedbyspringoilseed rape year 7. Analyses of the propagation of inoculums ofP. brassicae were performed byqPCR.ThesignificanthighestyieldofOSRwasmeasuredintreatmentswithWOSRgrowneverysixthyearandoutyieldedWOSRgrowneverysecondandeverythirdyearby37%and18%respecievely.P.brassicaewasnotdetectedinsoilsampledattheonsetofthestudy,whilea largemultiplicationisrecordedattheendofexperimentsfortreatmentsaandb.,whilethelevelwaslowintreatmentc.TheresultsclearlyshowthatP.brassicae (asmeasuredbyfgDNAg-1soil)propagatesinfieldswithhighintensityofOSR,despiteinoculumswerebelowdetectionlevel,andpointsattheadvantagetoregularlyusesoilanalysisasamanagementtoolinOSRproduction.Keywords:WOSR,yieldloss,Plasmodiophorabrasssicae,qPCR

Session2Poster39DNAextractionandPCRamplificationmethodsuitableforherbarium-storedspecimentsofPhoma-likefungi.MariaM.Gomzhina1,PhilippB.Gannibal2All-Russian InstituteofPlantProtection,Saint-Petersburg,Pushkin, shossePodbel’skogo3,196608,RussianFederationHerbarium collections are the enormously important biological resources for studies of fungalbiodiversity. The aim of this study was modification of DNA extraction method for herbariumspecimens of the pycnidial fungi Phoma s.l. for next sequencing and molecular-phylogeneticstudying.Plant samples for this studywereobtained fromHerbariumcollectionofphytopathogenic fungiatAll-Russian Institute of Plant Protection (LEP). The collection year of specimens ranged from 1897to1959.Topulverizeplant’stissuewithfungalstructuresandglasssand,twoball-mills(RetschMM400andFastPrep24)wereusedat30Hz/s.for10and6minutesrespectively.IsolationoftotalcellularDNAwascarriedoutusingCTAB-basedprotocolwithminormodifications.TheITSregionwasamplifiedaccordingtomethodofNested-PCRusingouter(NSA3/NLC2)andinner(NSL1/NLB4).TheNested-PCRwas successful formost samples, only for few samples reaction didn’t provide uswithanyPCR-products.SuccessfulPCRresultedin2bandsofapproximately600-700b.p.and900-1000b.p.Bandswith larger sizeareprobablynon-specificproductofamplificationofplant’sDNA.Bandswithsize600-700b.p.representspecificampliconsoffungalDNA.Particularnested-PCRandascomycete-specific primers helped to decrease amplification of plant’s DNA and has resulted inpreferenceofamplificationoffungalDNA.Sequencingof ITS-regionwaseffective for 8 samples andhas resulted in 600-700b. p. fragments.WithhelpofBLAST-toolwithobtaineddata,4samplesweredefinedtospecieslevel(Chaetopyrenapennicillata,Leptosphaeriamaculans,Boeremiaexigua),therest4samplesweredefinedtogenericlevel(Phomasp,Boeremiasp.).Theoldestsequencingspecimenwasdated-by1897(120years-old).The proposed modified CTAB-method to extraction DNA from herbarium material of Phoma-likefungiandoptimizedamplificationprotocolgenerallyareregardedtobereliableandusefultostudyherbarium-storedphomoidfungiwithhelpofmolecularmethods.PresentedworkwassupportbyRussianScienceFoundation(project14-26-00067).Keywords: mycological herbarium, biodiversity, phytopathogenic fungi, pycnidial fungi, DNAextraction,Phoma

Session2Poster40TomatoyellowleafcurlvirusdetectionfromcommerciallyavailabletomatoseedsEui-Joon Kil1, Jungho Park1, Eun-Young Choi1, Hee-Seong Byun1, Chang-Seok Kim2, Ji-Kwang Kim3,Hong-SooChoi2andSukchanLee1

1DepartmentofGeneticEngineering,SungkyunkwanUniversity,Suwon16419,Korea2Crop Protection Division, National Institute of Agricultural Sciences, Rural DevelopmentAdministration,Wanju55365,Korea3Research and Development Bureau, Chungcheongnam-do Agricultural Research & ExtensionServices,Yesan32418,Korea Tomatoyellowleafcurlvirus(TYLCV)isoneofthemostwell-knowntomato-infectingbegomovirusesand transmitted by sweet potato whitefly Bemisia tabaci. Seed transmission has previously beenreportedforsomeRNAviruses,butTYLCVhasnotpreviouslybeendescribedasaseed-bornevirus.In 2013 and 2014, without whitefly-mediated transmission, TYLCVwas detected in young tomatoplants germinated from fallen fruits produced from TYLCV-infected tomato plants in the previouscultivatingseason.Inaddition,TYLCVwasalsodetectedinseedsandtheiryoungseedlingsofTYLCV-infected tomatoplants thatwere infectedbybothviruliferouswhitefly-mediated transmissionandagro-inoculation.Theseed infectivitywas20-100%,respectively,andtheaveragetransmissionrateto seedlings was also 84.62% and 80.77%, respectively. Sequences were identical among TYLCVgenomes of inoculum and those isolated from seeds and their seedlings. When tomato plantsgerminated from TYLCV-infected seeds (as donor plants), non-viruliferous whiteflies (as insectvectors) and healthy tomato plants (as receiver plants)were placed in awhitefly-free insect cagetogether. TYLCV was detected from whiteflies as well as receiver tomato plants six weeks later.Infectivity tests for TYLCVwere also performed using 250 kinds of commercially available tomatoseeds on sale at overseamarkets from24 countries inAsia, Africa, America, Europe andOceania.Among them, TYLCVwas detected from 38 kinds of seeds by PCR, and TYLCV genome sequenceswereconfirmedfromtheiramplicons.Forthedisseminationtest,eachseedwasplantedinaninsect-free condition. Seedlings from 9 cultivars were identified as TYLCV-infected (showing 8-100%dissemination rates).Whitefly-mediated TYLCV transmission from tomato plants germinated fromTYLCV-infected commercial seeds to other healthy tomato plants was also confirmed. Takentogether, TYLCV can be transmitted via seeds and tomato plants germinated from TYLCV-infectedseedscanbeservedasaninoculumsourceofTYLCV(orasaTYLCVreservoir).Keywords: Begomovirus, geminivirus, reservoir, seed transmission, Tomato yellow leaf curl virus,whitefly

Session2Poster41PathotypingMelonNecroticSpotVirus(MNSV)inmelonSophie Perrot1, Carole Constant2, Mireille Buisson3, Isabelle Justafré4, Sylvia Chanéac5, ValérieGrimault1,FrançoisBertrand6,GrégoriBonnet7,DenisLosdat8,andThierryJaunet9

1GEVES,RueGeorgesMorel,49071Beaucouzé,France2SakataVegetablesEurope,DomainedeSablas,Ruedumoulin,30620Uchaud,France3GautierSemences,Routed’Avignon,13630Eyragues,France4HM.Clause,LeMasSaintPierre,LaGaline,13210,SaintRémydeProvence,France5TakiiFrance,660chemindelaCrau,13630Eyragues,France6Monsanto,ChemindeRoquemartine,13670Saint-Andiol,France7SyngentaFrance,346RoutedesPasquiers,84260Sarrians,France8Rijk-ZwaanFrance,LaVernède,30390Aramon,France9HM.Clause,1cheminduMoulindesRonzières,49800LaBohalle,FranceThe Carmovirus Melon Necrotic Spot Virus (MNSV) is seriously affecting melon greenhouseproductioninSpain.Sincetheearly2000s,recessiveresistancegenensvhasbeendeployedinmeloncommercial varieties and MNSV resistance declaration has been included in European varietyregistrationprocessformelon.Followingresistancedeployment,onlyfewresistancebreakingMSNVisolateshavebeenisolatedandcharacterized.However, inthe lastfewyears,an increasingnumbersofMNSVsymptomshasbeenreported in melon commercial fields grown with MSNV resistant varieties in Almeria region,suggestingthespreadingofresistancebreakingMNSV.Thisresistanceovercominghashadadirectimpact on MNSV resistance claim in commercial varieties, and emphasizes the need to definepathotypesforthisvirus.In order to confirm the overcoming of the resistance conferred bynsv gene,MNSV isolates havebeen isolated fromMNSV typical symptoms collectedon commercial resistant varieties inAlmeriaregionfrom2011to2013.Virulenceofsixrepresentativeisolateshasbeencomparedtothatoftheofficial reference isolate, on a panel including both the resistant (VIRGOS) and susceptible(VEDRANTAIS) controls used in the variety registration process, and seven commercial varietiescarrying the nsv resistance gene. This characterization has been carried out through an inter-laboratoryring-testandaccordingtotheofficialprotocolusedforvarietyregistration.All sixMNSV isolates expressed typical systemic symptomson all testedmelonmaterial, includingresistant control and the seven resistant commercial varieties, whereas the official MNSV isolatecouldinfectonlythesusceptiblecontrol.TheseresultsconfirmvirulenceofsomeMSNVisolatesonnsv resistancegene. It is thenproposed todefineaspathotype0 isolatesavirulent tonsv gene (inVIRGOS) and pathotype 1 isolates virulent to nsv gene. This MNSV pathotype classification willsupportamoretransparentclaimofresistancecarriedbycommercialvarieties.Keywords:MNSV,melon,resistance,pathotype

Session2Poster42Development of a resistance test of squash to Cucumber mosaic virus (CMV), Zucchini yellowmosaicvirus(ZYMV)andWatermelonmosaicvirus(WMV)SophiePerrot1,SandrineHoudault1andValérieGrimault1

1GEVES–Pathologylab,25rueGeorgesMOREL,CS90024,49071Beaucouzécedex,FranceThree viruses transmitted by aphids (no persistent transmission) are responsible of strong yieldlosses on squash crops in France and around the world: Cucumber mosaic virus (CMV), Zucchiniyellowmosaicvirus(ZYMV)andWatermelonmosaicvirus(WMV).Foreachvirus,differentbiologic,serologicandmoleculargroupsweredescribed.TheresistanceispolygenicforCMV,ZYMVandwithamajorgeneforWMVbutalwayswithanintermediateresistance.Theevaluationof squashresistance tovirusesand theassessmentof symptoms iscomplexdue tothe partial and not complete resistance and the quantitative results observed. It is important todescribe the different levels of virulence of virus strains and to identify the different levels ofresistanceof squashvarieties. Thereare currentlynoharmonizedprotocols todefine the levelsofpartialresistanceofsquashtotheseviruses.Theaimofthisproject, incollaborationbetweenGEVES,INRAandbreedingcompanies(HMClause,Gautier,Monsanto, Rijk Zwaan, Sakata, Syngenta) was to acquire a better knowledge of levels ofsquashresistanceandtodefinerobustresistanceteststoviruses.BasedontheINRA’scharacterizedcollectionof strainsof viruses, three strainsper virus (withdifferent virulencesand fromdifferentgroups) were selected. These strains and different protocols from INRA and partners (stage ofinoculation, optimal conditions of test, notation scale) were compared, on a panel made up ofcommercialvarietieswithdifferent levelsofresistance, todefineforeachvirusarobustresistancetest.Onestrain,allowing todifferentiate thedifferent levelsof resistanceofvarieties,wasselectedpervirus.Foreachvirus,aresistancetestandreferencematerials (strainandcontrols)werevalidated.ThereferencecollectionofGEVESwascharacterizedforthethreeviruses.Thesenewprotocols,usedforCTPSregistration,willbeproposedtoCPVOandUPOVforaharmonizedused.Keywords:squash,viruses,partialresistance,protocol

Session2Poster43CORKYRES:Developmentofaresistancetestoftomatoandroot-stocktoPyrenochaetalycopersiciSophiePerrot1,MathildeBuffard1,CélineAndro2andValérieGrimault1

1GEVES–Pathologylab,25rueGeorgesMOREL,CS90024,49071Beaucouzécedex,France2GEVES–BioGEVES,25rueGeorgesMOREL,CS90024,49071Beaucouzécedex,FrancePyrenochaetalycopersiciisthecausalagentofcorky-root,animportantdiseaseontomatoandroot-stockswhichcausesbrowncorkylesionsonrootsandisresponsibleofyieldlosses(from40%to70%incaseofstronginfection).Thecontrolofcorky-rootismainlybasedontheuseofresistantvarieties.The resistance recessive gene py-1 was introduced in tomato (L. esculentum) from Lycopersicumperuvianum.A protocol of evaluation of resistance to corky-root is present in the UPOV guidelines and in theCPVO technical protocols. But the described protocols rely on a sowing or transplantation incontaminatedsoilandnotationatfloweringorfruitsmaturitystage.Sothistestislonganddifficulttoperform.Moreovertheisolatesandcontrolsofthisprotocolhavenotbeenvalidated.The aim of this project, in collaboration betweenGEVES and six breeding companies (HM-Clause,Gautier,RijkZwaan,Sakata,Syngenta,Vilmorin)wastoacquireabetterknowledgeofthispathogenand todefinea robust resistance test.A strain collectionwasestablished from literatureand fieldisolates. Each strain was characterized for its morphology (appearance, growth), its physiology(adaptationtotemperatures)andgenotype.Apathogenicitytestwasdevelopedtodefinethelevelofvirulenceofeachstrainand to select reference isolate.Different inoculums, conditionsof tests,stagesof inoculationandnotationscaleswerecomparedtodefinearesistancetestontomatoandroot-stockscontrolsandonapanel,definedbythesteeringcommittee,ofcommercialvarietieswithdifferentlevelsofresistance.A collectionof16 characterized strainswasmadeupandconservationconditionsweredefined.Aresistance test and referencematerial (strain and controls)were validated. This newprotocolwasacceptedasanewcharacteristicbyCTPSforregistrationofvarietiesandwillbeproposedtoCPVOandUPOV.Keywords:tomato,root-stock,resistance,corkyroot

Session2Poster44SporetrapandmoleculardetectionoffungiAnnaBerlin1,JohannaBoberg1,andJonasOliva1

1SwedishUniversityofAgriculturalSciences,Dept.ForestMycologyandPlantPathology,Box7026,S-75007Uppsala,SwedenSeveralplantpathogenic fungiaredispersedbywind.Theaimofthisprojectwastocreateabasicunderstanding of the fungal communities present in air in agricultural and forest landscapethroughoutthegrowingseasons.Thiswasdonebycollectionofsporetrapcatchesinthefourmainplant production areas of southern Sweden. The spore sampleswere collectedusing two typesofsporetraps,oneactivetraprepresentedbytheionicsuctiontrapandonepassivefunneltrap.DNAwasextractedfromeachsampleandthefungalcommunitywasstudiedbasedontheITSregionby applying both IlluminaMiSeq and PacBio RS II sequencing techniques. The fungal communitiesstructures were investigated and the detected operational taxonomic units (OTUs) assigned to aspecieswherepossible.Thelistofspeciesdetectedwascomparedtoreportsofdiseasedevelopmentinthefieldcropsinthesurroundingsoftherespectivesporetraps.Theresultsformthisstudycouldbeusedtodevelopapredictionsystem,whichpotentiallycouldbeanimportanttoolindiseaseforecastingandriskmonitoring.Thetimingofthedetectionoftheplantpathogenic fungi in the traps depends on the biology of the fungus. As an example, rust fungi(Puccinia sp.) were first found in the elevated traps, and later at canopy leave, which may beexplainedbydispersalbylongdistancedispersal.Sporesofergot(Clavicepspurpurea)wasfirstfoundat thecanopy leveland later in theelevated traps,whichmaybeexplainedby theoriginof thesespores. C. purpurea sclerotia in the soil germinate via perithecia and air-borne ascospores. Weexpect that results form thisprojectwillbeuseful fordevelopmentofdiseasemonitoringand riskmanagementofplantdiseasesinthefuture.Keywords:ITSregion,fungalcommunity

Session2Poster45DescribingthecommerciallifecycleofbreadwheatvarietiestostudytheinfluenceofyellowrustepidemicsinFranceRémi Perronne1, Solène Diguet1, Emma Girard1, Marc Leconte2, Philippe du Cheyron3, Claude deVallavieille-Pope2,JérômeEnjalbert11UMRGQE–LeMoulon,INRA,UnivParis-Sud,CNRS,AgroParisTech,UniversitéParis-Saclay,91190,Gif-sur-Yvette,France2UMRBIOGER,INRA,AgroParisTech,UniversitéParis-Saclay,78850Thiverval-Grignon,France3Arvalis-InstitutduVégétal,routedeChâteaufort,ZAdesgraviers,91190Villiers-le-Bâcle,FranceYellowrustcausedbyPucciniastriiformisf.sp.tritici(Pst), isoneofthemostimportantdiseasesofbreadwheat in France, especially known to have led to the rapidwithdrawal from themarket ofsusceptiblevarietiesduringthelastdecades.However,nostudyhassofarexaminedhowpathogenpressurescouldimpacttheadoptionofvarieties.In this study, we proposed a set of indicators of the commercial life cycle of a variety, and weemphasized their relevance todistinguishbetween susceptible varieties and varieties presenting adurable resistance in several Frenchagricultural regionsover the lastdecades.We focusedon thesuccessive periods characterized by yellow rust epidemics, identified homogeneous agriculturalregionsintermsoflevelofyellowrustpressure,Pstpathotypesandlandscapevarietalcomposition,considering varieties sown on at least 10,000 ha. We applied principal component analyses andclusteringmethodsonasetofnon-redundantindicatorsofthecommerciallifecycleofvarieties.Wefirstconsideredtheepidemicperiod1987-1991characterizedbythebreakdownofYr6andYr9race-specific resistance genes in France. In thenorthwest coastline area, highly affectedby yellowrustpressures,weidentifiedeightvarietieswitharapidwithdrawalfromthemarket.Thesevarieties,either susceptible before the epidemic or concerned by the breakdown of resistance genes,appeared to be substitutable by highly resistant varieties available for the same end-use over theperiod.Wethenconsideredthewholeperiod1980-2015withtheaimofidentifyingvarietiescharacterizedbydurableresistanceagainstyellowrustbycross-checkinginformationonthecommerciallifecyclewithinformationonpostulationofresistancegenes,changesinadult-plantfieldresistancelevelandpedigrees.Following this procedure, a promising perspective for breeders could be a list of varieties withdurableresistancepromotingthelong-termdurabilityofyellowrustcontrol.Keywords:Pucciniastriiformis f. sp.tritici,Triticumaestivum, race-specificresistancegene,durableresistance,yellow(stripe)rust

Session2Poster46DiversityanalysisofXanthomonasaxonopodispv.manihotis(Xam)inMaliMoussaKante1,KarimDagno2,CarolinaFlores1,LaurenceBlondin3,IssaWonni4,RalfKoebnik1,ValérieVerdier1,OusmaneKoita5,ChristianVernière3andBorisSzurek1

1IRD,Cirad,UnivMontpellier,IPME,Montpellier,France2IER,CRRA‘‘AgronomicRegionalResearchCenterofSotuba’’AgronomicRegionalAgrogomicCenterofSotuba,SorghumProgramm,PhytopatholgyLaboratory,Sotuba,Bamako,Mali3Cirad, UMR BGPI ”Biology Genetic of Population Interaction”, TA A-54-K/Cirad-BIOS, CampusinternationaldeBaillarguet,34398Montpelliercedex5,Montpellier,France4INERA,NationalEnvironmentandAgronomicInstituteforResearch,Bobo-Dioulasso,BurkinaFaso5UniversitédesSciencesTechniquesetTechnologiques,FacultédesSciencesetTechniques, LBMA,Bamako,Mali

Understandingthediversityofpathogenpopulationsinaregionalscaleiscrucialinordertodevelopstrategies for sustainable control of plant diseases. Our work deals with the study of the plantpathogenic bacteriumXanthomonas axonopodispv.manihotis (Xam),which is the causal agent ofcassavabacterialblight(CBB).CBBoccursinthetropicsincludinginWest-Africawhereithasrecentlybeen reported inBurkinaFasoand IvoryCoast, sometimes imposing severeyield losses tocassavaproducers.ThestatusofCBBinMalistillremainsunstudied.Tofillthisgap,preliminarysurveyswereconductedinDec.2015intwocassavaproductionregions.Nearly80leafsampleswereprocessed,leadingtotheisolationofabout50Xamcandidateisolates.Of these, 40 strains of Xanthomonas axonopodis pv.manihotiswere validated throughmoleculardiagnostic tests and pathogenicity assays. This study represents the first demonstration for thepresenceofCBBinMaliandenabledtoinitiatetheestablishmentofafirstcollectionofnationalXamstrains. Secondly, aMLVA strategy (Multiple-Locus Variable number tandem repeat Analysis) wasdevelopedbasedontheanalysisof14microsatellites(VNTR)inordertoassessthegeneticdiversityof the40strains isolated fromour tworegionsunderstudy,BamakoandSegou,whichare250kmdistant.Ourresultsdemonstrateagreaterdiversity inpopulationsoftheBamakoregionrelativetothatofSegou,withreferencetothevaluesofdiversityindices.Also,comparisonofallstrainsofMaliwithacollectionof215strains fromborder regions inBurkinaFasosuggestedthat theMalianstrainsarelessdiversified.FurthersurveyscoveringmostareasofcassavaproductioninMaliareinprogresstobetterunderstandXaminvasionroutesinthiscountry.

Keywords:Cassava,Xanthomonas,cassavabacterialblight,Mali,moleculardiagnostic,MLVA-14

Session2Poster47Pantoeaananatiscausinggraindiscolorationiswidelydistributedintherice-growingfieldsoftherepublicofKoreaKimSang-Min1,RaJi-Eun,SeoSuJwa1,LeeBongChoon1,andChoiNakJung1

1CropFoundationResearchDiv.,NationalInstituteofCropScience,WanjuGun,Jeollabuk-do,55365,RepublicofKoreaKeywords:Pantoeaspp.,graindiscoloration,P.ananantis,distributuition,phylogeneticanalysis

Session2Poster48InfluenceofmaizegenotypesonFusariumspeciesfrequenciesandgrainmycotoxincontaminationindiverselocalitiesofPolandMonikaUrbaniak1,ŁukaszStępień1,ArturTrzebny1,SewerynFrasiński2,ElżbietaCzembor2,AgnieszkaWaśkiewicz31InstituteofPlantGenetics,PolishAcademyofSciences,DepartmentofPathogenGeneticsandPlantResistance,Strzeszyńska34,60-479Poznań,Poland2PlantBreedingandAcclimatizationInstitute-NRI,Radzików,05-870Błonie,Poland3UniversityofLifeSciences,DepartmentofChemistry,WojskaPolskiego75,60-625Poznań,PolandMaize has become one of the essential crops for Poland. It has several advantages compared tocommonlyusedwheat,mainly in termsofyieldanddiverseuse for foodsand feeds.Likeallothercereal crops, it suffers froma range of fungal diseases and Fusarium species are among themostdamaging pathogens, producing a plethora of toxic secondarymetabolites, accumulated inmaizegrain and, thus, introduced into the food chain. Humidity and temperature are the factorsdetermining the disease severity but geographical conditions, plant genotype and local pathogenpopulationsplayalsoimportantroles.We analysed Fusarium populations infecting several maize genotypes harvested in 2015 seasonundertheconditionsofelevenlocalitiesinPoland.Grainsfrominfectedcobswereplatedonwateragarmediumandall fungalstrainswererecovered.Over400Fusarium isolateswererecordedandidentifiedusingmoleculartechniques(tef-1αsequenceanalysisandspeciesspecificSCARmarkers).Consequently, grain contamination with typical mycotoxins was evaluated using liquidchromatographymethods.Species frequenciesdepended stronglyonmaize genotype (earliness group), geographical locationand weather conditions. Fusarium verticillioides was the most abundant species in all localities,followedbyF.proliferatum(47%and30,5%ofallFusariumisolates,respectively).Anumberofminorspecies was also present: F. temperatum (6,3%), F. graminearum (5,8%), F. poae (5,6%), F.subglutinans (1,9%),F.sporotrichioides (1,5%),F.equiseti (0,5%),F.oxysporum (0,5%),F.culmorum(0,2%)andF.tricinctum(0,2%).InallmaizesamplesFusariummycotoxins(fumonisins,deoxynivalenol,zearalenone)wereanalysed–afterextractionandpurification–usingchromatographictechniques(HPLC/PDA/FLD).OuranalyseshaveshownlocalvariabilityofFusariumcommunitiesinrelationtohostgenotypesandweatherconditions.Concentrationlevelsofanalysedmycotoxinswerealsovariedanddependedonlocationandmaizegenotype.Thepartof thisresearchwasfinanciallysupportedfromthePolishNationalScienceCentreproject2014/15/B/NZ9/02169Keywords:deoxynivalenol,fumonisins,Fusarium,maize,zearalenone

Session2Poster49One-stepmultiplexreversetranscription-polymerasechainreactionforthesimultaneousdetectionforbarleyvirusdiseasesBongChoonLee1,SooYeonChoi1,JuYoungBae1,SangMinKim1,Ji-EunRa1,NakJungChoi1andKiDoPark11NationalInstituteofCropScience,RuralDevelopementAdministration,Wanju55365,KoreaKeywords:BaYMV,BaMMV,BYDV,Simultaneousdetection

Session2Poster50InvestigatingtheAlternariadiseaseprogressionandspeciescompositiononpotatoinBelgiumM.Vandecasteele1,S.Landschoot1,M.Höfte2,S.DeSaeger3,K.Audenaert1andG.Haesaert1

1DepartmentofAppliedBioscienceEngineering,FacultyofBioscienceEngineering,GhentUniversity,ValentinVaerwyckweg1,9000Ghent,Belgium2DepartmentofCropProtection,FacultyofBioscienceEngineering,GhentUniversity,CoupureLinks653,9000Ghent,Belgium3DepartmentofBioanalysis,FacultyofPharmacologicalSciences,GhentUniversity,Harelbekestraat72,9000Ghent,BelgiumAlternaria species are a serious threat for potato cultivation since heavy infections can lead tosignificantyieldandqualitylosses.Theinfectioncausesnecroticsymptoms,whichcannotbevisuallydistinguished.Over thepast years,Alternaria onpotatohasbecome increasingly important inNWEurope.Althoughtheexactcauseforthisemergingproblemremainselusive,itmightbeattributedtothecombinedeffectofclimatechange,areduceduseof the fungicidemancozeb, the increasedspecificityofactiveingredientstocontrolPhytophtorainfestansandtheproductionofhigh-yieldingsusceptible cultivars. Furthermore, little is known about the Belgian Alternaria populationcompositionandthecontributionofbothA.solaniandA.alternatatothedisease.Themaingoalofthis research is to identify theprimarycausalagentsofAlternariadiseaseonpotato, todetermineinter- and intraspecific diversity within the Alternaria population in Flanders, and to unravel thecomplexinteractionbetweenstress-relatedhormonesandtheAlternariainfection.Toachievetheseobjectives, 22 locationsweremonitored throughout Flanders during the growing seasons of 2014and 2015. Results of this disease survey unequivocally show that the disease incidence and –pressureforbothseasonswaslowandthatcropsgrowninsandysoilsappeartobemorepronetoAlternariadisease.Inasecondpart,weidentifiedthepopulationcompositionatthespecieslevelondifferent time points during the growing season. Therefore, infected leaf samples were collectedfrom the field and using a microscopic and molecular approach, we concluded not only that A.arborescrens is the most abundant species present on infected leafs, but also that small-sporedspecieslikeA.alternataandA.arborescensratherthanthelarge-sporedA.solani,werepredominantat the beginning of the growing season. The disease escalated only when A. solani species wereaccumulating. Indeed, based on high-throughput virulence assays,we observed thatA. solaniwasmuchmore virulent thanA. alternata orA. arborescens.Next, a subset of isolateswill beused toinvestigate thecomplex interactionbetweenhost-specific toxinsandstress-relatedhormonessuchasethyleneorauxinsduringtheinfectionprocess.Indeed,previousresearchshowsthatethyleneisakeycomponentinupstreamsignalingofProgrammedCellDeathinducedbyhost-specificAAL-toxinintomato.

Session2Poster51PopulationstructureandhostspecializationinBotrytiscinereaAnne-SophieWalker1*,AlexMercier1,MargueriteCuel1, Jean-MarcPradier1,AdelineSimon1,MurielViaud1andPierreGladieux2

1UMRBIOGER,INRA,AgroParisTech,UniversitéParis-Saclay,Thiverval-Grignon,France2UMRBGPI,INRA,CIRAD,Montpellier,FranceTheBotrytisgenusencompassesmorethan30species,mostofthembeingabletoinfectarestrictedrangeofhostplants.Nevertheless,B.cinereaisconsideredageneralistpathogen,recentlyfoundtoinfect 586 plant genera. This makes B. cinerea an interesting candidate to study ecologicalspecialization.ThiswasfirstapproachedwhiledescribingpopulationstructureinFrance,fromstrainscollectedongreenhouse tomato, grapevine, blackberry, strawberry andhydrangea.Understandingthecausesofpopulationsubdivisionisoffundamentalimportance,asstudyingbarrierstogeneflowbetween populations may reveal key aspects of the process of adaptive divergence and, forpathogens, may help forecasting disease emergence and implementing sound managementstrategies.Populationgeneticsanalyses revealedaweakassociationbetweenpopulation structureand geography, but a clear differentiation according to the host plant of origin, and especially forgreenhouse tomato and grapevine. Host specialization was validated by cross-inoculationexperiments, carried out in vitro on detached leaves. Genomes from strains contrasting for theirspecialization on tomato, grapevine, blackberry and hydrangea were Illumina-sequenced andgenomicphylogenyanalysis confirmed the similaritiesbetweengenomes fromstrains collectedonthesamehost.Populationgenomicsanalysisusedtocomparevariationbetweenandwithinlinesofstrainsprovidedpromising setsofgenes thatmay inducehost specialization. Inaddition tocodingsequences,analysisofsmallRNAsfromstrainscollectedeitherontomatoorgrapevinewasinitiated.PreliminarysequencingdatasuggestthatthesetwopopulationshavecontrastedrepertoriesofsmallRNAs.FurtheranalysisandadditionalgenomesandsmallRNAsequencingarerequiredtodefinitelyidentifythedeterminantsofhostspecialization.OurfindingsopenupnewperspectivesfordiseasecontrolbymanagingBotrytishostsinagriculturallandscapes,andreinforcingprophylacticmeasures.Keywords: Botrytis cinerea, adaptation, population structure, host specialization, genomesequencing,cross-inoculationtest

Session2Poster52InvestigationofthefitnessofZymoseptoriatriticiinrelationshipwithfungicidetoleranceMyriamBomble1,AliSiah1,ConstanceTuffet2,PhilippeReignault3andPatriceHalama1

1ISA,InstitutCharlesViollette,EA7394-ICV,46bdVauban,BP41290,59014Lillecedex,France2BayerCropScience,16rueJean-MarieLeclair,CS90106,69266LyonCedex,France3ULCO,UnitédeChimieEnvironnementaleetInteractionssurleVivant(UCEIV),CS80699,62228Calaiscedex,FranceZymoseptoriatriticiiscurrentlythemainpathogenonwheatcrops,especiallyinNorth-WesternEurope,whereenvironmentalconditionsaresuitablefordiseaseestablishment.Diseasecontrolreliesmainlyontheuseoffungicidesandpartially-resistantcultivars,butthesemanagementstrategiesarefrequentlycircumventedinthefieldbythefungus,duetoitshighbiologicalfitnessdegree,resultinglikelyfromitsactivesexualreproduction.Althoughtheimportanceofthefitnessconcepttoperpetuatetheexistingfungicidesandtodevelopsustainableprotectionstrategies,itremainspoorlyunderstoodinZ.tritici.TheaimofthepresentprojectwasthustodecipherthefitnesscomponentsofZ.triticiinrelationshipwithfungicidetolerance.First,acollectionof240single-sporeisolatesofthepathogenwerecollectedin2016fromnorthernFranceandfingerprintedwithmicrosatellitemarkerstoselectuniquegenotypes.Onehundredhaplotypeswereselectedandareunderassessmentforresistancetosterol14α-demethylationinhibitor(DMI)andsuccinatedehydrogenase(SDHI)fungicidesusingphenotypicandmolecularassays.ResultswouldallowanupdateofresistanceleveltothesefungicidefamiliesinnorthernFrance.Furthermore,fungalstrainsbelongingtodifferentresistanceprofileswillbeselectedandexaminedfortheirfitnesscomponents(pathogenicitylevel,pathogenicitydeterminants,sporulationrate,etc.)andcompetitionability,usingcytological,biochemicalandmolecularapproaches.FindingswillprovidevaluableinformationaboutthefitnessofZ.tritici.Keywords:Zymoseptoriatritici,Mycospaherellagraminicola,wheat,fungicideresistance,fitnessdegree

SESSION3.EMERGINGDISEASESANDNEWINSIGHTSINTAXONOMYANDPHYLOGENYOFPLANTPATHOGENSINEVOLVINGGLOBALCONDITIONS

Session3Poster53

Genetic variabilitywithinSeptoria carviSyd. (Ascomycota) – a newpathogenof carawayCarumcarviL.EwaDorotaZalewska1,EwaDorotaKról1andAgnieszkaFurmańczyk11DepartmentofPlantPathologyandMycology,FacultyofHorticultureandLandscapeArchitecture,UniversityofLifeSciences,Leszczyńskiego7Str.20-069Lublin,Poland

Fungi of the genus Septoria (teleomorphMycosphaerella) belong to thewidespread pathogens ofcropsandwildplantsinallregionsoftheworld.ThespeciesSeptoriacarviisespeciallydangeroustocarawayplants. It causesseptoriosis.According to literature,S. cari, S.umbelliferarumandS. carviarerecognizedasthecauseofseptoriosisofcarawaycultivatedinEuropeancountries.ThepurposeofourstudywastoinvestigatewhetherS.carviisanewpathogenicspeciesofcarawayorperhapsitisoneof thedescribedspeciescolonizingotherplantsof the familyApiaceae. Identificationof thestudiedisolateswascarriedoutusingtheclassicalmethodsaswellasmolecularstudiesusingPCR.DNA was isolated from own isolates of S. carvi. The sequence of nucleotides rDNA of our ownisolates, obtainedduring the studyof four loci, i.e. regions ITS1- ITS2, actin (Act), calmodulin (Cal)andtranslationelongationfactor1-alpha(EF1-α)werecomparedwiththesequencesofreferenceisolates of Septoria species affecting other plants of the family Apiaceae. Phylogenetic analysisshoweddistinctdifferencesofthetestedisolates,whichallowedtotreatthemSeptoriacarvispeciesaffecting the above-ground organs of caraway Carum carvi L. The obtained results showeddifferences between the studied barcodes and indicated that translation elongation factor 1-αconstituted the best primer to identify S. carvi. This study is the first report on the geneticcharacteristics of the species S. carvi. The obtained sequences of S. carvi isolates have beensubmittedtotheGenBank.

Keywords:Septoria,caraway,PCR,geneticcharacteristic

Session3Poster54DiseasesofScotspine(Pinussylvestris)needlesinselectedareasofnorthernPolandMartaBełka1,FaizahN.Alenezi2,KatarzynaLisek1,RoksanaByszewska1,MałgorzataMańka1

1PoznańUniversityofLifeSciences,ul.WojskaPolskiego71c,60-628Poznań2UniversityofAberdeen,InstituteofBiologicalandEnvironmentalSciences,CruickshankBuilding,StMacharDriveAberdeen,AB243UUScotspine(Pinussylvestris) isacrutialecologicalbutalsoeconomiccomponentofPolishforests.Inrecent years incerased pine needle mortality rates have been observed around the country. Thisphenomenonisamatterofgreatconcernbecauseitappliestolargeareaofforeststands.Approximately 30-year-old pine stands have been chosen for the study and the area have beenexamined for two years, duringwhich symptomatic pine needles have been collected in differentconditionsasitisknownthatfoliarfungimayonlyproducefruitingbodiesduringaspecificseason.Duringthestudysymptomsconsistingofbrowndiscolorationofneedleshavebeenobserved inalltheareasstudied.Thecauseofthesymptomshasbeen identifiedasSphaeropsissapinea,and it isprobablethatclimateextremesinrecentyearshavecontributedtotheoccurrenceofthepathogento great extent. Other pathogens that have been isolated include: Lophodermium pinastri,Lophodermium seditiosum, Cyclaneusmaminus, Cenangium ferruginosum, Lophodermium pinastri,Sclerophomapythiophila,Alternariaalternata,Gremieniellaabietinaandmanyotherspecies.Keywords:Scotspine,Pinussylvestris,needles,disease,pathogen

Session3Poster55PathotypesofPlasmodiophorabrassicaeinPolandMalgorzataJedryczka1,WitoldIrzykowski1,JoannaKaczmarek1,NoorRamzi1,SzymonOrnatowski1

1InstituteofPlantGeneticsPolishAcademyofSciences,Poznań,PolandPlasmodiophora brassicae is a protist causing clubroot, which is a damaging disease of vegetablebrassicasandoilseedrape,alsofoundoncruciferousweeds.Stilllittleisknownaboutthepopulationofthispathogenandthedistributionofpathotypesworldwide.Comparisonbetweenthepathotypesdescribed in the literature are impossible due to different systems of classification. In this studypathotypesfoundinPolandhavebeenstudiedusingthreemostpopulardifferentialsets,describedby Williams (1966), Buczacki et al. (1975) and Somé et al. (1996). Moreover, we have used cv.Mendel–thefirstcultivarofoilseedrapeconferringthemajorresistancegenederivedfromBrassicarapa,whichwasadded to thestandarddifferential setofSomé.Thenumberofpathotypesvarieddependingon thedifferential systemand thresholdused. Itwas found thatmostof thepathogenpopulationinPolandiscomposedofnearlyequalnumbersofthepathotypesP1andP3,butasmuchas38%of isolatescouldbreakdowntheresistancepresent incv.Mendel.Basedontheremainingidentificationsystemsuptoninepathotypeshavebeenfoundbynow.SequencesoffragmentsITS1-5,8S-ITS2 of 22 isolates of P. brassicae belonging to different pathotypes showed no sequencepolymorphisms inregionsof theribosomalDNA,wholesequenceof5,8Sandstudiedfragmentsof18S and 28S. Asmall polymorphism was found in ITS1 and ITS2 sequences, but it only partiallycoincidedwiththedivisiontopathotypes.Keywords:clubroot,Plasmodiophorabrassicae,oilseedrape,InternalTranscribedSpacer

Session3Poster56NovellineageoftumorigenicbacteriacausingcanegallofblackberryNemanjaKuzmanovic1,JoannaPuławska2,KorneliaSmalla1

1JuliusKühn-Institut, Federal ResearchCentre forCultivatedPlants (JKI), Institute for EpidemiologyandPathogenDiagnostics,Messeweg11-12,38104Braunschweig,Germany2ResearchInstituteofHorticulture,ul.Konstytucji3Maja1/3,96-100Skierniewice,PolandTumorigenicbacteriabelonging to the familyRhizobiaceaeare responsible forcrowngallandcanegall disease of various fruit species and may cause significant economic losses in orchards andnurseries.While tumorigenicstrainsofAgrobacteriumtumefaciens speciescomplexandRhizobiumrhizogeneswerepredominantly isolated fromplantsshowingcrowngall symptoms,AgrobacteriumrubiisrecognizedasacausalagentofcanegalldiseaseofRubusspp.Theobjectiveofthisstudywastocharacterizeatypicalstrainsisolatedfromblackberryshowingcanegallsymptoms.Bacterial strains were isolated from tumor tissue of diseased plants collected in two localities inSerbiaduring2015-2016.BasedonPCRanalysis, itwasdeterminedthat theycarry tumor-inducing(Ti)plasmid.Tumorigenicityofstrainswasconfirmedbypathogenicityassayonseveral testplants.Phylogenetic reconstructionbasedon16S rRNAgeneplaced themwithin thegenusRhizobium,onthesamebranchasRhizobiumtubonense.Multilocussequenceanalysis(MLSA)basedonthepartialsequencesofatpD,recAandrpoBhousekeepinggenes,aswellaswhole-genomecomparisonsandcalculation of average nucleotide identity (ANI) indicated that these strains represent a newRhizobiumspecies,withRhizobiumtubonenseastheirclosestrelative.Overall, our results revealed the existence of additional taxonomic diversity within tumorigenicbacteria. Naturallyoccurring tumorigenic strains were so far detected only within the genusAgrobacterium, and species Allorhizobium vitis and R. rhizogenes. Additional investigations areongoinginordertofurthercharacterizeanovelgroupofatypicalstrains.Keywords:canegall,blackberry,tumorigenicbacteria,Rhizobiaceae,phylogeny,taxonomy

Session3Poster57EmergenceofXylellafastidiosainEurope:detectiononvectorsBrunoLegendre1,ValérieOlivier1,DimitriMolusson1,VirginieJuteau1,AntoineSainte-Luce1,FrançoisePoliakoff11Laboratoiredelasantédesvégétaux,Anses,7rueJeanDixméras,49044Angers,France.XylellafastidiosaisaxylemlimitedbacteriumoriginatingfromtheAmericaspresentonawiderangeof host plants. X. fastidiosa is now an emerging organism in Asia and Europe Since 2013, thesubspeciespauca ispresentonolive tree in Italyandsince July2015twostrainsof thesubspeciesmultiplexhavebeenidentifiedinCorsicaonPolygalamyrtifolia,thenonotherornamentalandwildplants. In October 2015 the first outbreak was identified in the French Riviera region with thepresenceof the subspeciesmultiplex. The subspeciespaucawasdetected inMenton in2016. Thesubspecies fastidiosa was found onOleander in Germany and the subspecies fastidiosa,multiplexandpaucaarepresentsonBalearicIslands.Since2015,aFrenchofficialmethod(MA039availableonwww.anses.fr)allowsthedetectionofX.fastidiosaonplantsbyrealtimePCRafterDNAextractionusingacommercialkitandanautomatedsystem.ThedeterminationofthesubspeciesiscarriedoutbyMLST(Yuanetal.,2010).Long-distance dispersal of X. fastidiosa depends mainly on the human-mediated movement ofinfected plants and propagatingmaterial but naturally spread on short distances ismade by sap-feeding insects. So, detecting the bacteria inside the vector is critical. Adaptation of the protocolused on plants was performed to get an efficient detection test, usable on the known vectorPhilaenus spumarius. This protocol allowed detecting infected Philaenus spumarius from Corsicanand Apulian outbreaks. Complementary works are in progress to validate the protocol on otherpotentialvectorsofX.fastidiosa.Moreover, a test performance study will be organized in 2017 as part of a European Euphrescoprojectinvolving12countriesonthedetectionofX.fastidiosaoninsectsinordertoassessdifferenttools(LAMP,realtimeandregularPCR)andtoselectthemosteffectivemethod.Keywords:Xylellafastidiosa,vector,PCRdetection,Philaenusspumarius

Session3Poster58

Variance in depsipeptide biosynthetic potential among phyto- and entomopathogenicrepresentativesofHypocrealesMonikaUrbaniak1,GrzegorzKoczyk1,ŁukaszStępień1,SilvioUhlig21InstituteofPlantGenetics,PolishAcademyofSciences,DepartmentofPathogenGeneticsandPlantResistance,Strzeszyńska34,60-479Poznań,Poland2NorwegianVeterinary Institute,SectionforChemistryandToxicology,Ullevålsveien68,0454Oslo,Norway

Fungi are among themost prolific producers of bioactive secondarymetabolites in the biosphere.Among the secondary metabolites, short cyclic or linear peptides are synthesized by modularmegasynthases (NRPS, non-ribosomal peptide synthases), originally of bacterial origin. Weinvestigated the multiple producers of different cyclic depsipeptides (enniatins, beauvericins,bassianolide) among entomopathogenic (Beauveria, Metarhizium, Lecanicillium, Isaria),phytopathogenic (Fusarium), aswell asmycoparasitic (Trichoderma) representativesofecologicallydiverseHypocrealesorder.

The resultsofprofilingover90potentialdepsipeptideproducing isolates throughchemicalanalyses (LC-MS), as well as degenerate PCR marker amplification and sequencing are shown inconjunction with bioinformatic and phylogenetic analysis. The investigation of extant geneticdiversityunderlyingdifferencesinchemotypesyieldsnovelquestionsastothemodeofinheritanceofdepsipeptidesynthasemodulesamongbeauvericin/enniatinvs.bassianolideproducingtaxa.

Ouranalyseshaveshownstructuressimilaritiesbetweenenniatin/beauvericinsynthaseandbassianolide synthase in fungi fromBeauveria and Lecanicillium genera. Although similarmoduleshavebeenfoundalsoinotherHypocreales,bassianolidesynthesiswasonlyconfirmedforBeauveriaand Lecanicillium. The accumulated results provide a better framework for future studies ofregulatorydivergencebetweensecondarymetabolismpathwaysofhigherfungioccupyingdifferentecologicalniches.

Keywords:cyclodepsipeptides,enniatnis,beauvericins,bassianolide,Hypocreales

ResearchsupportedbytheNationalScienceCentreOPUS8grant:NCN2014/15/B/NZ9/01544

Session3Poster59UnravellingthemechanismsofColletotrichumlupinihostspecializationRiccardo Baroncelli1, Sabrina Sarrocco2, Noemi Bonadies2, Daniele Da Lio1,2, Gaetan Le Floch1 andGiovanniVannacci2

1Université de Brest, EA 3882, Laboratoire Universitaire de Biodiversité et Ecologie Microbienne,IBSAM,ESIAB,TechnopôleBrest-Iroise,29280Plouzané,France2UniversitàdegliStudidiPisa–DipartimentodiScienzeAgrarie,AlimentarieAgro-Ambientali,PlantPathology&Mycologylab-ViadelBorghetto80–56124Pisa,Italy

Colletotrichum acutatum species complex is considered an important pathogen for severaleconomicallyimportantplantspeciesasitcausesanthracnose,adiseasethatcanresultindeathofthehost.SinceitsfirstdiagnoseonLupinin1939,anthracnose,causedbythesee-(andsoil-)borneColletotrichum lupini, has become a severe disease of Lupin worldwide, causing meaningful yieldlossesashighas100%andbecomingamain limiting factor forproduction.Severalmorphological,culturalandmoleculardataconfirmC. lupiniaspartof theC.acutatum speciescomplex.AlthoughtheC.acutatumspeciesareconsideredpolyphagous,C.lupinihasbeenshowntohaveastricthostspecializationthatmakesitafascinatingmodelforevolutionaryandbiomolecularstudies.

With theaim todeeply investigate the infectionprocessof LupinbyC. lupini, for the first timeanefficientartificialinoculationprotocolofLupinseedshavebeensetupbyusingthereferenceisolateIMI504893. Its genomewas sequencedandannotatedand the isolatewas submitted, for the firsttime,toAgrobacterium-mediatedtransformationinordertoobtainaGFPmarkedisolatetobeusedasatooltobetterunderstandtheinfectionprocessonLupinhypocotyls.

Five stable transformants have been morphologically and physiologically characterized and aphenotypicmicroarraytest(Biolog)wasperformedinordertodetectmetabolicdifferencesbetweentheGFPmarkedstrainsanditswt.Informationwerealsoexploitedtoselectonetransformedstrainto be used in host-pathogen interaction studies. Following the artificial inoculation by the GFPmarked strain, fluorescence microscopy observations allowed us to describe, step by step, theinfectionandcolonizationprocess. ThedefinitionofthetimingforhostpenetrationpavesthewayforfurtheranalysissuchasatranscriptomicanalysisoftheC.lupini/lupininteraction.

Keywords:Colletotrichumlupini,Lupinusalbus,anthracnose,GFPtransformant,artificialinoculation

Session3Poster60IdentificationofMeloidogynespeciesonorientalmeloninKoreaInsooChoi1,2,Eul-SooYoon2,DonggeunKim2,HeonilKang1andNamSookPark21PusanNationalUniversity,DepartmentofPlantBioscience,samrangjin-ro1268-50,50463,Miryang,Korea2PusanNationalUniversity, InstituteofLifeand IndustryConvergence,NematodeResearchCenter,samrangjin-ro1268-50,50463,Miryang,KoreaRoot-knotnematodeiswidespreadandcausemuchdamagetoorientalmelon(Cucumismelovar.makuwa) in Korea. When crop rotation is used as a control measure, accurate identification ofspecies ofMeloidogyne is crucial. This study was conducted to identifyMeloidogyne species onoriental melon in Korea. PCR-RFLP analysis of 47 Meloidogyne populations was performed usingsinglefemale.TheregionbetweenCOⅡand16SrRNAofthemitochondrialDNAwasPCRamplified.Forty six populations produced a single fragment at 1,700bp, and 1 population produced a singlefragmentat1,100bp.Fortheidentificationmorecorrectly,PCRproductsweredigestedwithHinfⅠ.ThirtyfourpopulationswereidentifiedasM.incognitabygenerating1300bpand400bpfragments.ThirteenpopulationswasnotdigestedwithHinfⅠ.ItisevidentthatM.arenaniaofasinglefragmentof1700bpwithnodigestionwithHinfⅠ iswidespreadatorientalmelonfield inKoreawhichmayposesses additional parts in the site of COII/16S rRNA and is genetically different from NorthAmericanisolatesofM.arenaira.

Keywords:orientalmelon(Cucumismelovar.makuwa),Root-knotnematodes,speciesidentification,morphology,PCR,RFLP

Session3Poster61PotentialofTrichodermaspp.strainsisolatedfromnaturalsourcestoproduceceratoplataninsCalinaPetrutaCornea1,FlorentinaIsrael-Roming1,MatildaCiuca2,CatalinaVoaides1,AglaieBurlacu1,DanielCristina2,FlorinOancea3

1UniversityofAgronomic SciencesandVeterinaryMedicineBucharest, FacultyofBiotechnologies,59,MarastiBld.,Bucharest07100,Romania2 National Agricultural Research and Development Institute Fundulea, N.Titulescu street; no.1,Calarasi;Fundulea;915200;Romania3 National Research andDevelopment Institute for Chemistry and Petrochemistry – ICECHIM, 202,SplaiulIndependenței,060021BucharestFungibelongingtoTrichodermagenusarenon-pathogenicsoil-bornemicroorganismsthatcolonizethe rootof plants as avirulentplant symbionts.Various speciesofTrichodermahave the ability toinhibit the development of plant pathogenic fungi and even to destroy such organisms throughmycoparasitism and other mechanisms. Moreover, they are able to induce the plant’s immunesystem by production of specific elicitors. Among these elicitors, cerato-platanins produced byspeciesofTrichodermaareintensivelystudiedinthelastyears.Theaimofthisworkwastoidentifynew isolates of Trichoderma spp. with antifungal properties, to detect the presence of the genesinvolvedincerato-platanin(CP)biosynthesisandtoevaluatetheirimpactonmycoparasitism.TheidentificationoftheTrichodermastrainsisolatedfromcompostwasrealizedwithspecie-specificprimers and allowed the identification of T.atroviride, T,asperellum, T.harzianum andT.longibrachiatum species. Intraspecific genomic variability was examined by RAPD and with ISSRprimers. Differences in electrophoretic patterns among the strains belonging to the same specieswere detected. The detection of epl1 gene involved in biosynthesis of CP was performed withprimersproposedinliterature.Directconfrontationassaysandmicroscopicallyanalysiswereusedtoevaluate theantifungalactionof selectedTrichodermastrainsagainstRhizoctonia solani, Fusariumoxysporum, Sclerotium bataticola, Botrytis cinerea and the influence of CP coding genes onmycoparasitism.Ampliconswithdesiredlength,correspondingtoepl1gene,wereobservedinstrainsofT.harzianum,T.atrovirideandT.asperellum.Differencesamongstrainswithorwithoutepl1generegarding the inhibition of plant pathogenswere revealed suggesting the possible involvement ofEPL1proteinintheinteractionsbetweentheantagonistandthetarget.These results contribute to the understanding of the complex mechanisms of the interactionsbetweenbiocontrolTrichodermaspeciesandplantpathogenicfungiandsustaintheuseofcompostasasourcesofbiocontrolmicroorganismsforplantprotection.Keywords:cerato-platanins,Trichoderma,moleculardetection

Session3Poster62FirstreportofAcidovoraxcitrullionmeloninGuadeloupeAmandine Cunty1, Valérie Olivier1, Corinne Audusseau1, Marie Bourgeais1, Christelle François1,FrançoisePoliakoff11Laboratoiredelasantédesvégétaux,Anses,7rueJeanDixméras,49044Angers,FranceAcidovoraxcitrulli(Schaadetal.,1978;Schaadetal.,2009),agram-negativebacterium,isthecausalagentofbacterialfruitblotch(BFB)ofcucurbitplants.Itsmainhostsarewatermelonandmelon,butcucumber,pumpkinandsquasharealsodescribedashostplants.As it isa seedborndisease, theseeds are likely to be responsible for spreading over long distances, mainly under warm climate.ThisbacteriumisnotaquarantineorganismforEUbutitwasaddedtotheEPPOA1listsince2014duetotheriskofemergencearoundtheMediterraneanBasin.InAmerica,ithasbeenpresentsincethe 1990s (first in Florida in 1989). Its impact on the production of marketable fruits can becatastrophicwithlossesofmorethan90%.Thenthediseasehasspreadtomanypartsoftheworld(Asia). Absent from Europe until 2005 (first description in Greece, Holeva et al., 2009), it wasdeclaredpresentwitharestricteddistributionorsporadicoutbreaksinGreece,Hungary(PalkovicsLet al., 2008), Turkey (Mirik et al., 2006), Israel (Burdman et al., 2005), Italy, Serbia (Popovic andIvanovic,2015);itisnowconsiderederadicatedinthesecountries.InMay2015,A.citrulliwasdetectedinGuadeloupe,inFrenchWestIndies,onsymptomaticmelon.Themelon presentedwater-soaked lesions on leaves and fruits and bacteriumwas isolated fromboth. Identificationoftheisolateswasperformedbybiochemicalandmoleculartests(Baharetal.,2008).ThepathogenicityoftheisolateswasverifiedandKoch'spostulatedemonstrated.Thisfirstreportshowstheriskofemergenceofnewpathogensunderhotandhumidclimateandtheimportanceofusinghealthyseeds.Keywords:Acidovoraxcitrulli,symptomaticplants,cucurbitseeds,isolation,PCR,emergence

Session3Poster63Newfoliarandsoil-bornepathogensrecentlyobservedonleafyvegetablesinEuropeMariaLodovicaGullino1,2,GiovannaGilardi1andAngeloGaribaldi11University of Torino, AGROINNOVA - Centre of Competence for Agro-Environmental Innovation,LargoPaoloBraccini2,10095Grugliasco,Torino,Italy2UniversityofTorino,DISAFA–DepartmentofAgricultural,ForestandFoodSciences,LargoPaoloBraccini2,10095Grugliasco,Torino,ItalyDuringthepastyearsnewdiseasescausedbysoil-borneandfoliarpathogenswereobservedforthefirsttimeinEuropeonlettuce,rocket,spinachandlamb’slettucegrownmainlyforthereadytoeatsector. Italy is the secondproducer in Europeof fresh-cut leafy vegetables, representing indeed averyinterestingcasestudyfromaphytopathologicalpointofview.Amongfoliardiseases,Fusariumequiseti on wild and cultivated rocket and lettuce, Allophoma tropica on lettuce, Colletotrichumkahawaeoncultivatedrocket,Myrotheciumroridumonlamb’slettuceandM.verrucariaonspinachandwildrocketwereobserved.Moreover,anewraceofFusariumoxysporumf.sp.lactucaehasbeenisolatedforthefirsttimeintheNetherlandsonaffectedlettuceplants,while,race1ofthispathogenhasbeenfoundrecentlyinFrance.Someofthesenewpathogenshavebeenisolatedfromseeds.Thesymptoms of the diseases caused by these fungal pathogens, the biology and physiologicalcharacteristics of the causal agents and some information concerning their epidemiology arepresented.Keywords:leafyvegetables,alienpathogens,seed-bornepathogens,Emphasisproject

Session3Poster64SeedtransmissionofTomatoyellowleafcurlvirusinsweetpeppers(Capsicumannuum)Eui-Joon Kil1, Jungho Park1, Eun-Young Choi1, Hee-Seong Byun1, Kangsan Roh1, Chang-Seok Kim2,Hong-SooChoi2,Ji-KwangKim3andSukchanLee1

1DepartmentofGeneticEngineering,SungkyunkwanUniversity,Suwon16419,Korea2Crop Protection Division, National Institute of Agricultural Sciences, Rural DevelopmentAdministration,Wanju55365,Korea3Research and Development Bureau, Chungcheongnam-do Agricultural Research & ExtensionServices,Yesan32418,Korea Thesweetpepper(Capsicumannuum)isapopularcropandanasymptomatichostofTomatoyellowleaf curl virus (TYLCV). A previous study showed that TYLCV could be transmitted by the seeds oftomatoplants,butthisphenomenonhasnotbeenconfirmedinotherplants.In2015,fourdifferentcultivars of sweet pepper (‘Super Yellow,’ ‘Super Red,’ ‘Sunnyez’ and ‘Cupra’) confirmed to besusceptible to TYLCV by the previous study were planted and inoculated with agrobacteriumcontainingaTYLCV infectiouscloneatan isolatedgreenhouse.Threemonthsafter inoculation, theleavesofthe‘SuperYellow’cultivarshowed80%(8/10)infectivityandtheotherthreesweetpeppercultivarsshowed30to50%infectivity.Allofthe‘SuperYellow’seedbunches(fiveseedsperbunch)fromplantswhoseleaveswereconfirmedtobeTYLCV-infectedwerealsoTYLCV-infected(8/8).Theseedsof other cultivars showed20 to40% infectivity.Virusdissemination rateswere also verifiedwith 10 bunches of seedlings for each cultivar (five seedlings per pool). Eight bunches of ‘SuperYellow’seedlings(8/10)wereconfirmedtobeTYLCV-infectedandonetothreebunchesofeachofthe other cultivar seedlings were also infected. Viral replication in TYLCV-infected seeds andseedlings was confirmed via strand-specific amplification using virion-sense- and complementary-sense-specificprimersets.ThisisthefirstreportofTYLCVseedtransmissioninsweetpepperplantsandamongnon-tomatoplants.Becausethesweetpepperplant isanasymptomatichostofTYLCV,sweet pepper seeds infectedwith TYLCV could act as a silent invader of tomatoes andother hostcrops;therefore,itisparticularlyimportanttoidentifyTYLCVseedtransmissioninnon-tomatohosts.Keywords:Begomovirus,geminivirus,seedtransmission,sweetpepper,Tomatoyellowleafcurlvirus

Session3Poster65FirstreportofseedtransmissionofTomatoyellowleafcurlvirusinwhitesoybean(Glycinemax)Eui-JoonKil1,JunghoPark1,Eun-YoungChoi1,Hee-SeongByun1,HeJiang1,HocheolShin1,Mi-KyeongKim2,Chang-SeokKim2andSukchanLee1

1DepartmentofGeneticEngineering,SungkyunkwanUniversity,Suwon16419,Korea2Crop Protection Division, National Institute of Agricultural Sciences, Rural DevelopmentAdministration,Wanju55365,Korea Tomatoyellowleafcurlvirus(TYLCV)causesdamagetoseveraleconomicallyimportantcropssuchastomatoes, peppers and cucurbits. TYLCV infection of common bean (Phaseolus vulgaris) wasreportedpreviously,butsoybean(Glycinemax)hasnotpreviouslybeenidentifiedasaTYLCVhost.In2015,fivecultivarsofwhitesoybean(‘Jinpung,’ ‘Taegwang,’ ‘Seonpung,’ ‘Neulchan’and‘Daepung’)were agro-inoculated using an infectious TYLCV clone in an isolated plant cultivation chamber inWanju. Thirty days post-inoculation, ‘Jinpung’ and ‘Taegwang’ showed 86% and 100% infectivity,respectively, and other soybean cultivars showed 25 to 66% infectivity. Typical TYLCV symptomswerenotobservedinanyinoculatedplants.ViralreplicationinTYLCV-infectedleaveswasconfirmedby the existence of double-stranded viral DNA with strand-specific amplification using the virion-sense-andcomplementary-sense-specificprimersets introduced in thepreviousstudy.Basedonaprevious study about seed transmission of TYLCV in tomato plants, seeds from TYLCV-inoculatedsoybeanplantsconfirmedtobeTYLCV-infectedwereharvestedandDNAwasisolatedfrombundlesof five randomly selected seeds and amplified with a TYLCV-specific primer set in order to verifyTYLCVseed transmission in soybean.Except foronebunchof ‘Jinpung’ seeds,allbundlesof seedsfrom infected soybean plants were verified to be TYLCV-infected (13 of 14 bunches). Virusdissemination rates were also confirmed from three bunches of seedlings out of 14 bunchesgerminated from infected seeds (five seedlings per bunch). Viral replicationwas also identified inseeds and seedlings confirmed to be TYLCV-infected via strand-specific amplification as previouslydescribed(Edgaretal.2014).This is the first reportprovingthatsoybean(G.max) isaTYLCVhostandthatTYLCVisaseed-transmissiblevirusinwhitesoybean.Keywords:Begomovirus,geminivirus,reservoir,seedtransmission,soybean,Tomatoyellowleafcurlvirus

Session3Poster66FirstreportofTomatoyellowleafcurlvirusinfectingmonocotyledonousplantsEui-JoonKil1,Hee-SeongByun1,Sang-HoCho1,SeungchanCho1,YongjunLee1,DongjunKim1,Hong-SooChoi2,Chang-SeokKim2andSukchanLee1

1DepartmentofGeneticEngineering,SungkyunkwanUniversity,Suwon16419,Korea2Crop Protection Division, National Institute of Agricultural Sciences, Rural DevelopmentAdministration,Wanju55365,Korea Tomato yellow leaf curl virus (TYLCV) is one of themost important plant viruses belonging to thegenusBegomovirusof thefamilyGeminiviridae.From2013to2014,to identifynaturalweedhoststhat could act as reservoirs of TYLCV, one hundred samples were collected at a TYLCV-infectedtomato-cultivating farm in Iksanand identifiedasbelongingto40species from18families.Amongthem, TYLCV was detected from 57 smaples belonging to 28 species by PCR using root samplesincluding five species (Eleusine indica, Digitaria ciliaris, Echinochloa crus-galli,Panicumdichotomiflorum and Setaria faberi) from the family Poaceae. Whitefly (Bemisia tabaci)-mediated TYLCV transmission from TYLCV-infected E. indica plant to healthy tomatoes wasconfirmed,andinoculatedtomatoesshowedtypicalsymptomssuchasleafcurlingandyellowing.Inaddition, TYLCVwas also detected in leaf and root samples ofE. indica plants inoculated by bothwhitefly-mediated transmission using TYLCV-viruliferous whitefly and agro-inoculation using theTYLCV infectious clone. The majority of mastreviruses infect monocotyledonous plants, but fewdicotyledonousplants infectedwithmasteviruseshavebeen reported.However,noexceptionwasreported among begomoviruses known as infecting dicots only. This is the first report ofbegomovirus-infectingmonocotyledonousplants.Keywords:Begomovirus,Eleusineindica,geminivirus,monocotyledonousplants,Tomatoyellowleafcurlvirus

Session3Poster67EmergingseabuckthorndiseasesinLatviaandassociatedfungalpathogensIngaMoročko-Bičevska1,OlgaSokolova1,JamshidFatehi11InstituteofHorticulture,LatviaUniversityofAgriculture,Graudustr.1,Dobele,LV-3701,LatviaSea buckthorn (Hippophae rhamnoides L.) is an important fruit crop in Latvia and its area undercultivationisrapidlyexpanding.Ingeneral,seabuckthorndiseasesandtheircausalagentshavebeenpoorlystudied,limitedtoonlyafewrecordsofstemcanker,wiltanddryshrinkdiseasescausedbyStigmina sp,Verticillium spp., andFusarium spp., respectively. In Latvia, concernsof growershaveraisedondiseasesspreadinginseabuckthornplantations.Toidentifyandcharacterizeseabuckthorndiseases prevailing in Latvia 55 locations including commercial orchards and wild habitats weresurveyed. The samples from branches, roots, and trunks with various disease symptoms werecollected. Fungi were isolated from surface sterilized plant tissues on potato dextrose agar, sub-culturedinpureculturesandpreservedforfurtherstudies.Theisolatedfungiwerecharacterizedandidentifiedbymorphologicalcharacters,sequencingoftheITSregionandcomparisontootherrelatedfungibyphylogenetic analysis.During the surveys,overalldecline,wilt, severe cankeranddiebacksymptoms often causing a death of the plants was observed. The fungal isolates obtained frominfectedtissueswereidentifiedasbelongingtothegeneraofDiaporthe,Eutypa,Fusarium,ValsaandVerticilliumwhicharethewell-knowncausalagentsofwilts,cankers,anddiebacksonvarioustrees.Besides,arangeofotherfungiwasobtainedwhichappearedtobe less-knownandthereforetheirrolesinseabuckthorndiseaseshavetobefurtherinvestigated.Thepathogenicityevaluationofthefungalisolatesonseabuckthornplantsinagreenhouseiscurrentlyunderdevelopment.

Keywords:Hippophaerhamnoides,fungaldiseases,canker,wilt

Session3Poster68Fusarium species associated with roots of leguminous plants grown in different environmentalregionsofEuropeAdnanŠišić1*,JelenaBaćanović1,PetrKarlovsky2,RaphaëlWittwer3,FlorianWalder3,EnioCampiglia4,EmanueleRadicetti4,HannaFriberg5,JörgPeterBaresel6,andMariaR.Finckh1

1Department of Ecological Plant Protection, University of Kassel, Nordbahnhofstr. 1a, 37213Witzenhausen,Germany2Department of Molecular Phytopathology and Mycotoxin Research, University of Göttingen,Grisebachstr.6,37077Göttingen,Germany3Plant-Soil-Interactionsgroup,InstituteforSustainabilityScience,Agroscope,Reckenholzstrasse191,CH-8046Zurich,Switzerland4Department of Agricultural and Forestry Sciences (DAFNE), University of Tuscia, via S. Camillo deLellissnc,Viterbo,Italy5Department of ForestMycology and Plant Pathology, Swedish University of Agricultural Sciences(SLU),Box7026,75007Uppsala,Sweden6Institute forPlantNutrition,Centerof LifeandFoodSciencesWeihenstephan,TechnicalUniverityMunich,Emil-Ramannstr2,85354Freising-Weihenstephan,GermanyGrowing legumes as cover crops, intercropped or under-sown with cereals can provide multiplebeneficial services to agro-ecosystems. However, agronomic benefits of their inclusion into therotations can only become effective if the pathological risks for crop rotations are known androtationsdesignedaccordingly.Thisstudybringstogether findingsfromsurveysconducted in2013and 2014 across five European sites in Switzerland, Italy, Germany, and Sweden. Incidence andcharacterization of Fusarium spp. associated with the roots of two clover and two vetch speciesgrownascovercropsorunder-sowninwheatweredetermined.Outof1480rootsanalyzedinbothyears,670Fusariumisolateswereobtained.Nostrongseparationamongthesitescouldbeobserved inFusariumcommunitystructuresuggestingthateachsitewasdominatedbysimilarspecies.ThespeciesrichnesscurveforbothcloverspecieswasmuchsteepercomparedtovetchspeciesshowingsomehosteffectsontheFusariumcommunitycomposition.ThemostfrequentlyisolatedspeciesinbothyearsfromallsitesandallfourhostswereF.oxysporumandF. avenaceum. In pathogenicity tests on peas isolates of F. avenaceum caused highest biomassreductionsandmostsevererootrotsymptoms, followedby isolatesofF.oxysporum,F.solaniandnontoweaklypathogenicisolatesofF.tricinctum,F.acuminatumandF.equiseti.TheprevalenceofF. avenaceum and the potential of isolates to cause severe yield reductions under favorableconditionssuggestthatitcouldemergeasapotentialriskunderlegumerichcroprotations.Keywords:clover,vetch,Fusarium,legumerootpathogens

Session3Poster69InteractionsofpeapathogenswithpotentiallyusefulleguminouscovercropspeciesAdnanŠišić1,JelenaBaćanović1,MariaR.Finckh1

1Department of Ecological Plant Protection, University of Kassel, Nordbahnhofstr. 1a, 37213Witzenhausen,GermanyModern cropping systemsmainly rely on growing a narrow range of crop species and genotypeswhilethepossibilityofusingnovelcrops,particularly leguminousspecies,whichhavethepotentialto play a major role in more diversified and sustainable food production systems, has beenneglected.Thesespeciescouldprovidemultiplebeneficialservicestoagroecosystemswhenusedascover crops (CC), such as green manures or living mulch. Before introduction, it is important,however,toassessifsuchspeciesshareimportantdifficulttomanagepathogensofmaincrops.Atotalof62accessionsbelongingtotenlegumegenerawerescreenedundercontrolledconditionsfor their susceptibility/resistance to selected pea pathogenic isolates of Fusarium avenaceum, F.oxysporum, F. solani, Peyronellaea pinodella (syn. Phoma medicaginis) and Didymella pinodes, allmajorpearootpathogens.Theplantswereinoculatedwith2x104sporesg-1substrateonedayaftertransplanting pre-germinated seeds. Three pea varieties, the resistant EFB 33 and the susceptibleSantana and a Brazilan accession, were included as additional controls. Five weeks after sowing,disease symptomswereassessedandplantgrowthparametersmeasured.Almostallplant speciesandaccessions testedwerehighly susceptible toFusariumavenaceum,withnotableexceptionsofCrotalaria ochroleuca, Lotus pedunculatus and a few Trifolium and Medicago accessions. F.oxysporum caused variable disease severity on some Trifolium species, otherwise, infectionswerelow,whileF.solanicausedoverallhigherdiseaseseveritywithsomevariationamongaccessions.Thissuggestsspecific interactions.PeyronellaeapinodellaandDidymellapinodesmostseverelyaffectedLathyrus,otherwiseinfectionswerelowwithD.pinodesapathogenthatapparentlyisrelativelyhostspecific. Integration of this knowledge is crucially important in the design of rotations with covercropsinordertointegratethemsuccessfullyandsustainablyintoagriculturalsystemsasrequiredbytheEUgreeningpolicy.Keywords:Pearootrot,covercrops,EUgreening,croprotation

Session3Poster70ReportandcharacterizationofbacterialdiseasescausedbyXanthomonasoryzaeinSenegalHamidouTall1,KandiouraNoba3,BorisSzurek2,SébastienCunnac2,CheickTekete4,ValérieVerdier2

1InstitutSénégalaisdeRecherchesAgricoles(ISRA),BelAir,DakarSénégal2IRD, Cirad, Univ. Montpellier, UMR Interactions Plantes Microorganismes Environnement (IPME),911AvAgropolis,Montpellier,France3Laboratoiredebotaniqueetdebiodiversité,DépartementdebiologieVégétale,UniversitéCheikhAntaDiop,Dakar,Sénégal4Laboratoire de Biologie Moléculaire Appliquée, Université des Sciences Techniques etTechnologiquesdeBamako,FacultédesSciencesetTechniques,Bamako,MaliOverthe lastdecadesricecultivationhasbeenincreasingdrastically inAfrica.Rice(Oryzasativa) isbecoming the number one crop produced for human consumption feedingmore than half of thepopulationinAfrica.AmongfactorsthatlimitorreducericeagriculturalyieldinAfrica,arebacterialdiseases that impact food security.Xanthomonasoryzaecause important yield lossesof rice.Bothbacterial blight (BB) and bacterial leaf steak (BLS) are reported in various countries in Africa.Xanthomonasoryzaepv.oryzae(Xoo,causalagentofBB)wasreportedinSenegalintheeightiesbyTrinh et al. Trinh did not isolate strains of Xoo and since no other report has been made.Xanthomonasoryzaepv.oryzicola(Xoc,causalagentofBLS)wasnotreportedinSenegalsofar.Theuseofricecultivarswithintrogresseddiseaseresistance(R)genesiscurrentlythebestwaytocontrolBBdiseasewithminimalenvironmentaleffectsandcost.NoriceresistancegenehasbeenreportedtocontrolBLSworldwide.Resistancebreedingdependsonharnessinggeneticdiversityofpathogenpopulations. The choice of BB and BLS resistance gene(s) should be made based upon theireffectivenessagainsttheprevalentracesofXooandmostvirulentstrainsofXocintheregion.Asnostrains of Xoo and Xoc were collected and characterized in Senegal so far, no strategy has beenpursuedtocontrolthediseases.Here,weaimatconfirmingTrinh'sobservations(BBpresence).TothatpurposewesurveyedthepresenceandprevalenceofBBindifferentregionsofSenegal.Atthesametimewe lookat thepresenceofBLS.Fields inmainareasof riceproduction inSenegalweremonitored between 2014 and 2016. Leaf samples were collected and analyzed. After bacterialisolation,aPCRmultiplexwasusedtoconfirmthepresenceofXanthomonasoryzaepv.oryzaeandoryzicolaindifferentsitesofeachregion.AsetofIRBBlineseachcarryingasingleRgenewasusedtoidentifyingtheraceofXooinpresenceinSenegal.WealsoassessedthevirulenceofXocstrainsonasusceptible rice cultivar. We will present and discuss the results obtained during this two year’ssurveyinSenegal.

Session3Poster71Characterization and pathogenicity ofBiscogniauxia mediterranea associated with cork oak(QuercussuberL.)inTunisia.Islem Yangui1,2, Meriem Zouaoui Boutiti1, Natalia Bruni3, Anna Maria Vettraino3, Andrea vannini3,MohamedLahbibBenJamaa2,ChokriMessaoud21National Institute for Research on Rural Engineering, Water and Forestry, Laboratory ofmanagementandvalorizationofforestresources,BP.N.2,Ariana2080,Tunisia2UniversityofCarthageNationalInstituteofAppliedScienceandTechnology(INSAT),DepartmentofBiology,LaboratoryofNanobiotechnologyandMedicinalPlants,BP676,1080TunisCedex,Tunisia3DIBAF,UniversityofTuscia,ViaS.CamillodeLellis,Viterbo01100,ItalyBiscogniauxiamediterranea is a virulent fungal pathogen involved in oak decline and caused highmortalityincorkoaktreesinthemediterraneanregion.Itisresponsableofcharcoalcankerdisease.TheoccurrenceofB.mediterraneaassociatedwithcorkoak inTunisiahasbeenobservedsincethesixties. Therefore, the present study was aimed to characterize B.mediterannea isolated fromdifferent organs of cork oak trees in a natural forest ecosystem in Tunisia and to evaluate itspathogenicity.B.mediterranea isolates were obtained from leaves, brunches and trunks of ten declining trees.Morphologicalcharacterizationwasbasedonthemycelialgrowthrateandthecolonypigmentation.Molecular identification was performed by PCR amplification of the fungal DNA with the specificprimersMED1/MED2.Pathogenicity test was conductedonyoungcorkoakplantationsof2yearsoldmaintainedinagreenhousefor2months.Atotalof70isolatesofB.mediterraneawasobtained.AlltheorganswererevealedtobeinfectedbyB.mediterranea. However, the statistical analysis showed a significant variation of the isolationfrequencybetweenthedifferentorgans.TheBruncheswereshowntobethemostinfected.Fortheobtainedisolates,PCRamplificationwiththeprimersMED1/MED2generatedonlyonePCRproductofapproximately380pb.Thus,allisolateswereconfirmedasB.mediterraneaspecies.Pathogenicitytest confirmed the virulence of the tested B.mediterranea isolates. These results emphasize thenecessityforfurtherin-depthstudiesonthefungusbiologyandtheepidemiologyofcharcoalcankerintunisianforests.Keywords:Biscogniauxiamediterranea,characterization,pathogenicity,corkoak

Session3Poster72AnewTobamovirus isolate infects tomatoplantsharboringTm-22 resistancegenes - apotentialglobalthreattotomatoproductionNetaLuria1,ElishevaSmith1,VictoriaReingold1,IlanaBekelman1,MosheLapidot2,IlanLevin2,NadavElad4,YehuditTam1,NoaSela1,AhmadAbu-Ras3,NadavEzra3,AmiHaberman3,LironYitzhak1,5,OdedLachman1andAvivDombrovsky11DepartmentofPlantPathology,ARO,TheVolcaniCenter,HaMaccabimRoad68,P.O.B15159RishonLeZion7528809,Israel2DepartmentofVegetables and field crops,ARO, TheVolcani Center,HaMaccabimRoad68, P.O.B15159RishonLeZion7528809,Israel3Plantprotectionandinspectionservices,P.O.B.78,Beit-Dagan,50250Israel4Electron Microscopy Unit, Departments of Chemical Research Support, Weizmann Institute ofScience,Rehovot,Israel5DepartmentofPlantSciences,GeorgeS.WiseFacultyofLifeSciences,TelAvivUniversity,TelAviv69978,IsraelAnoutbreakof a newdisease infecting tomatoes ocurred inOctober-November 2014 in SouthernIsrael. Symptomatic plants showed a mosaic pattern on leaves accompanied occasionally bynarrowing of leaves and yellow spotted fruit. The disease spread mechanically and rapidlyreminiscentoftobamovirus infection. Epidemiologicalstudiesshowedthespreadofthedisease invarious growingareas, in theSouthand towards theSoutheast andNorthernpartsof the countrywithin a year. Transmission electron microscope (TEM) analysis showed a single rod-like formcharacteristictotheTobamovirusgenus. WeconfirmedKoch’spostulatesforthediseasefollowedbypartialhostrangedeterminationandrevealedthattomatocultivarscertifiedtoharbortheTm-22resistancegene,aresusceptibletothenewviraldisease.Wefurthercharacterizedtheviralsourceofthediseaseusingarangeofantiseraforserologicaldetectionandanalyzedvariousvirusgeneraandfamilies for cross-reactivity with the virus. Next generation sequencing of total small RNA wasperformedontwocultivarsgrownintwodifferentlocations.Thecompletegenomesequenceofthenew Israeli tobamovirus showed high sequence identity to the Jordanian isolate of tomato brownrugosefruitvirus.Keywords:Tm-22,next-generationsequencing(NGS)

SESSION4.MICROBIALDETERMINANTSOFPATHOGENANDSYMBIOTICINTERACTIONSSession4Poster73Verticilliumwiltonfiberflax:symptomsandpathogendevelopmentinplantaAdrien Blum1,2, Mélanie Bressan1, Abderrakib Zahid2,3, Isabelle Trinsoutrot-Gattin1, AzeddineDriouich2,andKarineLaval1

UNILASALLE–CampusRouen,Agri’Terrunit,CS40118,3rueduTronquet,76134Mont-Saint-AignanCedex,FranceGlycobiologie et Matrice Extracellulaire végétale EA 4358, SFR Végétal-Agronomie, Université deRouen76821Mont-Saint-Aignan,FranceUniversitéduLittoralCôted’Opale,UnitédeChimieEnvironnementaleet Interactionssur levivant(UCEIV-EA4492),Univ.Lille-NorddeFrance,GISPhyNoPiCS80699,F-62229CalaisCedex,FranceFiberflax(LinumusitatissimumL.),animportantcropinNormandy(France),isincreasinglyaffectedbyVerticilliumwilt causedby the soil-borne fungusVerticilliumdahliae Kleb. This disease leads tonon-negligible yield losses and depreciated fibers that are consequently difficult to upgrade.Verticillium wilt is a major threat to a large broad range of agriculture; in this study, fiber flaxsusceptible cv. Adélie was infected by Vd05-833 (isolated on fiber flax, this study) or greenfluorescentprotein-taggedVdLs17 (isolatedon lettuce,ATCCaccessionMYA-4575).Between threeand four weeks post inoculation, wilting symptoms on leaves were first observed, with acropetalgrowth during the following weeks. Pathogen development was tracked by confocal microscopyduringtheasymptomaticandsymptomaticstages.First,conidiagerminationledtothedevelopmentofhyphaeonrootepidermismoreparticularlyonzoneofcelldifferentiationandaroundemerginglateralrootswhilethezoneofcelldivisionandtheroottipwerefreeofthepathogen.Atthreedayspostinoculation,thezoneofcelldifferentiationandlateralrootswereembeddedintoafungalmass.Swellingstructuressuchasappressoria,wereobservedatoneweekpostinoculation.Attwoweekspostinoculation,thepathogenhadcolonizedxylemvesselsinroots,followedbythestemandfinallyleavesduring thesymptomatic stage.Pathogenquantitywasassessedby real-timePCRduring thefirst 30 dpi. A significant decrease in fungal DNA was detected between the first days when thepathogenwasspreadontotheroot(asymptomaticphase)and30dpi,whensymptomsappearedandthepathogencolonizedvasculartissues.AlloftheseresultsprovideaglobalaccountoftheV.dahliaediseasecyclewheninfectingfiberflax.Keywords:fiberflax,Verticilliumwilt,GFPconfocalmicroscopy,real-timePCR.

Session4Poster74Comparative transcriptomic study of two grapevine fanleaf virus strains inducing contrastedsymptomsonVitisviniferacv.GewurztraminerIsabelle Martin1, Emmanuelle Vigne1, Jean-Michel Hily1, Corinne Schmitt-Keichinger2, CamilleRustenholz1andOlivierLemaire11UniversitédeStrasbourg,INRA,SVQVUMR-A1131,68000Colmar,France2UniversitédeStrasbourg,CNRS,IBMPUPR2357,67000Strasbourg,FranceGrapevine fanleaf virus (GFLV) is the major virus inducing fanleaf degeneration, one of the mostharmfulviraldiseasesofVitissppworldwide.IntheFrenchvineyard,thelossesduetothisendemicdisease are estimated between 350 and 850 million euros per year. Symptoms depend onenvironmentconditions,rootstock-scioncombinationsandvirusstrains.They includeleafandcanemalformations,yellowmosaic,smallfruitclusters,andaprogressivedeclinethateventuallyleadstodeath.LittleisknownaboutthedeterminismandthemechanismsofsymptominductionbyGFLVonitsnaturalhost.TogainknowledgeonvirusdeterminantsandpathogenesismechanismsweestablishedatrialwithGewurztraminerplantsinfectedwithsingleGFLVstrainF13orB844.GFLVinoculationwasperformedbyinvitroheterologousgrafting.Thetrialtookplaceinanexperimentalisolatedvineyardawayfromsuperinfection.Theseplantsweremonitoredfrom2012to2014forsymptoms,fruityieldandquality(Vigneetal.2015).GFLV-F13 infectedgrapevinesshoweddiscretefoliarsymptoms,whereasGFLV-B844infectedplantsdisplayedaseveredegenerationofthevinestock.Wetookadvantageofthesecontrastingsymptomsinanon-targetedapproach.Tothisend, leavesweresampledduringthe2016vegetativeseasonandtotalRNAswereextractedfromthreeB844-orF13-infected Gewurztraminer. A transcriptomic analysis by RNAseq was initiated to identifyderegulatedgenesanddeducethemajorpathwaysinvolvedinsymptomformation.Thefirstresultswillbepresentedanddiscussed.Keywords:symptomatology,Nepovirus,RNAseq,Vitis

Session4Poster75AnalysisofVenturiainaequalisgenesexpressionprofiles:searchingforthegenesassociatedwithapple-scab-resistancebreakdownMonikaMichalecka1,TadeuszMalinowski1,JoannaPuławska11Research InstituteofHorticulture,DepartmentofPlantPathology,Konstytucji 3Maja1/3Str, 96-100Skierniewice,PolandApplescabcausedbyVenturiainaequalis(Cooke)Wintisoneofthemostseriousdiseasesofapple(Malus x domestica). The naturally existing scab resistance in some apple cultivars has beenrecognizedasthepreferredsustainablealternativetochemicalcontrol.Twomainfactorsinvolvedingene-for-gene relationare: genesor gene ‘R’ at thehost sideand compatibleavr locuspresent inpathogen, responsible for avirulence toward the apple cultivars. Seventeen differentV. inaequalisraceshavebeenidentifiedsofar,eachovercomingonemajormonogenicresistancesourceinMalusspp. cultivars. It is expected that each different host-specificity of the race results not only fromdifferentgenecontentbutalsofromdifferentgeneexpressionprofileandinvolvementofdifferenteffectorproteins.Theaimofthestudywastoidentifycandidategenesassociatedwithapple-scab-resistance breakdown via transcriptome analysis with RNA-seq technique of two races of V.inaequalisintwosusceptiblecultivars.GeneexpressionprofileofV.inaequalisstrains,representingracesabletoinfect'Topaz'and'GoldenDelicious' cultivars of apple (containing Rvi6 and Rvi1 genes, respectively) has been analysed inplantaandinvitroduringthisstudy.Sporulating scab lesionsonapple leaves andmycelium respectively growingonPDAofboth raceswereexcisedand totalRNAwasextracted from them. cDNA librarieshavebeen constructed frompolyadenylatedRNAfractionandsequencedonIlluminaHiSeq™NGSplatform.ObtainedreadsweremappedtoMalusandV.inaequalisreferencegenomes.Thesubsetofreads,whichdidnotmappedtoMalusreference,hasbeenassembleddenovo.Morethan24000contigsobtainedasaresultofthe assembly have been used for calculation of expression profile for each race separately. Eachcontighasbeensortedtooneofthe15setsrepresentingallpossibleexpressionpatterns.Currently,about1500contigspotentiallyinvolvedinpathogenicityprocesshavebeenpreliminaryselectedforfurther analysis. Among them 122 contigs showed especially promising patterns, which will beverified using RT-rtPCR. Our long term goal is to identify V.inaequalis genes responsible forovercomingplantdefencebasedonRvi6andRvi1applescabresistancegenesofMalushost.ThisworkwasfinancedbyNationalScienceCentre,PolandgrantNo.UMO-2013/09/B/NZ9/02343.Keywords:geneexpressionprofiling,geneforgeneinteraction,RNAsequencing

Session4Poster76Genome-wideanalysisofCorynesporacassiicolaputativeeffectors involved intheCLFdiseaseofrubbertreeDavid Lopez1, Philippe Label1, Boris Fumanal1, Sébastien Ribeiro1, Annegret Kohler2, Robin Ohm3,JosephSpatafora4,IgorGrigoriev3,FrancisMartin2andValériePujade-Renaud5

1UniversitéClermontAuvergne(UCA),UMR547PIAF,BP10448,F-63000Clermont-Ferrand,France2INRA,UMRINRA-UniversitédeLorraine“InteractionArbres/Microorganismes”,54280Champenoux,France3USDepartmentofEnergyJointgenomeInstitute,WalnutCreek,California,94598,USA4DepartmentofBotanyandPlantPathology,OregonStateUniversity,Corvallis,Orgon97331,USA5CIRAD,UMR-AGAP,F-63000Clermont-Ferrand,FranceCorynesporacassiicolaisanAscomycetewithalargehostrange,mostlyintheplantkingdom.Itwasalsoreportedinotherfungi,nematodesandinrarecasesofhumandisease.Itwasmostlydescribedas anecrotroph,but also as anendophyteor a saprobe. The specieswaspreviouslyplaced in theFungal Tree of Life where it formed a distinct clade among the Pleosporales, together withCorynesporasmithii.Severalstudieshavedemonstratedthe importantgeneticdiversityamongthespecies.In rubber tree, it is responsible for the Corynespora Leaf fall (CLF) disease which causes massivedefoliationsonsusceptiblecultivars,thusimpairingrubberproduction.Wehavepreviouslypurifiedandcharacterizedasmallproteintoxin,secretedbythehighlyvirulentrubbertree isolateCCPandplaying a role in virulence. However the existence of other effectors, yet uncharacterized, wasevidenced.TheobjectiveofthepresentstudywastoidentifyinsilicoallpotentialeffectorsinvolvedinCLF.ThegenomeofourreferenceisolateCCPwassequencedandassembledbyDOE-JointGenomeInstitutein frame of the 1000 Fungal Genomeproject, and putative effectors identified. PCA based on thecomposition in effectors of 45 fungal species was only weakly related to phylogeny. However, C.cassiicolawasfoundassociatedwithspeciessharingcommonlifestylefeatures,ielargehostrangeand diverse trophic capacities. Transcripts profilingwas conducted to identify functional effectorsdifferentially expressed during the compatible interaction with rubber tree. Finally, intraspecificcomparative analysis of 37 newly-assembled C. cassiicola genomes was conducted in order tocompare their respective setsofputativeeffectors.These resultswillbediscussed in relationwiththegeneticdiversityandtheknownphysiologicalspecificitiesofthestudiedisolates.Keywords:Corynesporacassiicola,Heveabrasilensis,genomics,transcriptomics,effectors,Cassiicolin

Session4Poster77Comparative genomics identifies a new translocon candidate in the type III secretion system ofGram-negativebacteriaCélinePesce1,2,JonathanM.Jacobs1,2,EdwigeBerthelot1,TacaVancheva1,2,ClaudeBragard2andRalfKoebnik1

1Institut de Recherche pour le Développement, UMR 186 IRD-Cirad-Université Montpellier,InteractionsPlante-MicroorganismesEnvironnement(IPME),911avenueAgropolis,BP64501,34394Montpelliercedex5,France2Université catholique de Louvain, Earth and Life Institute, Applied Microbiology Phytopathology,Louvain-la-Neuve,BelgiumXanthomonas translucens is the causal agent of bacterial leaf streak, themost common bacterialdiseaseofwheatandbarley.Tocausedisease,mostxanthomonadsdependonahighlyconservedtype III secretion system,which translocates type III effectors intohostplant cells.MutagenesisoftheconservedtypeIIIsecretiongenehrcTconfirmedthattheX.translucenstypeIIIsecretionsystemis required to cause disease on the host plant barley and to trigger a non-host hypersensitiveresponse inpepper leaves.Type IIIeffectorsaredeliveredtothehostcellbyasurfaceappendage,the hollow Hrp pilus, and a translocon protein complex that inserts into the plant cell plasmamembrane. Homologs of the Xanthomonas HrpF protein, including PopF from Ralstoniasolanacearum and NolX from rhizobia, are thought to act as a translocon protein. Comparativegenomics revealedthatX.translucens strainsharboranoncanonicalhrpgenecluster,whichrathershares features with type III secretion systems from Ralstonia solanacearum, Paraburkholderiaandropogonis, Collimonas fungivorans and Uliginosibacterium gangwonense than otherXanthomonasspp.Surprisingly,noneofthesebacteria,exceptR.solanacearum,encodeahomologof the HrpF translocon. Here,we aimed at identifying a candidate translocon fromX.translucens.Notably,genomesfromstrainsthatlackedhrpF/popF/nolXinsteadencodeanothergene,calledhgiA(for hrpG-induced), adjacent to and likely co-regulated with the type III secretion system genecluster.AninsertionalmutantintheX.translucenshgiAgene,whichisthefirstgeneofatwo-geneoperon,hgiA-hpaH,wasnon-pathogeniconbarleyanddidnotcausethehypersensitiveresponseorprogrammedcelldeathinnon-hostpeppersimilartothehrcTmutant.ThehgiAmutantphenotypeswere partially complemented by either hgiA or the downstream gene, hpaH, which has beendescribedasa facilitatorof translocation inXanthomonasoryzae.These findingssuggest thatbothHgiAandHpaHmaycontributetotheinjectionoftypeIIIeffectorsintoplantcells.Keywords: Xanthomonas translucens, type III secretion system, type III effector, translocon, HrpFprotein,barley

Session4Poster78AdeepenknowledgeofColletotrichumlupini,amajorthreatforLupinculturesRiccardoBaroncelli1,GuillaumeDubrulle1,HenriLepilleur1,DanieleDaLio1,MorganeChalopin1,FloraPensec1,AdelinePicot1,PatriceNodet1,KarimRigalma1,StephanieMadec1,GaetanLeFloch11Université de Brest, EA 3882, Laboratoire Universitaire de Biodiversité et Ecologie Microbienne,IBSAM,ESIAB,TechnopôleBrest-Iroise,29280Plouzané,France.Colletotrichum is a fungal genus gathering numerous species causing anthracnose on a large hostrangeandthesepathogensarespreadworldwide.WithintheColletotrichumacutatumcomplex,thefilamentous fungusColletotrichum lupini is specific toLupins (Lupinusspp.)andresponsible for theanthracnosedisease,oneofthemajorthreatsforLupincrops.In order to get an in-depth understanding of this fungus, we built an extensive collection of 115strains including70strains isolated from infectedLupins inFranceand15 fromdifferentcountriesworldwide distributed, we also included 30 strains from closely related species as outgroups. Anextensivegeneticcharacterizationbasedon10locishowinghighresolutionhasbeencarriedoutinorder to investigate the population structure. Based on the results gained from the multilocusanalysis, we sequenced and annotated the genomes of two C. lupini reference genomes andresequenced three isolates along with 12 closely related acutatum species characterized by apolyphagouslifestyle.ThegenomesequenceswereusedtoperformacomparativeanalysiswiththeaimofexploringgenomicsignaturesassociatedwithhostpreferenceinC.lupini.Weprovidea first lookat thehostadaptationsat thegenomic level thatareassociatedwithhostspecialization in Colletotrichum spp. This study also demonstrates the flexibility of Colletotrichumgenomes,andshowsthatrecentchangesinthegenomesareassociatedwithmajorchangesinhostrangeandepidemiology.Thisstudyalsoexhibitstheneedforhigherresolutiontaxonomicsamplingin order to better understand the role of gene duplication and loss in the evolution of fungalgenomes.TogainabetterinsightinthemolecularprocessesinvolvedintheC.lupini/Lupininteractionand,toconfirm the computational results,wewill perform a comparative ‘omic approach (transcriptomicandproteomic)withthepolyphagouscloselyrelatedpathogenC.fioriniae.Keywords:Colletotrichum,Lupins,genomicanalysis,hostadaptation

Session4Poster79Cinnamylalcoholdehydrogenasedeficiencyimpairspoplar-root-knotnematodeinteractionFabienBaldacci-Cresp1,Pierre-YvesSacré2,AnnegrethKohler3,GeertGoeminne4,AdelineMol1,EricZiemons2, Philippe Hubert2, Gilles Pilate5, Mondher El Jaziri1, Janice de Almeida Engler6, WoutBoerjan4andMarieBaucher11Laboratoire de Biotechnologie Végétale, Université libre de Bruxelles (ULB), Rue des ProfesseursJeeneretBrachet12,B-6041Gosselies,Belgium2UniversityofLiege(ULg),CIRM,DepartmentofPharmacy,LaboratoryofAnalyticalChemistry,CHU,B36,B-4000Liege,Belgium3Interactions Arbres/Microorganismes, INRA-Nancy, INRA, UMR 1136 INRA-Université de Lorraine,Champenoux,54280,France4VIB Department of Plant Systems Biology, 9052 Gent, Belgium; UGent Department of PlantBiotechnologyandBioinformatics,9052Gent,Belgium5INRA,UR0588Amélioration,GénétiqueetPhysiologieForestières,CS40001Ardon,F-45075OrléansCedex2,France6INRA,UniversitéNiceSophiaAntipolis,CNRS,UMR1355-7254InstitutSophiaAgrobiotech,F-06900SophiaAntipolis,FranceAwell-characterizedbranchofthephenylpropanoidpathwayleadstothebiosynthesisofthe3mainmonolignols, which are the p-coumaryl, coniferyl and sinapyl alcohols. Altered expression ofpractically all the genes of the monolignol biosynthesis pathway has been analysed in transgenicplants and mutants, but essentially regarding their impact on lignification. Several studies, usingtranscriptomic,proteomicand/ormetabolomics approacheshave shown that thealterationof theexpressionofgenes involved inthispathwayaffectsnotonly themonolignolbiosynthesispathwaybut also the expression of genes and the accumulation of metabolites belonging to otherphenylpropanoid branches. Monolignols can be polymerized into lignins at the cell wall level orconverted in lignans,thataredi-oroligolignolsknownfortheirrole inthedefenseorduringplantmicroorganisms interactions. RKN belonging to the genusMeloidogyne are endoparasites causingconsiderable damages to both annual and perennial plants worldwide. Global analyses andparticularly transcriptomic analyses, in several interactions models have highlighted thephenylpropanoid pathway and particularlymonolignol pathway as to be altered during both earlyandlategalldevelopmentalstages.Perennialwoodyspeciesareasourceofamultitudeoffruitsbutalsoofahugebiomassfortheproductionofwood,pulpandpaperorbiofuels.Oneofthecurrentscientificchallenges inthecontextofclimatechange isontheonehandtobetterunderstandhowpathogensaffecttreesandinfluencetheirevolution,andontheotherhandtoidentifythemolecularbasis of the natural resistance of trees towards these pathogens. In this context, we studied theimpactofthedeficiencyofseveralmonolignolbiosynthesisstepsonthepoplar-root-knotnematodeinteraction. We show that cinnamyl alcohol dehydrogenase (CAD) deficiency impairs poplar-root-knot nematode interaction. Thanks to a multidisciplinary approach regrouping metabolomic,transcriptomicandRamanspectromicroscopywetriedtoexplainthemolecularoriginoftheimpactoftheCADdeficiencyforplant-root-knotnematodeinteraction.Keywords:Root-knotnematode,poplar,cinnamylalcoholdehydrogenase,metabolism

Session4Poster80StudyofaneffectorfamilyofPlasmoparaviticola,thegrapevinedownymildewpathogenMaudCombier1,FrançoisBerthold1,Marie-ChristinePiron1,FloraPensec1andPereMestre11UMR1131SantédelaVigneetQualitéduVin(SVQV),INRAGrandEst-UniversitédeStrasbourg,28ruedeHerrlisheim,BP20507,68000ColmarPlasmoparaviticola isanobligatebiotrophicoomycete responsible forgrapevinedownymildew,avery recentdiseaseon theEuropeancontinent,arriving fromAmerica in1878.Thisdiseasecausesheavy damage on traditional grapevines and it is currently controlled chemically. To obtain anoptimal health status, growers resort to many fungicide treatments. The excessive application ofphytosanitarymeasuresonvineyardsimposesaselectionpressureresponsiblefortheappearanceoffungicide-resistantstrains.Analternativesolutiontoexpensiveandenvironmentallyunfriendlypesticidesistheuseofresistantgrapevine varieties. Traditional European grapevine varieties are susceptible to P. viticola, theresistance needs thus to be introduced through breeding programs, from American and AsianresistantVitaceae.Inplants,most resistancegenes (R) recognizea specificproduct codedbyageneof thepathogencalled the avirulence (Avr) gene and activate defence responses. Almost all known oomycete AvrproteinsbelongtotheRXLR-EEReffector family.Theseproteinsaresecretedduringplant infectionand,thankstotheRXLRamino-acidmotif, internalizedintoplantcellswheretheymanipulateplantdefenceandpromotevirulence.To identify new resistance genes, our strategy is to search for RXLR effectors essential to thevirulence of the pathogen and use them to screen resistant grapevines. Using genomic resources,more than 250P. viticola effector candidates have been identified. Among the identified effectorcandidates,more than sixty are structurally similar to RXLR effectors from other phytopathogenicoomycetes but, surprisingly, they are devoid of RXLR motif. Two of these effectors held ourattention,becausetheyareexpressedduringtheinfectionandtheirtransientexpressioninleavesofNicotiana benthamiana leads to necrosis. Their study could lead to a better understanding of theevolution of the pathogen’s virulence andof themechanisms of effector’s internalization into theplantcells.Keywords:RXLReffectors,downymildew,grapevine,Plasmoparaviticola,oomycete

Session4Poster81Functionalanalysisofroot-knotnematode(Meloidogynejavanica)virulencegenesinriceMaíra Grossi-de-Sá1, Anne-Sophie Petitot1, Itamara M. Mezzalira1,2, Magda Beneventi1,2, MariaEugênia Lisei de Sá1,2, Deisy X. Amora2, Hugues Baimey3, Janice De Almeida-Engler2,4, Erika V.Albuquerque2,MariaFatimaGrossi-de-Sá2,DianaFernandez1,21IRD- InstitutdeRecherchepour leDéveloppement,UMR186IRD-UM2-Cirad"InteractionsPlante-MicroorganismesEnvironnement"(IPME),BP64501,34394Montpelliercedex5,France2EmbrapaRecusosGeneticoseBiotecnologia(CENARGEN),70770-917Brasília-DF,Brazil3UniversitédeParakou,Benin4INRA-InstitutNationaldelaRechercheAgronomique-UMRIBSV,06903SophiaAntipolis,FranceRoot-knot nematodes (RKN) are endo-parasiteswith awide host range, encompassingmono- anddicotyledonous plant crops. Several RKN species are responsible for rice (Oryza sativa) productionlossesinBrazil,AsiaorAfrica.Successfulinfectionislikelyachievedbyeffectorproteinsproducedinthenematodeesophagealglandcellsandreleasedinthehostplantcells.Theaimofthisstudywasto assess the functional role of threeMeloidogyne secreted proteins (MSP) in rice - nematodeinteractions.We show that theMeloidogyne sp. esophageal gland cell proteinsMSP7,MSP18 andMSP19areconservedinM.incognita,M.javanicaandM.graminicolaspeciesinfectingrice.RT-qPCRassaysshowedthatMSP2,MSP18andMSP19genesareover-expressedallalongtheinfectioncyclein rice roots. Subcellular localization experiments in onion cells showed that that MSP2 may beaddressedtothenucleus,andMSP18andMSP19tothecytoplasmandnucleus.Transgenicrice(O.sativaNipponbare)plantsexpressingthecandidateproteinsorartificialmicro-RNAs(amiRNAs)ableto silence the cognate genes in the nematode were produced. Assessment of nematodereproductiononhomozygoustransgeniclinesallowedtheidentificationofricelineageswithalteredsusceptibility, indicating that these proteins may be involved in Meloidogyne virulence.OverexpressionofMSP18inriceenhancedM.javanicaandM.graminicolareproduction,indicatingthat theMSP18 protein facilitates RKN parasitism. The role of these proteins in suppressing hostimmunity was investigated in tobacco cell death-induced transient assays.MSP18 suppressed theINF1-triggeredprogrammedcelldeath,suggestingthatMSP18can interferewiththeplantdefensepathways.Dataobtainedmayhelpdecipheringnematode-ricemolecular interactionsandhighlightMSP18asanovelRKNvirulenceeffectorabletomodulatehostimmunity.Keywords:Meloidogyne,amiRNA,functionalanalysis,immuno-modulator,rice,virulenceeffector

Session4Poster82UnrelatedvirulencefactorsencodedbynematodeandviralgenomesconvergeontoHUBproteinsinArabidopsisdeSaM.Grossi1,K.Lambou1,A.S.Petitot1,M.Koala2,J.F.Araujo3,M.F.Grossi-de-Sa3,R.Fragoso3,D.Fernandez1,F.Vignols1,4

1UMR 186 Interactions Plantes-Microorganismes-Environnement, Institut de Recherche pour leDéveloppement,Montpellier,France2INERA,Ouagadougou,BurkinaFaso3Embrapa-Cenargen,Brasília,Brazil4UMR 5004 Biochimie & PhysiologieMoléculaire des Plantes, Campus INRA-SupAgro,Montpellier,FrancePlants permanently face changing unfavorable environments against which they deploy adaptedresponses to maintain their life cycle. In turn, plant-invading pathogens need to take intoconsideration evolving defense responses from their host to counteract them efficiently. RecentstudiesinArabidopsisdemonstratedthatpathogensfromdifferentkingdomssuchphytopathogenicbacteria and eukaryote obligate oomycetes and ascomycetes, deploy independently evolvedvirulenceproteinsthatphysicallyinteractwithalimitedsetofhighlyconnectedproteinnodes(hubproteins).Hubsandtheirinteractorsconstituteprotein-proteininteraction(PPI)networkswithgivenarchitectureandbiologicalsignificance.Therefore,interferingwithplanthostPPInetworksthroughthetargetingofhubproteinsisthoughttoprovidetopathogensanefficientmeantomanipulatekeyhostfunctions.Here,weusedayeast-twohybridapproachonselectedplanthubproteinsandpathogeneffectorstoidentify two new evolutionary unrelated pathogen families, namely the rice yellow mottle andImperatayellowmottle sobemoviruses,and the root-knotnematode (RKN)Meloidogyne Incognita,that are able to converge onto plant hub proteins. We notably identify one hub most probablytargeted topromoteRKN feeding siteat theplant root,andotherhubproteins thatmightgovernplantsusceptibilitytovirusesattheleafsurface.Keywords: plant immunity, Hub proteins,Arabidopsis, protein-protein interaction, sobemovirus,root-knotnematode

Session4Poster83Both GacS-regulated lipopeptides and the type III secretion system contribute to PseudomonascichoriicausednecrosisinhostsChien-JuiHuang1,2,EllenPauwelyn1,3,MarcOngena4,PeterBleyaert3,andMonicaHöfte1

1DepartmentofCropProtection,LaboratoryofPhytopathology,GhentUniversity,CoupureLinks653,9000Ghent,Belgium2DepartmentofPlantMedicine,NationalChiayiUniversity,No.300,SyuefuRd.,ChiayiCity,60004,Taiwan(R.O.C.)3Inagrovzw,Ieperseweg87,8800Rumbeke,Belgium4WalloonCentre for Industrial Biology,Universityof Liège–GemblouxAgro-BioTech, PassagedesDéportés2,5030Gembloux,BelgiumPseudomonas cichorii SF1---54, the causal agent of lettuce midrib rot disease, produces severallipopeptides, including cichofactins and cichopeptins which are important virulence factors.Cichofactinsare linear lipopeptidesand involved inantimicrobial activityand surfacemotilityofP.cichorii SF1---54. Cichopeptins are phytotoxic cyclic lipopeptides and able to cause necroticsymptomsonchicory.TheGacS/GacAtwo---componentsystemiswellknowntoregulateproductionofsecondarymetabolitesincludinglipopeptidesinpseudomonads.Additionally,thefunctionsofthetypethreesecretionsystem(T3SS)inP.cichorii-plantinteractionsarenotclearlyclarified.In this study, we investigated the role of the GacS-regulated lipopeptides and the T3SS inpathogenicityofP.cichoriiSF1-54on twohostplants, chicory and lettuce,bydeletinggacSand/orhrpL,whichencodesthekeysigmafactortocontrolT3SSexpression.Pathogenicityandphenotypiccharacterizationofthewildtypeandconstructedstrainswereanalyzed.Comparedwiththewildtype,thehrpLmutantproducedlipopeptidesatasimilarlevelbutthegacSmutant was highly reduced in its lipopeptide production. The mutant deficient in hrpL did notsignificantlydifferfromthewildtypeininductionofnecrosisonchicoryandlettuce,whilethegacSmutantexhibitedsignificantlylesssymptomsonbothhostplantscomparedtothewildtypeandthehrpLmutant. Intriguingly, the gacS hrpL-doublemutant no longer produced lipopeptides and lostpathogenicityonchicorybutwasstillweaklyvirulentonlettuce.Thus,contributionofboththeGacS-regulated lipopeptidesandT3SStopathogenicity/virulenceofP.cichoriiSF1-54 ishighlyassociatedwiththehoststobeinfected.Keywords:gacregulon,cichopeptin,cichofactin,cyclic lipopeptides,LactucasativaL.var.capitata,midribrot

Session4Poster85ToxinproductionbySarocladiumoryzae,themajorsheathrotpathogenofriceKaatPeeters1,VincentdePaulBigirimana2,AshleyHaeck3,KristofDeMeestere3,MonicaHöfte11GhentUniversity,LaboratoryofPhytopathology,DepartmentofCropProtection,CoupureLinks653,9000,Ghent,Belgium2UniversityofRwanda,DepartmentofCropProtection,MusanzePOBox210,Busogo,Rwanda3Ghent University, Laboratory of Environmental Organic Chemistry and Technology (EnVOC),Department of Sustainable Organic Chemistry and Technology, Coupure Links 653, 9000, Ghent,BelgiumSheath rot is a very aggressive, emergingdiseaseof rice.During the last decades, thediseasehasspreadallovertheworldandhasthepowertodestroythetotalyieldofagrowingseason.Themajorpathogencausingthisdisease isa fungus,calledSarocladiumoryzae.This fungusproducesat leasttwotoxiccompounds,helvolicacidandcerulenin.Helvolicacidisnotonlytoxicfortheplant,butalsofor other bacteria, while cerulenin is toxic for other fungi. According to the literature, exogenousapplication of both toxins canmimic sheath rot symptoms, but a clear correlation of these toxinswithdiseaseseverityhasnotbeendemonstrated.ThegoalofthisresearchistostudythediversityofS.oryzaeisolatesfromRwandaandtheircapacitytoproducetoxins.Therefore,wemeasurehelvolicacidandcerulenininvitroandinplantausingLC-MSandstudythegeneticcapacitytoproducetoxinsusingPCRwith specificprimers. Toxin levelswerecorrelatedwith thepathogenicityofdifferentS.oryzaeisolatesafterriceinoculationusingthegraininoculumtechnique.ResultsshowthatS.oryzaeisolates differ tremendously in their toxin production. Ongoing researchwill further elucidate theroleofthesetoxinsinphytotoxicity.Keywords:Sarocladiumoryzae,helvolicacid,cerulenin,sheathrot

Session4Poster86Pathway-specificregulationofthebotcinicacidbiosyntheticgeneclusterinthegreymouldfungusBotrytiscinereaAntoinePorquier1, JavierMoraga2, BérengèreDalmais1,GuillaumeMorgant1, Adeline Simon1,HindSghyer1,Jean-MarcPradier1,IsidroG.Collado2andMurielViaud1

1UMRBIOGER,INRA,Grignon,France2DepartamentodeQuímicaOrgánica,FacultaddeCiencias,UniversidaddeCádiz,SpainBotcinicacid(BOA)isanon-hostspecificphytotoxinproducedbythepolyphagousphytopathogenicfungus Botrytis cinerea. Its biosynthesis relies on the two PolyKetide Synthase encoding genesBcboa6andBcboa9whichareclusteredtogetherwithotherco-regulatedgenesputativelyinvolvedinthepathway.InordertounderstandhowBOAanditsderivatives(botcinins)areregulatedduringthe life cycle of B. cinerea, we investigated the genomic environment of the BOA cluster andsearchedforputativeregulatorencodinggenes.Amongst the clustered genes, Bcboa13 was predicted to encode a Zn(II)2Cys6 transcription factor(TF).FusionofBoa13withGFPindicatedthatitlocalizesintonuclearfoci.InactivationoftheBcBoa13gene resulted in adrasticdiminutionof theexpressionof theBcboa genes, and in theabsenceofBOAandbotcinins.InadditiontoBcboa13,anothergene(Bcboa1)encodesaputativeregulator: thepredictedproteinhasaNmrA-likedomainthatmaybeinvolvedinprotein-proteininteractions.TheimpactofBcBoa1on botcinins production is investigated by gene inactivation, while its possible interaction withBcBoa13istestedbyBimolecularFluorescenceComplementation(BiFC).Finally, the BOA cluster is localized in a subtelomeric region in which the A+T/G+C-equilibratedregions that contain Bcboa genes alternate with A+T-rich regions (>85%) made of relics oftransposableelementsthathaveundergonerepeat-inducedpoint(RIP)mutations.TheoccurrenceofRIPraisesquestionsaboutpossiblechromatin-basedregulationofBOAsynthesis.Severalchromatinmodifiers(histonemethyltransferases)areunderstudiestotestthishypothesis.Identification of BcBoa13 as the major regulator of BOA synthesis is the first step toward acomprehensive understanding of the regulation network of toxin synthesis in B. cinerea. Ongoingworkmaypointouttherespectiveroleofpathway-specifictranscriptionalregulatorsandchromatinstructuremodifications.Keywords: phytotoxin, secondary metabolism, Zn(II)2Cys6 transcription factor, NmrA-like protein,histonemethyltransferases.

Session4Poster87Functionalanalysisofcassiicolin,effectoroftherubbertreepathogenCorynesporacassiicolaSébastien Ribeiro1,2,Marine Déon1,2, DinhMinh Tran2,3,Mouman Soumahoro 4, AurélienMasson5,AndréClément-Demange2,ValériePujade-Renaud1,2

1UniversitéClermontAuvergne(UCA),UMR547PIAF,BP10448,F-63000Clermont-Ferrand,France2CIRAD,UMRAGAP,F-34398Montpellier,France3RubberResearchInstituteofVietnam,HoChiMinhCity,Vietnam4MoumanSoumahoro,SociétéAfricainedePlantationsd’Hévéa,Côted’Ivoire5SociétédesCaoutchoucsdeGrand-Béréby,Côted’IvoireRubbertree(Heveabrasiliensis)isthemainsourceofnaturalrubberaroundtheworld.InAfricaandAsia, it is affected by the Corynespora Leaf Fall disease caused by the necrotrophic fungusCorynesporacassiicola. Somestrainsof thispathogensecretea smallprotein toxin, thecassiicolin,whichplayaroleintheearlystagesofdiseaseestablishment.Inthisstudy,wedemonstratedtheimportanceofcassiicolineforaggressiveness,bycomparingthewild-type virulent CCP strain and the same strain deleted for the cassiicolin-encoding gene Cas1(CCPΔCas1),ininteractionwithrubbertree.Deletion ofCas1 gene did notmodifymajor functions in the CCP strain: the growth rate, conidiaproduction and percentage of germinationwere found similar for both strains. Thewild-type andmutatedstrainswerecomparedfortheirvirulenceondifferentrubberclonesbyanalyzingtheextentof symptoms induced after application of conidia suspension.Only hardly detectable lesionswereobserved with CCPΔCas1, suggesting that cassiicolin Cas1 is an important factor of virulence inrubber tree.Moreover, the two strains were compared for filtrate toxicity over a larger range ofrubberclones,bymeasuringtheelectrolyteleakagesinducedondetachedleavesbytheapplicationof purified cassiicolin Cas1 or culture filtrate from several C. cassiicola strains (including CCP andCCPΔCas1). The filtrate of the mutated strain was on average less aggressive than the wild-typefiltratealthough itstillgeneratedseveresymptomsonafewclones,suggestingthe involvementoftoxicity factors other than cassiicolin. Hierarchical clustering (clones x treatments) based on thesetoxicityprofilesgroupedCCPΔCas1withstrainsofadifferentphylogeneticgroup(cladeA)comparedto CCP (clade C), without cassiicolin-encoding gene. These results demonstrate that, despite theproductionofothereffectorsbyCCP,cassiicolinCas1isamajordeterminantofthisstraincontrollingthevirulencelevelinrubbertree.Keywords:Corynesporacassiicola,rubbertree,cassiicolin,effector,deletionmutant

Session4Poster88InvestigationofinteractionsbetweeneffectorproteinsofBlumeriagraminisf.sp.hordeiandbarleyresistanceproteinsBjörnSabelleck1,RalphPanstruga11RWTHAachenUniversity,InstituteforBiologyI,UnitforPlantMolecularCellBiology,Worringerweg1,52074AachenTheobligatebiotrophpowderymildew fungusBlumeriagraminis f.sp.hordei (Bgh) coloniesbarley(Hordeum vulgare) epiphytically. Supposedly for nutrient uptake and the secretion of effectorproteins, the fungusestablishes its feedingstructure (haustorium) insideplantcells.Tomanipulatethe host defense, the fungus is believed to use an arsenal of around 530 candidates for secretedeffector proteins (CSEPs). Plants evolved with nonhost resistance (NHR) and R-gene mediatedresistance two different layers of defense to prevent a successful colonization. Through formerstudiesanumberofbarleygeneshavebeenidentifiedthatareassociatedwithNHR.Someofthesegenesbelongtothefamilyofreceptor-like-kinases(RLKs).R-genemediatedresistancereliesonNod-like receptors (NLR), which recognize specific effector proteins. Some of these NLRs evolvedintegrated decoy domains. These domains mimic target protein domains of effectors, possibly toattract them. Another potential target of effectors is the MLO protein, a negative regulator ofdefensetoBgh.TheaimofthisERA-CAPS/DURES-Trit-fundedPhD-Project istoanalyzethephysicalinteractionofselectedbarleyRLKswithBghCSEPs.Wealsoincludeasubsetofbarleydecoydomainsand the barley MLO protein to broaden the scope of this study. To discover protein-protein-interactions between CSEPs and RLKs, decoy domains and MLO proteins, we performed split-ubiquitinassays(amodifiedyeasttwo-hybridmethod)withthebarleyproteinsasbaits.Aspreysweused a constructed CSEP library. The most meaningful interactions wil be tested by in plantabimolecular fluorescence complementation (BiFC) analysis and transient gene expression assays inbarleysinglecells,whichallowtodeterminetheimpactoftheseinteractionsoninfectionsuccess.Toidentify amino acids/domains that are crucial for these interactions, we will use site-directedmutagenesisofbaitand/orpreyproteins.Ouranalysiswillfurtherimproveourunderstandingofthebarley-powderymildewinteraction.Keywords:barley,powderymildew,effectors,protein-proteininteractionstudy

Session4Poster89Susceptibility of sweet corn genotypes on Fusarium spp. and biosynthesis their secondarymetabolitesŁukaszStępień1,ElżbietaCzembor2,AdamPerczak3,BartoszCiorga3,AgnieszkaWaśkiewicz3

1InstituteofPlantGenetics,PolishAcademyofSciences,DepartmentofPathogenGeneticsandPlantResistance,Strzeszyńska34,60-479Poznań,Poland2PlantBreedingandAcclimatizationInstitute-NRI,Radzików,05-870Błonie,Poland3UniversityofLifeSciences,DepartmentofChemistry,WojskaPolskiego75,60-625Poznań,PolandAt present we are observing considerable progress in sweet corn breeding, manifested in theconsiderable number of new cultivars of high economic value, differing in the length of thevegetation period, yield, contents of sugars and quality of kernels. With the appearance of newsweet corn hybrids, a need has been recognised to conduct research on their susceptibility todiseases caused by fungal pathogens, including Fusarium spp., mainly F. proliferatum and F.verticillioides, which synthesised toxic secondary metabolites like fumonisins, beauvericin ormoniliformin.Theaimofthestudywastoestimatethesusceptibilityofvarioussweetcorngenotypes(Sweetstar,Shine Rock, Overland, GSS 8529) under natural infection and after inoculation two isolates ofFusarium spp. separately - Fusarium proliferatum and F. verticillioides – representative for thepopulationofthesespeciesinPoland,comingfromthecollectionofpathogenicfungiPASinPoznań.Inmaizesamplescollected fromthreeexperimentalgroups (control -natural infection, inoculationwith F. verticillioides and F. proliferatum) fumonisins from group B (FBs) – after extraction andpurification–wereanalysedusingHPLCmethods.Inourstudies, thedifferencesbetweentestedgenotypes inmycotoxins levelsweredemonstrated.ThehighestFBslevelshownforcv.GSS8529afterinoculationbothF.verticillioides(17.40µg/g)andF.proliferatum(18.11µg/g),whereasothergenotypeswerecontaminatedwithfumonisinsatsimilarlower levels of concentration (1.05-9.62 µg/g – inoculationwith F. verticillioides, 2.47-6.64 µg/g –inoculationwithF.proliferatum).OurresultsdependedstronglyontestedgenotypesandisolatesofFusariumspp.Thepartof this researchwasfinanciallysupportedfromthePolishNationalScienceCentreproject2014/15/B/NZ9/02169Keywords:beauvericin,fumonisins,Fusariumproliferatum,F.verticillioides,maize,moniliformin

Session4Poster90Autophagy indistant eukaryotic lineages is deregulatedby an effector from theoomyceteplantpathogenPhytophthoraparasiticaSerenaTesti1,4*,Marie-LineKuhn1,4,PascalineAuroy2,FantaoKong2,GillesPeltier2,NathalieZuccini-Pascal3,HaraldKeller1,4,FranckPanabières1,41UMR1355-7254InstitutSophiaAgrobiotech,06900SophiaAntipolis,France2UMR7265CEACadarache,13108Saint-Paul-lez-Durance,France3UMR1331TOXALIM,06903Sophia-AntipolisCedex,France4contributedequallyOomycetes from thegenusPhytophthora areplantpathogens,whichhavedevastating impactsonagriculture and natural ecosystems. Phytophthora parasitica is a root pathogen with ahemibiotrophic lifecycle:during the initial stagesof infection (biotrophy) theoomyceteestablishesanintimatecontactwiththelivingcellsofthehost,beforeinducingplantcelldeathtocompleteitslifecycle(necrotrophy).cDNAlibrariesthatwereobtainedfromP.parasitica-infectedtomatoplantsandonionepidermiscellsdisplayseveralsequencesthatencodesecretedproteinswithacanonicalRxLRtranslocationsignal,suchasAvh195.Avh195possessesthreepotentialbindingsitesforATG8,akeyproteinintheprocessofautophagy.HeterologousexpressionofAvh195intobaccoplantsslowsdown cell death responses such as those induced by proapoptotic BAX, and the HR inducerscryptogein and AvrPtoB. On this basis, we investigate the antagonism between death-inducingagentsandAvh195aimingatidentifyingthemanipulatedhostsignalingpathways,withaparticularfocusonthelinkbetweenAvh195andtheautophagymachinery.ToidentifythemoleculartargetsofAvh195,weinitiatedatrans-phylumanalysisonplants,humancells,andgreenmicroalgae.GeneticexpressionofAvh195dramaticallyaltersthecellularphenotypeinalltheseorganisms,indicatingthatthisproteintargetsanevolutionary-conservedmechanism.*S.TestiissupportedbytheLabex“Signalife”[#ANR-11-LABX-0028-01].Keywords:autophagy,Chlamydomonas,Arabidopsis,HeLa,oomycete,effector

Session4Poster91Transcriptomic and proteogenomic analysis of early gene expression profile of XanthomonasoryzaepathovaroryzaeinrecognitionofthehostJeong-GuKim1,SeunghwanKim1,Yong-JoonCho2,Eun-SungSong1,SangHeeLee3,Lin-WooKang4

1Genomics Division, National Institute of Agricultural Sciences, Rural Development Administration(RDA),Jeonju54874,Korea2Chunlab,Inc.,SeoulNationalUniversity,Seoul08826,Korea3 National Leading Research Laboratory of Drug Resistance Proteomics, Department of BiologicalSciences,MyongjiUniversity,116Myongjiro,Yongin17058,Korea4Hwayangdong,Gwangjin-gu,Seoul05029,KoreaGenome-widegeneexpressionofXanthomonasoryzaepathovaroryzae(Xoo)wasquantitativelyandtime-resolvedlyanalysedinrecognitionofthehost.Thepathogenicity-relatedgenesofthepathogenwere activated using rice leaf homogenate. The gene expressionswere quantitatedwith RNA-SeqandvalidatedwithqRT-PCRforsevendifferenttimepointswithin1hposttreatment.Globalanalysisof gene expression and regulation revealed themost dramatic changes in functional categories ofgenes related to inorganic ion transport and metabolism, and cell motility. Expression of manypathogenicity-related genes was induced within 15 min upon contact with RLX. hrpG and hrpXexpressionreachedthemaximumlevelwithin10and15min,respectively.Chemotaxisandflagellabiosynthesis-relatedgenesandcyclic-di-GMPcontrollinggenesweredownregulatedfor10minandwere thenupregulated.Genes related to inorganic ionuptakewereupregulatedwithin5min.Weintroducedanon-linear regression fit togeneratecontinuous time-resolvedgeneexpression levelsand tested the essentiality of the transcriptionally upregulated genes by a pathogenicity assay oflesion lengthusing single-geneknock-outXoo strains. The in vitro systemcombinedwithRNA-Seqgeneratedagenome-widetime-resolvedpathogenicgeneexpressionprofilewithin1hofinitialrice-Xoo interactions, demonstrating the expression order and interaction dependency of pathogenicgenes.Thesynthesisandsecretionofproteinsfollowedlate.Keywords:Xanthomonasoryzae,time-resolved,geneexpression,interaction

Session4Poster92PathogenicityandgenomecharacteristicsofBurkholderiaspp.causingricegrainrotinKoreaJi-EunRa1,2,Sang-MinKim1,SuJwaSeo1,BongChoonLee1,NakJungChoi1,Man-YoungChoi1,KiDoPark1,Ill-MinChung2

1NCIS–NationalInstituteofCropScience,RuralDevelopmentAdministration,Wanju,55365,Rep.ofKorea2DepartmentofAppliedLifeScience,CollegeofLifeandEnvironmentalSciences,KonkukUniversity,Seoul,05029,Rep.ofKoreaRice is an important cereal crop globally and the important rice diseases have been studiedwell.However, theclimatechangeandcroppingof restrictcultivarscauseapprearanceofnewdiseases.Therefore,itisnecessarytoconductcontinuousdiseasemonitoring.Burkholderiaspp.areknownasapathogencausingricegrainrotandblightinseedling.Thisdiseaseoutbreakresultsinyieldlossandafallingoffingrainquality.Grainrotgenerallyspreadwidelyduringtheheadingstagewithhotandhigh humidity weather condition. As global warming progresses, the important of this disease isincreasing.From the discolored grain collected in the fields Korea, we screened 66 isolates by colonymorphology and PCR-based screening. Then, 3 pathogens, B. glumae (GRbb 6), B. gladioli (GRBB15041,GRBB15043),andB.plantarii (GRBB15061,GRBB15061-1),were identifiedwith16SrRNAand phylogenetic marker genes sequencing. From phylogenetic analysis of 16S rRNA, these aredifficulties ingroupingbetweenreferencestrainandKorean isolates.The isolatesGRBB15041andGRBB15043werenot assigned to cluster togetherwith reference strainB.gladioli. These isolatesweredifferentiatedearlierthanB.glumaeandB.plantariiisolates,andformedagroup.Ontheotherhand, the isolates GRBB 15061 and GRBB 15061-1weremore near toB. glumae thanB. gladioli.AftersequencinganalysisofrpoDandgyrBgenes,theseisolateswereclassifiedaccordingtospecies.Therefore, to differentiate groups among Burkholderia spp. accurately, it is necessary to analysisother phylogeneticmarker genes. Asmajor factors showing pathogenicity against rice plant weredetectedastoxoflavinandtropoloneusingultra-highperformance liquidchromatographyequippedwithmass/mass.Theamountofthesephytotoxinsdifferedbetweenthe isolatesaccordingtotheirdegree of pathogenicity. Results from this study, we revealed the relationship between Koreanisolates,andevaluatedtheirpathogenicity.Keywords:Burkholderia,grainrot,rice

Session4Poster93Genome sequencing of isolates of the emerging pathogen Ramularia collo-cygni improvesunderstandingoffungallifestyleHind Sghyer1, Remco Stam1,2, Aurelien Tellier2, Martin Münsterkötter3, Ulrich Güldener3, RalphHückelhoven1,MichaelHess11Technical University of Munich (TUM) School of Life Sciences; Chair of Phytopathology, 85354Freising-Weihenstephan,Germany2TUMSchoolofLifeSciences,SectionofPopulationGenetics,85354Freising-Weihenstephan3InstituteofBioinformaticsandSystemsBiology,HelmholtzZentrumMünchen,D-85764NeuherbergRamulariaLeafSpot(RLS)hasemergedtoamajorthreatforbarleyproductioninmanyregionsoftheworld. Due to the late appearance of unspecific symptoms there were especially moleculardiagnostics thatdetected the fungusRamularia collo-cygni (Rcc) throughout the lifecycleof barleyandinsamplesworldwideasthebioticfactorofthecomplexdisease.TheeconomicimpactofRLSisdependingontheavailabilityandeffectivenessofcontrolmeasures.Since thereareno sufficiently resistant varieties, control is relyingon theuseof fungicides. In thepastyearsRcchasquicklyadaptedtothelimitednumberofeffectivefungicidecompounds.ThestudyofthebiologyofRccasabasisforsustainablecontrolhasbeencomplicatedbydifficultieswithtraditionalapproachesbyinvitrogrowth,sporulation,andinoculation.To address urging questions, especially the uprise to a major disease, the relevance of seedtransmission,andquickadaptationtocontrolmeasures,thegenomeofRcc (urug2isolate)wasde-novosequenced.FungalRNAfromsixdifferentconditionswassequencedtosupportannotationandto uncover putative genes of interest. The assembled genome is about 32 Mb and the overallannotation enabled the prediction of 12346 genes. RNAseq data identified genes differentiallyexpressedbetweenthedifferentconditionsandenrichedinthefunctionalcategory“plant-pathogeninteractions”. Many of those are homologues of genes expressed in planta by closely relatedZymoseptoriatritici.To evaluate the genetic diversity, whole genome sequencing of 19 Rcc isolates from multiplegeographic locations andhostswasperformedandmapped to the reference genome.Preliminaryanalysisindicatedsubstantialgeneticdiversityandapossiblepopulationsizeexpansion,whichmightexplaintherecentemergenceofthisfungus.Theanalysis isongoingandrecentconclusionsonthepathogenbiologywillbepresented.Keywords:leafspots,barley,Ramulariacollo-cygni,genome,populationgenetics

Session4Abstract94(requestfororal)Multiplemetabolites produced byPseudomonas sp. CMR12a are involved in the suppression ofPythiumrootrotdiseaseFeyisara EyiwumiOlorunleke1,Oluwatoyin FaithOlorunleke1,NamKieuPhuong1,OlumideOwolabiOmoboye1,MarcOngena2andMonicaHöfte11Laboratory of Phytopathology, Faculty of Bioscience Engineering,GhentUniversity, Coupure Links653,B-9000Gent,Belgium2GemblouxAgro-BioTech,UniversityofLiège,Gembloux,Belgium.RootrotdiseasecausedbyPythiummyriotylumisthemostdevastatingdiseaseofthetropicaltubercrop cocoyam (Xanthosoma sagittifolium) with production losses of up to 90%. This study wasconducted inorder todetermine the roleofphenazines (PCN),and twocyclic lipopeptidesnamelysessilins and orfamides, produced by the biocontrol strain Pseudomonas sp. CMR12a, in thesuppressionoftheCocoyamRootRotDisease(CRRD).PreviouslygeneratedbiosynthesismutantsofCMR12a that were deficient in either one, two or all three metabolites were used in plant andmicroscopic experiments. Plant experiments revealed that mutants, which produced sessilins,orfamidesorphenazinesaloneoratleastanytwoofthemetabolites,couldeffectivelysuppressthecocoyamrootrotdiseasecausedbyP.myriotylumalbeit invaryingcapacities.Microscopicanalysisshowed that 1 nM, 10 nM, and 25 nM of crude sessilins, purified orfamides, and phenazines,respectively, resulted in hyphal damage of P. myriotylum including vacuolization and lysis.Interestingly, during plant experiments, the null mutant, which did not produce any of the threemetabolites,wasable togive some levelofdisease suppression, indicating thatCMR12aproducesothercompound(s)whichcouldbeantagonistic towardsour targetpathogen.SubsequentgenomeminingoftheCMR12adraftgenomerevealedthatatabtoxingeneclusterissituatedonthegenomicisland thatharbors thephenazine-sessilinbiosynthesisgeneclusters.Furthermore,weshowthatadoubledeletionmutantwhichlostboththegenomicislandandtheorfamidegeneclustercouldnolonger suppress CRRD. In summary, our study reveals that multiple metabolites produced byPseudomonas sp.CMR12a,canplay independentandadditiveroles in thesuppressionofPythium-mediatedCRRD.Keywords:Pythiumspp,Pseudomonas,orfamides,sessilins,tabtoxin,cocoyam

Session4Poster95Antioxidant responses in ‘Braeburn’ and ‘Golden Delicious’ apple fruit to post-harvest storagediseasecausedbyBotrytiscinerea.Tuyet T. A. Bui1,4*, Sandra A. I. Wright4*, Anders B. Falk5, Tanja Vanwalleghem2, Wendy VanHemelrijck2,MaartenL.A.T.M.Hertog3,BartM.Nicolai3,7,JohanKeulemans1,MarkW.Davey1,61Lab. of Fruit Breeding and Biotechnology, Department of Biosystems, Faculty of BioscienceEngineering,K.U.Leuven,Leuven,Belgium2ProefcentrumFruitteeltvzw,Sint-Truiden,Belgium3Division of MeBioS, Department of Biosystems, Faculty of Bioscience Engineering, K. U. Leuven,Leuven,Belgium4SectionofBiology,UniversityofGävle,SE-80176Gävle,Sweden5DepartmentofPlantBiologyandForestGenetics,SwedishUniversityofAgriculturalSciences(SLU),75007Uppsala,Sweden6Currentaddress,BayerCropScience,Technologiepark38,9052,Zwijnaarde,Belgium7FlandersCentreofPostharvestTechnology,Heverlee,BelgiumApplesareusuallystoredforaperiodofupto6-9monthstoensureasteady,year-roundsupplyofhigh-qualityfruit.Duringthistime,majorlossesmayoccurfrompost-harveststoragedisordersanddiseases.Botrytiscinereaisapost-harvestpathogen,whichattacksover200differentplantspecies,includingapple.Thepresentworkexaminestheinvolvementofantioxidantmetabolismofapplefruitin their susceptibility to artificial inoculation with B. cinerea. Apple fruit of two cultivars withcontrasting susceptibility, ‘Braeburn’ and ‘Golden Delicious’, were inoculated on the sun-exposedand shaded sides. The antioxidant enzyme activity and antioxidant content of apple fruit weremeasuredovertimeinpeelandfleshsamplestakenfrombothsidesofthefruit.Overall,‘Braeburn’was more susceptible than ‘Golden Delicious’ to B. cinerea, even though sun-exposed tissues of‘Braeburn’hadhigher initial levelsof totalvitaminC inpeelandphenoliccompounds in flesh,andhigher of superoxide dismutase (SOD) activity and flavonoid peroxidase (POX) activity in all fleshtissues as compared to those of ‘Golden Delicious’. In ‘Braeburn’, inoculation with B. cinereaprovoked an antioxidant response, involving an increase superoxide dismutase (SOD) activity andascorbateperoxidase(APX)activity,whichwereaccompaniedbytheprogressiveoxidationofvitaminC,andadecreasePOXactivity.Diseasetendedtodevelopmorerapidlyontheshadedthanonthesun-exposed side of ‘Braeburn’ fruit, involving a decrease of total phenolic content. Pre-harvestexposuretohighlight/hightemperaturestressisproposedtoreducethesusceptibilityofapplestosubsequent postharvest pathogen.Disease susceptibility dependedon different factors in the twocultivars. In ‘GoldenDelicious’, vitaminC level remained low throughout,andantioxidantenzymeswerenotsignificantlyinducedasaresultofinoculation.Thus,‘GoldenDelicious’appearstoutilizeadifferentlineofdefencetofendoffB.cinerea.Keywords:MalusXdomestica,postharveststorage,Botrytiscinerea,antioxidantmetabolism

Session4Poster96IlluminatingtheroleofextracellularvesiclesintheinteractionbetweenmicrobesandplantsEgidioStigliano1,LuciaGrenga2,3,FrankMenke1,JanSklenar1,JacobMalone2,3,SilkeRobatzek11TheSainsburylaboratory2JohnInnesCentreNorwichResearchPark,Norwich,NR47UH,UK3School of Biological Sciences, University of East Anglia Norwich Research Park NorwichNR47TJUKMany microbes colonize the extracellular spaces between plant cells. A physical barrier is thuspresent between the two organisms. Yet, the signal exchange between plants andmicrobes overtheir cell surfaces remains poorly understood. Here we elucidate the role of microbe-derivedextracellular vesicles, with their proteins, in the cell-to-cell communication with plants for theoutcome of immunity. Our research focused on the bacterial model Pseudomonas and theinteractionwithArabidopsisthaliana.Webiochemicallypurifiedoutermembranevesicles(OMVs)oftwo Pseudomonas species and examinedA. thaliana anti-bacterial immunity upon induction withOMVs.Surprisingly,plantswerenotprotectedagainst infectionwhenpre-treatedwithOMVs fromthe bacterial pathogen P. syringae pv tomato DC3000 but exhibited increased susceptibility.However, Pto DC3000 OMVs are immunogenic inducing typical defence responses known formicrobe-associated molecular patterns (MAMPs). To elucidate the mechanism by which pre-treatmentwith PtoDC3000OMVs supports disease susceptibility,we determined their proteomiccontents.Ofthese,158proteinswerespecifictoPtoDC3000OMVswhile351proteinswerefoundinbothOMVsandthesupernatantfractions.InagreementwiththeimmunogenicactivityofOMVswefound bacterial EF-Tu, a knownMAMPs for A. thaliana, in the OMV proteome. Furthermore, theOMV proteome was enriched in a putative effector with a cellulase activity. The correspondingbacterialmutantinthiscellulasewaslessefficientinbindingCongo-Red,indicatingapotentialroleinbacterialcellsurfacemodifications.Insummary,ourresultsprovideevidenceoftheinvolvementofOMVsfromphytopathogenicbacteriatopromotetheirvirulence.Keywords:outermembranevesicles,plantimmunity,MAMP

SESSION5.FROMPLANTIMMUNITYTOINNOVATIVEPLANTBREEDINGSession5Poster97ElucidatingthemolecularandchemicalresponsesofresistantricetoFusariumfujikuroiDavide Spadaro1,2, Slavica Matic1, Ilenia Siciliano1, Paolo Bagnaresi3, Chiara Biselli3, Luigi Orru’3,GiampieroValé3,MariaLodovicaGullino1,21AGROINNOVA – Centre of Competence for the Innovation in the Agro-environmental Sector,UniversityofTurin,LargoBraccini2,10095Grugliasco,TO,Italy2DepartmentofAgricultural,ForestryandFoodSciences(DISAFA),UniversityofTurin,LargoBraccini2,10095Grugliasco,TO,Italy3Council foragriculturalresearchandeconomics (CREA),GenomicsResearchCentre,viaS.Protaso,302I−29017,Fiorenzuolad’Arda,PC,ItalyThemechanismsofricedefenseagainstFusariumfujikuroihavenotyetbeenfullyclarified.Inorderto elucidate the factors involved in rice resistance against bakanae disease, an RNA-seqtranscriptomestudywasperformed.Themoleculareventsthattakeplaceduringtheresponseoftheresistant ‘Selenio’ cultivar and susceptible ‘Dorella’ cultivar were identified at 21 days postgermination. Thebasic rice resistancemachineryagainstF. fujikuroi involvedPRgenes, glucanasesand peroxidases, since theywere upregulated in both the resistant and susceptible cultivars. Thespecialized and evolved resistance mechanisms in the resistant cultivar included WRKYtranscriptionalfactors,MAPKcascades,andsomecytochromeP450genes.ThesemechanismswerefurtherconfirmedbyKEGG identificationofCa2+-dependentproteinkinasegene, JASMONATEZIM-DOMAIN-likegenes,CEBiP,CERK1,andMYC2genes,foundonlyin‘Selenio’.Thesegenesparticipatein one of the molecular patterns: response to chitin, jasmonic acid biosynthesis, and planthypersensitive response. When the gibberellin production was controlled, the ‘Selenio’ plantsactivated the jasmonic acid metabolic pathway. In this way, ‘Selenio’ maintained its bakanae-resistancelevel.Increasedconcentrationsoffourricephytoalexinswereonlyfoundin‘Selenio’.ThegreatestincreaseinphytoalexinbiosynthesiswasobservedforsakuranetinandmomilactoneA.Thefungal pathogen in the resistant cultivar acts locally, at lower concentrations, and causes a ricehypersensitiveresponsewithoutanyfurtherdamagetotheplants.Keywords:bakanae,jasmonicacid,Oryzasativa,phytoalexins,RNA-seq

Session5Poster98Moniliasp.infectionweakensresistanceoftransgenicplumPrunusdomesticaL.,cv.HoneySweettotheplumpoxvirusJ.Polák1,J.K.Kundu1,P.Komínek1,E.Beoni1,andT.Neubauerová11DivisionofCropProtectionandPlantHealth,CropResearchInstitute,Drnovská507,16106Prague,CzechRepublicSharka caused byPlum pox virus (PPV) is themost harmful disease of stone fruits in Europe andelsewhereintheworld.ThereisnohighlyPPVresistantcultivarofplum.Biotechapproachhasledtothedevelopmentofresistancethroughgeneticengineering.Inthisstudy,weevaluatedatransgenicplumPrunus domestica L., clone C5 (cv. HoneySweet),where the PPV resistance is based on RNAinterference(RNAi).ResistanceinC5plumshasbeenevaluatedforPPV,Prunedwarfvirus(PDV),andApplechloroticleafspotvirus(ACLSV)inaregulatedfieldtrialintheCzechRepublicforfifteenyears(2002-2016).Nonatural infectionofPPVbyaphidswasrecorded inC5plums.Co-infectionsofPPVwithPDVand/orACLSVhadpracticallynoinfluenceonstabilityofresistancetoPPVinC5trees.Evenunder high and permanent infection pressure introduced through graft inoculation of the viruses,PPV has been detected in C5 trees only in several leaves situated close to the point of inoculumgraftinginthefirstnineyears.MildsymptomsofPPVdisappearedyearbyyear.NoPPVsymptomswere observed in the following three years (2011-2013) and results of ELISA detection testswerenegative. Similar results were obtained, when RT-PCR was used for PPV detection. There was asevereattackoftransgenicplumtreesbyMoniliasp. inthetwelfthyear.MildPPVsymptomshaveappearedagaininseveralleavesinthenexttwoyears(2014-2015)aftertheMoniliasp.infectionanddisappearedinthefiteenthyear.ThepresenceofPPVwasconfirmedbyELISAandRT-PCRnotonlyinsymptomatic leaves,butalso inseveral fruitsshowingnosymptoms.The lowpresenceofPPVwasconfirmedinseveralasymptomaticleavesandfruitsinthefifteenthyearofevaluation,too.Keywords: Plum pox virus, resistance, transgenic plum, cv. HoneySweet, Monilia sp., infection,resistanceweakening

Session5Poster99Developmentofamycorrhizationmethodofgrapevineinsemi-hydroponicsasatoolforstudiesofdefenseresponsesagainstpathogensLaurence Deglène-Benbrahim1, Pascale Maillot1, Melha Kendel1, Réjane Hussenet2, Christine LeJeune1,PaulSchellenbaum1andLaureValat11LVBE-LaboratoireVigne,Biotechnologies&Environnement,UniversitédeHauteAlsace,33ruedeHerrlisheim,F-68008ColmarCedex,France2IUTdeColmar,UniversitédeHauteAlsace,29ruedeHerrlisheim,BP50568,F-68008ColmarCedex,FranceMycorrhization by arbuscular mycorrhizal fungi (AMF) has been demonstrated to have positiveeffectsonplantgrowthandresistancetomanybioticaggressors (Pozo&Azcon-Aguilar,2007)andcouldbeaninterestingwaytoreduceorreplaceuseofpesticides.Studies of AMF impact on grapevine defense responses during pathogen challenge would befacilitated by a procedure to obtain rapidly a large number of mycorrhizal plants in controlledconditionsofnutritionandenvironment.AccesstotherootsystemshouldbealsosimplifiedtocheckthepresenceofAMfungusand/orsoil-bornepathogensafterinoculation.Forthispurpose,wedevelopedamethodtoobtainmycorrhizalgrapevineplantsacclimatizedfrominvitrocultureinsemi-hydroponics.Six-old-weeks invitroplantletsoftherootstock41Bweretransplantedinindividualpotscontaining1:1 sterilized sand/vermiculite and acclimatized in growth chamber (25°C/20°C day/night, 16hphotoperiod) during five weeks. They were then inoculated with an AMF strain of Funneliformismosseae spore suspension. Mycorrhization level was checked six weeks later according to themethodofTrouvelotetal(1986).Root systemofplantswasmycorrhizedwitha frequencycomprisedbetween90and100%andanintensityof51.3-69.8%.Proportionofarbusculeswasfrom70to95%inmycorrhizalpartsandfrom44 to 60% in whole root system. AM fungus root colonization is thus particularly efficient in thissystem comparing to others results reported in literature. For instance, Eftekhari et al (2012)reportedmycorrhizal frequency of 70% for this AMF species in grapevine plants from hard-woodcuttings.Withthismethodalotofhomogeneoushighlymycorrhizalgrapevineplantscanbeobtainedrapidly,allovertheyear,instandardizedgrowingconditionsandwithoutgrowthinhibition.Thistoolwillbeusedforanalysisofgrapevinedefensereactionsagainstabovegroundorsoil-bornepathogens.Keywords: grapevine, mycorrhization, arbuscular mycorrhizal fungi, semi-hydroponic culture,acclimatizedinvitroplants

Session5Poster100StudyofasoybeaninduciblepromoterbyPhakopsorapachyrhiziLisaCabre1,4,LaurineGilles2,LauraVelazquez3,StephanePeyrard4,CatherineSirven4,SophieDucerf4,BernardPelissier4andNathaliePoussereau1

1Equipe de Génomique Fonctionnelle des Champignons Pathogènes des Plantes UMR 5240MicrobiologieAdaptationPathogénie,CNRS-UCB-INSA-BayerSAS,14ImpassePierreBaizetBP99163,69263LyonCedex09,France2Laboratoire de Reproduction et Développement des Plantes équipe développement de la graine,ENSdeLyon46,alléed'Italie,69364LYONCedex07,France3Faculty of Life Sciences, Michael Smith Building, The University of Manchester, Oxford Road,ManchesterM139PT,UK4BayerSAS,CropScienceDivision,14impassePierreBaizet,BP99163,69263LyonCedex09,FrancePhakopsorapachyrhiziisabiotrophicfungusresponsibleoftheAsianSoybeanRust(ASR),adiseasecausingimportantyieldlossesinsoybeanproducingcountries.IthasbeenshownthatP.pachyrhiziinfectiontriggersimportanttranscriptionalchangesinsoybean(Tremblayetal.,2010and2011)withmore than hundreds of genes up or down regulated as a consequence of promoter modulation.Study of the response of promoters to pathogens is central to understand the gene regulation inplant which is important for biotechnological applications. In this work we focused on theidentification and characterization of a soybean promoter induced by P. pachyrhizi. Analyze ofinternal andexternalRNAseqdataallowed to select a chitinasegene stronglyup-regulatedat24hpost infection for 10 days, but not by chitin treatment. We first validated and investigated theinduction of this promoter during P. pachyrhizi infection by qRT-PCR. Then stable soybean linesexpressing a GFP under the control of this promoter were generated. GFP fluorescence wasquantified (+/-) after biotic and abiotic stresses to determine the promoter specificity. We alsolookedattheactivityofthispromoterafteractivationofdifferentsignalingpathways implicatedinplant defense response. The results showed that this promoter induced byP. pachyrhizi, was notinduced by a necrotrophic fungus and physical wounding. Activation of Jasmonate, Ethylene andSalicilatesignalingdidnot trigger inductionof thispromoter.Wedemonstratedthat thispromotercouldbespecificallyinducedbyP.pachyrhizitothecontraryofotherpathogen-induciblepromotersoftenactivatedbyseveralstimuli.Keywords:soybean,Phakopsorapachyrhizi,induction,promoter,GFPfluorescence

Session5Poster101Resistancetofusariumearrotinmaize:heritabilityandtraitassociationsElzbietaCzembor1,SewerynFrasinski1,KrzysztofWojcik2andJozefAdamczyk31ForageGrassesandLegumesDept.,PlantBreedingandAcclimatizationInstitute-NRI,Poland2PlantBreedingSmoliceLtd,Poland3MałopolskaPlantBreedingRaisingHBPLtd,PolandRed and pink ear rots caused by Fusarium spp. are important factors affecting the yield and itsquality,mainlybecauseofcontaminationwithmycotoxinsproducedbythefungi.Thedevelopmentofresistanthostgenotypesstronglydependsonavailabilityofsourcesofresistanceandinformationon host pathogen interactions. Themode of inheritance of resistance appears to differ: additive,possibly non-additive effects, digenic and polygenic patterns have been identified. It depends onseveral components such as resistance to initial infection, resistance to fungal degradation of silktissues, resistance to fungal spread by through a wax layer in the grain or grainmorphology andchemicalcompoundsof thepericarp.Theaccumulationof toxinscanalsobeaffectedby theplantgenotype. Although selection is effective to reduce disease severity after inoculation with F.graminearum, additional genes seemed to affect grain DON concentration (i.e., ratios DON/DS ingrainsdependedongenotype),indicatingthatspecificmechanismsarepresentintheplantaffectingDONproductionbydefungusandadditionalgeneticprogresswouldbeachievedbyincludinggrainDON concentration as a selection parameter. The present research was conducted to estimateheterosis, heritability and correlation coefficients to ear rot in set of F1 crosses generated fromresistantandsusceptibleparents.ThepositiveheterosisforearrotresistanceandDONcontentwasobserved.When the twoparents componentswere susceptibleorhighly susceptible theheterosiseffectfordiseasesymptomswasmorethan36%andforDONmorethan86%.Additionally,narrow-senseheritabilitiesh2nsforearrotandDONcontentwereveryhigh–dependofcombinationmorethan90%.ThisworkwasfundedbyPolishMinistryofAgricultureandRuralDevelopmentProj.No.4-1-06-3-01(33).Keywords:maize,earrot,heterosis,heritability

Session5Poster102Effectiveresistancetopowderymildew(Blumeriagraminisf.sp.hordei)inwinterbarleyinPolandJerzyH.Czembor1andAleksandraPietrusińska11Plant Breeding and Acclimatization Institute - National Research Institute (IHAR-PIB), NationalCentreforPlantGeneticResources-KCRZG,Radzików,05-870Błonie,PolandThe presented investigation describes the introduction of themlo gene for resistance to powderymildew(B.graminisf.sp.hordei)intowinterbarleycultivarsusingMASstrategy.ThesecultivarsarecharacterizedbyhighandstableyieldunderPolishconditions.FieldtestingoftheobtainedlineswithMlo resistance for theiragriculturalvaluewasconducted.Fourcultivars (Souleyka,Titus,SUVireniandMetaxa) ashighyieldingparentswereused. Inaddition,existing resistancegenes topowderymildew in these cultivars were preserved. Two lines (BKH 735 and line 42) as parents with Mloresistancewereused.LineBKH735wasobtainedinIHAR-PIBRadzikówin2002-2011.Selectionforpresence of the mlo gene was conducted in backcross populations by phenotyping in the field(naturalinfection)andundergreenhouseconditions(differentialbarleylinesforresistancegenesforpowderymildewanddifferential fungus isolates). Inaddition, to confirm thepresenceof themlogene in backcross populationsMAS strategywas applied using SSRmarkers HVmlo1 andHVmlo3.Field trials with backcrossed lines were conducted during 2016/17 in 3 locations: in Central(Radzików)andWesternPoland(Szelejewo,Wiatrowo).Theparentallineswereusedascontrol.Theaim of these trials was to obtain information on agricultural value of obtained lines. Our resultsdemonstratethepracticaluseoftheintroductionofMloresistanceintobackgroundofwinterbarleygermplasmwithvaluableeconomicalcharacteristics inPolishagriculturalconditions.Thisworkwasconducted in the project: Interaction between powdery mildew (Blumeria graminis f.sp. hordei)resistancedeterminedbymlogeneandeconomicalvaluecharacteristicsinwinterbarley.2014-2020.Prog.: Basic Research for Biological Progress in Crop Production; Funded by the Ministry ofAgricultureandRuralDevelopmentProj.No.41-04-3-01(27).Keywords:winterbarley,pre-breeding,powderymildew,Mloresistance,MAS,SSRmarkers

Session5Poster103NitrogenlimitationmoderatelyaffectsplantresponsetobioticstressM.Farjad1,M.Rigault1,L.Taconnat2,Marie-laureMartin-magniette2andM.Fagard1

1InstitutJean-PierreBourgin,UMR1318,INRA,Versailles,France2InstituteofPlantSciencesParis-Saclay(IPS2),CNRS,INRA,FranceInordertodeterminehowdefenseactivationisaffectedbymildnutrientlimitation,weperformedamicroarray analysis. Plants were grown in full or limiting nitrogen and subjected to biotic stress.Firstly,theeffectofabioticstress(Nlimitation)wasobservedonglobalgeneexpressionpatternsinresponsetobioticstress(bacteria).Inresponsetothebioticstress,wefoundthatNlimitationhadamild impact on the set of genesmodulated in response to infection although the limitationhad avisible effect on plant growth. However, some related-defense genesweremodulated by plant Nstatus. We then considered that Arabidopsis plants were subjected to two individual abiotic andbioticstresstreatments“Nlimitation”and“bacteria”respectively,aswellasacombinationofboth.We further presented and analyzed the homogeneous dataset of plant responses to single andcombined stresses according to Rasmussen et al, 2013. When two individual stresses werecombined,33%oftranscriptsresponded innon-predictablemanner.Onlyasmall fraction(3%)ofthe transcripts prioritized between potentially antagonistic responses. Moreover, several knowndefence-related genes were regulated in the prioritized mode reflecting an alteration in plantdefense program under N limitation treatment. Analyze the microarray data indicated thatArabidopsis genes were regulated to a greater extent by bacteria than N limitation, suggesting adominanceinapairofabioticandbioticstresstreatmentintermsofgenesmodulation.Keywords:abiotic,biotic,defense,plant,stress

Session5Poster104PromisingsourcesofresistancetopathogensofwinterwheatinUkraineLisovaGalina1,GolosnaLesya1,AfanasievaOksana1,KucherovaLyudmila11InstituteofPlantProtectionNAASUkraine,03022str.Vasilkivska33,Kyiv,UkraineInthelaboratoryofimmunityofagriculturalcropstodiseasesofNAASInstituteofPlantProtectionwas developed the technology of creation of artificial complex infectious background (ACIB)pathogens Puccinia recondita f. sp.tritici Rob. et Desm., Septoria tritici Rob. et Desm. andPseudocercosporella herpotrichoides (Fron.) Deighton on provocative background of BlumeriagraminisDCSpeersp.triticiE.M.Marchalonwinterwheatinthefield.Theresyntheticpopulationofpathogens,basedon theannual surveysofwinterwheatcrops indifferent regionsofUkraineanddata on the racial composition and internal population structure of powdery mildew pathogens,brownrust,septorialeafblotch,eyespotpathogen,wasformed.Therewereevaluatedtheresistanceof34varietiesofwinterwheatfromdifferentecologicalandgeographicaloriginduring2014-2016inForest-SteppezoneofUkraineusingACIBandseparateartificialbackgroundof infectiouspathogenTilletiacaries(DC)Tul.We have allocated sources of resistance to powdery mildew: Vluchna, L168-27, L155-03KH,Zolotoverha, Benefis, Niva, Hvulya (Ukrainian selection), MV Laura (HUN), Shahriar (IRN), Nikifor(ROU);torootrot–SofiaKyivs'ka(UKR),Mukhran(GEO),MV17/Zrn(IRN).Thesystemicresistanceagainst pathogens of powdery mildew and bunt had next varieties Syaivo (UKR), Miranda (ROU),F94578G3-1 / BUCUR // DELABRAD (ROU). Variety Midas (AUT) showed resistance to powderymildewandrootrot.Varietieswithresistancetoseptoria leafblotchandbrownleafrustwerenotfound.Search effective sources of resistance using artificial backgrounds of infectious pathogens enablesbreeders get promising starting material for the creation of new productive varieties, which canovercomeoftheinfectionpathogens.Keywords:artificialcomplexinfectiousbackground,pathogen,sourcesofresistance,winterwheat

Session5Poster105Comparative analysis of resistance of oats genotypes to contamination by different FusariumspeciesandmycotoxinsOlgaGavrilova1,AlexandraOrina1,TatianaGagkaeva1andIgorLoskutov21All-RussianInstituteofPlantProtection(VIZR),St.-Petersburg–Pushkin,Podbelskogosh.,3,196608,Russia2N.I.VavilovAll-Russian InstituteofPlantGeneticResources (VIR), St.-Petersburg,B.Morskaya st.,40-42,190000,Russia

TheaimofourstudywastoevaluatetheamountofDNAandmycotoxincontentof24oatscultivarsaftertheirartificialinoculationwithFusariumculmorumandF.sporotrichioidesseparately.Thecharacteristicswerebasedonsumoftwofeatures:theDNAcontentoftrichotecenesproducingFusarium fungi (Tri-Fusarium) and the amount of mycotoxins in grain of cultivars: deoxynivalenol(DON)orT-2toxin.Thegenotypes inoculatedwithaggressiveF.culmorum strainscontainedDNAofTri-Fusarium from0.03to3.64pg/gofmilledgrain,theamountofDONrangedfrom20to1179ppb.ThequantityofDNAofTri-FusariuminthesamplesinoculatedbyF.sporotrichioidesstrainsvariedfrom0.03to1.93pg/g,theamountofT-2toxinrangedfrom0to133ppb.Thehighpositivecorrelationbetweentheamounts of Tri-Fusarium DNA and DON was observed (r=+0.84, p<0.001), while the strongcorrelationbetweentheTri-FusariumDNAandT-2toxinamountswasnotfound.ThegroupofrelativelyresistantgenotypestoFusariuminfection(bothFusariumpathogens)includedcultivarsBisuandorodu(Russia),Geszty(Hungary)andGehl(Canada).Thegroupofhighsusceptiblecultivars consisted of Medved and КSI 432/08 (Russia), Bessin and Hurdal (Norway). Relationshipbetween DNA contents of Tri-Fusarium in the grain of genotypes inoculated by two species wasrelatively weak (r=+0.21). The most reliable results were obtained under inoculation of plantgenotypesbythemostaggressivepathogen.Abreeding strategy to combine the resistances topathogens andmycotoxins accumulationwouldprobablyleadtothedevelopmentofnewoatsvarieties,whichabletoefficientlylimittheproblemofmycotoxiningrain.TheinvestigationwassupportedbytheRussianScienceFoundation(No.14-16-00072).Keywords:Avena,genotypes,fungi,Fusarium,real-timePCR,mycotoxins

Session5Poster106Combined transcriptome and metabolome analyses to understand the response of flax (Linumusitatissimum)tothepathogenicfungusVerticilliumdahliaeSylvain Lecomte1,2, Mehdi Cherkaoui1,3, Ophélie Fliniaux1, Roland Molinié1, Jean Xavier Fontaine1,StéphanieGuénin3,HervéDemailly3,GaëlleMongelard3,DominiqueCailleu4,LucianeZabijak5,PauloMarcelo5, Christine Rusterucci1, Olivier VanWuytswinkel1, Reynald Tavernier2, Laurent Gutierrez3,FrançoisMesnard1,ChristophePineau21Laboratoire Biologie des Plantes et Innovation (BIOPI) EA3900, Université de Picardie Jules VerneAmiens.2Linéa–SemencesdeLin,Grandvilliers.3CentredeRessourcesRégionalesenBiologieMoléculaire,Amiens.4PlateformeAnalytique,Amiens.5PlateformeICAP,AmiensVerticillium dahliae is a major flax pathogen (Linum usitatissimum). The current increase of itsfrequency leads to significant economic loss in flax cultivation. This plant disease remainsmisunderstoodandtheversatilityofthepathogenaswellasthelackofflaxresistantcultivarsmakeitspropagationdifficulttocontrol.Thisstudyconsistsinidentifyingtranscriptomicandmetabolomiccharacteristicsinplantsdisplayingvariousbehaviorstowardsthediseaseinordertodevelopmarkersformarker-assistedbreeding.Anartificialinfectionprotocolwasoptimizedundercontrolledconditions.Usingthissystem,severalflaxcultivarswerescreened,allowingidentificationoftwocultivars,onepartiallyresistant(Évea)andonesusceptible(Violin),whichweresubsequentlyusedasmodelsinthisstudy.The pathogen progression in these two flax cultivarswas studied using qPCR quantification and acorrelationwas found between the amount of pathogen in plants and the severity of symptoms.These results showed that during their growth, despite a rapid colonization of the whole plant,tolerantflaxcultivarswereabletomaintainaloweramountoffungiwithintheirstissues.Transcriptomicandmetabolomicanalyseswereperformedoninfectedandnon-infectedplantsfromthe twoselectedcultivarsusingmicroarrays,RT-qPCRandNMRtechniques.Foreachcultivar,RNAandmetaboliteswereextractedatdifferentstagesofdevelopmentbetweensevendaysandthreeweeksafterinfection.Microarraysdatashowedaclearresponseinthemostresistantcultivarwhileno significant change was observed in the susceptible one. This response was characterized byexpressionchangesforgenes involved inreceptionandtransductionofpathogensignals,hormonesignaling, and activation of pathogenesis-related, chitinase and β-1,3-glucanase biosynthesis. NMRanalysesrevealedthatthepathogenaffectsthemetabolomeofthetwoflaxcultivars,inparticularinaminoacids,sugarsandpolyaminescontents.Keywords:flax,Verticillium,transcriptome,metabolome,microarrays,NMR

Session5Poster107"Omics-"foramappingofgrapevineresponsetoelicitorsandidentificationofinducedresistancemarkersChristelleLemaître-Guillier1,Marie-ClaireHéloir1,XavierDaire1,AgnèsHovasse2,ChristineSchaeffer-Reiss2, Philippe Schmidt-Kopplin3 Marianna Lucio3, Chloé Roullier-Gall3, Régis Gougeon4, SophieTrouvelot1,BenoîtPoinssot1andMarielleAdrian11Agroécologie,AgroSupDijon,CNRS,INRA,Univ.BourgogneFranche-Comté,17ruedeSully,F-21000Dijon,France2 Laboratoire de Spectrométrie de Masse BioOrganique, Université Strasbourg, CNRS, IPHC UMR7178,F-67000Strasbourg,France3HelmholtzZentrumMuenchen,ResearchunitAnalyticalBioGeoChemistry,IngoltaedterLandstrasse1,85764Neuherberg,Germany4Institut Universitaire de la Vigne et du Vin, Jules Guyot, UMR A 02.102 PAM, Université deBourgogne,RueClaudeLadrey,21078DijonCedex,FranceMostcultivatedgrapevinecultivars(Vitisviniferacvs)aresusceptibletoseverecryptogamicdiseasessuchasdownymildewcausedbytheobligatebiotrophicoomycetePlasmoparaviticola.Fungicidesare usually employed to control these diseases. However, in a context of sustainable viticulture,complementary and / or alternative strategies, such as elicitor-induced resistance, are expected.Despite numerous elicitors from diverse origins have been identified, their mode of action andimpactonplantmetabolismremainoften largelyunknown.Wethereforeusedamultipleanalysesapproach encompassing Proteomics, Metabolomics, Transcriptomics and Volatilomics to generateand cross-compare data to gain insight into the mode of action of elicitors (impact on the plantmetabolicpathways)andtouncoverputativemarkersofinducedresistance(IR).Methods : Sulfated laminarin (PS3), a sulfated b1,3-glucan affording a high level of grape downymildewcontrol(90%) ingreenhouseconditions,wasthemainelicitorusedforthisstudies.Elicitor-treated leaves were collected and extracts were prepared for transcriptomic (using the Roche-NimbelgenGrapearray),proteomic(2DEseparationandnanoLC-MS/MSanalysis)andmetabolomic(FT-ICR-MS) analyses. For volatilomic,VolatileOrganicCompounds (VOCs)were caughtonto SPMEfiberspriortoGC-MSanalysis.Results : Thesedifferent -omics approaches allowedus topoint out themainmetabolic pathwaysinvolved in PS3-IR and to identify possiblemarkers of PS3-IR. Among themwere PR10, farnesene,anderythritolphosphate.Conclusion :Theseglobalomics-analysesprovided largeamountsofvaluabledata thatenrichourknowledge of elicitor-IR. Moreover, they allowed the identification of IR-markers that would beuseful tools for further screening of putative elicitors and to follow the plant response to elicitortreatmentinfieldconditions.Keywords:-omics,elicitor,inducedresistance,grapevine

Session5Poster108Theeffectivenessoftranslocation1AL/1RSinresistancetoleafrustinUkraineGalinaLisova1,TetianaSobko11InstituteofplantprotectionNAAS,Kyiv,UkraineIn2011-2015theeffectsoftranslocation1AL/1RSonresistancetothenaturalpopulationofleafrustraces,was investigated. It isspentanassessmentofresistanceofcarriersof translocation1AL/1RS(12 cultivars - Amigo, TAM107, TAM201, Nekota, Century the American selection and domesticselection-Monologue,Colombia,Zolotokolosa,Eritrospermum26221,KnyaginyaOl'ga,Smuglyanka,Spasivka) at a various infectious load. In 2011 and 2015 observed average level of diseasedevelopment.In2014-highinfectionlevel,andin2012and2013recordedepidemics.StableresistancetolocalpopulationofleafrustwerecvsNecota,Century,Monologue,Smuglyanka,Spasivka. At recorded epidemics they showed high resistance. It is known, that in these cultivarscontainresistantgenesLr24,andatcvCenturyalsoLr42.Consideringtheirhighstableresistanceitisnot thewhole setofgenesof resistance.CvsAmigoandColombiahave shownvariable resistance(decreaseinayearofepidemics,followedbyrestorationwithatreducingof infectionlevel).Othercvs Zolotokolosa and Eritrospermum 26221 under epiphytotic 2013 lost stability, but regained itwhen reducing of infection load in 2014 and 2015. The cv KnyaginyaOl'ga for all years of studiesshowed the resistance -moderate susceptibility. It contains gene Lr34,which in the conditions ofUkraine are not effective. Lines TAM107, TAM201 are resistant only in years with an averageinfectiouslevelofdevelopment.Whenepiphytoticstheyaresusceptible,andinthesubsequentyeararemoderatesusceptible.Thus, exhibiting of resistance of carriers of wheat-rye translocation 1AL/1RS in the conditions ofUkrainevariously.Incaseofepiphytoticdisease,itdoesnotfullyprovideresistanceofcultivars.Thepresence background of translocation 1AL/1RS other resistant genes increases the resistance ofcultivars.Such featuresmustbeconsideredwhenattractingofcarriersof translocation1AL/1RS inbreedingprocess.Keywords:winterwheat,translocation1AL/1RS,leafrust,resistance,genes

Session5Poster109SWEET»sugartransportersingrapevine:roleintheinteractionswithbioaggressorsEloïseMeteier1,SybilleFarine1,FloreMazet1,DidierMerdinoglu2,ChristopheBertsch1,PereMestre2andJulieChong11LaboratoireVigne,BiotechnologiesetEnvironnement (LVBE,EA3991),UniversitédeHauteAlsace,33ruedeHerrlisheim,68000Colmar,France2UMR 1131 Santé de la Vigne et Qualité du Vin, INRA-Université de Strasbourg F-68000 Colmar,FranceGrapevine is susceptible to diverse pathogens including oomycetes, fungi, bacteria and viruses.Carbonallocationandsugarpartitioningplaykey roles inplant-bioagressor interactions.There isacompetition for sugar at the plant-pathogen interface which is controlled by membrane sugartransportersastheSWEETtransporters(Chenetal.2010).Theseproteinsareatargetofextracellularpathogenswhichmodifytheirexpressiontoacquirethenecessarysugarstotheirgrowth(Chuetal.2006).Theaimofthisstudy is tocharacterizethefunctionofgrapevineSWEETtransporters intheinteractionwithpathogenswithdifferentlifestyles.In previous work, we characterized the SWEET family of sugar transporters in Vitis vinifera, andshowed that VvSWEET4 could be involved in plant cell death and resistance to Botrytis cinerea(Chong et al. 2014).Wealso identified two additional grapevine SWEET transporters up-regulatedafter inoculation with Plasmopara viticola and Erysiphe necator. To assess the sugar transportspecificityofVvSWEET4,thegenewasoverexpressedingrapevinehairyroots.Ourresultsshowthathairy rootsoverexpressingVvSWEET4haveahigher growth than controlson sucroseandglucose-containingmedia.TransportexperimentswithradiolabeledsugarswillconfirmthetransportactivityofVvSWEET4.We obtained different knockout mutants of AtSWEET2 and AtSWEET17 and overexpressors ofVvSWEET2aandVvSWEET17dgenesinArabidopsis.Westudiedthesusceptibilitylevelsoftheselinesto the bacteria Pseudomonas syringae pv. tomato and the oomycete Hyaloperonosporaarabidopsidis.PreliminaryresultssuggestaneffectofAtSWEET2onP.syringaeinfection.To validate our results in grapevine,we are creating plantswhere our candidate genes are eitheroverexpressedor knocked-down.Thismaterialwill help toelucidate the involvementof grapevineSWEET transporters in the interaction with pathogens and provide information concerning sugartransportandpartitioning inthoseinteractions.Finally,the involvementofgrapevineSWEETgenesinplant-pathogeninteractioncouldleadtotheidentificationofnewrecessiveresistancegenes.Keywords:grapevine,SWEETtransporter,sugartransport,pathogenresistance

Session5Poster110TraditionalandModernPlantBreedingMethodswithExamplesinwheatAleksandraPietrusińska1,JerzyHenrykCzembor11NationalCentreforPlantGeneticResources,PlantBreedingandAcclimatizationInstitute,NationalResearchInstituteatRadzików,05-870Błonie,PolandTheaimofpresentedresearchispyramidingofleafrustandpowderymildewresistancegenesinaonegenotype.Asthedonorofresistancetoleafrustseverallineswereused.ThelineKS90WGRC10,which carries the Lr39 (= Lr41) gene derived from the diploid wild wheat Triticum tauschii (syn.Aegilops squarrosa). The line KS04WGRC45, that carries the Lr55 gene derived from the Elymustrachycaulus.ThelineHRSYecoraRojo,whichcarriestheLr47genederivedfromTriticumspeltoides.Asthedonorofresistancetopowderymildewtwolineswereapplied.A6VS/6ALtranslocationlineof Yangmai5 that carries the Pm21 gene derived from the wild cv. Dasypyrum villosum. LineNC99BGTAG11, which carries the Pm37 gene derived from the Triticum tiomopheevii and lineNC97BGTD7, which carries the Pm34 gene transferred from the Aegilops tauschii. To detectresistance genes (foreground selection) severalmolecularmarkers for Lr41, Lr47, Lr55,Pm21 andPm37wereapplied.Inaddition,plantmaterialswereinoculatedinthegreenhouseatthethree-leafstagewithanaturalpathogenpopulationofP.reconditaandB.graminis.Themarker-assistedselectionandresistancetestsallowedtoobtainhomozygouslinescarryingthreeor four resistance genes to leaf rust and powderymildew in winter wheat.Moreover, the newlyproducedhomozygouswheatlines,canbeusedasthesourceofeffectiveresistancefortheleafrustandpowderymildewbygeneticists,breedersandplantpathologists.Keywords:Blumeriagraminis,genepyramiding,marker-assistedselection,Pucciniarecondita

Session5Poster111Identificationofcandidategenesinvolvedininnateimmunityandeffector-triggereddefenceHUStotz1,GTValente2,ROliveira2,NDavey1,VSteuber1

1SchoolofLife&MedicalSciences,UniversityofHertfordshire,Hatfield,Herts.AL109AB,UK2BiotechnologyandBioprocessDepartment,UniversidadeEstadualPaulista,Botocatu,BrazilThe innate immune system of plants consists of two layers. The first layer of defence relies onrecognition of pathogen-associated molecular patterns (PAMPs) by pattern recognition receptors(PRRs).Host-adaptedpathogenssecreteeffectorproteinstodampenorevadehostPAMP-triggeredimmunity(PTI).Resistance(R)genesencodeproteinsthatareinvolvedinrecognitionofeffectorsortheirhosttargetstoactivateeffector-triggeredimmunity(ETI)ordefence(ETD)againstintracellularor extracellular fungal pathogens. Whereas R genes that encode cytosolic receptors are wellcharacterised,thosethatencodetransmembranereceptorsarenot.WediscusstheuseofadvancedcomputationalmethodstobetterdefinetheRgenecomplementingenomesofplantsandtoassistthe identification of genes involved in PTI that may contribute to partial or quantitative diseaseresistance.Thesetoolscanbeanassetforplantbreedingandcropprotection.Keywords:computationalbiology,diseaseresistance,machinelearning

Session5Poster112EvaluationofselectedcultivarsCucurbitamaximatypeUchikiKuriforresistancetoZucchiniyellowmosaicvirusJiriSvoboda1,PetrKominek1andLeonaLeisova-Svobodova1

1CRI-CropResearchInstitute,Drnovska507,16106,Prague6,CzechRepublicZucchiniyellowmosaicvirus(ZYMV)causesconsiderablelossesofcucurbitaceousvegetablesgrownnearly all over the world. Commonly planted cultivars are highly susceptible to ZYMV especiallyorangecultivarsofpumpkinsCucurbitamaximatypeUchikiKuri.Fruitscollectedfrominfectedplantsareoftendark-greenstainedandtheirshelflifeisrapidlyshortened.Thereforelookingforresistantcultivars Cucurbita maxima has started. Three cultivars from South Africa, three from the CzechRepublic and two from theNetherlandswere selected for evaluation their resistance to themostvirulent Czech ZYMV-H strain (GenBank Acc. No. DQ144054). Zucchini squash (Cucurbita pepo)‘Zelena’wasaddedto thisgroupasan internal standardofasusceptiblecultivar toZYMV.Testedplantsweresowninagreenhouseandcomingupplantsweremechanically inoculatedbyZYMV-H.Their resistance was assessed by comparison of the virus protein relative concentration andobservedsymptomsfourweeksaftertheinnoculation.TherelativeconcentrationofZYMVproteininleaves was calculated from the virus titer determined by ELISA. The South African pumpkins‘Invincible‘and‘Star‘wereevaluatedashighlyresistant;thevirusconcentrationintheir leaveswasnearlyzeroandtheydidnotshowanyviralsymptomontheirleavesandfruitsbuttheywereZYMVpositive in PCR. The Dutch recent hybrids E 30R.080 F1, E 30R.079 F1 and South African ‘Babypumpkin‘weremediumresistant;thevirusmultipliedintheseplantsinalowrate.Czechpumpkins‘Blue Kuri‘, ‘Grey Queen‘ and ‘Hokkaido‘ were evaluated as highly susceptible; the virusconcentrationinplantswasthesameorhigherthanintheinternalsusceptiblestandard‘Zelená’andtheirleavesandfruitswereseverelydamagedbyZYMVinfection.Inconclusion,pumpkins‘Invincible‘and ‘Star‘ are appropriate for planting in ZYMV infected areas and we can recommend them forbreedingnewresistantcultivars.Keywords:virusconcentration,virustiter,ELISA,PCR,ZYMV

Session5Poster113Pharmacologicalscreenforsuppressorsofmlo-mediatedpowderymildewresistanceHongpoWu1,MarkKwaaitaal3,RoxanaStrugala2,UlrichSchaffrath2,RalphPanstruga1

1UnitofPlantMolecularCellBiology,InstituteforBiologyI,RWTHAachenUniversity,52056Aachen,Germany2DepartmentofPlantPhysiology,RWTHAachenUniversity,52056Aachen,Germany3Department of Molecular Plant Pathology, University of Amsterdam, 1098 XH Amsterdam,NetherlandsPowdery mildew resistance conferred by absence of MLO gene featured with broad spectrumpenetrationresistance.Themolecularmechanismunderlyingthisresistanceisstillelusive.Wetrytoidentify new molecular components involved in mlo-mediated resistance via pharmacologicalapproach.We screened 65 chemicalswith viarantmolecular targets by peelingmethod in barley-Blumeria graminis f. Sp. Hordei interaction system, out of which twelve chemicals were able toinducesusceptibilityofbarleymlomutantaccordingtoresultsofpenetrationcounting. Targetsofthe positive chemicals include cAMP biosynthesis and vesticle trafficking, and targets of variantpolyaminespeciesandalloxanwhichshowedpositiveeffectarenotyetidentified.Keywords:MLOgene,pharmacologicalscreen,barley,powderymildew,cAMPbiosynthesis,vesicletrafficking

Session5Poster114InvestigatinginductionofSARduringgene-for-geneinteractionsbetweenArabidopsisthalianaandPseudomonassyringaeTruptiP.Gaikwad1,MartadeTorres-Zabala1,StephenGreen2,PeterWinlove2,DavidHorsell2,MurrayGrant31Biosciences,CollegeofLifeandEnvironmentalSciences,UniversityofExeter,UK2Physics,CollegeofEngineering,MathematicsandPhysics,UniversityofExeter,UK3Biosciences,SchoolofLifeSciences,UniversityofWarwick,UKPlantsdeploytwokeyactivedefensivestrategiestocombatmicrobialpathogens;(i)RecognitionofPathogen-AssociatedMolecular Patterns (PAMPs) by extracellular surface receptors leading to theactivationof PAMP-Triggered Immunity (PTI); (ii) Recognitionof pathogeneffector activity, usuallyintracellularly, by host Resistance (R) proteins leading to Effector-Triggered Immunity (ETI). ETI ischaracterised by a rapid localised Hypersensitive Response (HR). HR induces Systemic AcquiredResistance(SAR)throughtheproductionofaninducibleimmunesignal(s),leadingtobroadspectrumsystemic resistance.We investigated the earliest events associatedwith SAR signalling using plantelectrophysiology,SARmutantsandauniquepromoter-luciferasefusionthatcapturesearlysystemictranscriptional events underlying initiation of systemic immune signal(s). We describe thetranscriptionaldynamicsofA70 (At5g56980), ageneofunknown function (Trumanetal. 2007), inlocal and systemic tissue following challenge with different elicitors and virulent or avirulentpathogenchallenges.WeprovideevidencethatA70respondstoajasmonate(JA)relatedsignalthatis rapidly generated following ETI recognition. We further evaluate A70::LUC reporter activity inresponsetoJAstimulusandcorrelateactivitywithhistologicalexpressionofaJArepressorreporter(JAZ10::GUS)andA70::GFP reporter insystemically responding leaves followingavirulentpathogenchallenges. Finally, we examine changes in electrophysiological signals following ETI in local andsystemic leaves. Focussing on events underpinning initiation, propagation and perception of SAR-inducing signals within the first 6-8 h of pathogen challenge we provide new insight into theintegrated signalling mechanisms, dynamics and connectivity underpinning systemic immuneresponses.Weconcludethattheirmulticomponentsignalsthat linkETI inducedtranscriptionalandelectricalsignals,withaCOI1receptorpropagativetranscriptionalwavetheleadstorapidtemporalspatialtranscriptionalactivationofjasmonateresponsivegenesinsystemicrespondingleaves.

Session5Poster115Nextgenerationcrossprotection:singleviruswithmultipletargetsAimingWang1andHonggunagCui11LondonResearchandDevelopmentCenter,AgricultureandAgri-FoodCanada1391SandfordStreet,London,Ontario,N5V4T3,CanadaPlantvirusesareobligate intracellularparasitesthat infectmanyagriculturally importantcropsandcauseseverelosseseachyear.Geneticresistanceisconsideredthemosteffectivemeanstocontrolthese viruses. However, extensive screening of germplasms for natural genetic resistance onlyidentifies a few resistance genes. Cross-protection is the practice of protectionof plants against aviral infectionbypriorinoculationofthemwithamildversionofthevirus.Thisapproachhasbeenused to controlmany important viral diseases such asCitrus tristeza virus (CTV),Cucumber greenmottle mosaic virus (CGMMV) and Pepino mosaic virus (PepMV) in the world. The possiblemechanisms underlying cross-protection include RNA silencing and superinfection exclusion. Thisraisesthepossibilitythatanattenuatedvirus(servingasaviralvector)maybemodifiedtocontaingenomic fragments of different viruses and of a host factor gene of multiple viruses. Thus, thismodifiedvirusmaybeusedforcross-protectionagainstseveralviruses.Here,wepresentourdataonthedevelopmentofaviralvectorderivedfromPrunusnecroticringspotvirus(PNRSV),awidespreadfruittreevirusthatisendemicinallPrunusfruitproductioncountriesandregionsintheworld.Themodified PNRSV vector, harboring the sense-orientated target gene sequence of 100~200-bp inlengthingenomicRNA3,couldefficientlytriggerthesilencingofatransgeneoranendogenousgenein themodelplantN.benthamiana. ThePNRSV-basedvector couldalsobemanipulated to silenceendogenousgenesinpeachsuchaseukaryotictranslationinitiationfactor4Eisoform(eIF(iso)4E),ahost factorofmanypotyviruses includingPlumpox virus (PPV).Moreover, theeIF(iso)4E-knocked-downpeachplantswereresistanttoPPV.Ourdataopenapotentialavenueforthecontrolofvirusdiseasesinperennialtreesviaviralvector-mediatedsilencingofhostfactors.

Session5Poster116BurkholderiaphytofirmansPsJNconfersgrapevineresistanceagainstBotrytiscinereaviaadirectantimicrobialeffectcombinedwithabetterresourcemobilizationLidianeMiottoVilanova1,Cédric Jacquard1,BarbaraCourteaux1, LaurenceWortham2, JeanMichel2,ChristopheClément1,EssaïdA.Barka1andLisaSanchez1

1LaboratoiredeStress,DéfensesetReproductiondesPlantesURVVC-EA4707,UFRSciencesExactesetNaturelles,UniversityofReims-Champagne-Ardenne,Reims,France2LaboratoiredeRechercheenNanosciences,EA4682,DepartmentofPhysics,UFRSciencesExactesetNaturelles,UniversityofReims-Champagne-Ardenne,Reims,FrancePlantgrowth-promotingrhizobacteria(PGPR)areofgreatinterestsincetheyarebeneficialnaturallyoccurringsoilbacteriathatcolonizeplantrootsandconferbeneficialeffects.Theycanincreaseyield,stimulate plant growth, reduce pathogen infection, and reduce biotic and abiotic stresses. AmongthesePGPRs,endophytesaredefinedasthosebacteriathatareabletocolonizetheinternaltissueoftheplantwithoutcausingexternalsignsof infectionornegativeeffectsontheirhost.Burkholderiaphytofirmans PsJN, classified as an ePGPR, was first isolated from surface-sterilized onion rootsinfected with the mycorrhizal fungus Glomus vesiculiferum. This rhizobacterium significantlypromotesgrowthandprotects thegrapevineagainstbiotic (greymoulddisease)andabiotic (cold)stresses. Ifmechanisms implied in cold tolerance inducedby PsJNwere elucidated, the protectiveeffectinducedbythePGPRagainstB.cinereahoweverremainselusive.Tounravelthemechanisticofpathwaysinvolvedintheobservedresistance,differenttraitsofthetripartiteinteractionbetweenVitis vinifera L., Botrytis cinerea and Burkholderia phytofirmans were highlighted. Among theseaspects,directantimicrobialactionofPsJN,theabilityofthebacteriumtoprimedefenseresponsesandcarbohydratemetabolismofgrapevinewillbepresented.Keywords:grapevine,ParaburkholderiaphytofirmansPsJN,Botrytiscinerea,defense,priming

SESSION6.EMERGINGTOOLSFORTHEMANAGEMENTOFPLANTDISEASES:AGROECOLOGYANDDISEASEMANAGEMENTSession6Poster117ImprovementofNorwegianpotatoproductionthroughbetterseedpotatohealthMerete Wiken Dees1, Simeon Rossmann1, Vinh Hong Le1, Arnaud Lefrançois1, AbdelhameedElameen1,BorghildGlorvigen2,AlisonLees3,JanvanderWolf4andMayBenteBrurberg1

1NIBIO,NorwegianInstituteofBioeconomyResearch,Aas,Norway2NorskLandbruksrådgivning,Aas,Norway3TheJamesHuttonInstitute,Dundee,ScotlandUK4PlantResearchInternational,Wageningen,theNetherlandsPotatoesarethemost important foodcropwithrespect to firsthandvalue inNorway.Particularlychallengingisthefactthatmanyoftheimportantpotatopathogenscanbedifficulttodetectastheygive rise to latent infections. A first step towards management of plant diseases is appropriateidentification and detection methods. Consequently, one of the major goals of this study was toimplementqPCR for rapidandsensitivedetectionandquantificationof latent infectionofbacteriaandfungi inseedtubers.Theworkwasfocusedonbacterialsoftrotdiseases (Pectobacteriumspp.and Dickeya spp.), late blight (Phytophthora infestans), gangrene (Boeremia foveata) and skinblemishdiseases suchas silver scurf (Helminthosporium solani), skin spot (Polyscytalumpustulans)andblackdot (Colletotrichumcoccodes).During the first twoyearsof theproject,qPCRtesting fortheabovementionedpathogenswereimplementedand15seedlotsweretestedeachyear.Samplesfromtheseed lotswere testedat three timepoints,beforeplanting,afterharvestandafter threemonthsinstorage.Skinspotwasfoundinallseedlotsexceptfromthepre-basicseedlots.Gangrenecouldbedetectedinall lots,while lowlevelsof lateblightwasfoundinaquarteroftheseedlots.Lowlevelsofblackdotcouldbedetectedinsomeoftheseedlotsandsilverscurfwasfoundinalllots.Softrotbacteria(Pectobacteriumsp.)werefoundinallsamplestested,butinlowerlevelsinthepre-basicseedlotscomparedtothecertifiedseedlots.Dickeyasolaniwasnotdetectedinanyofthelots.Thelevelsofbacteriaweresignificantlyhigherinthesamplestestedafterharvestincomparisontotheseedpotatoesandsamplestakenafterstorage.Therewasnocorrelationbetweenblack legobserved in the field throughout the season and the amount of bacteria found in the lots afterharvest.Keywords:diagnostics,potatodiseases,skinblemishdiseases,softrotbacteria

Session6Poster118ShiftinsensitivityofDanishZymoseptoriatriticipopulationstotriazolefungicidesThiesMartenHeick1andLiseNistrupJørgensen11AarhusUniversity,InstituteforAgroecology,Forsøgsvej1,4200Slagelse,Denmark ZymoseptoriatriticiisthecausalagentofSeptoriatriticiblotch(STB)onwheatandoneofthemostdevastatingfoliardiseasesworldwide.Thediseaseiscommonlycontrolledtwotothreetimesduringthe seasonwith strobilurin, triazole and SDHI fungicides. In recent years, however, a gradual shifttowards more strains insensitive to these three fungicide groups has been observed in severalcountriessuchas IrelandandtheU.K.Thedeclineof fungicidesensitivityhasbeenassociatedwithmolecular changes in the different targets of the different fungicide groups. Sensitivity towardstriazole fungicides has been seen on an acceptable level in Northern Europe for many years,however, in recent years a gradual decline has also been witnessed here. In 2016, results of thenational sensitivity testing showed that EC50 values for triazole epoxiconazole were significantlyhigherthanintheyearsbefore,indicatingamajorshiftintriazolesensitivity.ThisdevelopmentgoeshandinhandwiththedetectionofmorediversCYP51Z.triticivariantsthatmightexplainthislossinsensitivity. The number of actives available in Denmark for control wheat diseases is low onlycovering fewmultisite inhibitors (folpetandmancozeb), fewtriazole fungicides, fewstrobilurinsaswell as a single SDHI (boscalid). Therefore, it remainsdifficult to combineefficient disease controlwithanti-resistance strategies. Ithashoweverbeenshown thatawell thought throughsprayplanhelpsminimisingselectionformoreresistantZ.triticivariants.Keywords:epoxiconazole,Septoriatriticiblotch,diseasecontrol,CYP51

Session6Poster119Effect of preparations containing nanoparticles towards chosen fungal pathogens of carawayCarumcarviL.EwaDorotaZalewska1,EwaDorotaKról1,RenataNurzyńska-Wierdak2andGrażynaZawiślak2

1DepartmentofPlantPathologyandMycology,FacultyofHorticultureandLandscapeArchitecture,UniversityofLifeSciences,Leszczyńskiego7Str.,20-069Lublin,Poland2Department of Vegetable Crops and Medicinal Plants, Faculty of Horticulture and LandscapeArchitecture,UniversityofLifeSciencesinLublin,Leszczyńskiego58Str.,20-068Lublin,PolandIn recent years interest has increased in the possibility of using preparations containing activesubstancesintheformofnanoparticlestoreducetheoccurrenceofbacteriaandfungipathogenictohumansanddifferentcrops.Thefertilizerscontainingnanoparticlesofcopper,silver,andchitosan:Viflo®copper,VifloChitosol®Silver®VifloCalSandisolatesofSeptoriacarviSyd.wereusedtothestudy.Thepercentofgrowthinhibition,developmentofS.carvicoloniesandabilityofViflo®Caltocontrol plants against infection by S. carvi was adopted as the criterion of evaluation of thenanoformulations.InvitroithasbeenshownthatthetestedformulationswithnanoparticleslimitedthegrowthanddevelopmentoftheS.carviandVifloCal®Sattheconcentrationof1g·cm-3wastheinhibitorofmycelialgrowth.InclimaticchamberVifloCal®Sprotectcaraway`sseedlingsagainsttheinfectionbyS.carvi.InvivoVifloCalS®significantlylimitedtheseverityofseptoriosisandpowderymildewErysipheheracleiD.C.,whichcontributedtoincreasethesizeandqualityofthecarawaycrop.Actually,thestudyoftheeffectofnanoformulationsonthecontentandcompositionofbiologicallyactive substances and on the quality of the raw pharmacopeias material of chosen plants werecarriedout.Keywords:caraway,fungi,control,nanoformulations

Session6Poster120EmergingframeworkforadjustingIPMresearchtocustomervalueJanBuurma1,ErikdeBakker1andNicovanderVelden1

1WageningenEconomicResearch,P.O.Box29703,2502LSTheHague,TheNetherlandsIntegratedPestManagement(IPM)researchersarechallengedtoincreasetheadoptionrateofIPMknowledgeintopractice.ManyIPMresearchleadersbelievethesolutionis infurtherimprovementof innovativeIPMtoolsandknowledgesupply. Inouropinion,however,thesolutionis inattentionforthemotivationsoffarmers&growersandforthevaluesandincentivesofvaluechainpartners.Therefore,inthispaperweshiftthefocusfromtechnologypushtomarketpull.A flow chartwill be presentedwhich shows the dynamics of innovations in food chains. The flowchart startswith social unrest and (5 – 8 years later) results in customer value. The secret of thisresult isa coordinatedactionofknowledgepartners, farmers&growersandvaluechainpartners.Involvement of civil society organisations, farmers’ organisations and retail and food servicecompaniesiscrucialforbringingthiscoordinatedactiontoasuccessfulconclusion.Theeconomicdriver in thisprocess is integrating IPMsolutions innewproduct concepts thatgiveaccesstoexpensivemarketsegments.Newproductconcepts(e.g.residue-freesnacktomatoesinaplasticcup)provideawin-winsituationforallpartnersinthevaluechain.Ananalysisofexportdatarevealed10-20%higherexportpricesforexportsoffreshtomatoes,sweetpeppersandapplestothehighlydemandingGermanmarket.Thetake-homemessageforIPMresearchersistofocustheirworkonco-creationofcustomervaluetogetherwithinputsuppliers,farmers&growersandvaluechainpartners.Thetrendsettersamongfarmers&growersarealreadyapplyingthecustomervaluestrategy.Accordingtofarmaccountancydata,thesetrendsettersmanagetorealisehighgrossyieldsagainstlowcropprotectioncosts.Ontheothersideofthespectrumcropprotectioncostsarehighandgrossyieldsarelow.Keywords:adoptionrate,dynamics,motivations,incentives,co-creation,valuechain

Session6Poster121GreenmanurebasedmulchhelpssuppressPhytophthorainfestansMariaR.Finckh1,StephanJunge1,HannesSchulz1,ChristianBruns1

1Kassel University, Department of Ecological Plant Protection, Nordbahnhofstr. 1a, D-37213,Witzenhausen,GermanyNutrientsupplyandlateblight,causedbyPhytophtorainfestansarethemajorchallengesinorganicpotatoproduction.Copperfungicidesareharmfultotheenvironmentinhighdosagesandtheirbanin Europe is imminent. Someorganic farmers reported to us that the use of greenmanure basedfresh mulch would considerably reduce late blight while supplying nutrients to the potatoes.Experiments were conducted in 2014-2016 under organic conditions with natural inoculum usinginversionandnon-inversiontillageandvariousgreenmanuremulchsourcestoverifytheseeffects.No P. infestans occurred in 2015 and Data from one experiment in 2014 and two in 2016 arepresented.Theexperiment in2014combinedreducedtillagewith10-12cmfreshwinterpearyebasedmulchapplied right at potato emergence compared to a regular plough based system. In the firstexperiment in 2016, triticale-vetch and grass clover mulches were compared ton o mulch in aregularlyploughedfield.Inthesecondexperiment,triticalevetchwasgrownaspre-croptopotatoesandrototilledtoadepthof5cmpriortopotatoplanting.HalftheareawassubsequentlyhilledandcoveredwithaC-richmulchoffreshtriticale,theotherhalfwasleftunmulched.Inallthreeexperiments,theepidemiconsetwithP.infestanswasdelayedbyafewdaysandplantsdied of late blight between 10 to 20 days later whenmulched compared to non-mulched plants.Areasunder thediseaseprogresscurvewere reducedby27 to38%dependingonmulch typeandyear.Microclimaticconditionsinthefieldsweresubstantiallydrierduetothehygroscopicpropertiesofthedryingplantmaterials.However,dependingontheweatherconditions,mulchingincreasedorreduced potato yields by affecting soil moisture and temperature pointing to a need for systemoptimizationbeforerecommendingthetechnologyinpractice.Keywords:lateblight,potatoes,mulch,reducedtillage

Session6Poster122ImpactofUV-CradiationsofstrawberryplantsonitssensitivitytoBotrytiscinereaM.Forges1,2,H.Vàsquez1,F.Charles1,L.Urban1,Y.Lizzi1,M.Bardin2,J.Aarrouf11UnitéMixte de Recherche Qualisud, Laboratoire de Physiologie des fruits et Légumes, Universitéd’AvignonetdesPaysdeVaucluse,301RueBaruchdeSpinoza,BP2139–84916,Avignon,France2PathologieVégétale,INRA,84140Montfavet,FranceIncreasing the resistanceofplantsbyusingphysicalmethods remainspoorly studied compared totheuseofbiologicalorchemicalelicitors.TheobjectiveofthestudyistotestthehypothesisthatitispossibletoreducethesensitivityofstrawberryplantstoBotrytiscinereabytheapplicationofnon-deleteriousdoseofUV-Cradiationontheplant.Testswere carriedouton three cultivars of strawberry.We first showed that these cultivars havedifferent levelsof basal sensitivity toB. cinerea on leaves.Various treatments combiningdifferentdoses of UV-C and frequency of application resulted in the selection of non-deleterious doses forstrawberryplants. Measuresofchlorophyllfluorescence, leafcolorandplantgrowthrevealedthatdoses comprised between 0.4 and 1.70 kJ/m² had not any significant effect on these plantphysiologicalparameters,regardlessofthefrequencyofapplicationandofthecultivartested.It also resulted in the identification of hormetic doses that can improve the resistance of leavesagainstgreymold.UV-Ctreatmentsappliedonplantsat0.85and1.70kJ/m²4timeseverytwodays,resulted in a systematic decrease in the sensitivity of the cultivar Candiss to the strain Bc1 of B.cinerea (leafprotectionbetween11and27%).ButtheeffectobservedisdifferentaccordingtothestrawberrycultivarandtothestrainofB.cinereaused. In futureexperiments,weenvisagetotesttheeffectofthesetreatmentsonfruitqualityandintheirpost-harvestsensitivitytoB.cinerea.Keywords:strawberry,Botrytiscinerea,UV-Cradiations,hormeticdose

Session6Poster123R2E:aparticipatoryexperimentationnetworkforabetterintegrationofbiocontrolsolutionsClaudeMaumené1,CélineDrillaud2

1ArvalisInstitutduvégétal,StationExpérimentale,91720Boigneville2ArvalisInstitutduvégétal,DomaineexpérimentalduMagneraud,17700SaintPierred’Amilly

The Experimental Excellence Network (R2E) is a collaboration between various research anddevelopmentorganizations,allinclosecontactwithfarmers,workingtogethertodevelopagronomicreferences inorder todevelopamulti-efficientagriculture.ThemainmotivationsofR2Emembersare:- evaluate agronomic innovations in the broad sense (crop protection, nutrition, cultivationtechniques,etc.,whichcanopenuppathwaysforagriculturalprogress,-topooltechnicalexpertiseandexperimentalmeansinordertocarryoutacommonR&Dprogram,-tosharereferencesonagronomicinnovations,mainlyfromfieldexperiments,-ensurehomogeneityandexcellenceofmethodologies,- strengthen the representativeness of acquired references by valuing the diversity of agronomicsituations(networkdynamics).Foritslaunchyearin2015,theR2Enetworkhaschosentoevaluatetheavailableorindeveloppmentbiocontrol solutions for the control Septoria (Zimoseptoria tritici) onwheat. In 2016, the thematicfieldwasextendedtothecontrolofFusariumheadblight(Fusariumgraminearum)onsoftwheatbybiocontrolproducts.Thispaperwillattempttopresentthisexperienceofparticipatoryresearch,aswell as the first results obtained in this frameworkwith sulphur andwithPythium oligandrum tocontrolrespectivelySeptoriaandFusariumheadblight.Keywords: participatory experimentation network, wheat, Zimoseptoria tritici, Fusariumgraminearum,biocontrol,sulphur,Pythiumoligandrum

Session6Poster124Exploring theeffectof combining cropgenetics and fungicidal chemistry to control amajor rootpathogenofwheatJoseph Moughan1, Vanessa McMillan1, Rodger White2, James Melichar3, David Ranner3, and KimHammond-Kosack11Department of Plant Biology and Crop Science, Rothamsted Research, Harpenden, Hertfordshire,AL52JQ,UK2Department of Computational and Systems Biology, Rothamsted Research, Harpenden,Hertfordshire,AL52JQ,UK3SyngentaUKLimited,Fulbourn,Cambridge,CB215XE,UKGaeumannomyces tritici is the world’s most destructive root pathogen of wheat, causing take-alldisease.Anewlydiscoveredgenetictraitinelitehexaploidwheat,calledLowTAB(lowtake-allbuild-up), limits take-all inoculumbuild-up in the rhizosphere. Thus, LowTABcultivars reduce the riskoftake-alldiseasedamagingaconsecutivewheatcrop, thereby improvingcrophealthand increasinggrainyields.Thisresearchhas investigatedtheeffectofcombiningLowTAB,withfoliarapplicationsof take-all active fungicide,Amistar (active ingredient: azoxystrobin),on take-all inoculumbuild-upand subsequent disease in second wheats. Nine chemistry regimes were applied to two cultivartreatmentsinthreereplicated,fullyrandomisedfirstwheatfieldtrialsinHertfordshire,UK,between2013-2015. A post-harvest soil core bioassaymeasured the resulting amount of take-all inoculumbuild-up.All three trialswere thenoversownwitha take-all susceptiblesecondwheatcultivaranddiseaseresultingfromthefirstwheattreatmentswasquantifiedduringgrainfilling.EarlyfirstwheatspraysofAmistarwerefoundtoreducetake-alldisease inthesubsequentsecondwheatcrops,whenmeasuredusingthetake-all index ina threeyearcombinedREMLanalysis (P=0.009).Amid-seasonfirstwheatsprayhadanimmediatereductiveeffectontake-all inoculum(P=0.014), but no significant effect on second wheat take-all index was observed.We conclude thatAmistar,appliedinearlysprayregimestofirstwheats,effectivelyloweredsecondwheatdisease,butwhenappliedasmid-late firstwheat sprays, altered thenormal take-all diseaseprogression cycle.TheLowTABgenetic trait consistently loweredboth inoculum (P<0.001)andsecondwheatdisease(P<0.001),howevernosynergywithAmistarwasidentified,indicatingtheeffectsofthegeneticsandchemistry will work additively. The LowTAB trait is currently being genetically mapped in severalbreeding populations and findings from this research should help towards providing on-farmsolutionstotake-alldisease.Keywords:Gaeumannomycesgraminis,take-all,integrateddiseasemanagement,Triticumaestivum,azoxystobin,wheatgenetics

Session6Poster125Towards a more sustainable agriculture: arbuscular mycorrhizal fungi protect wheat againstpowderymildewG.Mustafa1,N.G.Khong1,B.Tisserant1,B.Randoux1,J.Fontaine1,M.Magnin-Robert1,Ph.Reignault1,A.Lounès-HadjSahraoui11UnivLittoralCôted’Opale,EA4492–UCEIV–UnitédeChimieEnvironnementaleetInteractionssurleVivant,SFRCondorcetFRCNRS3417,F-62228Calaiscedex,FranceThe use of arbuscularmycorrhizal fungi (AMF) as resistance inducers and therefore as biocontroltoolscouldbeaninnovativealternativetochemicalsforcontrollingplantdiseases.Ourworkaimedat studying the possible protective effect of arbuscular mycorrhization in bread wheat (TriticumaestivumL.)againstBlumeriagraminisf.sp.tritici(Bgt),theobligatebiotrophicAscomycetefungusinfectingwheataerialorgansand responsible for thepowderymildewdisease.Wheatmycorrhizalinoculation by Funneliformismosseae (Fm), under controlled and nutritional optimized conditions,allowedustoobtainconcomitantlyamycorrhizalrateof38%,asignificantincreaseofplantbiomassandaprotectionlevelagainstBgtestimatedat78%.Theseresultssuggesttheinductionofsystemicwheat defense reactions in response towheatmycorrhization, i.e.Mycorrhiza-Induced Resistance(MIR).ThisprotectionislinkedtoanaccumulationofphenoliccompoundsandhydrogenperoxideattheBgtpenetrationsites inepidermal leafcellsofmycorrhizedplants.Up-regulationsofPOX,PAL,NPR1andCHI1genesencodingfordefensemarkerswerealsopointedout in leavesofmycorrhizalwheat in the absence of Bgt infection. Our study also highlighted the importance of taking intoaccount various environmental parameters to optimize the potential use of AMF as biocontrolagents. The highest protection against powdery mildew was obtained with a 5-fold reducedphosphorusinputcomparedtothatrecommendedinthefield,bothinamoderatelycultivarandinamore resistant cultivar. Moreover, the level ofwheat protection dependsmore on the inoculumtypethanonthemycorrhizalrateobservedinwheatroots.Keywords: arbuscular mycorrhizal fungi, powdery mildew, wheat, mycorrhiza-induced resistance,biocontrol

Session6Poster126ZnOnanoparticlesforpostharveststrawberrygreymouldcontrolNeringaRasiukevičiūtė1,AlmaValiuškaitė1,NobertasUselis1,ŽivilėLukšienė2

1Institute of Horticulture, Lithuania Research Centre for Agriculture and Forestry, Kaunas st. 30,Babtai,Kaunasdistrict,Lithuania2InstituteofAppliedResearch,VilniusUniversity,Saulėtekio10,10223Vilnius,LithuaniaIt is well known that most of the pathogens can develop high resistance to chemical fungicides.Researchers all over the world are looking for new technologies reducing pesticide consumption.Newimplementsareneededtoreducepostharvestlossesandcontrolfoodpathogens.Thismethodare based on measures of zinc oxide (ZnO) nanoparticles. Photosensitization is an innovativeapproach for eliminating fruit pathogens based on simultaneous use of photoactive compound(photosensitizer)andvisiblelight.Thisstudyaimedtoevaluatetheefficacyofnewmeasuresforreducingcontaminationofpathogenicfungi in strawberry. The research was carried out at the LAMMC Institute of Horticulture. Thephotosensitization (PH) evaluated in LED-based light source prototype. Experimental treatmentsincluded1)control,2)ZnO+PH,and3)sterilewater+PH.Visuallyhealthystrawberrycv.“Darselect”fruitswereincubatedwithphotosensitizerZnOfor1handilluminated30min.withlight(λ=400nmwith an energy density of 20 mW/cm2). Decontaminated fruits assessed after 4 and 8 days ofsimulatedstorageat5-7oC.Theexperimentalresultsrevealedthat4and8daysaftertheZnOtreatment,thenumberofBotrytisinfectedfruitsreducedby10%and13%.Inwatertreatment,theincidenceofgreymoldaftera4-daystorage was 9% and 19 % higher compared to the control and ZnO. Data indicated that ZnOnanoparticles reduced strawberry contamination with Botrytis cinerea. Such ZnO nanoparticlespropertiesareapromisingtoolforthedevelopmentofeffectivenonchemicalfungicides.Acknowledgement. This work was carried out within the framework of the long-term researchprogram“Harmfulorganismsinagroandforestecosystems”implementedbyLAMMC.Keywords:Botrytiscinerea,contamination,photosensitizer,ZincOxide

Session6Poster127Are sown flower strips an efficient tool to limit viral epidemics and aphid colonization inmeloncrops?AlexandraSchoeny1,NathalieBoissot2,CatherineWipf-Scheibel1,KarineNozeran1,PascaleMistral2,PatrickGognalons1,PaulineMillot1andHervéLecoq11PathologieVégétale,INRA,84140,Montfavet,France2GénétiqueetAméliorationdesFruitsetLégumes,INRA,84140,Montfavet,FranceThemelonaphidAphisgossypii is a seriouspestonmelon crops causing leaf-curling, stuntingandevenplantdeathwhencolonizationisintense.Itisalsoanefficientvectoroffourvirusesfrequentlyobserved on melon crops in France: Cucurbit aphid-borne yellows virus (CABYV, Polerovirus,Luteoviridae),Cucumbermosaicvirus (CMV,Cucumovirus,Bromoviridae),Watermelonmosaicvirus(WMV,Potyvirus,Potyviridae)andZucchini yellowmosaicvirus (ZYMV,Potyvirus,Potyviridae).Themanagementofaphidsandviruses is all themorechallenging that theevolutionof the regulation(planEcophyto2018) imposesaprogressive reductionof theusageof thephytosanitaryproducts.Innovative strategies areneeded to control thesebioagressors. Thebibliography suggests that themanagement of fieldmargins could contribute to regulate the populations of aphids and/or theirviralload.Indeed,flowerstripscanparticipateinaphidbiologicalcontrolbyfavoringnaturalenemiesand stripsofnon-hostplants canprotect crops fromnon-persistentlyaphid-transmittedvirusesbyallowing aphids to probe on healthy plants and thus to lose their virus load before reaching thecrops. A pluriannual (2011-2015) and multidisciplinary experiment allowed an evaluation of thepotential roleof sown flower strips todecrease the riskof aphid colonization and the riskof viralepidemicsinmeloncrops.Keywords:Aphisgossypii,conservationbiologicalcontrol,Cucumismelo,integratedpestanddiseasemanagement

Session6Poster128BacillusamyloliquefaciensGA1aspotentialbiocontrolagentofSeptoriatriticiblotchAliSiah2,AliceRochex1,SégolèneArnauld1,3,AmelElArbi4,DaphnéPerret3,GaëlHuet3,AloïsBahu3,ThibaudChabanon3NéjiGharsallah4,PatriceHalama2,PhilippeJacques11Univ.Lille,EA7394,USC1281-ICV-InstitutCharlesViollette,F-59000Lille,France2ISA,EA7394-ICV-InstitutCharlesViollette,F-59000Lille,France3Univ.Lille,Plate-formeagro-alimentaire,PolytechLille,F-59000Lille,France4FacultédesSciencesdeSfax,LaboratoiredeBiotechnologieMicrobienneaffiliéauLaboratoiredeBiotechnologies Végétales Appliquées à l’Amélioration des Cultures in vitro B.P. 802, 3018 Sfax,TunisiaZymoseptoria tritici is the causal agent of Septoria tritici blotch (STB), one of themost damagingfoliar disease of wheat. Conventional fungicides are widely used to reduce STB and related yieldlosses but chemical pesticides are increasingly controversial. Thus, research efforts are needed toidentifyalternativescontroltoolssuchasbacterialcontrolagentsortheirmetabolites.Amongthem,strains belonging to the genus Bacillus and particularly B. subtilis and B. amyloliquefaciens areefficientforthebiocontrolofmultiplefungaldiseases.TheaimofthisworkwastodetermineifthefiltrateofaB.amyloliquefaciensGA172hculturepossessestheability toprotectwheatagainstZ.tritici. Invitrobioassays showeda strongantifungaleffectof the filtrate,withaminimal-inhibitoryconcentrationof6.25%v/v(inmixturewithpotatodextroseagarmediumusedforfungalgrowth).Inplanta experiments, carriedout in thegreenhouseon the susceptible cv.Alixan, showed strongdiseasereductions(upto98.5%)onplantstreatedwiththeculturefiltrateat100%,50%or25%v/v(indistilledwater).Cytological investigationsrevealedasignificant impactofthetreatmentsonboth spore germination (at day one post-inoculation) and fungal growth (at day five post-inoculation), thusagreeing the invitro findings.Furthermolecular investigationsare inprogress todeterminetheabilityoftheculturefiltratetoelicitplantdefensepathways.Inconclusion,thisstudydemonstrated a significant potential ofB. amyloliquefaciensGA1 to be used as a bio-pesticide onwheatagainstZ.tritici.Keywords:Zymoseptoriatritici,Bacillusamyloliquefaciens,biocontrol,biopesticides

Session6Poster129Oligosaccharide-based inducer exhibits high protection against Zymoseptoria tritici and inducesvariousrelated-defensepathwaysinbreadanddurumwheatLamia Somai-Jemmali1, Ali Siah2, Beatrice Randoux3, Maryline Magnin-Robert3, Patrice Halama2,PhilippeReignault3andWalidHamada11Laboratoire Génétique et Amélioration des Plantes, Institut National Agronomique de Tunis, 43AvenueCharlesNicolle,1082Tunis,Tunisie2InstitutCharlesViollette(EA7394),SFRCondorcetFRCNRS3417,InstitutSupérieurd’Agriculture,48BoulevardVauban,BP41290,59014LilleCedex,France3Univ.LittoralCôted'Opale,UCEIV-EA4492,UnitédeChimieEnvironnementaleetInteractionssurleVivant,SFRCondorcetFRCNRS3417,F-62228CalaisCedex,France

Theaimofthisworkwastoinvestigatetheelicitingandprotectiveeffectsofpreventivetreatmentswith an oligosaccharide-based new formulation (Oligos) against Zymoseptoria tritici, a majorpathogen of both bread wheat (BW) and durum wheat (DW). Oligos treatment led to stronglyreduced fungal hyphal growth on leaf surface, penetration attempts, mesophyll colonization andfructificationduringZ.triticiinfectiononbothtestedBW(cv.Premio)andDW(cv.Karim)cultivars.ItalsodrasticallydecreasedCWDEactivitiesbyZ. tritici, suchasendo-β-1,4-xylanaseandendo-β-1,3-glucanase during the necrotrophic phase of infection, suggesting their correlation with diseaseseverity and their potential use as markers of resistance inducer efficiency. The expression ofdefense-related genes such asGLUC, Chi4, POX and LOX, the activities of GLUC and LOX and thecontentoftotalsolublephenoliccompoundswereenhancedinbothnon-inoculatedandinoculatedplants. However, induced PAL activity, H2O2 accumulation and polyphenol deposition inmesophyllcells were observed only in inoculated context but not to the same way between both wheats.Overall,Oligos treatment induced the samedefensepathways in testedBWandDWcultivars,butnottothesameextent,suchasGLUC,CAT,LOXandPALenzymaticactivities.Furtherinvestigationsusing additional susceptible and partially-resistant BW and DW cultivars are required to confirmtheseresults.TheOligosresistanceinducerpresentsaninterestingactivitycharacterizedbyhighandstable protection efficiency when it is used inappropriate conditions, and therefore could beintegratedintoalternativecontrolstrategiesagainstZ.tritici.Keywords: Zymoseptoria tritici, Mycosphaerella graminicola, oligosaccharides, resistance inducer,wheat

Session6Poster130Evaluationofleafextractsoffourplantspeciesagainstriceblastpathogen(Magnaportheoryzae)M.O.Adebola1,O.B.Ayeni1andM.B.Aremu2

1FederalUniversityofTechnology,DepartmentofPlantBiologyMinna2NationalCerealResearchInstitute,Badeggi,Nigeria Rice (Oryza sativa) is oneof themost popular food crops inNigeria. Its successful productionhasbeen drastically affected by blast disease caused byMagnaporthe oryzae. In vitro control of thepathogen by four medicinal plants (Carica papaya, Azadirachta indica, Calotropis procera andAnacardiumoccidentale)wasassessedinthisstudy.Theextractsoftheplantswerepreparedusingaqueous and methanol, and agar well diffusion method was used to assess the toxicity of eachextract. The pathogenwas isolated from rice infectedwith blast disease. The results revealed thepresenceofoneormorephytochemicalsineachoftheplantextracts.Amongthesewerealkaloids,tannins,flavonoids,saponin,anthocyaninandphenol.AlltheextractsinhibitedmyceliagrowthofM.oryzae. The potency of all the extracts increased with increasing concentration in the order;50mg/ml<100mg/ml/150mg/ml. The inhibitionsbymethanol extractswerehigher and significantlydifferent (P>0.05) from aqueous extracts. At the highest concentration tested (150mg/ml), A.occidentale andC. procera gave the highest inhibitions (99.0mm and 98.6mm respectively) whichwere not significantly different (P<0.05) but different fromC. papaya andA. indica (89.1mmand90.4mm respectively). However, in all, A. occidentale aqueous and methanol extracts gave thehighestpercentagegrowthinhibitionofthepathogenatall levelsofconcentrationstestedwhileC.papaya aqueous andmethanol extracts though effective were the least. Therefore, field trials ofthesefourmedicinalplantsonthecontrolofriceblastdiseasearerecommendedsincetheyareeasytoobtainandtheextractscouldeasilybemadeviaasimpleprocessofmacerationorinfusion,theycouldbecheapersubstituteforconventionaldrugsincontrollingriceblastdisease.Keywords:blast,potency,extract,phytochemicals,pathogen

Session6Poster131Saltandthermos-tolerantTrichodermaasperellum isolatefromOmanisapotentialantagonistofrootrotpathogensM.S.Alansari123,J.Bacanovic1,A.M.Al-Sadi3,M.Deadman3,M.R.Finckh11EcologicalPlantProtection,UniversityofKassel,Germany,2RoyalCourtAffairs,P.O.Box17,Muscat111,Oman3DepartmentofCropSciences,CollegeofAgriculturalandMarineSciences,SultanQaboosUniversity,OmanOneofthebiggestthreatstomaizeinOmanarerootrotdiseases.Trichodermaspeciesareknowntobenaturalantagonistsofmanypathogensandarewidelyused inbiologicalcontrolof fungalplantdiseases.Therearecommercialbiocontrol fungibut theymightnotperformoptimallyunderhighsalinityandhightemperatureconditions.TwoTrichodermaasperellum isolatesfromOmaniorganiccompostandamaizerhizospherewereevaluatedforantagonisticeffectsagainst fourknownplantpathogenscausingmaizerootrotinOman:thefungiFusariumfujikuroi,Botryodiplodiatheobromae,and Rhizoctonia solani and the oomycete Pythium arrhenomanes. A commercially availablebiocontrolisolateofTrichodermaharzianumwasusedascontrol.The local isolates showedantagonisticeffectsandparasitismagainst the three fungi similar to thecommercialspecies.Incontrast,biocontrolactivityagainstP.arrhenomaneswassignificantlyhigherthan forT.harzianum. Scanningelectronmicroscopyshowedthat theT.asperellum isolatescoiledaround the pathogens and led to hyphal collapse at the interaction zone suggesting directhyperparasiticinteractions.Effects of different salinity concentrations (24µS/cm-50 dS/m) and temperatures (5-40°C) on thelocal T. asperellum and the commercial T. harzianum in vitro were assessed. All isolates werenegatively affected by increasing salinity. However, the growth rates of the local isolates weresignificantlyhigherthanthatofthecommercialT.harzianumatallsalinityconcentrations.Growthofthe local isolateswas comparable to the commercial agent at temperaturesup to30°Cbut itwassignificantlyhigherat35°C.These results demonstrate that the local Trichoderma local isolates are more thermo- and salt-tolerant than the commercially availableproduct and, therefore, couldbepotentially effectivebiocontrolagentsforlocaluse.Keywords:bio-control,Trichodermaasperellum,MaizeRootrot,antagonism

Session6Poster132AspergilluswesterdijkiaewithstrongactivityagainstPythiaceaecausingdiebackofappletreesYosraBenM’henni1,2,DanieleDebieu2,StéphaneMann3,SlimTounsi4,NaimaBoughallebM’hamdi1andSabineFillinger21Department of Biological Sciences and Plant Protection, High Institute of Agronomy of ChottMariem,UniversityofSousse,Tunisia2UMRBIOGER,INRA,AgroParisTech,UniversitéParis-Saclay,ThivervalGrignon,France3CNRS, Muséum National d’Histoire Naturelle, Chimie des Produits Naturels Fongiques – MCAMUMR7245MNHN-CNRS,63,rueBuffon75005Paris4LaboratoryofBiopesticides,CenterofBiotechnologyofSfax,P.B.1177,3018Sfax,TunisiaDiebackdiseaseofappletreescausedbyPythiaceae (Pythiumsp.,Phytophthora sp.;oomycetes) isan important soil-borne disease reducing apple (Malus communis) production in Tunisia. It causesseveredamagesand tree losses innumerousappleorchards.Thisdiseasecausescankers,necrosisandrotinthecollarsandroots.The present study aimed at investigating the biocontrol potential of antagonistic fungal (e.g.,Trichoderma spp., Aspergillus spp., Penicillium spp.) or bacterial isolates (Bacillus spp.) againstPythium and Phytophthora sp, to determine their phylogenic position, and to characterize theantifungalcompoundsproduced.Theantagonisticeffectsofselectedbiocontrolagents(BCAs)wereevaluatedbydifferentscreeningmethodsinvitro.Usingthedualculturetechniqueonsolidmediumaswellasusingculturefiltrates,weconcentratedourstudyonthose isolates forwhichwewereable todetectsecretedantifungalactivityinhibitinginvitrogrowthoftestedPythiaceaebyover50%.TheresultsofthisstudyshowedinvitroefficacyoftwoisolatesofTrichodermaspp.,oneisolateofAspergillusspp.andtwostrainsofBacillussppagainstPythiumandPhytophthoraspp.ThehighestactivitywasfoundforanAspergilluswesterdijkiaeisolatewhoseculturefiltrateinhibitedthemycelialgrowthofPythiumundulatum,Phytophthoraunidataupto100%. Inorderto identifytheactivemolecule(s)wepurifiedandcharacterizedtheculturefiltratebydifferentchromatographicandspectroscopictechniquesincludingHPLCandLC/MS.We selected a unique fraction from culture filtrates with high inhibitory activity. Its chemicalcharacterizationisongoing.Theseencouragingresultsinciteustoproceedthrough invivoassaysoftheselectedAspergilluswesterdijkiaeisolateagainstPythiumsp.onappleplants.Keywords:Maluscommunis,Pythiaceae,BCAs,antifungalactivity,HPLC,LC/MS

Session6Poster133AntifungalpotentialofhopsextractsagainstthewheatpathogenZymoseptoriatriticiLaetitiaBocquet1,AliSiah2,JenniferSamaillie1,Jean-LouisHilbert1,PatriceHalama2,SevserSahpaz1,CélineRivière11 Univ. Lille, INRA, ISA, Univ. Artois, Univ. Littoral Côte d’Opale, EA 7394 - ICV - Institut CharlesViollette,F-59000Lille,France2 ISA, Univ. Lille, INRA, Univ. Artois, Univ. Littoral Côte d’Opale, EA 7394 - ICV - Institut CharlesViollette,F-59000Lille,FranceResearchonalternativemethodstoconventionalpesticidesagainstcroppathogensisanimportantchallenge. Here, we tested the potential of extracts and compounds from hops to be used asbiofungicidestowardsZymoseptoriatritici,themainpathogenonwheatcropstodayinEurope.Hopsis a food and medicinal plant, rich in prenylated phenolic compounds which are responsible fornumerous biological activities including antiviral, antibacterial and antifungal properties. Hops hasnotyetbeenstudiedforthebiocontrolofplantpathogens.Wefirstassessedtheantifungalpotentialof crude hydro-alcoholic extracts of different parts of hops plant (leaves, stems, roots and femalecones), aswell as theessential oil of female cones. Theantifungal assayswereperformedusing aspotting test carried out in Petri dishes containing potato dextrose agar medium amended withdifferentconcentrations(1.25,0.62,0.31,0.15and0.07g.L-1)ofeachextract.Dose-responsecurveanalyzesrevealedthatonlythehydro-alcoholicextractoffemalecones,aswellastheiressentialoil,significantlydecreasedfungalgrowth,withhalf-maximalinhibitoryconcentrations(IC50)of0.73g.L-1and0.36g.L-1, respectively. Furthermore, a fractionationof thehydro-alcoholic crudeextract fromfemale cones using centrifugal partition chromatography allowed the purification of phenoliccompounds. Antifungal assays using the purified products revealed that the prenylated chalconedesmethylxanthohumol, as well as the acylphloroglucinol derivative co-humulone, are responsibleforpartoftheantifungaleffectoffemalecones.Theessentialoil isbeingcharacterizedinordertohighlighttheantifungalterpeniccompoundsinvolvedintheactivityagainstZ.tritici.Keywords:biofungicides,hops,wheat,Zymoseptoriatritici

Session6Poster134MultiplescreeningapproachfortheselectionofefficientbiologicalcontrolagentsagainstBotrytiscinereaontomatoYousraBouaoud1,2,AbdelhamidFoughalia1,ClaireTroulet2,KamelAissat1,MarcBardin21MicrobialEcologyLaboratory.UniversityofAbderrahmaneMira,Bejaia06000,Algeria2PlantPathology,INRA,84140Monfavet,FranceOne hundred and twenty one bacterial isolates collected from tomato stems and from soil in theBejaiaregion(Algeria)werescreenedfortheirpotentialtocontrolgraymoldontomato.Thebacteriawere initially tested for theirdirect invitroeffectagainst twostrainsofB.cinerea.Basedontheseresults,37isolatesshowinginvitroinhibitionofB.cinereamycelialgrowth(rangingfrom0%to69%inhibition) were chosen for further analysis. These isolates were identified based on 16S rDNAsequencing. Those identifiedaspotentiallypathogens forhumans,mammalsorplants andable togrow at 37°C were discarded. Twenty five were then selected to evaluate their effectiveness asbiological control agents againstB. cinerea using a tomatoplant bioassay. Comparisons of in vitroandinplantascreeningmethodshighlightedtheabsenceofcorrelationbetweenthesemethodsandconfirmthatasinglescreeningstrategyisnotsufficienttoselectefficientbiologicalcontrolagents.Among the 25 isolates, 8 exert high and significant antagonistic activity against two strains of B.cinerea.Amongthem,threebacteriabelongingtothePseudomonasgenuswerefinallyselectedfortheir significant and stable efficiency at 3 concentrations (107, 108 and 109CFU/mL) to reduce thelesiondevelopmentofB.cinereaontomatostem(%protectioncomprisedbetween60and100%),theirabilitytogrowat15°Candtheirinabilitytogrowat37°C.ThesethreeisolatesarealsoabletoreducesignificantlythesporulationofB.cinereaontomatostem.This study is a first step in the selection of biological control agents able to protect the tomatoagainstB.cinereaandadaptedtothegreenhouseconditionsinAlgeria.Keywords:Botrytiscinerea,tomato,biologicalcontrolagents,screening,Pseudomonasspp.

Session6Poster135InvitroeffectofessentialoilsontheColletotrichumgloeosporioidesdevelopmentDanieleMariadoNascimento1,PaulaLeitedosSantos1,AdrianaZaninKronka11UniversidadeEstadualPaulista (Unesp),FaculdadedeCiênciasAgronômicas,CâmpusdeBotucatu,SP.RuaJoséBarbosadeBarros,nº1780,CEP18.610-307,Botucatu,SãoPaulo,BrasilColletotrichum gloeosporioides, causal agent of sweet pepper anthracnose, is one of the mostimportant pathogens of this crop. The present study was carried out in order to identify analternativeoptionforthisfunguscontrol,evaluatingtheeffectofessentialoils(EO)onthefungusinvitro development. The experiment, performed in duplicate, was carried out in a completelyrandomizeddesigninafactorialscheme8x3[8essentialoils(rosemary,citronella,clove,copaiba,eucalyptus,peppermint,basilandteatree)x3concentrations (0.25%;0.5%and0.75%)andonecontroltreatment(potato-dextrose-agarmedium-PDA),with3replications.TheoilswereaddedtoPDAmediumatpredeterminedconcentrations.Adiscof0.5cmdiameterofPDAmediumwiththefungus was placed in the center of each petri dish containing the treatments. The plates weremaintainedat22°Cand12hour-photoperiod.Toevaluatetheinhibitoryeffectoftheoils,mycelialgrowthandsporulationforeachtreatmentweredetermined.Inbothexperiments,clove,citronella,eucalyptus, mint and basil essential oils inhibited the fungus mycelial growth and sporulationcompletely in all concentrations. Tea tree EO showed the same inhibitory effect in concentrationshigherthan0.50%,whilerosemaryEOprovidedtotalinhibitionat0.75%;forbothEOtherewashighsporulation.CopaibaEOhadnon-satisfactory results.According to the results, it canbe concludedthatclove,citronella,eucalyptus,mintandbasilessentialoilshaveapotentialuseinthealternativecontrolofC.gloeosporioides.Keywords:Anthracnose,naturalfungicide,sustainabledevelopment

Session6Poster136AntagonismofTrichoderma.spwithPestalotiopsis.spinTunisiaSawssenHlaiem1,MeriemZouaouiBoutiti1andMohamedLahbibBenJemâa11NationalInstituteforresearchinengineering,Ruralwaterandforests,BP.N.2,Ariana2080,TunisiaThefungiareoneofthemaincausesofthediseasesoftrees.Thesymptomsofdiebackincludeafallandayellowingoffoliage,adryingandnecrosisatthelevelofbranches,cankers,deformations,ofablackish fluidandflowofrotsat the levelof thetrunks (BenJamâa,et.Al.,2005;Hasnaouiandal.,2008; Barky & Abourouh, 1996; Barky et al. 1999; Franceschini et al., 1999, 2002). The forest ofHenchir Kort, (northeast of the Tunisia) has suffered heavy infestation since 2012. Symptoms ofwiltingwerenotedonthepine(Pinuspinea)andseveralotherspeciesofscruboakscale (Quercuscoccifera). In October 2016, attacked samples with symptoms of necrosis and dryness have beencollectedshrubsofoakscale.ThepathogenicfungusPestalotiopsis.sphasbeenisolatedfromtheselesions. TheKoch’s postulate has been verified. The antagonistic actionwas assessed in vitro. Theresults showthat inmixedculturewithPestalotiopsis. sp. isolateofTrichoderma.sphasshownaneffect from the inhibition of Mycelial growth (fungistatic effect) to the degradation and thedisappearance of themyceliumofPestalotiopsis (mycoparasitism and fungicidal effect). The studycontinueswithabenchmarktestoftheeffectivenessofthenaturalantagonistObjective:Thepresentworkisthesearchforabiologicalcontrolmethodusinganaturalantagonist.Materialsandmethods1.Isolationandidentificationoffungi(IsolationweremadeaccordingtothetechniqueconductedbyFranceschinietal.2005).2. Test of pathogenic pathogenesis test was conducted using the method of inoculation ofLinaldedduetal.(2014).3.Testofconfrontation(BenhanouandChet,1996).4.AssessmentofthesurvivalrateofPestalotiopsis.spResultsandconclusion1. The koch’s postulate has been verified: all of the inoculated twigs were infected withPestalotiopsis.sp.2.ThetestofdirectconfrontationinvitrobetweenPestalotiopsis.spandTrichodermasp.hasshowafongistaticeffectandfungicidaleffect.3.TheassessmentofthepercentageofsurvivalofpathogenicfungustestrevealsthatPestalotiopsis.sp.hasarateof0 %.Keywords:kermesoak,necrosis,Pestalotiopsis,Trichoderma,antagonistic

Session6Poster137Fitness competitiveness and sensitivity to fungicides ofMonilinia laxa and Monilinia fructicolaisolatesfromSerbiaJovanaHrustić1,MilicaMihajlović1,MilaGrahovac2,BrankicaTanović11InstituteofPesticidesandEnvironmentalProtection,Banatska31b,Belgrade,Serbia2UniversityofNoviSad-FacultyofAgriculture,TrgDositejaObradovića8,NoviSad,SerbiaBrown rot is oneof themost severe pre- andpostharvest diseaseswith aworldwide distribution.Since M. fructicola, the most destructive pathogen of the genus Monilinia, has recently beenintroduced into Serbia and many other European countries, many studies were conducted toevaluate differences in characteristics of Monilinia species that could have an impact onestablishment and survival of thepathogen in newareas. The aimof this studywas to assess thecapacityofM. fructicola to repressand replaceM. laxa inSerbia,basedon thecomparisonof theisolates in terms of fungicide sensitivity (iprodione, tebucanozole, prochloraz, chlorothaloni,azoxystrobin, fluopyram, and boscalid), growth rate and virulence at different temperatures. TheresultsshowedthattheisolatesofM.fructicolaweresignificantlylesssensitivethanM.laxatothefollowingfungicides:iprodione,tebucanozole,chlorothalonil,azoxystrobin,fluopyram,andboscalid.Moreover,M. fructicola isolates displayed a wide range of sensitivity, whereas M. laxa isolatesexhibited little variation in sensitivity to all the tested fungicides. In addition, growth rate andvirulence of both species were significantly affected by the temperature. The lowest testedtemperature(5°C)wasfavorableforM.laxagrowthrateandvirulence,whereasat30°CM.fructicolagrewfasterandhadhigherlesionexpansionrate.TheseresultsprovideanadditionalexplanationforsignificantchangesinthepopulationstructureofMoniliniaspp.inSerbiaaftertheintroductionofM.fructicola,esspeciallyinpeachesandnectarinesthatarereadyforharvestduringhotsummertime.ProjectIII46008.Keywords:fungicidesensitivity,competitiveness,speciesdisplacement,stonefruit

Session6Poster138Controlofpowderymildew(Blumeriagraminis)incerealsusingSerenade®ASOLiseNistrupJørgensen1&NielsMatzen1

1DepartmentofAgroecology,AarhusUniversity,Flakkebjerg,4200Slagelse,DenmarkPowdery mildew (Blumeria graminis) is a serious pathogen of cereal crops in many parts of theworld. Traditionally this disease has been controlled by using resistant cultivars or by applyingfungicides. In recent years more focus has been put on biological control agents (BCA’s) asalternatives to chemical fungicides for control of diseases in various crops, including cereals. Thebiofungicide Serenade® (Bacillus subtilis strainQST713 fromBayerCropScience)was investigatedfor itspotential for controlofpowderymildew in cereal crops.Bacillus subtilis is a rhizobacteriumthatcanformendospores,produceseveraldifferentantibioticsandproducemicrobialdisruptersofpathogen cellmembranes. The product is approved for use in the EuropeanUnion, and themaintargetssofarhavebeenBotrytiscinereaandErysiphespeciesinvegetablesandstrawberries.FieldtrialscarriedoutinwheatandbarleycropsinDenmarkhaveshownmoderatetogoodcontrolof powdery mildew using Serenade® when repeated treatments have been applied. Data fromgreenhousetrialshaveshownthatcorrecttimingisveryimportantforgoodcontrol.Bestcontrolisobtained if treatments are applied just around the time of infection. Only minor or no doseresponseswereseenwhenusing2,4or6lSerenade®perha.Bothinthefieldandthegreenhousetrials the traditional fungicides (triazoles) provided superior control compared to treatments withSerenade®. The challenge for BCA’s in cereal crops is to findmeans of integrating their use withtraditional chemistry and understand their role in a combined diseasemanagement scenario. Theresults so far indicate that the BCA’s can’t be stand-alone solutions for conventional farmers, buttheir use needs to be exploited further also as a means of reducing the risk of development offungicideresistance.Fororganicfarmersthemainchallengewillbetoknowwhenthecropisatriskofattackasadelayedsprayinghasbeenseentoprovidelesseffectivecontrol.Keywords:biologicalcontrolagents,efficacy,wheat,barley

Session6Poster139InductionofpotatodefenseresponsesandreductionofsymptomsfromPhytophthorainfestansbytwodifferentelicitorsRafaelaLopesMartin1,2,CécileThomas1,2,JeanClaudeYvin3,EricNguema-Ona3andFlorenceVal1,21AGROCAMPUSOUEST,UMR1349IGEPP,F-35000Rennes,France2UniversitéEuropéennedeBretagne,France3CentreMondiald’InnovationRoullier,LaboratoiredeNutritionVégétale,SaintMalo,35400,FrancePhytophthora infestans isanoomycete responsible forpotato lateblight.Due toaggressivenessofthis disease, the only way to control is withmany fungicide applications. Thus, a major scientificchallenge is to develop alternative methods as plant defense induction to reduce pesticidetreatments. In this aim, our study focus on the potential efficiency of defense elicitors to protectpotatoagainstP. infestans. Thena concentratedculture filtrated (CCF) fromP. infestans (endogenelicitor) and a green algae’s (Ulva spp.) extract (exogenous elicitor) were tested on two potatogenotypes:BF15,moderatesusceptibletoP.infestansandDésirée,moderateresistant.Potatoplantshavebeentreatedwiththeelicitorsatdifferentconcentrationsand48hlaterdetached-leaveswereinoculated with P. infestans at 5.104 sporangia.mL-1. Induction of defense responses has beenevaluatedafterelicitationonnon-inoculatedleavesbyatranscriptanalysisoftwelvepotatodefensegenes.Thequantityof symptomshasbeenevaluateduntil sevendaysafter inoculationontreatedanduntreatedleavesbythemeasureofnecroticareas.OurresultsshowedthatCCFinducedmorestrongly defenses genes in BF15 than in Désirée genotype but none reduction of symptoms wasmeasuredafterCCFapplication.However,algalextractinducedtheexpressionofgenesdifferentiallyinbothgenotypes.ButasignificantreductionofnecroticareawasonlyobservedonBF15genotype.ThisreductioncouldbeexplainedbythestronginductionofPhenylalanineAmmonia-LyasegeneandtoalesserdegreebytheinductionofTyramineN-hydroxycinnamoylTransferasegene.Indeed,thesetwo genes are implicated in the phenylpropanoid and salicylate pathways, which produceantimicrobial components. To verify this hypothesis, metabolic and proteomic analysis are beingtested an attempt to correlate defense induction and effective protection in different interactionsbetweenplantsandpathogens.Keywords:biocontrol,plantimmunity,Potatolateblight,Solanumtuberosum

Session6Poster140EffectofessentialoilsontheapplescabcausativeorganismVenturiainaequalisMarieMaňasová1,JanaWenzlová1,MiloslavZouhar1,JanaMazáková1,PavelRyšánek11Czech University of Life Sciences Prague, Department of Plant Protection, Kamýcká 129, 165 00,Prague6,CZCausalagentofApplescabVenturiainaequalisitistheoneofthemostimportantpathogenlimitingapple fruit production. Decreasing number of fungicides and active substances, as well as theoccurrence of resistant pathogen populations and increasing demand for biological fruitproduction,leadingustodevelopnewfungicidesonanaturalbasis.Inthisworktheefficiencyof13essential oils fromMentha spicata, Cymbopogon citratus, C. winterianus, Litsea cubeba, Thymusvulgaris, Pelargoniumgraveolens, Rosmarinus officinalis, Eugenia caryophyllus,Ocimumbasilicum,Cinnamomum ceylanicum, Lavandula hybrida, Origanum majorana and Cin. camphora ontheseveralisolatesofVenturia inaequalishavebeentested.BestresultsarereachedessentialoilsfromplantsofT.vulgarisandE.caryophyllus,whichsignificantlyreducedthegrowthoftheorganismandthusgiveabasisforfurtherworkinvivo.ThisstudywassupportedbytheCzechMinistryofAgriculture,projectnumbersQJ1510353.Keywords: Apple scab,Venturia inaequalis, essential oils, biofungicides, Thymus vulgaris, Eugeniacaryophyllus

Session6Poster141TheprotectionofcerealsbyfungicidalessentialoilsextractedfromhigherplantsPavelMatusinsky1,MiloslavZouhar2,RomanPavela3,PavelNovy4,51Agrotestfyto,Ltd,Havlickova2787,76701Kromeriz,CzechRepublic2Czech University of Life Sciences Prague, Faculty of Agrobiology, Food and Natural Resources,DepartmentofPlantProtection,Kamycka129,16521Prague6–Suchdol,CzechRepublic3CropResearchInstitute,Drnovska507,16106Prague6Ruzyne,CzechRepublic4Czech University of Life Sciences Prague, Faculty of Agrobiology, Food and Natural Resources,Department of Quality of Agricultural Products, Kamycka 129, 165 21 Prague 6 – Suchdol, CzechRepublic5Czech University of Life Sciences Prague, Faculty of Agrobiology, Food and Natural Resources,DepartmentofCropProduction,Kamycka129,16521Prague6–Suchdol,CzechRepublicBotanicalpesticidesaresubstancesderivedfromplantswithawiderangeofutilization.Wetestedeffect of essential oils extracted from aromatic plants on inhibition of mycelium growth ofphytopathogenic fungiattackingcereals. Inourstudy (i) the laboratory testswithphytopathogenicfungiinvitro,(ii)growthchamberbiotestswithplantsand(iii)thefieldsmallplotexperimentsusingofartificialinoculationwereconducted.Inthelaboratoryteststheinhibitoryeffectofessentialoilsonthemycelialradialgrowthoffungiwastested by the agar dilutionmethod. The effect of essential oils extracted fromPimpinella anisum,Thymus vulgaris,Pelargonium odoratissimum,Rosmarinus officinalis and Foeniculum vulgare weretestedon the fungiOculimaculayallundae,Microdochiumnivale, Zymoseptoria tritici,Pyrenophorateresand Fusarium culmorum. The best antifungal activitywas demonstrated byThymus vulgaris.Themost prevalent compounds of T. vulgaris were thymol (44.60%), p-cymene (21.94%) and γ-terpinene(7.80%).The biotest with artificially inoculated (by F. culmorum) wheat seed were conducted in growthchamber.Testplantswerecultivatedinhydroponicmediaamendedbyessentialoils,thesameasinlaboratory test above (0.1% emulsion). The best antifungal activity was demonstrated again byessentialoilextractedfromThymusvulgaris.EfficacyofspraymicroencapsulatedformulationoftheessentialoilfromT.vulgaristotheearswastestedinthefieldexperiment.Wheatplantswereartificially inoculatedbyF.culmorum.Mycotoxincontent in harvested grain was analysed using ELISA. Level of deoxynivalenol was significantlyreducedaftertreatmentwithoilfromT.vulgaris.WorkwassupportedbyMinistryofAgricultureoftheCzechRepublicprojectsQJ1310226,QJ1310091andRO0211.Our results suggest that the essential oil from T. vulgaris have great potential for safe andenvironmentallyfriendlytreatmentcerealsagainsttofungalpathogens.Keywords:botanicalpesticides,Thymusvulgaris,phytopathogenicfungi,wheat,barley

Session6Poster142EfficacyoffungicidesandbiofungicidesinthecontrolofVerticilliumWiltonpepperMilicaMihajlović1,EmilRekanović1,JovanaHrustić1,MilaGrahovac2,BrankicaTanović1

1InstituteofPesticidesandEnvironmentalProtection,Belgrade,Serbia2UniversityofNoviSad,FacultyofAgriculture,NoviSad,SerbiaVerticillium dahliae is considered to be one of the most destructive soilborne plant pathogens.Managementof thepathogen isdifficult,becauseof itsendophyticgrowthand longpersistence insoil.TheobjectiveofthisstudywastoexaminethepossibilitiesofVerticilliumwiltcontrolinpepperby using conventional fungicides (thiophanate-methyl, difenoconazole, fluopyram, azoxystrobine,prochloraz) and two commercially available biopesticides based onBacillus subtilis strain and tea-treeoil. InvitrosensitivityofV.dahliae isolate,derivedfrominfectedpepperplantand indentifiedbased on pathogenic and morphological features, to the tested fungicides and tea-tree oil wasdetermined in radial growth experiment on PDAmedium supplementedwith a range of fungicideconcentrations. The concentration that inhibitedmycelial growthby 50% compared to the control(EC50)wascalculated.Theefficacyofthetestedproductsinvivowasevaluatedoninoculatedpepperplants under greenhouse conditions. The results of the in vitro experiment showed thatdifenoconazoleandthiophanate-methylwerethemosttoxicfungicidestotheisolate(EC50=0.02mg/land0.03mg/l,respectively),while,undergreenhouseconditions,thehighestefficacywasrecordedfor thiophanate-methyl (83.1%). The essential oil and B. subtilis – based product exhibited lowefficacyincontrollingthedisease(46.2%and35.4%,respectively).Theobtainedresultsrevealedthatneither fungicide nor biofungicide treatments could provide adequate supression of the diseasewithoutcombinationwithothercontrolstrategies.ProjectTR31043.Keywords:Verticilliumdahliae,pesticidecontrol,Bacillussubtilis,tea-treeoil

Session6Poster143SpotIT–IT-solutionsforuserfriendlyIPM-toolsinmanagementofleafspotdiseasesinbarleyandwheatBerit Nordskog1, Andrea Ficke1, Tor-Einar Skog1, Asbjørn Veidal1, Jonathan Yuen2, Annika Djurle2,Björn L.-E. Andersson2, Hanna Bergeå3, Helena Nordström Källström3, Antanas Ronis4, RomaSemaskiene4, Lise Nistrup Jørgensen5, Jens Grønbech Hansen6, Egon B. Noe6, Marja J. Jalli8, JussiNikander7,AnneNissinen8,JensErikØrum91NIBIO–NorwegianInstituteofBioeconomyResearch,Pb115,N-1431,Ås,Norway,2SLU–SwedishUniversityofAgricultural Sciences,Dept.ofForestMycologyandPlantPathology,Box7026,S75007,Uppsala, Sweden, 3SLU - SwedishUniversity of Agricultural Sciences,Dept. ofUrban andRuralDevelopment,Box7012,S75007,Uppsala,Sweden,4LithuanianResearchCentreforAgricultureandForestry,InsitituteofAgriculture,InstitutoAve.1,Akademija,Kedainiaidistrict,LT58344,Lithuania,5Aarhus University, Faculty of Agricultural Sciences, Dept. of Crop Protection, Forsøgsvej 1,Flakkebjerg,DK-4200Slagelse,Denmark,6AarhusUniversity,FacultyofAgriculturalSciences,Dept.ofAgroecology,ResearchCentreFoulum,DK-8830Tjele,Denmark,7LUKE–NaturalResourcesInstituteFinland, Vakolantie 55, FI-03400 Vihti, Finland, 8LUKE – Natural Resources Institute Finland, PlantHealth,Tietotie2C,FI-31600Jokioinen,Finland,9UniversityofCopenhagen,SectionforEnvironmentandNaturalResources,Rolighedsvej25,1958Frederiksberg,DenmarkLeafdiseasesareamajorthreattocereals intheNordic-Balticareaandfungicidesarewidelyusedfor reducing yield losses. Theproject SpotIT is aNordic-Baltic initiative started in2017, toprovidecereal farmerswithbettermodels forpredicting leafspotdiseases inwheatandbarley,aimingforuser-friendly dissemination of model outputs through locally adapted IPM-tools. The NorwegianOpen Source technology platform VIPS will provide a trans-national facility formodel testing andvalidation,usinginputdatafromallparticipatingcountries.VIPSwillalsoprovideabasisforefficient,userfriendlyandlow-costexpansionoflocallyadapteddecisionsupportsystems(DSSs)withatrans-nationalsharingofknowledgeandmethods.Despite development of national DSSs, farmers do not always use the novel tools. Data fromprevious studies in Denmark, Norway and Finland suggest that relatively few farmers use DSSdirectly, while agricultural advisors are themain target group. Themain approach of SpotIT is toidentifyuserneedsandexpectations,understandfarmers’decision-makingandsuggestsolutionsfordisseminationofplantprotectionrisksineachcountryorregion.End-userpreferenceswillbe investigated to identify factors topromote increaseduseof localDSSandimplementationofIPMtools.Thechoiceofriskmodelsforleafdiseasesinwheatandbarleywillbebasedontheuserneedsaswellastheperformanceofthemodelsunderlocalconditions.Wearecurrentlycompilingadatabaseofmodelcandidates,whichwillbepresentedduringtheconference.Theaimistotestthembasedonhistoricaldatainthedifferentregions.Fieldtestswillbemadeincooperationwithfarmersandextensionservicetovalidatethemodelsindifferentcountries.Whilethisprojectwillfocusonleafspotpathogens,theresultingDSSplatformcaneasilybeusedforotherhost,pestorpathogensystems.Keywords:integratedpestmanagement,DSS,VIPS,cereals

Session6Poster144IntegratedcropsolutionstoofferdifferentiatedsolutionsBernardPélissier1andFrédéricDerolez2

1CropScienceDivision,BayerSAS,14impassePierreBaizet,BP99163,69263LyonCedex09France2CropScienceDivision,BayerSAS,16rueJean-MarieLeclair,69266LyonCedex09FranceManydiseaseswhichincludefungi,oomycetesandbacteriasignificantlyimpactcropproductionandquality. In addition growers are continually facing new emerging or re-emerging diseases, such assoybeanrustinBrazil(Phakopsorapachyrhizi),citrusgreeninginCalifornia(CandidatusLiberibacter),EscainEuropeanvineyards(complexoffungi)orryeergot(Clavicepspurpurea)inFrance. Intopofthattheindustryisfacingincreasinglychallengingregulatoryandsocietaldemandsinachangingworld (eg increasing population, changing consumption patterns, climate change, unsufficientstorage)makingfoodsafetyatrisk.Inthiscomplexenvironment,innovationinamodernagricultureiskeytosupportthesenewchallengesandintegratedcropsolutionswillbenecessarytoreachtheworldwidedemand.AtBayer, tohelp farmers secure their harvest andenhance thepublic perceptionof their job,wefocus our research and development towards a sustainable approach using all tools of amodernagriculture: small molecules, biologicals, biotechnologies including native or GM traits, SMARTbreeding,precisionagricultureandservices,systematicallyaccompaniedbyfullstewardshipsupport.Diverseexamplesoftheseapproacheswillbepresentedanddiscussed.Keywords:diseases,integratedcropsolutions,farmers,foodquality

Session6Poster145Use of compost to partially substitute non renewable growing media and suppress soil-bornepathogensonpottedvegetableplantsMassimoPugliese1,2,MariaLodovicaGullino1,2,AngeloGaribaldi2

1AgriNewTechsrl,ViaG.Quarello15/A,Torino,Italy2University of Torino, Centro Agroinnova and DISAFA Largo P. Braccini 1, I-10095 Grugliasco (TO),ItalyCompostsareexpectedtosuppressplantdiseases,accordingtothetypeofwastes,thecompostingprocess,thechemicalandmicrobiologicalcomposition.Suppressivecompostsaregenerallyappliedas soil improvers, while it is necessary to develop specific compost based growing media forapplicationsonpottedplants.The aim of this research was to evaluate the suppressiveness of compost/peat growing mediacomparedtopeat.Acommercialgrowingmedia,“Hortofan”,madeof20%v/vcompost,peatandpumicewastested.SuppressivenesswastestedingreenhouseonpottedplantsagainstPythiumultimumoncucumber,Phytophthora nicotianae and Fusarium oxysporum f. sp. lycopersici on tomato. Pathogens weremixed into the substrate at 1 g of biomass on wheat kernels L-1 7 days before seeding or withchlamydosporestalcat1x104UFC/gofsubstrate.Seedsofcucumberandtomatoweresowninto2Lpots ingreenhouseandfivepotswereusedforeachtreatment.Acommercialpeatbasedgrowingmediawasusedascontrol. Thenumberofaliveplantsandweightofabovegroundbiomassweremeasured20-30daysafterseeding.Cucumber and tomatobiomass significantly increasedup to 40-50%with “Hortofan” compared tocontrol. The number of diseased tomato plants in substrates inoculated with P. nicotianae wassignificantlyreducedby40%andthenumberofdiseasedcucumberplantsby30%comparedtothepeatsubstrate.Fusariumwiltoftomatowasreducedby60%inplantsgrownwith“Hortofan”.Thegrowingmedium“Hortofan”,thankstoitscompositionbasedonhighqualitycompost,improvedplant development and controls Pythium ultimum on cucumber, Phytophthora nicotianae andFusariumoxysporumf.sp.lycopersiciontomato.Compostisavaluablecomponentofgrowingmediathatcanpartiallysubstitutenonrenewablesourcesandsuppresssoil-bornepathogens.

Keywords:tomato,cucumber,peat,Pythium,Fusarium,Phytophthora

Session6Poster146AttemptstouseCoriandrumsativumessentialoiltoreduceseedpathogensNeringaRasiukevičiūtė1,EditaDambrauskienė1,RasaKarklelienė1,AlmaValiuškaitė11Institute of Horticulture, Lithuania Research Centre for Agriculture and Forestry, Kaunas st. 30,Babtai,Kaunasdistrict,LithuaniaThe increasing resistance to chemical fungicides drawn attention to the natural productsantimicrobial activity. Essential oils arebiodegradable andeco-friendly botanical products that areaccumulatedinvariousplants.Theaimofthisstudywastoevaluateantimicrobialeffectofcorianderessentialoil againsthorticultural crops seed-bornepathogens.The researchwascarriedoutat theLAMMC Institute of Horticulture. The Coriandrum sativum essential oil was extracted from localmaterial. The C. sativum essential oil efficacy against seed-born fungi was evaluated by plantingnaturally contaminatedonionandcucumber seedsonPetriplatescontainingpotatodextroseagar(PDA).ThePDAwascooledto45oC,andessentialoilwasadded:200µl/l,400µl/l,600µl/l,800µl/land1000µl/l.TheantifungalactivityofC.sativumessentialoiloncucumberandonionrevealedthatseveral concentration differently inhibited bacterial and fungal pathogens. Evaluation of theefficiencyofessentialoilonseed-bornepathogensatconcentrationof1000µl/lcompletelyinhibitedthe pathogens. The concentration of 200 µl/l least inhibited bacterial and fungal pathogens. Theessential oils as bio-fungicides would help to control seed-borne pathogens without increasingchemicalfungicidesresistance.Acknowledgement. This work was carried out within the framework of the long-term researchprograms“Horticulture:agro-biologicalbasicsandtechnologies”and“Harmfulorganismsinagroandforestecosystems’implementedbyLAMMC.Keywords:antifungal,corianderoil,inhibition,pathogens

Session6Poster147InvestigationstocontrolOculimaculayallundae,theeyespotcausingpathogen,usingvarietyresistanceandfungicideapplicationBerndRodemann11InstituteforPlantProtectioninFieldCropsandGrassland,JuliusKühn-Institute,Messeweg11/12,D-38104Braunschweig,GermanyAsaresultoftheexpansionofthecerealcultivationarea,inparticularofwinterwheat,anintensifiedattack with stem diseases is increasingly being identified. In paticularOculimacula yallundae andOculimacula acuformis, damages thehosts and inhibit thenutrient uptake,which leads to lodgingandyieldlosses.Thisdangerofcontaminationisstronglyinfluencedandpromotedbythechangingweatheroverwinter.Forthisstudy,approachestocontrolhavebeentestedbythecultivationofresistantvarietiesandbytheuseofeffectivefungicides.Inthiscontext, itwasalsonecessarytoinvestigatethesensitivityofthepathogenanditsalterationagainsttheactivesubstances.In laboratory tests, the active substances boscalid, fluxapyroxad, cyprodinil, prothioconazole,prochloraz,metrafenoneandpyriofenoneweretestedinvitroagainstconcentrationsof0.01,0.11.0and 10.0 ppmagainstOculimacula yallundae. After 31 days, the highest efficiencies of about 80%wereachievedbyboscalidandfluxapyroxad.Theefficacyoftheactivesubstancesboscalid,fluxapyroxad,cyprodinil,prothioconazole,prochloraz,metrafenone and pyriofenone in the wheat cultivars Atomic, Partner, Ritmo and Tobak wereinvestigatedinadplantaexperimentswithartificial infection.ThespeciesAtomicandPartnerwereequippedwiththeresistancegenePch1.Inyoungplant tests, theactive ingredientsBoscalid>Fluxapyroxad>Cyprodinil showedthehighestefficacywithapproximately55%.OnlyindividualsymptomscouldbedeterminedfortheAtomicandPartner,whereasasignificantrootrotwasassessedinthehighlysusceptiblecv.Tobak.The investigations show that in practice the cultivation of resistant varieties is the basis for theprevention of primary attack and the secondary spread in the stalk. By combining with effectivefungicidestheefficacyinthecontrolofOculimaculayallundaecanbefurtherincreased.Keywords:Oculimaculayallundae,varietyresistance,fungicidecontrol,inoculationtest

Session6Poster148EffectsoforganicfertilizerandleafnitrogencontentonLeafspotdiseasedevelopmentingrapecv.‘CampbellEarly’causedbyPseudocercosporavitisRyu, YoungHyun1, SuGon Bae1, IlKwon Yeon1, KwangSup Kim1, JeongMin Park1, SangSeok Lee1,JongHoPark2

1Organic Agriculture Research Institute, Gyeongbuk Agricultural Research and Extension Service,Uisung-eup,Uisung-gun,Gyeongbuk,Korea2Organic Agriculture Division, National Academy of Agricultural Science, Rural DevelopmentAdministration,Wanju,KoreaThe table grape cultivar “Campbell Early”,widely cultivated inmost SouthKorea region and someJapanregion,hasauniqueflavorandtastecomparedtoEuropeantablegrapecultivarsandaccountsformore than70%of tablegrapemarket inSouthKorea. “CampbellEarly” isvery sensitive to soilnutrients and needs efficient soil nutrient management from early growth stage to harvestingseason,somostofcultivatingtechniquesarefocusedonwinterandsummerpruningandselectionof fruiting number before harvesting for efficient nutrient uptake and most of disease controlmeasuresweredependonbordeauxmixture.Oneofcharacteristicsof“CampbellEarly” isthatthiscultivar ishighlyresistanttopowderymildewandwhitemildew,whilemostofEuropeancultivarsaresusceptibletothem.However,oneofmajorweaknesses in “Campbell Early” is its susceptibility to grape leaf spot disease caused byPseudocercosporavitis.Grapeleafspotdiseaseisamajorissueespeciallyinorganicgrapecultivationpractice, due to limited supplement of organic fertilizer during growing season and shortage oforganic-allowedmaterialfordiseasecontrolbeforeharvesting.Toexaminetherelationshipbetweensoil nutrient supply and disease development, some kinds of organic fertilzers applied during“CampbellEarly”cultivationandchemicalcompositionsuchasnitrogenandcarboncontentsinleafduringmid-growthseasonwereanalyzedinthisstudy.Leafnitrogencontents inearly July seasonhad lesseffectsondisease spotnumberbutcorrelatednegativelywithdiseasespotarea,whiletheratioofcarbontonitrogenshowedpositivecorreationwithit.InFishmealorganicfertilizerapplyingplot,thenumberofappearedspotandareawashigherthanExpellercakefertilizerandBloodmealfertilzerplotbeforegrowingseasonandevenworsenedthedegree of disease in without organic fertilizer plot. When applying Expeller cake or Blood mealfertilizer, leaf nitrogen contents were 2.26∼2.48% and spot area was 76%, but without organicfertilizer,leafnitrogencontentswere1.91∼1.94%andspotareaincreasedto94%.Inconclusion,wesuggestthatforpropercontrolofgrapeleafspotdiseaseincv.“CampbellEarly”,itis necessary to take appropriatemeasures not only on the use of control agent but also on vinenutrients.Keywords:CampbellEarly,Pseudocercosporavitis,leafspot,orgnicfertilizer,nitrogen

Session6Poster149Biofungicide activity of Juncus maritimus extracts and effusol against Zymoseptoria tritici, thecausalagentofSeptoriatriticiblotchofwheatRamlaSahli1,2CélineRivière1,AliSiah3,AbderrazakSmaoui2,JenniferSamaillie1,ThierryHennebelle1,VincentRoumy1,RiadhKsouri2,PatriceHalama3,SevserSahpaz11Univ. Lille, INRA, ISA, Univ. Artois, Univ. Littoral Côte d’Opale, EA 7394 - ICV - Institut CharlesViollette,F-59000Lille,France2The Laboratory of Aromatic and Medicinal Plants, Biotechnology Centre of Borj-Cédria (CBBC),Hammam-lif,Tunisia3ISA, Univ. Lille, INRA, Univ. Artois, Univ. Littoral Côte d’Opale, EA 7394 - ICV - Institut CharlesViollette,F-59000Lille,FranceZymoseptoriatriticiistodaythemostfrequentlyoccurringpathogenonwheatcropsinEurope.Thewide use of synthetic fungicides caused an emergence of fungal resistant strains and potentiallynegative impacts on the environment. Searching for eco-friendly biofungicides is an importantchallenge and a promising alternative strategy to such products. In this context, we assessed thebiocontrolpotentialofextractsfromeightextremophileplantspeciesfromTunisiaagainstZ.tritici.Crudemethanolicextractsfromdifferentpartsoftheseplantswerepreparedandassessedfortheirantifungal activity using a spotting test performed in Petri dishes containing potato dextrose agarmediumamendedwithdifferentconcentrationsofeachextract.Extractfromahalophyteplantfromtherushfamily,Juncusmaritimus,showedthehighestactivitywhencollectedonaspecificsubstrate.In extremophile plants, the production of secondary metabolites is often influenced by abioticconditions. Thus, we collected J. maritimus rhizomes at different vegetative stages, at differentperiods and from different substrates to compare their antifungal activities. HPCL-UV analyses ofextracts showed differences in chromatogram profiles, suggesting that the plant environment,especiallythesubstrateofthesoil,shouldbetakenintoaccounttoidentifygreatsourcesofnaturalantifungalproducts.Fromthemostactiveextract,a9,10-dehydrophenanthrenederivative,effusol,absent from other J.maritimus samples, was purified. This product showed a strong antifungalactivity against the pathogen, with aMIC of 19mg.L-1 and an IC50 of 9.98mg.L-1. Therefore, thisphenanthrene derivative could be a promising biocontrol molecule against Z. tritici. Furtherinvestigations would allow a deeper understanding of the modes of action of effusol against thepathogen.Keywords:biofungicides,extremophileplants,Juncusmaritimus,wheat,Zymoseptoriatritici,effusol

Session6Poster150Seedcoatingwithnaturalbiostimulants:apracticaltoolforcontrolofwheatdiseasesMaissaBenJabeur1,ZaynebKthiri1,EssaidAitbarka2,WalidHamada1

1LaboratoryofGeneticsandplantbreeding,NationalinstituteofAgronomyofTunis,Tunisia2Laboratory of Stress, Defenses and Reproduction of Plants, University of Reims-Champagne-Ardenne,FranceSeptorialeafblotchandFusariumheadblightaretwomajordiseasescausingseverelossesindurumwheat.Coatingseedswithbeneficialmicro-organismsandplantextractsappearstobeapromisingapproachtomaintain theproductivityofplantsunderstresscondition. In thisstudy,weevaluatedtheendophyticbacterium,BurkholderiaphytofirmansstrainPsJN,thymeessentialoil,theantagonistTrichoderma harzianum, the yeastMeyerozyma guilliermondii, and their different associations fortheirabilitytocontrolthediseasesmentionedbothundercontrolledconditionsandinfield.SeedsofasensitiveTunisiancultivarofdurumwheat“Karim”werecoatedwithB.phytofirmans(10⁸CFU/ml)and T. harzianum (10⁶ spores/ml), thyme essential oil (5 ppm),M. guilliermondii (108 spores/ml).Undercontrolledconditions,SeptorialeafblotchwasmonitoredinpotswhileFusariumheadblightwas assessed in hydroponic system. Treatments reduced pycnidial coverage of septoria to 10%comparedtocontrol(40%).Cytologyandenzymaticanalysisshowedthatthesetreatmentsenhanceplant resistancewith increasedcatalasesactivity, reducedperoxidasesactivityandH2O2 levelsandreduced fungal colonization and development in leaf cells. In field, coated seedlings showed areducedseptorialeafblotchattacktoanaverage10-20%comparedtocontrol(43%).AsforFusariumhead blight, under controlled conditions, severity was reduced of about 30%, with reducedperoxidasesactivityinroots,enhancedphenoliccompoundscontentinleavesandrootsandreducedcolonizationandmacroconidiaabundanceinrootcells.Keywords:Septoria,Fusarium,Burkholderia,Trichoderma,thyme,Meyerozyma

Session6Poster151BiologicalactivityofselectedessentialoilsagainstAlternariadauciEvaZusková1,MartinaPačesová1,JanaMazáková1,VeronikaŘíčařová1,EvženieProkinová11Plantprotectiondepartment-CzechUniversityofLifeSciencesPrague.Kamýcká129,16500Praha6-Suchdol-Praha-SuchdolEssential oils in plant protection have got already thousands years old history, but theirmode ofaction and all the possibilities of their use are themain object of the researchers in last decades.Thesedaysisaneedtofindalternativemethodtosyntheticpesticidesbecausefinalcustomerscallfor ecological commodities and also for methods of harvesting, which are friendly to nature.Pesticidesonthebaseofessentialoilscanbeecologicalformofsyntheticpesticideswhichcangetatqualitiesofconventionalproduction.ObjectsoftheresearchareAlternariadaucioncarrot.Onthepathogenweretested16essentialoils(EO) fromdifferentplants.Theexperimentswerecarriedat laboratoryconditions,butalso in fieldconditions.Thebiggestpotential showessentialsoils fromEugeniacaryophyllus andCinnamomumzeylanicum.A. dauciwas 100% inhibited in 0,1% concentrationby EO fromEugenia caryophyllus, alsoby EOfromCinnamomumzeylanicum.Otheressentialoilsalsoshowssomeinhibitionpotential,butnotin100% (Litseacubeba,Pelargoniumgraveolens,Thymusvulgaris).Results fromfieldexperiments infirstyeararenotsoclearandneedmoreresearchforcorrectinterpretation.Keywords:fungalpathogensonvegetable,Alternariadauci,carrot,essentialoils

Session6Poster152Steamingandbiologicalcontrol:potentialstrategiesforIPMofFusariumwiltoflettuce?FrançaSC1,DeBusscherJ2,VandeveldeI3,HöfteM21Microbials,R&Ddepartment,BiobestBelgium,IlseVelden18,2260Westerlo,Belgium.2LaboratoryofPhytopathology,Dep.CropProtection,FacultyofBioscienceEngineering,GhentUniversity,Coupurelinks653,9000Gent,Belgium.3ProefstationvoordeGroenteteelt(PSKW),Duffelsesteenweg101,2860Sint-Katelijne-Waver,BelgiumFusariumwiltisanincreasingproblemingreenhouse-grownlettuceinBelgiumandtheNetherlands.Thediseaseiscausedbythesoil-bornefungusFusariumoxysporumf.sp.lactucae(fol).SinceitsfirstreportinJapanin1955,thediseasehasbeendescribedcausinglossesinlettuceproductionareasworldwide.Oncethepathogenisestablishedinsoil,itisverydifficulttomanageit.Folformsrestingstructuresthatcansurviveinsoilandplantdebrisforyears.UrgentdevelopmentofeffectivecontrolmeasuresisnecessarytoensurethefutureofintensivelettucecultivationinsoilinBelgianandDutchgreenhouses.Commerciallyavailablechemicalsoilfumigantsseemnottobeabletocontrolthepathogen.Inaddition,non-chemicalcontrolmethodsaremoreinlinewiththeIPMapproachsupportedbytheEuropeanUnion.InSeptember2016,steamingwasappliedinaninfestedgreenhouseinBelgium.Soilsamplesweretakenfrom0-20cmand20-40cmdepthbeforeandaftersteaming.FusariumpopulationsweredeterminedbyplatingonKomada’smedium.Clearreductionofpopulationswasfoundinbothsoillayersaftersteaming.However,remainingpopulationswerehigherinthedeepestlayer.Nodiseasesymptomswereobservedduringthefollowingtwocropcycles.Fusariumpopulationsandcropdevelopmentwillbemonitoredinthespringandsummerof2017.UseofabiologicalcontrolagenthasshownpotentialagainstfolinapotexperimentintheNetherlands,butfailedtocontrolthediseaseinapottrialinBelgium.Differentcultivars,growthconditionsandinoculumdensityprobablyexplainthedifferentresults.Currently,moretrialsarebeingperformedconcerningbiocontrol.Keywords:Fusariumoxysporumf.sp.lactucae,IPM,biocontrol,soildisinfestation

Session6Poster153EssentialoilsontheFusariumoxysporumf.sp.capsiciinvitrocontrolPaulaLeitedosSantos1,DanieleMariadoNascimento1,AdrianaZaninKronka11UniversidadeEstadualPaulista (Unesp),FaculdadedeCiênciasAgronômicas,CâmpusdeBotucatu,SP.RuaJoséBarbosadeBarros,nº1780,CEP18.610-307,Botucatu,SãoPaulo,BrasilFusariumwilt on pepper, caused by the fungus Fusarium oxysporum f.sp. capsici, is an importantdisease thathascaused increasing losses in this crop. This studyaimed toevaluate theactivityofdifferentessentialoilsonthemycelialgrowthandsporulationofF.oxysporumf.sp.capsici,isolatedfrom sweet peppers. The experiment was carried out in duplicate, on a completely randomizeddesignaccordingtothe8x3factorialscheme[8essentialoils(clove,copaiba,eucalyptus,teatree,basil, rosemary, citronella andpeppermint) x 3 concentrations (0.25%0.50%and 0.75%)] andonecontroltreatmentPDAmediumonly),withthreereplicationspertreatment.Theoilswereaddedtothe PDA culturemedium at the predetermined concentrations. A disc of 0.5 cm diameter of PDAmediumwiththefunguswasplacedinthecenterofeachpetridishcontainingthetreatments.Theplatesweremaintainedat25 °Cand12hour-photoperiod.Toevaluate the inhibitoryeffectof theoils,mycelialgrowth(diameterofcolony)andsporulation(numberofspores/mL)foreachtreatmentweredetermined.Inbothexperiments,citronella,cloves,eucalyptus,peppermintandbasilessentialoils inhibited completely themycelial growth, in all concentrations, differing from the control andavoiding the sporulation. For rosemary and copaiba essential oils, high fungal sporulation wasobserved in both trials, not differing from the control, showing that both essential oils did notpresentagoodinhibitoryeffect.Wecanconcludethatcitronella,clove,eucalyptus,peppermintandbasilessentialoilscanbeusedfortheFusariumoxysporumf.sp.capsicialternativecontrol.Keywords: alternative control, mycelial growth, sporulation, natural fungicide, sustainabledevelopment

Session6Poster154COSTActionCA16107EuroXanth: integrating scienceonXanthomonadaceae for integratedplantdiseasemanagementinEuropeRalfKoebnik1andEstelleEmeriau2

1Institut de Recherche pour le Développement, UMR 186 IRD-Cirad-Université Montpellier,InteractionsPlante-MicroorganismesEnvironnement(IPME),911AvenueAgropolis,BP64501,34394Montpelliercedex5,France2COSTAssociation,AvenueLouise149,1050Brussels,BelgiumBacteriaofthefamilyXanthomonadaceae, includingspeciesofXanthomonasandXylellafastidiosa,belongto themostdevastatingplantpathogenscontinuallychallenging foodsecurity.Manyof thepathogensarelistedasquarantineorganismsintheEUandtheirstudyisofuttermostimportance.Theconcernedpathogens infectallkindsofcropplants,suchascereals, foragecrops forruminantfeed,vegetables,fruits,shrubsandtrees.COST (European Cooperation in Science and Technology; www.cost.eu) is a funding agency forresearchandinnovationnetworks.COSTActionshelpconnectresearchinitiativesacrossEuropeandenable scientists to grow their ideas by sharing themwith their peers. This boosts their research,careerandinnovation.This COST Action will bring together some of the brightest and best minds to join in aninterdisciplinary network to develop strategies for sustainably protecting plants and significantlyreducing yield losses. Specifically, this initiative will address several key aspects of the pathogen-vector-host interactions from the cellular to the population level. A better insight into populationstructures and virulence mechanisms of the pathogens, together with the exploration of themolecularmechanisms underlying disease resistance to the pathogen,will enable development ofdurably resistantplantcultivarsandexploitationofbio-controlschemestailoredtopopulationandpathogen.This COST Actionwill generate a platform that gathers experts from different disciplines, such asmolecular diagnostics,molecular host-microbe interactions, plant resistance breeding, and appliedmicrobiology. Joining their efforts will help to develop and implement effective plant protectionschemes,beitviaresistantcropcultivarsorviaothercontrolmechanisms.Thisgoalwillbeachievedby mobilizing and training scientists from major European institutions, regulatory bodies andcommercialcompaniesworkingonthevariousaspectsofthiscomplexofproblems.Please follow our COST Action at Twitter (https://twitter.com/EuroXanth), Scoop/it!(http://www.scoop.it/t/xanthomonadaceae-plant-diseases) and ResearchGate(https://www.researchgate.net/project/EuroXanth-Integrating-science-on-Xanthomonadaceae-for-integrated-plant-disease-management-in-Europe).Keywords: pathogen detection, genetic diversity, population structure, genetic resistance, hostdefense,diseasemanagement

Session6Poster155Enhancing plant disease resistance through synthetic re-engineering of ABA signalling andcatabolismSatishKulasekaran1,MartadeTorresZabala1,DeboraShoemark2,RobSessions2andMurrayGrant1

1SchoolofLifeSciences,TheUniversityofWarwick,Coventry2DepartmetofBiochemistry,UniversityofBristol,BristolUsing a high resolution, time-resolved microarray dataset of Pseudomonas syringae pv. tomatoDC3000infectedArabidopsisthalianaleavesweidentifiedearlyresponsegenesthatarespecificallytargetedbyoneormoreofthe28DC3000effector,whileremainingunresponsivetoabioticstresses,wounding and a disarmed DC3000hrp mutant. We designed strategies to re-engineer hormonesignalingpathwaysutilizingthese“effectorresponsive”promoters tocreateconditionallyactivatedsynthetic constructswhich canneutralizepathogenvirulence.Herewepresent results focusingonsynthetic constructs designed modulate ABA signaling and ABA catabolism during susceptibleinteractions.Usingmodelling informedapproaches,wemutatedkeyresidues inthePYL5andPYL9ABA receptors to enhance binding of pathogen induced ABA without activating the downstreamsignalingcomponents.TransgenicplantscarryingthemutantPYLproteinsunder thecontrolof theeffector responsive promoters showed markedly reduced symptom development and were moreresistanttoDC3000.Moreover,inre-engineeredPYL9expressingplants,thelevelsofABAresponsivetranscript,theproteinphosphatase2C,HAB1,remainedsignificantlylessincomparisontowildtypeplantsat6hafterDC3000infection.Tovalidatetheseresultswedemonstratethat35SCaMVoverexpression the mutated PYL5 generated plants which were more insensitive to exogenous ABAapplication, indicating that the ABA signaling pathway is disrupted in these mutant PYL5 lines.Concurrently,wegeneratedtransgeniclinesdesignedtocatabolizepathogengeneratedABAdrivingtheABAcatabolicenzyme,CYPA3,underthecontrolofaneffectorresponsivepromoter.Transgenicplants carrying the conditionallyactivatedCYPA3 geneweremore resistant toDC3000 infection incomparisontowildtypeCol-0.Keywords:ABA,PYL,effector,DC3000,CYPA3

Session6Poster156The BacPlant project: towards a sustainable agriculture by increasing plant tolerance to bioticstressunderclimaticchangeMélodie Sawicki1,MarylineMagnin-Robert1, Béatrice Randoux1, Lisa Sanchez2, Ali Siah3, Essaïd AitBarka2,PatriceHalama3,CédricJacquard2,andPhilippeReignault11Univ.LittoralCôted'Opale,UCEIV-EA4492,UnitédeChimieEnvironnementaleetInteractionssurleVivant,SFRCondorcetFRCNRS3417,F-62228CalaisCedex,France2Univ.ReimsChampagne-Ardenne,URVVC-EA4707,UFRSciencesExactesetNaturelles,LaboratoiredeStress,DéfensesetReproductiondesPlantes,SFRCondorcetFRCNRS3417,F51687ReimsCedex2,France3SA,EA7394-ICVInstitutCharlesViollette,SFRCondorcetFRCNRS3417,F-590007Lille,FranceWith7.27billiontodate,theworldpopulationisprojectedtoreach9.1billionpeopleby2050andthedemandfor food isalready increasing.Meanwhile, theclimate is likelytobecomewarmerandwetterduringwintersandhotteranddrierduringsummers.Climatechange(CC)willinducechangesinagriculturalpractices,thereforemultiplyingthethreatstosustainablefoodsecurity.Tocounteract,mitigateorcontrolplantdiseases,growersoftenrelyheavilyonchemical fertilizersandpesticides,butarangeofnegativeimpactsforhumanhealthandenvironment.Itisthereforecrucialtodeveloprelevant and innovative control strategies toprotectplants fromvariousdiseases. In recent years,promisingalternativestrategiesfor improvingplanthealthemergedthankstotheuseofbeneficialmicroorganisms(calledBCAsforBiologicalControlAgents)thatpromoteplantgrowthandimmunity.TheseBCAs can indeedenhanceplant resistanceand counteract environmental biotic stress. Theyoffernovelpossibilitiesinincreasingthesustainabilityofproductionsystems.Wheatplaysanessentialroleinsocietiesandtheirfoodsupplyissues.Asothercropplants,wheatissubjected to several fungaldiseases,whicharewidelydistributedwherever thecrop is grownandalterwheatyieldandquality.Powderymildew(PM)andSeptoriatriticiblotch(STB),areamongthemostprevalentandthemostdamagingfoliardiseasesonwheatinducingimportantyieldlosses.With the BacPlant research project, the impact of the climate change (mainly temperature anddrought stresses)on the toleranceofwheat to thedisease-causingagents responsible forPMandSTBinthepresenceofBCAswillbeanalyzed.Atastartingpoint,weusedtheBurkholderiaphytofirmansPsJN,arhizobacterialstrain,todevelopacolonizationprotocol.PsJNisanendophyticbacteriumwhichisabletocolonizetheinternaltissuesof the plant without causing external signs of infection or negative effects. In our protocol, weinoculatedPsJNbysoakinggrainsinbacterialsolutionfor6hours.Withthismethod,wefoundPsJNin aerial parts and in roots severalweeks after inoculation using determination on culturemedia.BacterialdetectionbyPCRmethodiscurrentlybeingperformed.The impactof thepresenceofPsJNondifferent traitsof the tripartite interactionbetweenwheat,BurkholderiaphytofirmansandPM/STBwillbeinvestigated.Keywords:biologicalcontrolagents,climatechange,Powderymildew,Septoriatriticiblotch,wheat

Session6Poster157Non-leguminouscovercropscontributetoPratylenchuscontrolinconservationagricultureJanH.Schmidt1,JohannesHallmann1,2,MariaR.Finckh1

1Kassel University, Department of Ecological Plant Protection, Nordbahnhofstr. 1a, D-37213,Witzenhausen,Germany2Julius Kühn-Institut, Federal Research Center for Cultivated Plants, Institute for Epidemiology andPathogenDiagnostics,Toppheideweg88,D-48161,Münster,GermanySoilconservation isoneof themajorchallenges foragriculture in the21stcenturyemphasizingtheneedtodevelopsustainablefarmingsystemsbasedonconservationtillageandpermanentsoilcoverbylivinganddeadplantsaswellastheuseofcroprotations.However,thesemeasuresmayfosterplant-parasitic nematodes with a broad host range (Pratylenchus spp.). These are especiallyproblematiconlegumesandneedtobekeptbelowdamagethresholdstoensureecologicalservicessuchasbiologicalnitrogenfixation.The dynamics of the indigenous fauna of plant-parasitic nematodes were studied in eightcoordinatedMulti-EnvironmentfieldExperiments(MEEs)underfouragro-environmentalconditionsinEurope(Continental,Nemoral,AtlanticNorthandMediterraneanNorth).Ploughedsystemswerecomparedwithnon-inversiontillageanddifferentcroppingsequences.TheMEEsconsistedofatwo-yearsequenceofwheatgrownwithorwithoutacloverlivingmulchandfollowedbyleguminousornon-leguminous cover crops or weedy fallow. The second main crops were maize, potatoes, ortomatoes,dependingonsite.Wheat grain yields were not affected by up to 900 Pratylenchus individuals 100 ml soil-1, thusindicating that densities were generally below the economic damage threshold. Initial populationdensitiesofPratylenchuswere19-311nematodes100mlsoil-1dependingonsite.Theyincreasedby32%inwheat-maizerotationsanddecreasedby37%inwheat-potatoandwheat-tomatorotations.Finalpopulationswere74%higherundernon-inversiontillagecomparedtoploughing.Leguminouscover crops and livingmulches increased the numbers by 89% compared to theweedy fallow. Incontrast,oilseedradish(Raphanussativus)andblackoat(Avenastrigosa)covercropsreducedfinalpopulationsofPratylenchusby12%comparedtotheweedyfallow,thushighlightingtheirvaluefornematodecontrolparticularlyforsoilswithhighsandandlowhumuscontents.The studywas supported by the EU7th Framework programprojectOSCAR (289277) (www.oscar-covercrops.eu)Keywords:plant-parasiticnematodes,conservationtillage,croprotation,subsidiarycrops

Session6Poster158PreharvesthormeticdosesofUV-CradiationcandecreasesusceptibilityoflettucetoBotrytisHilarion Vàsquez1, Jawad Aarrouf1, Laurent Urban1, Yves Lizzi1, Marine Forges1,3, ChaymaOuhibi1,2,NasseraAzzouz1,MarcBardin3andPhilippeNicot31LaboratoirePhysiologiedesFruitsetLégumes(UMRQualisud),Universitéd'AvignonetdesPaysduVaucluse,301RueBaruchdeSpinoza,84916Avignon2Unité de Physiologie et Biochimie de la Réponse des Plantes aux Contraintes Abiotiques,DépartementdeBiologie,FST,UniversitéTunisElManar,1068Tunis,Tunisie3PlantPathology,INRA,F-84140,Montfavet,FrancePostharvestapplicationsofUV-Cradiationhaveprovenveryefficient inreducingthedevelopmentofpost-harvestdiseasesinmanyspeciesincludinglettuce(LactucasativaL.).SeveralstudiessuggestthatUV-C radiation is effective not only because of its disinfecting effect but also because itmaystimulate plant defences. Preharvest treatmentwith UV-C radiationmay thus offer an interestingpotential for lettuce protection, provided that application doses are effectivewhile excluding anyharmfuleffectson theplants.Hereweprovideevidence that0.85kJ.m-2and1.70kJ.m-2 representdoses of UV-C radiation that are not deleterious for lettuce plants. We used several criteria toevaluate the effect of UV-C radiation on the plant, including histological observations; theconcentration of malondialdehyde, an indicator of membrane integrity, as well as parametersderivedfrommeasurementsofchlorophyllfluorescence,suchasmaximalefficiencyofphotosystemII(Fv/Fm)andthePerformanceIndex(PI)ofStrasser.Weobservedthatasingledoseof0.85kJ.m-2slightlyincreasedplantresistancetogreymould(BotrytiscinereaL.)whileasingledoseof1.70kJ.m-2hadtheoppositeeffect.Whena0.85kJ.m-2dosewasapplied4times,attwo-dayintervals,therewasanincreaseinthetotalphenolcontentofleaves,andinphenylalanineammonialyase,catalase,andMDAHR activities. Leaves inoculated 2 days after the last UV-C treatment showed significantlyincreasedresistance(-30%)whencomparedtothecontrol.Keywords:UV-C,Botrytis,fluorescence,resistance,PAL,Lactuca

Session6Poster159Characterizationofplantdefense stimulators (PDS)effect according todevelopmental stageandnitrogensupplyCamille Verly1,2, Marie Emmanuelle Saint-Macary3, Martine Rigault4, Frédéric Giraud2, Loïc Rajjou4andAliaDellagi41INRA,InstitutJean-PierreBourgin(IJPB),UMR1318INRA-AgroParisTech,ERLCNRS3559-UniversitéParis-Saclay, Laboratoired’Excellence"SciencesdesPlantesdeSaclay" (LabExSPS),RD10,F-78026Versailles,France2Staphyt-Biorizon,SitedeMontesquieu,AvFrançoisMagendie,33650Martillac,France3Staphyt,LaPaluzette,RoutedesMas34590Marsillargues,France4AgroParisTech,InstitutJean-PierreBourgin(IJPB),UMR1318INRA-AgroParisTech,ERLCNRS3559-UniversitéParis-Saclay,Laboratoired’Excellence"SciencesdesPlantesdeSaclay"(LabExSPS),RD10,F-78026Versailles,FrancePlantspossessaninnateimmunesystemthatallowsthemtodefendthemselvesagainstpathogens.The use of Plant Defense Stimulators (PDS) is based on this ability of plants to defend against apathogen.Theseproductscanactivatedifferentplantdefensepathwaystherebylimitinginfectionbythe pathogens. The use of conventional crop protection products can thus be reduced. Thedevelopment of SDP capable of eliciting plant defenses is therefore a promising strategy in thecurrent agro-ecological context. The effectiveness of these SDPs in activating systemic defensesdependson several parameters suchas the stageof developmentof theplant andenvironmentalconditions.Morespecifically,mineralnutrition,andmainlynitrogennutrition,playsamajorrole inthe establishment of defenses. The response to SDPs also depends on the genotype considered(varietaleffect).The impact of nitrogen anddevelopmental stageon the efficacy of theseproductswas studied inordertoidentifytheoptimalconditionsfortheiruse.TheeffectsofthestageofdevelopmentoftheplantaswellastherateofnitrateavailablewerestudiedonArabidopsisthalianaCol0sprayedwiththecommercialSDPBion®.TheactivationofdefensesinresponsetoSDPwasmeasuredbythestudyof2typesofresponses:-The level of expression of marker genes of the 2 systemic defense pathways (PR1 and PR5 forsalicylicacidpathway,PDF1.2forjasmonicacidpathway)usingqRT-PCR.-The levelofprotectionagainst thepathogenPseudomonas syringaeDC3000wasmonitoredusingcountingcolonyunit.Theresultsofthesetwointerrelatedanalyzeswillbepresentedatthisseminar.Keywords:plantdefensestimulator,nitrogen,stage,plantpathogen

Session6Poster160Effect of seed treatment with novel strains of Trichoderma spp. on establishment and yield ofspringwheatAllenG.Xue1,WeiGuo1,2,YuanhongChen1andGenevieveMarchand1

1OttawaResearchandDevelopmentCentre,AgricultureandAgri-FoodCanada,960CarlingAvenue,Ottawa,ONK1A0C6,Canada2College of Plant Sciences, Heilongjiang Bayi Agricultural University, 5 Xinfeng Road, Daqing,Heilongjiang163319,China

Fusarium head blight (FHB) is themost important disease ofwheat in Canada. FHB reduces grainyield and quality and results in seed contamination with Fusarium spp. that is associated withreducedseedvigorandpoorstandestablishment inwheat.Theeffectof seed treatmentswithsixstrainsrepresentingthreespeciesTrichoderma,selectedbasedontheirsuperiorantagonisticabilityonmyceliumgrowthofF.graminearumindualcultureassays,onwheatseedlotscontaminatedwithFusariumspp.(28-43%)wasexaminedinfieldtrialsin2008,2009,and2011.NoneofthesixstrainsofTrichoderma spp. showeda significant seed treatment effect for all parametersmeasuredeachyear, but over the three years, all six strains significantly reduced root rot severity and increasedyield,threestains(Trich12,TrichC70andTrichPine) increasedemergenceandfourstrains(Trich06,TrichC39, TrichC70, and TrichMM7) increased plant dry weight, compared with the untreatedcontrol.TrichC70wastheonlystrainthatshowedasignificantimprovementtoallfourparameters,increasing emergence by 10.9%, dry weight by 51.7%, and yield by 11.0% and reducing root rotseverityby51.7%.TheseeffectswerelessbutnotsignificantlydifferentfromthatoftheregisteredfungicideVitaflo-280(carbathiin+thiram)usedasthepositivecontrolinthefieldtrials.TheresultsindicatethatTrichodermastainTrichC70maybeusedasanalternativetofungicideseedtreatmentstoalleviatethedetrimentaleffectoftheseed-bornephaseofFHBinwheat.Keywords: Fusarium graminearum, Trichoderma spp., biological control, seed treatment, springwheat

Session6Poster161Chemicalscreenofnaturalsmallmoleculesidentifiedasteroidalalkaloid,holaphyllamine,abletoinduce defense responses inArabidopsis thaliana and increase resistance against PseudomonassyringaeAbderrakibZahid1,5,FerdousseLaggoun1,MaïtéVicré1,ArnaudLehner1,IsabelleRemy-Jouet2,OlivierPamlard3,SandraBeaupierre3,JéromeLeprince4,XavierLatour5,VincentRichard2,CatherineGuillou3,PatriceLerouge1,AzeddineDriouich1andJean-ClaudeMollet11Laboratoire Glycobiologie et Matrice Extracellulaire végétale (Glyco-MEV) EA4358, NormandyUniversity,University of Rouen, Cell imaging platform (PRIMACEN-IRIB), 76821Mont-Saint-Aignan,France and the research network VASI-Haute Normandie region, 2Laboratoire nouvelles ciblespharmacologiques du traitement de la dysfonction endothéliale et de l'insuffisance cardiaque,INSERM1096, IRIB, UFR Médecine Pharmacie, Normandy University, University of Rouen, 76183RouenCedex1, 3InstitutdeChimiedesSubstancesNaturelles,CNRS,LabExLERMIT,91198Gif-sur-Yvette, France. 4Laboratoire de Différenciation et Communication Neuronale et NeuroendocrineINSERM982, IRIB, Normandy University, University of Rouen, 76821 Mont-Saint-Aignan, France.5Laboratoire de Microbiologie Signaux et Microenvironnement (LMSM) EA4312, IRIB, NormandyUniversity,UniversityofRouen/ IUTEvreux,27000Evreux,France,andtheresearchnetworkVASI-HauteNormandie.5Currentaddress:Univ.LitorralCôted’Opale,UnitédeChimieEnvironnementaleet Interactions sur le vivant (UCEIV-EA4492),Univ. Lille-Nord de France,GIS PhysNopi CS80699, F-62229CalaisCedex,FrancePlantsrelyontheirinnateimmunesystemtocombatmicrobialinfections.Usingachemicalscreenof1,600plant-derivedcompounds,wehaveidentifiedasteroidalalkaloidthatisabletoinducedefenseresponses inArabidopsisthalianawithoutalteringgrowth.Theidentifiedalkaloid isholaphyllamine(HPA)whose chemical structures is similar to steroid pregnanesofmammals.WhileHPAwasnotdetectedinuntreatedArabidopsisseedlings,thesteroidalalkaloidisabletotriggertheformationofreactiveoxygenspecies,accumulationofcalloseandexpressionofanumberofpathogenesis-relatedgenes of the salicylic and jasmonic acid pathways. In addition, we show that pre-treatment ofArabidopsis seedlings with HPA before infection with the pathogenic bacterium Pseudomonassyringae DC 3000 results on a significant reduction of symptoms (i.e., reduction of necrosis andpreservation of pigment content). Our findings demonstrate that plant-derived HPA is able toactivatetheplantimmunesystemandimproveresistanceagainstpathogenicbacterialinfection.Keywords: Arabidopsis thaliana, chemical screen, holaphyllamine, plant defense, Pseudomonassyringae,steroidalalkaloids

Session6Poster162The effect of fungicides and plant growth regulators application on the severity of Phoma stemcankerandseedyieldofwinteroilseedrapeNazaninZamani-Noor11JuliusKühn-Institut,InstituteforPlantProtectioninFieldCropsandGrassland,Messeweg11-12,D-38104Braunschweig,GermanyAtpresentstudy,multifactorialfieldexperimentshavebeenconductedoverthreegrowingseasons(2013 to2015) toevaluate theeffectof five fungicidesandplantgrowth regulatorsapplicationongrowthparameters,yieldparametersandseedyieldof fourdifferentwinteroilseedrapevarieties.CultivarsElektra,Genie,PR46W20andVitarawerechosenbecauseoftheirdifferencesinresistanceto winter hardiness, lodging and blackleg disease caused by Leptosphaeria maculans. Theexperimental design was a completely randomized block design, comprising four replications foreachplant varietyper treatment for each growing season. Fungicides andplant growth regulators(Ampera: prochloraz and tebuconazole; Carax:mepiquat andmetconazole; Folicur: tebuconazole;Tilmor: tebuconazole and prothioconazole and amixture of Imbrex plus Folicur: fluxapyroxadplustebuconazole)wereappliedtwice;inautumn(BBCH14-18)andinspring(BBCH30-55).Resultsshowedthat fungicideandplantgrowthapplicationaffectedoilseedrapegrowthandyieldparameters during growing season, but effect depended onmeteorological conditions in researchyear,aswellasusedoilseedrapevarieties.Plantgrowthregulatorshadsignificanteffectsonplantheightandwinterkilling(%).PlotstreatedwithCaraxhadtheshortestheightandplotstreatedwithFolicur had the lowestmortality rate of plants afterwinter. Significant differences in L.maculans-diseaseseveritywereobservedamongtreatmentandthevarieties.Diseaseseveritywasthehighestinoilseedrapecv.PR46W20.Thefungicidemixture ImbrexplusFolicurdecreasedsignificantlythestem canker development on stem bases than that of those from untreated control and othertreatments. Thousand grain weight (g) and yield (t/ha) of oilseed rape varieties were improvedsignificantlywhenTilmororamixtureofImbrexplusFolicurwereapplied.Keywords: Brassica napus, winter hardiness, lodging, Leptosphaeria maculans, disease severity,winterhardiness

SessionSMARTBIOCONTROLPoster163Bioinformatics tools dedicated to secondary metabolite analysis: contribution to the screeningplatformofBioscreenInterregVaprojectMickael Chevalier1, Qassim Esmaeel1,2, Maude Pupin3,4, Essaid Aït Barka2, Philippe Jacques1,5 andValérieLeclère1

1Univ.Lille,INRA,ISA,Univ.Artois,Univ.LittoralCôted’Opale,EA7394-ICV-InstitutCharlesViollette,F-59000Lille,France2LaboratoiredeStress,DéfensesetReproductiondesPlantesURVVC-EA4707,UFRSciencesExactesetNaturelles,UniversityofReims-Champagne-Ardenne,Reims,France3Univ.Lille,CNRS,CentraleLille,UMR9189-CRIStAL-CentredeRechercheenInformatiqueSignaletAutomatiquedeLille,F-59000Lille,France4Inria-LilleNordEurope,Bonsaiteam,F-59655Villeneuved’AscqCedex,France5TERRA Research Centre, Microbial Processes and Interactions (MiPI), Gembloux Agro-Bio TechUniversityofLiege,B-5030Gembloux,BelgiumBioscreen is one of the fifth constitutive projects of the SmartBioControl portfolio, granted byInterreg Va program. The aim of this project is to build a platform to enhance the screening forbiocontrol agents. The bioinformatics approach will be presented, describing the diverse toolsdeveloped by University of Lille-Sciences and Technologies partner, dedicated to nonribosomalanalysis. Their use topredict bioproductionof secondarymetaboliteswithbiocontrol applications,andtheircontributiontotheefficientscreeningwithintheplatformwillbeexplained,supportedbyrelevantexamples.Keywords:bioinformatics,Norine,Interreg,SmartBioControl,bioscreen

SessionSMARTBIOCONTROLPoster164BioProtect; Biological crop protection in practice: optimization of the efficacy of biological cropprotectionproductsin(field)trialsJennyNeukermans1,BernardBodson2,NathalieCap1,SofieDarwich3,StephanDeclerck4,AbdelnasserElghazouani5, Monica Höfte6, Mark Lateur7, Sandrine Oste8, Philippe Reignault9, Ali Siah10 andDominiqueWerbrouck11

1 PCG - ProvinciaalProefcentrumvoor de Groenteteelt Oost-Vlaanderen vzw, Karreweg 6, 9770,Kruishoutem,Belgium.2ULG-UniversitédeLiège–GemblouxAgro-BioTech,PassagedesDéportés2,5030Gembloux,Belgium.3Inagrovzw,Ieperseweg87,8800Roeselare,Belgium4UCL - Université catholique de Louvain, place de l’université 1, 1348 Louvain-La-Neuve, Belgium.5Lipofabrik, Bâtiment Polytech, Avenue Paul Langevin, 59655 Villeneuve d’Ascq, France. 6GhentUniversity,LaboratotyofPhytopathology,DepartmentofCropProtection,CoupureLinks653,9000,Ghent, Belgium. 7CRA-W - Centre Wallon de Recherches Agronomiques, Rue de Liroux 4, 5030Gembloux,Belgium.8FREDON–FédérationRégionaldeDéfensecontrelesOrganismesNuisibles,rueBecquerel 265 BP74, 62750 Loos-en-Gohelle, France. 9Université du Littoral Côte d’Opale (ULCO),Unité de Chimie Environnementale et Interactions sur le Vivant (UCEIV), CS 80699, 62228 Calaiscedex, France. 10Institut Supérieur d’Agriculture, Boulevard Vauban 48, 59046 Lille Cedex, France.11Pôle Légumes Région Nord, Route d’Estaires 209, 62840 Lorgies, France & CAR – Chambred’AgriculturedeRégionNordPas-de-Calais,BoulevarddelaLiberté140,59013LilleCedex,FranceTheuseofbiocontrolproductstocombatortocontrolpestsanddiseasesplaysanimportantroleinboth organic and conventional farming. The InterregV project BioProtect which is part of SmartBiocontrol, the European Platform of research and development of tools for biocontrol ofphytopathogens, contributes to this. Currently, biocontrol products often provide variable efficacyresultsorinsufficientefficacyincomparisontochemicalpesticides.Forthisreason,growersarenotconvincedtousebiopesticides.Therefore,theprojectaimstogainmoreinsightinthemodeofactionofsomebiocontrolproductsthroughaliteraturestudy,anagriculturalpracticesurveyand(field)trials.Experimentsinthefieldaswell as small scale tests under more controlled conditions will be conducted focusing onpathosystemsthatareofgreatimportanceforthecrossborderregionFrance-Wallonia-Flanders.Notonly recognized biological crop protection products, but also new biocontrol products that arecurrentlyunderdevelopment,willbeincluded.Afterdeterminationoftheoptimalapplicationtiming,applicationmethod and application techniques, this knowledgewill be transferred to the farmersandgrowersonthebasisofdemonstrationplatforms.Keywords:biocontrolproducts,biologicalcropprotection,diseasemanagement,smartbiocontrol

SessionSMARTBIOCONTROLPoster165Alternative methods for crops protection: interest on natural substances, microorganisms andplantsS. Oste1, J. Bruyere1, L. Culiez1, T. Delannoy1, L. Durlin1, K. Leleu1, K. Petit1, A. Quennesson1, S.Quennesson1,L.Tournant11Fédération Régionale de Défense contre les Organismes Nuisibles (Regional Federation for pestcontrol)NordPas-de-Calais21et265,RueBecquerel-BP74-62750Loos-en-Gohelle-FranceFREDONNordPas-de-Calaisknownas“OrganismeàVocationSanitaire(OVS)/PlantHealthOrganisation”inplantproductionisspecialistincropprotection.Withinofheractivitiesof“Researchanddevelopment”,FREDONdevelopsandsharesaboutalternativemethodssuchasnaturalsubstancesandmicroorganismshavingrepulsiveeffectorstraightinfluenceonbioagresseurstomajor crops in North of France. Also, FREDON evaluate different plants producing volatileorganiccompounds(VOC)witharepulsiveeffectoninsects.Somereferenceswerelearnedabouttheseparticulartargets:-Chicory/wireworms,-Chrysanthemum/thrips,-Wheat/commonbunt,-Leek/flyandthrips,-Potatoes/lateblight,wireworms,-Appletree/applescab,applesawfly,appleweevil,aphids,-Speedwell/fusariumwilt.Naturalsubstancesthathavebeenevaluatedincontrolledconditions(inClinicoftheplant(CliniqueduVégétal®)atFREDON)and/oronfieldtrialsand/oronfieldproductionsare:essentialsoils, lipopeptides, basic substances and plants extract such as, for example, decoction andinfusion.Microorganismstestaremadewithentomopathogenicfungusantagonistorstimulatingofplanthealth.StartingnewprojectlikeBioprotect(withintheprojectportfolioSMARTBIOCONTROL)giveusprospects for improve evaluation skills with Interreg V Flanders and Wallonia cross-bordercollaboration.Keywords:naturalsubstances,microorganisms,biocontrol,crops,protection

SessionSMARTBIOCONTROLPoster166BIOPROD FWVL Interreg Project: New strategies for production and formulation of low toxicitybiopesticidesF.Krier1,F.Coutte1,D.Lecouturier1,A.Rochex1,R.Ravallec1andP.Dhulster11UniversitéLille,INRA,ISA,Universitéd’Artois,UniversitéLittoralCôted’Opale,EA7394-ICVInstitutCharlesViollette,F-59000Lille,FranceInthecontextofmorerespectfulutilisationofpesticides intheenvironment, lipopeptidesareverypromisingmolecules.TheprincipalobjectivesoftheFWVLInterregBIOPRODprojectaretoremovethe constraints related to the industrial production of these new bio-pesticides and themicroorganismsthatproducethem.Themulti-disciplinaryandcollaborativeworkswillbeconductedon both sides of the border by operators specialized in their respective fields. Thus, the actionsconductedinthisprojectwillbe:•Optimisationofproductionandpurificationconditionsofmoleculesbydeveloping,amongothers,newinnovativeproceduresimplementinghigh-throughputscreeningmethods•Sizingandscalingupof installations toachieve future industrialproductionof lipopeptidesThesetwotechnicalapproacheswillcontributetodeterminationofacostforeachmolecule,whichwillbeoneofthefundamentalsforthemarketanalysishereaftermentioned.•Detailed studies of biodegradability and toxicity of molecules on different models in order todemonstratethepositiveimpactofthesenewbio-pesticidescomparedwiththechemicalscurrentlyusedinagriculture.•Realisationofmanyformulationstriesinordertogetthesemoleculescommercialized,morestable,moreactiveandeasierinutilization.Thesedifferentactionswillbecomplementedbyacross-bordermarketstudytodeterminethepracticesandexpectationsofphytosanitarydistributorsandfarmers.The results will be compared in order to have a territorially adapted communication aimed atpromotingthesenewphytosanitarymolecules.Theultimateresultof theprojectbeingtheplacingonthemarket inaclosefutureoftheseproductsandthus improvetheprotectionofenvironmentthroughtheuseofrespectfulnewbio-pesticidesthataremorerespectfulfornatureandhuman.Keywords:biopesticides,highthroughputscreeningmethods,innovativeprocess

SessionSMARTBIOCONTROLPoster167Fungiasbiocontrolagents:screeningstrategytowardsvariousplantpathogens-promisingresultsonpotatoplantschallengedbyPhytophthorainfestansArnaudSipel1,MarylineCalonne-Salmon1,StéphaneDeclerck1

1UniversitécatholiquedeLouvain,EarthandLifeInstitute,Mycology,CroixduSud,2boxL7.05.06,B-1348Louvain-la-Neuve,BelgiumPotato plants are facing numerous pathogens. Among these isPhytophthora infestans, the causalagent of late blight. Currently, chemical treatments are themost recommended strategies for thecontrol of this disease, despite their detrimental impacts on environment and human health. Thedevelopmentofalternativestrategies isthusbecomingessential forreducing/controllingpesticidesapplications.Inthatcontext,theuseoffungiandfungalbio-effectorsrepresentsapromisingstrategytocontrol,at least partially, below and above-ground pathogens, and the screening of collections opens theperspectiveofdiscoveringnewenvironmental-friendlyactivesmolecules.ThemainobjectiveofourresearchwastodevelopanefficientscreeningmethodtoidentifyefficientfungalstrainsforthebiocontrolofP.infestansandalternativelyofotherfungaldiseases.One hundred fifty fungal strains were first selected from the literature for their potential asbiocontrol agent. These strainswereprovidedby theMycothèqueof theUniversité catholiquedeLouvain(BCCM/MUCL),oneoftheworldlargestcollectionoffungiofagro-foodandenvironmentalinterest.A96-wellmicroplateinvitrotestwasusedtoscreencrudeextractsofthefungalcollection.Sixty-nineorganisms(i.e.crudeextracts)displayedastronginhibitoryonthegrowthofP.infestans.Their potential activity was then evaluated in vitro on potato plantlets grown in microboxes.Seventeen strains showed a strong activity against P. infestans. These strains mostly belong toAspergillus, Chaetomium, Fusarium, Hyphomyces, Penicillium and Trichoderma species. Furtherisolation,purificationandcharacterizationoftheactivemoleculesareon-going.This screening strategy allows the fast track identification of fungi as well as fungal bio-effectorseffectiveagainstP. infestansandwillbeextendedtoothermajorrootandshootpathogens in theSMARTBIOCONTROLproject.Keywords:fungi,screening,biocontrolagents,Phytophthorainfestans,potatolateblight,microplateinvitrotest

Session6Poster167MosaicsofplantdiseaseresistancegenesareamoreversatilemeansofachievingdiseasecontrolthanpyramidsinmostagriculturallandscapesRamsesDjidjou-Demasse1,BenoîtMoury2andFrédéricFabre11IRDINRA,UMR1065,F-33882Villenaved'Ornon,France2INRA,UR407,PathologieVégétale,F-84140Montfavet,France

Thebreakdownofplantvirusresistancegenesisamajorissueinagriculture.Weinvestigated

whether a set of resistance genes would last longer when stacked into a single plant cultivar(pyramiding)orwhendeployedindividuallyinregionalmosaics(mosaicstrategy).

Wemodeledthegeneticandepidemiologicalprocessesshapingthedemo-geneticdynamicsof viruses under a multi-locus gene-for-gene system, from the plant to landscape scales. Thelandscape consisted ofmany fields, was subject to seasonality, and of a reservoir hosting virusesyear-round.

Strategy performance depended principally on the fitness costs of adaptive mutations,epidemic intensitybeforeresistancedeploymentandlandscapeconnectivity.Mosaicswereat leastas good as pyramiding strategies in most production situations tested. Pyramiding strategiesperformed better only with slowly changing virus reservoir dynamics. Mosaics are more versatilethanpyramidingstrategies,andwefoundthatdeployingamosaicofthreetofiveresistancegenesgenerallyprovidedeffectivediseasecontrol,unlesstheepidemicsweredrivenmostlybywithin-fieldinfections.

We considered the epidemiological and evolutionary mechanisms underlying the greaterversatilityofmosaicsinourcasestudy,withaviewtoprovidingbreedersandgrowerswithguidanceastothemostappropriatedeploymentstrategy.

Keywords: Durable disease resistance, Pyramids of plant resistance, Mosaic of plant resistance,Landscapeepidemiology,Regionaldeploymentstrategy