University of Nottingham

11-13 September 2017

BSPP & BSM

FungalControl &

Exploitation

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Contents    

Page    2………………………………………………..President’s  welcome    4………………………………………………..Programme      10………………………………………………..Awards    12……………………………………………..  Opening  plenary  talk  abstracts    15……………………………………………..  BMS  talk  abstracts  –  Exploitation  1  (Tuesday  12th)    19……………………………………………...BMS  talk  abstracts  –  Control  1  (Tuesday  12th)    23………………………………………………BMS  talk  abstracts  –  Exploitation  2  (Wednesday  13th)    26………………………………………………BMS  talk  abstracts  –  Control  2  (Wednesday  13th)    28………………………………………………BSPP  talk  abstracts  (Tuesday  12th)    38………………………………………………BSPP  talk  abstracts  (Wednesday  13th)    44………………………………………………Closing  plenary  talk  abstracts    46………………………………………………Poster  abstracts    82………………………………………………Date  of  next  BSPP  presidential  meeting    

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BritishMycologicalSocietyandBritishSocietyforPlantPathologyJointPresidentialMeeting2017

FungalExploitationandFungalControl

UniversityofNottingham

11th–13thSeptember2017

Presidents’WelcomeOnbehalfoftheBritishSocietyforPlantPathologyandtheBritishMycologicalSociety,itisourpleasuretowelcomeyoutothisjointannualmeetinginNottinghamUniversity.Despitethefactthatthetwosocietiessharealonghistoryandhavemuchincommon,asfarastheOrganisingCommitteeisaware,thisisthefirsttimeajointmeetinghasbeenheld.Thethemesofthisconferencearethecontrolofplantpathogens(mainlyfungi)andtheexploitationoffungalspecies.Plantpathogenscontinuetocausesubstantiallossestofarmersinallpartsoftheworld.Themainmethodsusedtocontroldiseasesaregeneticdiseaseresistanceincropcultivars,fungicides,biologicalcontrolagentsandculturalpractices.Thisconferencewillhighlightsignificantadvancesinthefirsttwomethods.Cultivarswithimproveddiseaseresistance,whichcombineseffectivenesswithlongevity,arebeingreleasedthroughpartnershipsbetweenacademicscientistsandplantbreedingentities.Newfungicidesarebeingdevelopedandbettermethodstodelayandreducetheimpactoffungicideresistancearebeingdeployed.Aparticularlypleasingaspectofthisworkisthatitisnowbecomingclearthatthelongevityoffungicidesisextendedbygoodgeneticdiseaseresistanceandviceversa;fungicidesprotectthelongevityofgeneticdiseasegenes.AremainingfrustrationisthatGMdiseaseresistancehasnotbeenwidelydeployedevenaswepass20yearssincethefirstcloningofR-genes.Somefungiandoomycetescanalsoinfectanimalsandcreateseriousproblems,oftenresultinginfatalities.Newmethodstodetectsuchpathogensatanearlystagewouldresultinbettertreatmentpractisesandbettersurvivalrates.Inmanyanimal–fungalinteractions,basicbiologyquestionssuchas“howdoesthepathogeninfect?”and“whatdoesthehosttodefenditself?”cannotbeansweredatpresent.Forsomeanimaldiseases,controloptionsareverylimitedandalternativemethodsareurgentlyneeded.Besidesbeingpests,fungicanalsobeverybeneficialtous.Obviously,weliketoeatsomeofthem,butwecanalsomanipulatethemandusethemforbiotechnologicalpurposestodevelopmedicines,forexample;oralternatively,usethemforbiomassdegradation,insectorfungalbiocontrolandenhancingproductivitythroughbetterunderstandingofsymbiosisinmycorrhizalinteractions.

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Itpromisestobeanexcitingannualmeetingandwewouldliketothanktheorganisingteamfrombothsocieties:PaulDyer,SimonAvery,MatthiasBrockandElizabethOrton,forputtingtogethersuchastimulatingprogramme.WealsothankAlisonTorfortheabstractbookletandMoiraHartwhoassistedwiththeorganisation.WewishyouafruitfulandenjoyabletimeinNottingham.RichardOliver,PietervanWestandNickRead(formerBMSpresident)TheBSPPandBMSaregratefulforthesupportfromApolloScientific,CABI,ElsevierandWiley.

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Monday  11th  September    

 13.00   Lunch  and  registration  14.00    Welcome  14.15   Plenary  -­‐  BSPP  President  Richard  Oliver     Translating  understanding  of  plant  pathogen  control  genetics  into  

grower  profits      15.00   Plenary  -­‐  BMS  President  Pieter  van  West     Invasive  oomycetes  are  a  serious  threat  to  our  natural  and  man-­‐made  

aquatic  ecosystems      15.45   Coffee        16.15    Plenary  -­‐  Richard  Michelmore     The  impact  of  new  technologies  on  strategies  to  increase  the  

durability  of  disease  resistance      17.00   Plenary    -­‐  Mark  Lynas     Anti-­‐science  and  post-­‐truth:  why  we  got  it  so  wrong  about  GMOs      17.45   Poster  session  and  drinks  reception  (sponsored  by  Elsevier)     18.15  BSPP  AGM  Small  lecture  theatre  19.30   Dinner                

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Tuesday12thSeptember

ConcurrentBMS(Exploitation1) ConcurrentBSPP

9.00 BMSInvited-VeraMeyer BSPPInvited-BruceMcDonald Movingfromdescriptivetopredictivebiology:Rationalrewiringof

thecellfactoryAspergillusnigerUsingpopulationgenomicstodeterminethegeneticbasisofpathogenadaptationinagro-ecosystems

9.30 BMSInvited-ScottBaker BSPPInvited-LiseNistrupJorgensen Fungiforbiotechnology WhydoesIPMnotleadtomorereductionsinfungicides? 10.00 MuhammadAkbar:Anovel inoculationmethodofmycorrhizae in

wheat fields has remarkable effects on crop yield and soilproperties

VanessaMcMillan:Identifyingrootresistanceintake-alldisease

10.20 ElenaGeib:AspergillusnigerversusAspergillusoryzae:Expressionplatformsforheterologoussecondarymetaboliteproduction

YongjuHuang:Ignoreduntilrecently-thestoryofthefungalpathogenLeptosphaeriabiglobosa

10.40 Coffee Coffee11.10 BMSInvited-EckhardThines PHGregoryStudentCompetitionTalks

UnravellingpyriculolbiosynthesisanditsregulationinthericeblastfungusMagnaportheoryzae

1)CorinneArnold(JIC):PowderyMildew:ataleoftwofungicides

2)ShaoliDasGupta(RHUL):Blumeriametallo-proteaselikeeffector;Azincophoreactingasauniversalvirulencefactorinfungi?

11.40 KristiinaHildén:Biochemicalcharacterizationofmannan-acting 3)KathrynHales(Warwick):Understandingtheecologyand

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enzymesfromthewhite-rotfungusDichomitussqualens epidemiologyofPythiumviolaetoenablediseasemanagementincarrotcrops.

4)SaschaJenkins(Warwick):Examiningidentity,phylogenyandpathogenicityfactorsinFusariumspeciesaffectingpea.

12.00 MichaelaNovodvorska:PigmentbiosynthesisinPenicilliumroqueforti,whybeblue?

5)ChinthaniKarandeniDewage(Hertfordshire):FinemappingofamajorgenelocusforresistanceagainstPyrenopezizabrassicae(lightleafspot)inBrassicanapusandidentificationofcandidateresistancegenes

12.20

Lunch

6)JosephMoughan(Rothamsted):Fromthefieldtothelabandbackagain:exploringlegacyeffectsofrootgeneticsandfungicidefortake-allrootdiseasecontrolinwheatrotations.

13.30 Elsevierauthorworkshop ConcurrentBMS(Control1) ConcurrentBSPP

14.30

BMSInvited-AxelBrakhage BSPPInvited-VivianneVleeshouwers

Melanin-dependentpathogenicityofAspergillusfumigatus ExploitingeffectorsinbreedinganddeploymentofRgenesinpotato

15.00 JanetQuinn:FungalSAPKpathwaysasmediatorsofvirulence FranziskaTrusch:TheRxLRMotifoftheHostTargetingeffector

AVR3aofPhytophthorainfestansiscleavedbeforesecretion

15.20 CindyVallieres:Synergisticcombinationstargetingtranslationfidelityanddiversefungalpathogens.

SarahSchmidt:Knowyourenemy-engineeringresistanceagainstFusariumwiltinbanana.

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15.40 Coffee Coffee 16.10

BMSInvited-AlBrown

BSPPInvited-Karl-HeinzKogel

Candidaalbicansisamovingtargetfortheimmunesystemasitadaptstohostsignals

TheAgronomicPotentialofGeneSilencingApplication

16.40 Rhys Farrer: Targeting infection mechanisms for two amphibian-

infectingchytridspecies.LaurenceBindschedler:Combiningproteomicsofhaustoriaandgenesilencingtodiscoverkeyplayersofvirulenceduringbarleypowderymildewinfection

17.00 Posterssessionanddrinksreception(sponsoredbyWiley)

BMSAGMSmalllecturetheatre

19.30 Conferencedinner

Wednesday13thSeptember

ConcurrentBMS(Exploitation2) ConcurrentBSPP9.00 BMSInvited-RonalddeVries BSPPInvited-SanderSchouten Diversityinplantbiomassdegradationapproachesoffungi Directandindirecteffectsoffungalendophytesonplant-parasitic

nematodes 9.30 IolyKotta-Loizou:Mycovirusesasprospectiveenhancersof

biocontrolagentsChristianVoigt:Improvingplantresistancetofungalpathogensbycallosemodification

9.50

Stefany Solano:Genomic and genetic analysis of a basidiomyceteyeastfocusedonthecharacterisationofMELbiosynthesis

AndrewTaylor:UnderstandingthegeneticcontrolofpathogenicityandresistanceforFusariumoxysporuminOnion

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10.10 Sara Casado López: Intraspecies diversity and induction of thewooddecaybythewhiterotfungusDichomitussqualens

RyanAmes:UnderstandingappressoriumdevelopmentinMagnaportheoryzae

10.30 Coffee Coffee Concurrent.BMS(Control2) ConcurrentBSPP

11.00 BMSinvited-AlanGange BSPPInvited-FrankVandenBosch Exploitingunspecialisedendophytesforbiologicalcontrolofinsects

andplantsIntegrateddeploymentofhostresistanceandfungicides:managingevolutionofvirulenceandinsensitivity

11.30 BMSInvited-MikeBromley BSPPInvited-NeilPaveley RegulationofDrugResistanceinAspergillusfumigatus Integrateddeploymentofhostresistanceandfungicides:

economics,efficacyandevolution

12.00 TakanoriFurukawa:SystematicfunctionalannotationoftranscriptionfactorsinthemajormouldpathogenAspergillusfumigatus

AndrewLeader:Fenpicoxamid(InateqTM,activefungicide),anewfungicidewithutilityincerealsforcontrolofZymoseptoriatritici(Septorialeafblotch)andotherdiseases

12.20

Endofsession

AviceHall:ControllingStrawberrypowderymildewsrequiresallyear-roundvigilance

12.40 Lunch

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PlenarySessionandClose

13.30 Plenary-RobJohnson TheScienceofQuorn

14.00 Plenary-NataliaRequena Fungalweaponstomanipulateplants

14.45 PrizesandClose

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Awards  The  Boards  of  the  BSPP  and  BMS  wish  to  encourage  young  plant  pathologists  and  mycologists  to  present  their  work.  There  are  two  BSPP  prizes  and  one  BMS  prize  awarded  at  the  presidential  meeting:      The  P  H  Gregory  prize,  awarded  for  the  presentation  of  an  oral  paper.  The  competition  is  open  to  (a)  members  of  BSPP  who  have  not  previously  presented  a  paper  to  a  meeting  of  a  learned  society  and  (b)  all  registered  postgraduate  students,  whether  or  not  they  are  members  of  the  society,  or  have  presented  a  paper  before  to  a  meeting  of  a  learned  society.  Contestants  should  not  have  entered  the  P  H  Gregory  competition  previously.  The  winner  receives  a  certificate  and  a  cheque  for  £250.    Philip  Gregory  (1907–1986)  pioneered  aerobiology  as  a  topic  for  research,  combining  many  disciplines  to  contribute  to  better  understanding  of  fungal  spore  dispersal  and  plant  disease  epidemiology.  He  developed  theories  of  spore  dispersal,  published  in  his  classic  paper  on  the  dispersion  of  airborne  spores  (Gregory  PH.  (1945)  Trans.  Br.  Mycol.  Soc.  28:  26–72).  He  became  Head  of  the  Plant  Pathology  Department  at  Rothamsted  in  1958  where  he  further  developed  his  research  on  spore  dispersal  and  sedimentation  velocities.  In  his  retirement,  Philip  Gregory  continued  work  on  elucidating  the  epidemiology  of  black  pod  disease  of  cocoa  in  Nigeria,  and  maintained  enthusiasm  for  his  wide  range  of  scientific  interests.  He  proudly  and  enthusiastically  showed  his  garden  to  visitors  and  they  often  regarded  this  as  the  highlight  of  their  visit  to  Harpenden  (Source:  Lacey  et  al.  (1997)  Ann.  Rev.  Phytopathol.  35:  1—14).      The  John  Colhoun  Prize,  awarded  for  a  poster.  The  work  presented  in  the  poster  must  form  part  of  a  research  project  conducted  by  the  entrant  in  support  of  a  PhD  or  Masters  degree,  and  the  entrant  must  not  have  been  awarded  the  degree  prior  to  the  deadline  for  abstracts  date.  Students  need  not  be  members  of  BSPP.  The  winner  receives  a  certificate  and  a  cheque  for  £100.    John  Colhoun  (1913–2002):  cryptogamist  and  plant  pathologist.  John  Colhoun  was  awarded  a  MAgr  in  1937  at  Queens  University,  Belfast,  and  then  moved  to  Imperial  College,  London,  working  on  fungal  pathogens  of  apples  for  his  PhD,  awarded  in  1940.  He  returned  to  Northern  Ireland  where  he  worked  on  flax,  leading  to  a  definitive  text  (Muskett  A.  E.  &  Colhoun  J.  (1947):  The  Diseases  of  the  Flax  Plant).  He  became  reader  at  Queen’s  University  in  1954.  Subsequently,  he  took  up  the  Chair  of  Cryptogamic  Botany  at  the  University  of  Manchester  in  1960,  where  he  worked  on  Fusaria,  Phytophthora,  Septoria  and  Phoma,  with  hosts  ranging  from  cereals  to  chrysanthemum,  yam,  oil  palm,  and  banana.  In  1968,  he  was  elected  Chairman  of  the  Federation  of  British  Plant  Pathologists,  forerunner  of  the  British  Society  for  Plant  Pathology.  He  retired  from  Manchester  University  in  1980  as  Professor  Emeritus,  having  occupied  the  Barker  Chair  of  Cryptogamic  Botany  for  20  years.  (Source:  Epton  H.  (2003)  Mycol.  Res.  107:  377–381).      The  BMS  Student  Poster  Prize:  Awarded  to  the  best  mycology  student  poster  at  the  meeting.  

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TALKS

*PHGregoryCompetitionentrant

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OpeningPlenaryTalksTranslatingunderstandingofplantpathogencontrolgeneticsintogrowerprofitsRichardP.OliverCentreforCropandDiseaseManagement,CurtinUniversity,Perth,WA,AustraliaEmail:Richard.oliver@curtin.edu.auPlantfungalpathogenscausemajoravoidablelossestocropinallpartsoftheworld.Reliabledataonthedegreeoflossesisrare.ThatAustralia,withitslowinputsandfrequentdroughts,isthecountrywiththehighestdocumentedoverallpercentagelossesinbroadacreagricultureiscertainlyduetothepaucityofdatafromotherregions.ThemajordiseasesofgraincropsinAustraliathattheCCDMworksonarecausedbyarangeofnon-obligatefungi;thewheatdiseasestanspotcausedbyPyrenophoratritici-repentis,andseptorianodorumblotch(SNB)causedbyParastagonosporanodorum,thebarleynetblotchdiseasescausedbyPyrenophorateres,thelegumeleafblightdiseasescausedbyvariousAscochytaspp.andthecanoladiseasecausedSclerotiniasclerotiorum.DiseasescausedbyrustshavebeenreasonablywellcontrolledbythedeploymentofmajorR-genes.Suchmajorgeneswerenotavailablefortheseblightdiseases.Thestrategydeployedtocontrolthemhadtoawaitthedevelopmentofsomeunderstandingabouthowthepathogensinteractwiththehost.AcollaborativeefforthasshownthatatleastfortanspotandSNB,thekeywastodeterminethatthediseaseswerecontrolledbytheinteractionofpathogengeneproductsnamednecrotrophiceffectors(NEs).NEsinteractwithcorrespondinghostsensitivitygenes.ThekeytoimprovingdiseaseresistancewastotransferthisknowledgetobreederssothattheycouldeliminategermplasmcarryingrelevantNEsensitivitygenes.FungicidesremainthebackstopoffungaldiseasecontroleveninlowintensitycountrieslikeAustraliaandespeciallysince2005whenpricesdropped.FungicideresistancewassoondiscoveredinmanypathogensinAustralia.Thepatternoffungicideresistanceconfirmedthedevelopingunderstandingthatfungicideresistancecanbedelayedbyapplicationofcoreevolutionaryprinciplesthatguideustouseminimumdoses,minimumnumbersofapplicationsandmixturesoralternationsofmodesofaction.Itisalsobecomingclearprudentuseoffungicidesextendsthelifeofgeneticdiseaseresistanceandviceversa.

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Invasive oomycetes are a serious threat to our natural and man-made aquaticecosystemsPietervanWestAberdeen Oomycete Laboratory, International Centre for Aquaculture Research andDevelopment at the University of Aberdeen, College of Life Sciences and Medicine,InstituteofMedicalSciences,Foresterhill,Aberdeen,AB252ZD,Scotland,UK.Email:p.vanwest@abdn.ac.ukOomycetesarefungal-likeorganismsthatareclassifiedasChromalveolatesandphylogeneticallygroupedwithdiatomsandbrownalgae.Theyareamongthemostimportantgroupsofdisease-causingorganismsinbothagricultureandaquacultureandthusrepresentahugethreatforglobalfoodsecurity.Someaquaticspeciescancauseseriousenvironmentaldisasters,wipingoutournativeaquaticanimals,forexampleEuropeancrayfishandalsoseveralamphibianshavebeenseverelyaffectedorhavebecomeextinct.Oneparticularpathogen,Aphanomycesinvadans,isnowapproachingEuropeanwatersandrepresentsadevastatingpathogen.Itisatremendousproblemincountrieswhereithasalreadyarrivedandhasestablished(e.g.Bangladesh,IndiaandpartsofAfrica).TherearealsootheraquaticoomycetepathogensknownincludingHaliphthoros,Halioticida,Lagenidium,Atkinsiellaspp.thatinfectmarineandbrackishanimalsincludinglobsters,langoustines,abaloneandprawns.Saprolegniaparasiticaisoneofthemostdestructivefishpathogensandisfoundinmostfreshwaterenvironmentsaroundtheworld.Remarkably,verylittleisactuallyknownaboutthebiologyoftheseaquaticpathogens.However,whatwedoknowisthattheyare:1)potentialinvadersofmarine&freshwaterhabitats,2)theycancauseseriouseconomicorenvironmentaldamageandmostimportantly,3)theyarealluncontrollableatpresent.Anoverviewofthesevariousanimalpathogenicoomycetesispresented.Furthermore,ourcurrentknowledgeaboutthecellularandmolecularinfectionstrategiesofS.parasiticaandA.invadansisbeingdiscussed.Inordertosuccessfullyinfecttheirhoststheseparasitesrelyontheireffectorrepertoires.Effectorsaresecretedproteinsofthepathogenthatcanalterthehostandwhichadvancetheinfectionprocess.Alargenumberofeffectorsislocatedattheinterfacebetweenpathogenanditshostandfulfilafunctionontheoutsideofthehostcell.Sucheffectorsareoftenclassifiedasextracellulareffectors.Whileseveralotheroomyceteeffectorsareabletotranslocateintotheirhostcellswheretheycan,forexample,interferewithdefenceresponsesofthehost.Theseeffectorsarecommonlyreferredtoasintracellulareffectors.Themechanismoftranslocationordeliveryoftheintracellulareffectorsisunderintenseinvestigationandseveralroutesofentryhavebeenproposed.Herewewilldiscussourrecentadvancesinunderstandingsecretion,deliveryandfunctionsofeffectorsfromanimalpathogenicoomycetes.

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The impact of new technologies on strategies to increase the durability of diseaseresistance.RichardW.MichelmoreThe Genome Center & Department of Plant Sciences, University of California, Davis,Californa,95616,USA.Email:rwmichelmore@ucdavis.eduGreatadvanceshaverecentlybeenmadeintheunderstandingofthemolecularandgeneticbasisofdiseaseresistanceinplants.Itisnowtimetodeploythisknowledgetoprovidemoredurablediseaseresistance.Manyadvanceshavebeenenabledbytechnologicalimprovements.Inparticular,highthroughputDNAsequencingenablesdetailedanalysisofcropsandtheirpathogens.Itisnowpossibletocharacterizevariablepathogenpopulationsandusethisinformationfortherationaldeploymentofresistancegenessoastomaximizetheevolutionaryhurdleforthepathogentobecomevirulent.Muchofourworkoverthepastthirtyyearshasfocusedonresistancetodownymildewinlettuce.Lettuce(Lactucasativa)isoneofthemostvaluablevegetablecropsanddownymildew,causedbyBremialactucae,isitsmostimportantfoliardiseaseworldwide.Theuseofresistantvarietiesisthemosteffectivemethodforcontrollingthisdisease;however,pathogenvariabilityhasledtotherapiddefeatofindividualresistancegenes.Over50resistancegeneshavebeenidentifiedandlettucedownymildewisoneofthebestgeneticallycharacterizedplantdiseases.Wholegenomesequencingofmultiplegenotypeshasallowedtheidentificationofcandidateresistancegenesinthehostandvirulencefactorsinthepathogen.Knowledge-drivendeploymentofeffectiveresistancegenesasgenepyramidsprovidestheopportunityformoredurableresistance.Genestackingusinggenomeeditinghasthepotentialmakingthisprocessmoreefficient.Inaddition,host-inducedgenesilencingofvitalpathogengenespresentspotentiallyinsurmountableevolutionaryhurdlesforthepathogentoovercome.Anti-scienceandpost-truth:whywegotitsowrongwithGMOsMarkLynas@mark_lynasEmail:marklynas36@gmail.comWith“post-truth”theOxfordEnglishDictionarywordoftheyearfor2016,itseemsasifanewzeitgeistischallengingthewholebasisofevidence-basedthinkingandthescientificmethod.Buthownewisthisreally?MarkLynas,areformedformeranti-GMOactivist,tracessomeofthedeeperrootsoftoday’s“post-truth”attitudesintheenvironmentalmovement’srejectionofgeneticallymodifiedcropsandothertechnologies-likenuclearpower-thatwereseenasbeing‘unnatural’.

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BMSTalksMoving from descriptive to predictive biology: Rational rewiring of the cell factoryAspergillusnigerVeraMeyerDepartmentAppliedandMolecularMicrobiology,TechnischeUniversitätBerlin,Berlin,13355,GermanyEmail:vera.meyer@tu-berlin.deFungalbiotechnologyiscurrentlyundergoingarenaissancewithimportantimplicationsforitsroleasplatformtechnologyforthesustainableproductionofproducts,goodsanddrugs.AlliedtothisaretherecentadvancesinfungalSystemsandSyntheticBiologywhichcanbeseenastwocomplementaryapproachestoinvestigatethecomplexityofbiologicalsystemsincludingfungi.WhereasSystemsBiologyanalyzescellularsystemsinaniterativecycleofhigh-throughputgenerationof‘omics’dataandmodeling,SyntheticBiologytakesaconstructiveapproachtoreengineerbiologicalnetworksandtodesignnovelbiologicalpartsandcircuitswithnon-naturalfunction.Bothdisciplines–systemsanalysisanddesign–ideallycomplementeachotherand,notsurprisingly,aimtodiscoverthedesignprinciplesof(fungal)Life.OurgoalistounderstandandrationallyrewiretheindustrialcellfactoryAspergillusniger.Ontheonehand,wegenerateandevaluatedifferentomicsdatatypesforA.nigerandusetheseBigDatatopredictgenefunctionaswellasgeneregulatorynetworks.Ontheotherhand,weengineertitratableandtightlyregulatedconditionalmono-andpolycistronicgeneswitchesandtargetthemtospecificgenomicloci.Bycombiningthesetwoactivities,wenotonlysuccessfullyimproveA.nigerasproducerforproteins,newnaturalproductsorevennew-to-naturecompounds,wearealsoenteringanewerainAspergillusnigerbiology:wearemovingfromdescriptivetopredictivebiology.FungiforbiotechnologyScottE.Baker1,21Joint BioEnergy Institute, Emeryville, California; 2Environmental Molecular SciencesLaboratory,PacificNorthwestNationalLaboratory,Richland,Washington,USAEmail:scott.baker@pnnl.govFungihavebeenlongbeenexploitedforproductionofsmallmoleculesandenzymesforcenturies.Theunderlyingbiologyoffungalbioprocesses,however,isnotwellcharacterized.Usingavarietyofcellbiological,geneticandgenomicmethodswehaveassessedthebiologicalpathwaysunderlyingbothenzymeandsmallmoleculeproduction.Wehaveexploredenzymeproductioninthreephylogeneticallydiverse

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filamentousascomycetesfindingsharedgeneticdeterminantsofhighenzymeproduction.Inaddition,wehaveusedavarietyofcellbiologicalandgeneticmethodstobetterunderstandfactorsimportantforlipidproductionintheoleaginousyeast,Yarrowialipolytica.AnovelinoculationmethodofmycorrhizaeinwheatfieldshasremarkableeffectsoncropyieldandsoilpropertiesMuhammadAkbar1,SafeerA.Chohan1,MuhammadS.Iqbal1,NazirAslam1,AqeelAhmad2andTayyabaKhalil11DepartmentofBotany,UniversityofGujrat,Gujrat,50700,Pakistan;2InstituteofAgriculturalSciences,UniversityofthePunjab,Lahore,54000,Pakistanemail:makbarpu@gmail.comIn recent years there is growing trend towardsorganic farming tomeet food securityand food safety. Inorganic fertilizers are available to boost yield but there areenvironmentalandpublichealthconcerns.Amongstvariousorganicalternatives,usingmycorrhizal species is one of the most promising options. Therefore, the presentresearchwasconductedtoevaluatetheinfluenceofindigenousmycorrhizalspeciesonthe growthofwheat and soil properties under field conditions. In total 11 species ofmycorrhizaewere identified fromtheexperimentalareas, themostprominentgenerabeing,Claroideoglomus,RhizophagusandFunneliformis.Forinoculationofmycorrhizae,theirnativedensitywasmaintainedwithanovelideathatthesespeciesworkbetterinconsortia when their native population density was maintained. There were eightdifferent treatments having plot sizes as 6meters x 2meters. Thewhole experimentwasrepeatedattwodifferentsites.Bio-inoculationofconsortiaofdifferentmycorrhizalspeciesshowedasignificant increaseinallgrowthparametersstudiede.g.,numberoftillerperplant(upto39.53%),plantheight(upto13.66%),drybiomass(upto15.84%),grainyield(upto18.69%)andhayweight(upto15.69%).Moreover,remarkableeffectswereencounteredonsoilfertilitysuchassoilorganicmatter,availablephosphorusandpotassiumwere increasedup to92.30%,12.24%and8.83%, respectively.Thepresentstudyconcludesthatincreasingthemycorrhizalinoculumincropfieldsbykeepingtheirdensityasnativehastremendouseffectsoncropproductivityandsoilfertilitystatus.Aspergillus niger versus Aspergillus oryzae: Expression platforms for heterologoussecondarymetaboliteproductionElenaGeibandMatthiasBrockFungalGeneticsandBiology,UniversityofNottingham,Nottingham,NG72RD,UKEmail:mbxeg3@nottingham.ac.ukTo exploit fungal secondary metabolite production a strong and specific expressionsystemisrequired.WedevelopedsuchanovelexpressionsysteminAspergillusniger,

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which bases on regulatory elements from theAspergillus terreus terrein biosyntheticgenecluster.Weconfirmeditssuitabilitybyheterologousproductionofpolyketides(e.g.lecanoricacid),non-ribosomalpeptidesynthetase-likeproducts(e.g.aspulvinoneE)andusedittocharacterisetheAsp-melaninbiosynthesispathwayfromA.terreus.ThelatterstudiesledtoaninterestintheunderstandingofthechemistryofNRPS-likeenzymesthatproducemetaboliteswithantifungal,cytotoxic,antitumorigenicandantiviralactivity.WhiledifferentNRPS-likeenzymesmayacceptthesamesubstratetheresultingproductdependsonthebiochemistryofthethioesterasedomain.Tostudythesedomains,wecomparedtheaspulvinoneEsynthetaseMelAfromA.terreuswiththeatromentinsynthetaseInvA5fromPaxillusinvolutus.WhilerecombinantexpressionofmelAinA.nigerresultedinaspulvinoneEproduction,expressionofinvA5ledtoarangeofproductsthatwerealldistinctfromatromentin.WethereforespeculatedthatthephysiologyofA.nigermightleadtoamodificationoftheInvA5-derivedmetabolite.Consequently,invA5expressionwasstudiedinthealternativehostAspergillusoryzaeand,indeed,atromentinwasproduced.Inconclusion,ourrecombinantexpressionsystemisperfectlysuitedforheterologousproductionofawiderangeofdifferentsecondarymetabolites.However,asthemetabolicphysiologyoffungidiffers,atleasttwodifferentexpressionplatformsshouldbeselectedwhenaiminginthecharacterisationofnovelsecondarymetabolitebiosynthesisgenes.Unravelling pyriculol biosynthesis and its regulation in the rice blast fungusMagnaportheoryzaeEckhardThines1,2andStefanJacob21Institute of Molecular Physiology, University of Mainz, D55128 Mainz, Germany;2InstítuteofBiotechnologyandDrugResearch(IBWF),D-67663Kaiserslautern,GermanyEmail:thines@uni-mainz.deFungalsecondarymetaboliteshaveinthepastbeenverysuccessfulleadstructuresforpharmaceuticaloragrochemicalapplications.Manyofthosecompoundshavebeendiscoveredinaxenicculturesinartificialmedia,whicharepoorsurrogatesformimickinganorganism’snaturalhabitat.Asaconsequenceonlyasmallnumberofgenespotentiallyinvolvedinthebiosynthesisofsecondarymetabolitesareexpressedundertheseconditions.Magnaportheoryzaeisthecausalagentofblast,themostdevastatingfungaldiseaseoncultivatedrice.Thefungusisknowntoproducesecondarymetabolitesinvolvedinplant/pathogeninteraction,suchaspyriculol/pyriculariol.Withinthegenome20genesencodingpolyketidesynthaseshavebeenidentified,whereasqPCRexperimentsrevealedthatonlyfourofthemaretranscribedinaxenicculture.Apartfromcultivationvariationapproachesmolecularbasedtechniqueshavebecomesuitablemethodsinordertoexploretheproductsofsilentsecondarymetabolitepathways.

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Inordertoinvestigateregulatoryelementsofsecondarymetabolismandtoidentifynewnaturalproductsmanytechnologiescanbeapplied.Forthosestudiesgenedeletion/genedisruptionstrainsweregenerated,promoterswerechangedandtranscriptionfactorsorotherpleiotropicregulatorswereover-expressed.Furthermoreepigeneticstrategiesweresuccessfullyappliedinordertoelucidatenewsecondarymetabolites.TheapplicationofdemethylaseinhibitorsledtotheidentificationofnovelpolyketidesfromM.oryzaethathavenotbeenisolatedfromextractsofaxenicculturesbefore.Apartfromtheidentificationofnewsecondarymetabolitesandtheirpotentialroleinpathogenicitythestudieswillprovideinsightsintotheregulationofphytotoxinbiosynthesisinplant/pathogen-interactions.Biochemicalcharacterizationofmannan-actingenzymesfromthewhite-rotfungusDichomitussqualensAngelZhang1,JaanaKuuskeri1,JohannaRytioja2,RonalddeVries3,MiiaR.Mäkelä1andKristiinaHildén11DivisionofMicrobiologyandBiotechnology,DepartmentofFoodandEnvironmentalSciences,UniversityofHelsinki,Helsinki,Finland;2DepartmentofChemicalTechnologyandBiotechnology,AaltoUniversity,Espoo,Finland;3FungalPhysiology,WesterdijkFungalBiodiversityInstitute&UtrechtUniversity,Utrecht,TheNetherlandsEmail:kristiina.s.hilden@helsinki.fiWhite-rotfungihaveabilitytodecomposeallcomponentsofplantcellwallduetotheirwideenzymerepertoire.Theseenzymescouldbeusedforbioconversionofvariouslignocellulosesinindustrialapplications.Asatypicalwhite-rotfungus,Dichomitussqualenshasanextensivesetofgenesencodingenzymesforplantpolysaccharidedegradation.Thepreviousstudy[1]indicatedthatD.squalensexpressesandsecretesvarietyofmannanolyticgenes/enzymeswhengrowingondifferentplantbiomasses.Inthecurrentstudy,wecharacterizedfourmannan-actingenzymesderivedfromD.squalens.Inparticular,twoβ-1,4-endomannanases(GH5_7),oneβ-1,4-mannosidase(GH2)andoneα-1,4-galactosidase(GH27)wereclonedandexpressedinPichiapastoris.Theoptimaltemperaturesoftheexpressedrecombinantenzymeswereintherangeof50to60°CandoptimalpHfrom3to5.Themolecularmassofthecharacterizedproteinsrangedfrom47to110kDa.Thehighestthermaltolerancewasobservedfromβ-1,4-endomannanase(MAN1).MAN1retainedover50%ofitsenzymeactivityafter30minutesofincubationat70°C.Themeasuredenzymeactivitiesfromtwoofthecharacterizedenzymes,namelyMAN1andα-1,4-galactosidase,exhibitedhighenzymaticactivitiestowardsthetestedsubstrates.Overall,theresultssuggestthehighpotentialofD.squalensasasourceofindustrialenzymes.

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[1]J.Rytioja,K.Hildén,M.DiFalco,M.Zhou,M.V.Aguilar-Pontes,O.-M.Sietiö,A.Tsang,R.P.deVries,M.R.Mäkelä,Environ.Microbiol.(2017)doi:10.1111/1462-2920.13652PigmentbiosynthesisinPenicilliumroqueforti,whybeblue?MichaelaNovodvorska,MatthewCleere,JackWhittaker,PaulS.DyerSchoolofLifeSciences,NottinghamUniversity,UniversityPark,Nottingham,NG72RD,UKEmail:michaela.novodvorska@nottingham.ac.ukPenicilliumroquefortiisanindustriallyimportantfungusthatisusedasastarterculturefortheproductionofblue-veinedcheese.Thefungusproducesvolatilecompoundsthatare responsible for theuniquebluecheesearoma, thesecompoundsbeinggeneratedfromtheextensivecatabolismofmilk lipidsandproteins.Thebluecolourcomesfrommelanin, and possibly other, pigments covering the surface of spores formed duringgrowth in the cheese. It is known that a DHN-melanin biosynthetic pathway isresponsibleforconidialmelaninpigmentbiosynthesisincloselyrelatedspeciessuchasAspergillus fumigatusandPenicilliummarneffei. In thisstudywe investigatedwhetherthesamepathwayispresentinP.roquefortiandassessedtheimpactofthepathwayonresistanceagainstenvironmentalfactors,mycotoxinproductionandvolatileprofile.Weidentified six homologous genes of a canonical DHN-melanin biosynthetic pathway,developed a novel transformation protocol for P. roqueforti, and generated deletionstrains.Deletionof eachof these geneshadan impactonpigmentbiosynthesis,withdeletion mutants producing conidia exhibiting colour differences to the wild-typeparentalstrain.DeletionmutantsshoweddecreasedviabilitywhenexposedtoUVlightandelevated temperatures, indicating the importanceofpigmentproduction in sporesurvival under varying environmental conditions. Production of mycotoxins such asmycophenolic acid and roqueforine C was greatly reduced when the pigmentbiosynthesispathwaywasnot fully functional.Deletionofeachof thepathwaygenesalso resulted in a change in production of flavour volatiles. These findings are ofrelevanceforcheesemanufacture.Melanin-dependentpathogenicityofAspergillusfumigatusAxelA.Brakhage1,21Department,MolecularandAppliedMicrobiology,LeibnizInstituteforNaturalProductResearch and Infection Biology (HKI), Jena, Germany and 2Institute for Microbiology,UniversityofJena,GermanyEmail:axel.brakhage@leibniz-hki.deThefilamentousfungusAspergillusfumigatusisthemostimportantair-bornefungalpathogencausing90%ofallsystemicAspergillusinfectionsinhumans.Previously,we

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showedthattheconidialpigmentconsistingofdihydroxynaphthalene(DHN)melaninrepresentsavirulencedeterminant.ConidialackingDHN-melaninshowreducedvirulenceinamouseinfectionmodel.WefoundthatthepresenceofDHN-melaninontheconidialsurfaceinfluencestheresponseofimmunecellsatvariouslevels.Forexample,DHN-melanininhibitsphagolysosomalacidificationandcausesincreaseddamagetomacrophagescomparedtonon-pigmentedpksPmutantconidia.WediscoveredanovelmolecularmechanismbywhichDHN-melaninleadstothedisturbanceoflipidraftmicrodomainsinthemembraneofphagolysosomes,therebypreventingtheassemblyandactivityofvATPaseandthusinhibitingtheacidificationofthephagolysosome.Also,assemblyofNADPHoxidasecomplexwaslipidraft-dependentandreducedbymelanizedconidia.Furtherclassificationofthelipidraftmicrodomainsshowedthatflotillin-enrichedlipidraftsco-localizedwithsphingolipidsonphagolysosomalmembranescontainingpksPconidiabutnotwild-typeconidia.FungalSAPKpathwaysasmediatorsofvirulenceAlisonM.Day1,CarmenM.Herrero-de-Dios2,AlistairJ.P.Brown2andJanetQuinn11Institute for Cell and Molecular Biosciences, Faculty of Medical Sciences, NewcastleUniversity,Newcastle upon Tyne,NE2 4HH,UK; 2Aberdeen FungalGroup, Institute ofMedicalSciences,UniversityofAberdeen,AberdeenAB252ZD,UK.Email:janet.quinn@ncl.ac.ukRobuststressresponsesarecommonvirulenceattributeswithinabroadrangeoffungalpathogens,includingthehumanfungalpathogenCandidaalbicans.Centraltosuchstressresponsesarethestressactivatedproteinkinases(SAPK),whichareevolutionarilyconservedeukaryoticsignallingmodulesthatareessentialforthevirulenceofhumanpathogenicfungi.TheHog1SAPKisessentialforC.albicansvirulence,howeveritsstructuralandfunctionalconservationwithanalogoushumanSAPKs(p38/JNK)makesthedevelopmentofantifungaldrugsthatwillspecificallytargetfungal,andnothuman,SAPKschallenging.Becauseofthis,thereismuchinterestinidentifyingfungal-specificregulatorsofSAPKsaspotentialdrugtargets.Suchtargetsincludetwo-componentrelatedphosphorelaysystems,whichareusedbyfungi,butnotmammals,tosenseandrelaystresssignalstoSAPKmodules.Suchphosphorelaysystemscompriseofastress-sensinghistidinekinase,anintermediaryphosphorelayprotein,andaterminalresponseregulator.HereIwillpresentrecentfindingsfrommygroup,whichhavedissectedtheroleofkeyphosphorelayproteinsinregulatingHog1,andexploredtheirpotentialasantifungaldrugtargets.

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SynergisticcombinationstargetingtranslationfidelityanddiversefungalpathogensCindyVallieres,ElenaMoreno-Martinez,AshleighGilliot,SaraL.HollandandSimonV.Avery.SchoolofLifeSciences,UniversityofNottingham,UniversityPark,NottinghamNG72RD,UK.Email:C.Vallieres@nottingham.ac.ukAwiderangeoffungicides(orantifungals)areusedinagricultureandmedicine.Unfortunately,thegrowingresistancetocurrentantifungalshasbecomeamajorissue,andthereforenewagentswithnovelmodesofactionareurgentlyneeded.Combinationtreatmentsofferanefficientmethodformanagingfungalgrowthbyenhancingefficacycomparedtoindividualcompounds,decreasingthedoseofsingleagentusage,andsubsequentlyloweringthedrugtoxicityandcost.Moreover,combinatorialinhibitioncanslowtheevolutionofresistance.Astudyconductedinourgroupshowedthatdifferentaminoglycosideantibioticscombinedwithdifferentsulphate-transportinhibitorsproducedstrong,synergisticgrowth-inhibitionofseveralhumanandplantpathogenicfungimainlybycausing,viadifferentmechanisms,errorsinmRNAtranslation.Theresultsindicatethattranslationfidelityisapromisingnewtargetforcombinatorialtreatmentofundesirablefungi.Inlinewiththisstudy,wescreenedawiderrangeofcompoundsknowntoaffecttheavailabilityoffunctionalaminoacidsincombinationwitheitheri)anotheragentthataffectstheavailabilityoffunctionalaminoacidsorii)anaminoglycosideantibiotictouncovernewsynergisticdrugcombinationstargetingtranslationfidelity.Screenswereperformedinthreefungalphytopathogens(i.e.,Rhizoctoniasolani,SeptoriatriticiandBotrytiscinerea)andthreefungalpathogensofhumans(i.e.,Cryptococcusneoformans,CandidaalbicansandAspergillusfumigatus).43combinationsoutof172wereabletoabrogatefungalgrowthwith9combinationsbeingrepresentedinatleastthreepathogenicfungi.Thus,thisstudyunveilsatargetprocessandchemicalcombinationswiththerapeuticpotentialagainstdiversefungalpathogens.CandidaalbicansisamovingtargetfortheimmunesystemasitadaptstohostsignalsGabrielaM. Avelar, Delma S. Childers, Judith Bain, Arnab Pradhan, Daniel Larcombe,LarsErwig,ElizabethR.Ballou,NeilA.R.GowandAlistairJ.P.BrownAberdeenFungalGroup,MRCCentreforMedicalMycology,InstituteofMedicalSciences,UniversityofAberdeen,AberdeenAB252ZD,UKEmail:al.brown@abdn.ac.ukThepotencyoftheimmunesystemplaysakeyroleinfungalcolonisationandinfection.MoststudiesofCandidaimmunologyhavebeenperformedonfungalcellsgrownunderstandardisedconditionsinvitro.ByexaminingtheimpactofkeyhostsignalsuponCandidaalbicansinvitro(suchaschangesincarbonsources,pH,temperature,oxygenation,stressesandmicronutrients),wefindthat,inadditiontoaltering

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resistancetohoststressesandantifungaldrugs,somehostsignalstriggerchangesinthefungalcellwall.ThisleadstosignificantmodulationofkeyPathogenAssociatedMolecularPatterns(PAMPs)onthefungalcellsurface,turningC.albicansintoamovingtargetfortheimmunesystem.InparticularwehavefoundthatphysiologicallevelsofL-lactatetriggerβ-glucanmaskingviaaspecificlactatesignallingpathwaythatincludesthereceptorGpr1andthetranscriptionfactorCrz1.Lactate-inducedβ-glucanmaskingleadstoareductioninphagocyticrecruitment.ThiscorrelateswithanincreaseinthevirulenceofC.albicans.Therefore,wearguethattheadaptationofC.albicanstohostmicroenvironmentscanaffectitssusceptibilitytoantifungaltherapy,itsdetectionbytheimmunesystem,anditspathogenicity.ThesefindingsmayberelevanttootherpathogenicCandidaspecies.Targetinginfectionmechanismsfortwoamphibian-infectingchytridspeciesRhysA.Farrer1,2,AnthonyNash3,AnMartel4,ElinVerbrugghe4,AmrAbouelleil1,RichardDucatelle4,JoyceE.Longcore5,TimothyY.James6,FrankPasmans4,MatthewC.Fisher2andChristinaA.Cuomo11 Genome Sequencing and Analysis Program, Broad Institute of MIT and Harvard,Cambridge, Massachusetts 02142, USA; 2 Department of Infectious DiseaseEpidemiology,SchoolofPublicHealth,ImperialCollegeLondon,LondonW21PG,UK;3Department of Chemistry, University College London, London WC1H 0AJ, UK; 4

Department of Pathology, Bacteriology and Avian Diseases, Faculty of VeterinaryMedicine,GhentUniversity,Salisburylaan133,B-9820Merelbeke,Belgium;5SchoolofBiologyandEcology,UniversityofMaine,Orono,Maine04469,USA; 6DepartmentofEcologyandEvolutionaryBiology,UniversityofMichigan,AnnArbor,Michigan48109,USAEmail:rfarrer@broadinstitute.orgNearlyhalfofallamphibianspeciesaredeclininggloballyduetofactorssuchashabitatlossanddisease.Tworelatedfungalpathogens(Batrachochytriumdendrobatidis;BdandtherecentlyidentifiedB.salamandrivorans;Bsal)havebeenattributedtotheseglobaldeclinesandextinctions.Bycomparingthegenomecontentandgeneexpressionduringinfection,weidentifiedfundamentaldifferencesinthemolecularbasisofpathogenesis.Notably,weidentifiedanewgroupofsecretedgenesinBsalthathavenosequencesimilaritytopreviouslycharacterizedgenes,butarehighlyexpressedinthehost.Usingstructuralpredictiontools,wearestartingtogaincluestoitspossiblefunctions.Additionally,weidentifiedalargeexpansionofmetalloproteasesinBsalcomparedtoBd,geneswhichhavebeenalsobeenreportedinavarietyofdermatophytes.PreliminaryworktosilencethesegenesusingRNAiisbeinginvestigated.Wefoundthatbothofthesecandidatepathogenicityfactorsweredifferentiallyexpressedinthepresenceofthehost,andthatthehostresponseishighlydistincttothetwopathogens.Together,theseanalysesdemonstratethedivergent

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infectionstrategiesandimmuneresponsetowithinthisamphibian-killinggenus,andtwopossibleroutestofurtherunderstandtheirfunctionalmechanisms.DiversityinplantbiomassdegradationapproachesoffungiRonaldP.deVriesFungal Physiology, Westerdijk Fungal Biodiversity Institute & Fungal MolecularPhysiology,UtrechtUniversity,Uppsalalaan8,3584CTUtrecht,TheNetherlandsEmail:r.devries@westerdijkinstitute.nlTherearefewaspectsoffungalbiologythatareasdiverseastheirstrategiestoobtaincarbonandenergyfromtheirnaturalsubstrates.Plantbiomass,andinparticularlignocellulose,isamajorsubstrateformanyfungiduetoitsabundanceinnatureanditshighcontentofmonosaccharides.Theavailabilityof(post-)genomicdatasetshaveprovidedanunprecedentedviewonthestrategiesoffungitodepolymerizeandconvertplant-basedpolysaccharides.Comparativegenomicsrevealedahugevariationingenome(CAZome)contentwithrespecttolignocelluloseactiveenzymesandinmanycasesthesevariationscouldbelinkedtothenaturalbiotopesofthespecies.However,thedifferenceinstrategygoesbeyondthegenomicvariations.RecentstudiesinourlabrevealedthatevencloselyrelatedspecieswithsimilarCAZomesproducesignificantlydifferentenzymesetswhenexposedtothesamesubstrate.Thissuggestssignificantdifferencesintheregulationofthegenesencodingtheseenzymesaswellasapreferencefordifferentcomponentsoflignocellulosebythedifferentspecies.Inthispresentation,majordifferencesobservedacrossthefungalkingdomwillbepresentedandplacedintocontextofbiotopeandlifestyle.Secondly,differencesbetweenspeciesofthegenusAspergilluswillbepresentedasashowcaseforpost-genomicvariation.Finally,Iwilladdresshowtheseinsightscanbeusedtoimprovecommercialenzymecocktails.MycovirusesasprospectiveenhancersofbiocontrolagentsIolyKotta-Loizou1andRobertH.A.Coutts21Department of Life Sciences, Imperial College London, London, SW7 2AZ, UK;2Department of Biological and Environmental Sciences, University of Hertfordshire,Hatfield,AL109AB,UK.Email:i.kotta-loizou13@imperial.ac.ukThecurrentworldpopulationisca.7.5billionandapproximately2billiontons/yearofcropsareneededtocoverournutritionalrequirements.Pesticidesareusedtoprotectthecropsfromdamageandincreaseagriculturaloutput;however,theyareconsideredoneofthetopsixglobalthreatsgreatlyimpactingthebiodiversityoftheenvironmentandputting7millionpeopleatrisk.Environmentallyfriendlyalternativestopesticides

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arebiologicalcontrolagents,includingcommerciallyavailableentomopathogenicfungiwhosegrowthandpathogenicitymaybeenhancedbymycoviruses.Theultimateaimoftheprojectistoinvestigatemycovirus-inducedhypervirulenceinentomopathogenicfungi.TothisendpanelsofBeauveriabassiana,MetarizhiumanisopliaeandLecanicilliummuscariumwerescreenedforthepresenceofdouble-stranded(ds)RNAelements.Amongothermycovirusesdiscovered,membersofthenovelvirusfamilyPolymycoviridaewereidentifiedasthemostpromisingenhancers,sincetheynaturallyconfermildhypervirulencetotheirhosts,arenotconventionallyencapsidatedandareinfectiousasnakeddsRNAfacilitatinggeneticmanipulations.Themechanismscontrollingmycovirus-inducedhypervirulencearestillunclearbutsuchknowledgewouldfacilitategeneticengineeringofknownmycovirusestoincreasehypervirulence.Off-targetRNAsilencingisexpectedtoplayarolegiventhedsRNAnatureofthesevirusesthatactasbothtriggersandtargetsoftheantiviraldefencesystem.Additionally,host-virusprotein-proteininteractionsviaEukaryoticLinearMotifsleadingtodisruptionofhostpathwaysmayalsobeimplicatedinaccordancewithotherhost-pathogensystems.Inconclusion,thisstudydiscussesthepotentialofmycovirusesasenhancersofbiocontrolagents.Genomic and genetic analysis of a basidiomycete yeast focused on thecharacterisationofMELbiosynthesisStefanySolano1,2,AnnaSobolewska3,DougCossar3,MarkCaddick1&AlistairDarby11Institute of Integrative Biology, University of Liverpool, UK; 2Escuela de CienciasBiologicas,UniversidadNacional,CostaRica;3CrodaEurope,Ditton,Widnes,UKEmail:S.Solano-Gonzalez@liverpool.ac.ukThisworkfocusesonabasidiomyceteyeastknowntobeagrassendophyte.Ofspecificinterestisthestrain’sabilitytoproduceMannosylerithritollipids(MELs).Theseareofinteresttoindustryasbiosurfactantsandareclassifiedintofourgroups(A–D)accordingtotheirstructure.ThebasidiomyceteinthisstudyisprimarilyaMELCproducer.Theregulationandproductionofthesesecondarymetabolitesandtheunderpinningbiologyremainunclear,despitethemanystudiescarriedoutwithbasidiomycota,Ustilagomaydis,andPseudozymaaphidis.CurrentlytherearefewannotatedgenomesforMELproducingfungi.Wethereforeundertookthegenomesequencing,assemblyandannotationofourisolate,asafirststeptoinvestigatingitspotentialforindustrialexploitation.UsingthePacBiosequencingplatformwehaveproducedagenomeassembly,composedof34contigs.Transcriptomesequencinghasbeenutilisedtofacilitategenecalling,withatotalof6603genesbeingidentified,ofwhich4877havebeenautomaticallyannotatedusingprimarydatabases.AmongstthesearethefivegeneswhichformtheputativeMELbiosyntheticcluster.Phylogeneticanalysisshowsthatamongstsequencedfungi,ourstrainismostcloselyrelatedtoS.reilianumandU.maydis(bothplantpathogens)andsurprisinglynotPseudozymaspecies,ashadbeenexpected.Wehaveundertakentranscriptomeanalysisto

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investigategeneexpressionassociatedwithMELproductionoverafivedaytimecourseandwearecurrentlyusingdirectedgenereplacementtoinvestigatethefunctionofspecificgenesinthebiosyntheticclusterandtheroleofputativeregulatoryproteinsinMELproduction.IntraspeciesdiversityandinductionofthewooddecaybythewhiterotfungusDichomitussqualensSaraCasadoLópez1,WilliamB.Chrisler2,SamuelPurvino2,AliceDohnalkova2,GalyaOrr2,ErrolRobinson2,ErikaZink2,ScottBaker2,KristiinaHildén3,MiiaR.Mäkelä1,3andRonaldP.deVries1,31 Fungal Physiology, Westerdijk Fungal Biodiversity Institute & Fungal MolecularPhysiology, Utrecht University, Utrecht, 3584CT, The Netherlands; 2 EnvironmentalMolecular Sciences Laboratory, Department of Energy, Richland, WA 99354, USA;Department of Food and Environmental Sciences, University of Helsinki, Helsinki, FI-00014,Finland.Email:s.casado@westerdijkinstitute.nlDichomitussqualensisawhiterotfungusthatpreferablygrowsandcolonizesdeadsoftwood,butisalsoabletoutilizehardwood.ItiscommonlyfoundinthenorthernpartsofEurope,NorthAmericaandAsia.Asaresultofsexualcrossingoftwocompatiblemonokaryons,afertiledikaryonisformedandfruitingbodiescanbeproduced.Inthisstudyweaimedtodissectthedifferencesinwoodcolonizationbyasetofmono-anddikaryoticstrainsofD.squalens,toanalyzehowvariedthisprocesscanbewithinonespecies.Forthispurpose,weintegratedacombinationoftranscriptomics,proteomics,metabolomicsandhighresolutionmicroscopyenabledbythecombinedfacilitiesoftheJGIandEMSLinstitutes.Allstrainsweregrownonsolidsprucesticks,andsamplingwasperformedaftertwoandfourweekstoevaluatethevariationbetweenthestrainsduringthewooddecayprocess.Moreover,weperformedtranscriptomicsoftwoofthestrains(mono-anddikaryon)onplantpolysaccharidederivedsugarstostarttounraveltheinductionmechanismofcarbohydrateactiveenzymes(CAZymes)encodinggenesinD.squalens.

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ExploitingunspecialisedendophytesforbiologicalcontrolofinsectsandplantsAlanC.Gange,AmandaF.Currie,NadiaAbRazakandRuthChittySchool of Biological Sciences, Royal Holloway, University of London, Egham, SurreyTW200EXUKEmail:a.gange@rhul.ac.ukHerbaceousplantsharbourdiversecommunitiesofendophyticfungiintheirrootsandshoots.However,comparedtoendophytesingrassesorwoodyplants,almostnothingisknownabouttheecologyofthesefungi.Ithasbeensuggestedthatastheseendophyteareunspecialisedandubiquitous,thentherelationwiththeirhostsmaybealooseone,withlittleeffectsontheplantitself,orotherorganismsthatattackthehost,suchasinsects.Oneparticulargroupoffungithatcanoccurasendophytesinplantsaretheentomopathogensandthereisincreasinginterestintheuseofthesefungiasinternalplantprotectants.Herewereporttheresultsofameta-analysis,toexaminetheeffectsofunspecialisedendophytes,includingentomopathogens,oninsectherbivores.Wewillcombinethiswithsomeofourownexperimentaldatatoshowthatthesefungimayhavegreatpotentialinbiologicalcontrolofinsectpests.Contrarytoexpectations,someunspecialisedendophytescanalsohavedirecteffectsonthegrowthoftheirhostsintheabsenceofantagonists,andwewillexplorehowsucheffectsmaybeexploitedinthebiologicalcontrolofweeds.RegulationofDrugResistanceinAspergillusfumigatusMichaelJ.BromleyManchester Fungal Infection Group, School of Biological Sciences, University ofManchester,Manchester,UnitedKingdom.Email:Mike.bromley@manchester.ac.ukAspergillusfumigatusisthemostimportantairbornemouldpathogenandallergenworldwide.Estimatessuggestthatover3millionpeoplehaveinvasiveorchronicinfectionsthatleadtoinexcessof600,000deathseveryyear.Veryfewdrugsareavailabletotreatthevariousformsofaspergillosisandwerelypredominantlyontheazoleclassofagents(Itraconazole,Voriconazole,PosoconazoleandtherecentlylicensedIsavuconazole).Resistancetotheazolesisemerging.Forindividualsthatareinfectedwitharesistantisolatethemortalityrateexceeds88%.Therapyfailureisinpartattributedtodelaysinadministeringalternativetherapies,somethodstorapidlydetectresistancearecritical.Whileresistanceinaround50%ofclinicalisolateshasbeenlinkedtomodificationofthegeneencodingthetargetoftheazoles,cyp51A,ourunderstandingofwhatleadstoresistanceintheremainingstrainsislacking.Wehaveundertakenaprogrammeofworktoidentifytranscriptionalandpost-transcriptionalregulatorsofazoleresistance.Wehavediscoveredacohortoftranscriptionfactors,includingtheCCAAT-bindingcomplexandNegativeCofactor2,

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whichmodulateazoletolerability.Withaviewtounderstandingwhichgenesaredirectlyregulatedbythesetranscriptionfactors,wehaveperformedgenome-wideprotein-DNAinteractionanalysisusingChIP-seq.Wehaveidentifiedthatthesetranscriptionfactorsbindthepromotersofgenesknowntobeassociatedwithazoletoleranceincludingcyp51Athedrugtransportercdr1B.OurChip-seqdataprovidesevidencetosuggestthattheseregulatorsbindthepromotersofmanygeneshoweverdonotalwaysmodulatetheirexpression.SystematicfunctionalannotationoftranscriptionfactorsinthemajormouldpathogenAspergillusfumigatusTakanoriFurukawa,NormanvanRhijn,PanosPapastamoulis,SayemaKhan,LeaGregson,PaulBowyer,MagnusRattray,MichaelBromley,ElaineBignellManchester Fungal Infection Group, Division of Infection, Immunity, and RespiratoryMedicine,UniversityofManchester,Manchester,M139NT,UKEmail:takanori.furukawa@manchester.ac.ukAspergillusfumigatusis,globally,themostimportantairbornefungalpathogenofhumansthatcausesover90%ofallAspergillus-relatedinfections.Ithasbeendemonstratedthatthevirulenceofthisfungusismultifactorialandseveralvirulence-determiningfactorshavebeenidentified.Inparticular,theabilityofthefungustorespondtovarioushost-imposedstresseshasrepeatedlybeenproventobeessentialforpathogenicity,andthereforetranscriptionalregulationplaysacriticalroleinestablishinginfection.However,weonlyhaveaverylimitedunderstandingofthemolecularbasisofA.fumigatuspathogenicity.Inthisstudy,wecarriedoutasystematicphenotypingofatranscriptionfactorknockout(TFKO)collectionofA.fumigatusmutantstodefinethegenomiccohortoftranscriptionfactorsrequiredforpathogenicity.Ahigh-throughputanalyticalpipelinewasdevelopedina96-wellplatebasedliquidcultureformattoanalyzerelativefitnessof401A.fumigatusTFKOstrains.GrowthtraitsoftheTFKOstrainswerethenanalyzedinparallelundereachofthefollowinghost-imposedstressconditions:ironstarvation,temperaturestress,andpHtolerance.Dataanalysiswasautomatedtoconvertopticaldensityreadingscollecteddirectlyfromthespectrophotometerintofitnessindices.Ourstudyidentifies,forthefirsttime,thefullcohortofA.fumigatustranscriptionfactorsthatgovernresponsestosignificanthost-imposedstressesandprovidesafirstgenome-scalefunctionalannotationoftranscriptionfactorsinA.fumigatus.

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BSPPTalksUsingpopulationgenomicstodeterminethegeneticbasisofpathogenadaptationinagro-ecosystemsBruceA.McDonaldPlant Pathology Group, Institute of Integrative Biology, ETH Zurich, 8092 Zurich,SwitzerlandEmail:bruce.mcdonald@usys.ethz.chSeptoriatriticiblotch(STB)isthemostdamagingwheatdiseaseinEurope.ThepathogencausingSTB,Zymoseptoriatritici,isgloballydistributedandhasevolvedtobecomeresistanttofungicidesandvirulentonresistantwheatcultivars.WecoupledQTLmappingbasedon700progenyfromtwocrosseswithanalysesofgenome-wideassociations(GWAS)basedon150strainsdrawnfromfourglobalfieldpopulationstoelucidatethegeneticarchitectureofseveraltraits,includinghostspecialization,virulenceandfungicideresistance.FinishedgenomesequencesandextensiveRNA-seqprofilesobtainedforallfourparentsenabledidentificationofcandidategenesaffectingeachtrait.Severalofthesegeneswerefunctionallyvalidated.Whilegenotypinghasbecomerelativelyeasy,phenotypingremainsdifficultandlimitsprogressinmostmarker-traitassociationstudies.Wedevelopedhigh-throughputphenotypingmethodstoacceleratebreedingforSTBresistance.Automatedanalysisofdigitalimagesgenerated2.7millionphenotypicmeasurementsassociatedwithmelanization,virulence,hostspecialization,fungicidesensitivityandthermaladaptationintheQTLmappingpopulations.Thisapproachwasextendedtoanaturallyinfectedfieldplantedto335eliteEuropeanwheatcultivars,yielding>15millionphenotypicmeasurementsassociatedwithhostdamage(percentleafareacoveredbySTBlesions)andpathogenreproduction(numberofSTBpycnidiapercm2ofleaflesion)thatrankedtherelativeSTBresistanceamongthesewheatcultivars.AGWASusingthesedatasetsallowedustogeneticallyseparateresistanceaffectinghostdamagefromresistanceaffectingpathogenreproduction.OurepidemiologicalanalysesindicatedthatresistanceaffectingpathogenreproductionoffersthegreatestpotentialforlimitingdamageduetoSTB.WhydoesIPMnotleadtomorereductionsinfungicides?LiseNistrupJørgensenDepartmentofAgroecology,AarhusUniversity-Flakkebjerg,DK-4200Slagelse,DenmarkEmail:lisen.jorgensen@agro.au.dkCerealcropsaccountforthemajoruseoffungicidesinEurope.Despitethepoliticallydrivendesiretoreducedependenceonpesticidesingeneral,ithasproveddifficulttoreducetheconsumptionoffungicidessignificantly.Severalnon-chemicalmethodsare

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knowntohelpreducediseasesincerealcrops.However,farmers’willingnesstogeneratemajorchangesintheircroppingsystemssuchasincludingnewcropsoradoptingnewtillagemethodsissteeredbyeconomicrationaleandfarmers’abilitytoprovideflexiblemanagement.Croppingvarietieswithgoodresistanceareknownasthemostwidelyadoptedandadvocatedmethodofreducingpathogenattack,andvarietyresistancehasinseveralcasesprovidedgoodpossibilitiesofreducingtherelianceonfungicides.Evenso,incerealcrops,nocultivarswithcompleteresistancetoallrelevantdiseasesexistandfungicidesarecommonlyusedtoensureagainstmajorcroplossesfromdiseases.Changesinpathogenvirulencehaveinmanycasesledtolossofeffectivecultivarresistance,whichagainrequiredadjustedinputoffungicides.AsanelementofIPM,farmersinmanycountriesoftenapplyreduceddoses,althoughtheymostlikelystillapplyhigherdosesthantheoptimumtoensureagainstlossesinseasonswithahighdiseaselevel.Unfortunately,reliabletools,whichcanimprovepredictionoffuturediseasedamageandaccountforthecombinedeffectofintegratedcontrolmeasuresarestilllacking.Ideally,reliablemodelsshouldpredicttheoptimumnumberoftreatmentsandtheoptimumdoseneededtominimizebotheconomiccostsandenvironmentalimpactinagivenseason.Identifyingrootresistancetotake-alldiseaseVanessaE.McMillan,GailCanning,RichardJ.GutteridgeandKimHammond-KosackDepartmentofBiointeractionsandCropProtection,RothamstedResearch,UKEmail:vanessa.mcmillan@rothamsted.ac.ukTake-all(Gaeumannomycestritici)isamajorrootdiseaseofwheatwhichisaseriousconstraintinshortwheatrotations,reducingyieldsof2ndand3rdwheatcropsby1to3tonnes/ha.Distinctivepatchesofdiseasedevelopinthefieldwithstuntedandprematurelyripeningplantswheretherootsystemisseverelyinfected.Moderateandseverediseaseoutbreaksalsoputfieldsatriskofnitrateleachingasexcessunusednitrogenfertiliserisleftbehindinthesoil.AspartoftheWheatGeneticImprovementNetwork(WGIN,www.wgin.org.uk)andWheatImprovementStrategicProgramme(WISP,www.wheatisp.org)protocolshavebeendevelopedtophenotypearangeofdiversegermplasmcollections,includingelitewheatvarieties,landracesandwheatrelativesforrootresistancetotake-alland/ortheireffectonfungalinoculumbuild-upinthesoil.Phenotypingwasprimarilycarriedoutunderfieldconditionswiththeaimofidentifyingresistancethatwasconsistentlyexpressedacrossmultiplefieldseasonsandsites.Since2008over700Watkinshexaploidwheatlandraceshavebeenphenotypedfortake-allresistance.Moderateresistancetotake-alldiseasewasidentifiedforasubsetofgenotypeswithinthecollection.AParagon(moderatelysusceptible)xWatkins777(moderatelyresistant)mappingpopulationiscurrentlybeingphenotypedtodissectthegeneticbasisofresistance.ResultsfromthefirstyearoffieldphenotypingrevealtwoQTLsconferringresistanceinalowdiseasepressuresituation.Workisnowunderwayto

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confirmandvalidatethesegenomiclocationsacrossfurtherfieldseasonsandtoexploretherootresponseinmoredetail.Ignoreduntilrecently–thestoryofthefungalpathogenLeptosphaeriabiglobosaYongjuHuang,AsnaJavaid,LakshmiH.Gajula,ChinthaniS.Karandeni-Dewage,GeorgiaMitrousia,BruceD.L.FittSchoolofLifeandMedicalSciences,UniversityofHertfordshire,Hatfield,Hertfordshire,AL109AB,UKEmail:y.huang8@herts.ac.ukManyplantdiseasesarecausedbymorethanonepathogenandtherearemanyfactorsaffectingtheirco-existenceontheirhostthatleadtochangesintheirpredominance.Phomastemcankerisagloballyimportantdiseaseofoilseedrape,causedbytworelatedpathogens,LeptosphaeriamaculansandL.biglobosa.SinceL.maculansisoftenassociatedwithstembasecankerwhileL.biglobosaisassociatedwithsuperficialupperstemlesions,L.maculansisconsideredmoredamagingthanL.biglobosa.AlthoughL.maculansandL.biglobosaco-existontheirhost,previousworkhasmainlyfocusedonL.maculansandtheimportanceofL.biglobosainphomastemcankerepidemicshasbeenignored.ResultsofrecentworkshowthatL.biglobosahasincreasedinimportanceintheUK,causingbothdamagingupperstemlesionsandstembasecankers.Furthermore,L.biglobosaislesssensitivetosometriazolefungicidesthanL.maculans.PreviouscontrolofphomastemcankerintheUKbycultivarresistanceorfungicideshastargetedonlyL.maculans,disregardingL.biglobosa.RecentstudieshaveshownthatcultivarsresistantagainstL.maculansareoftenmoresusceptibletoL.biglobosa.Foreffectivecontrolofphomastemcanker,thereisaneedtotargetbothL.maculansandL.biglobosa.Inthistalk,thereasonswhyL.biglobosahasrecentlyincreasedinimportanceinphomastemcankerepidemicsintheUKwillbediscussed,basedonresultsfromfieldexperimentsoverthreeseasonsandcontrolledenvironmentexperiments.*PowderyMildew:ATaleofTwoFungicidesCorinneJ.Arnold,DianeG.O.SaundersandJamesK.M.BrownCropGenetics,JohnInnesCentre,Norwich,NR47UH,UKEmail:Corinne.Arnold@jic.ac.ukBlumeriagraminisisapowderymildewpathogenthatinfectsmanywildgrassesandcerealsacrosstheworld.Ifleftuncontrolled,itcancausesignificantyieldlossesincereals.OnemainmethodforcontrolistheapplicationoffungicidesbutresistancetostrobilurinandoldertriazolefungicidesalreadyoccursinB.graminis,andfewotherfungicidesareavailableformildewcontrol.Ihavecharacterisedisolatesfroman

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outbreakofB.graminisf.sp.tritici(Bgt)onwheatwheretwoofthemajorfungicidesusedagainstBgtwereineffective:cyflufenamid(Cyflamid),aneradicantfungicidewithanunknownmodeofaction,andfenpropimorph(Corbel),amorpholinefungicidetargetingasterolreductase.AllBgtisolatesfromthestudysitewerecompletelyresistanttoCyflamidsprayedattherecommendedfieldrate.However,overthefollowingtwoyears,theBgtpopulationrecoveredsensitivitytoCyflamid,suggestingthatresistancemayhaveasignificantfitnesspenalty.Additionally,followingtwospraysofCorbel,theseBgtisolateshadsignificantlylowersensitivitytofenpropimorphthanfieldisolates.Overuseoffenpropimorphwouldalmostcertainlyleadtostrongresistanceinmildew,soitshouldbeusedaspartofarotationoffungicideswithdifferentmodesofaction.Likewise,Cyflamidcancontinuetobeusedaspartofanintegrateddiseasecontrolprogramme.Geneticandgenomicanalysisisinprogress,withtheaimofidentifyingcandidategenesforresistancetocyflufenamidandtofenpropimorph.*Blumeriametallo-protease-likeeffector;azincophoreactingasauniversalvirulencefactorinfungi?ShaoliDasGupta1,KateOrman1,MartinUrban2,GabrielScalliet3,KimHammond-Kosack2andLaurenceBindschedler11School of Biological Sciences, Royal Holloway, University of London, Egham, SurreyTW200EX,UK;2DepartmentofPlantBiologyandCropScience,RothamsteadResearch,Harpenden, Hertfordshire AL5 2JQ, UK;3Syngenta Crop Protection, Research Biology,Schaffhauserstrasse,Stein,SwitzerlandEmail:Shaoli.DasGupta.2015@live.rhul.ac.ukBlumeriaeffectorcandidate19(BEC1019)isavirulencefactorofthebiotrophicpathogenBlumeriagraminisf.sp.hordei(Bgh),acausalagentofbarleypowderymildew.BEC1019silencingledtoreducedBghgrowthanddiseasedispersion.BEC1019hashomologuesinonethirdofthesequencedfungalgenomes,includingthezincscavengingvirulencefactorsPRA1(Candidaalbicans)andAspf2(Aspergillusfumigatus).BEC1019homologueslackproteaseactivitybutshareaconservedzincbindingHRXXHdomain,relatedtotheHEXXH-containingM35zincmetallo-proteasesuperfamily.Therefore,BEC1019homologuesarepotentialfungal“zincophores”withauniversalvirulencerole.ThisprojectisinvestigatingthepotentialinvolvementofBEC1019homologuesinzincsequestrationforthevirulenceofeconomicallyimportantwheatfungalpathogens.WewillshowthatBEC1019silencinginB.graminisf.sp.tritici(Bgt)ledtoreducedinfectionofwheatpowderymildew,thusindicatingitsroleinvirulence.Fusariumgraminearum(Fg)andZymoseptoriatritici(Zt)BEC1019knockoutsarebeingproducedtoestimatetheroleofBEC1019duringFusariumheadblightandSeptoriatriticiblotchinfection.SinceBEC1019wasshowntobehighlyinducedduringZtgrowthunderzincdeficiency,theroleofBEC1019andotherfactorsinvolvedinzincsequestrationforpathogensuccessandcounteringplant

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nutritionalimmunitywillbefurtherinvestigated,inthequestoffindingnoveltargetsforcropprotection.*Understanding the ecology and epidemiology of Pythium violae to enable diseasemanagementincarrotcropsKathrynR.Hales1,GaryDBending2,TimRPettitt3andJohnP.Clarkson11WarwickCropCentre,UniversityofWarwick,Wellesbourne,CV359EF,UK;2SchoolofLife Sciences, University of Warwick, Coventry, CV4 7AL, UK; 3National Pollen andAerobiologyResearchUnit,UniversityofWorcester,Worcester,WR26AJ,UKEmail:k.r.hales@warwick.ac.ukCavityspotisamajordiseaseofcarrotprimarilythoughttobecausedbythesoilborneoomycetepathogenPythiumviolae,althoughotherPythiumspeciesmayalsobeassociatedwiththelesions.Diseasemanagementischallenging,asfungicideefficacyisvariableandlongrotationsnotalwayspossible.NewapproachestocontrolhavebeenhamperedbyalackoffundamentalknowledgeconcerningtheepidemiologyofP.violaeandbasictoolsforresearch.ThisPhDprojectaimstoaddresssomeoftheseissuesbydevelopingmoleculardetectiontechniquesandartificialinoculationmethods.TodeterminetherangeofPythiumspp.associatedwithcavityspotsymptoms,acollectionof150isolateswasassembledfromdiseasedcarrots.FollowingDNAextractionandsequencing,61%wereidentifiedasP.violaewithP.sulcatumandP.intermediumbothaccountingfor14%.TheremaindercomprisedotherPythiumspecies.TodetectP.violaeinrelativelylargesamplesofsoil,asucroseflotation/filtrationmethodwasdevelopedtocaptureoosporesfrom10gsoil.WhencombinedwithaspecificPCRtest,initialresultssuggestedthatdetectionoflessthan10oosporesispossible.ThisapproachisbeingdevelopedfurtherforRT-PCRapplicationtoenableaccuratequantificationofP.violaeandmonitoringofpathogendynamicsinsoilandonroots.AmethodofinoculumproductionwasdevelopedforP.violaeandusedtoinoculatepot-growncarrotsintheglasshouseandsmallfield‘macrocosms’.Initialresultsrevealedeffectsoncarrotgrowthandthedevelopmentoftypicalcavityspotlesionsonroots.*Examining identity, phylogeny and pathogenicity factors in Fusarium speciesaffectingpeaSaschaJenkins1,RichardJ.Harrison2andJohnP.Clarkson11WarwickCropCentre,UniversityofWarwick,Wellesbourne,CV359EF,UK;2Genetics,Genomics&Breeding,NIAB-EMR,EastMalling,ME196BJ,UKEmail:sascha.jenkins@warwick.ac.uk

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PeaisanimportantlegumecropfortheUK,worthanestimated£50millionperyear.TheUKisalsothelargestproduceroffrozenpeasinEurope,duetoidealconditionsalongtheEasternseaboard.Fieldsarelocatedclosetoprocessingfactories,whichreducesthepotentiallandavailabletogrowpeas.Soilborneplantpathogensinthe‘footrot’complexareoneofthemaincausesofcroplossinpeas,resultinginwilting,reductionsinpodnumbersandplantdeath.SeveralFusariumspeciesaremajorcomponentsofthiscomplexwithF.oxysporumf.sp.pisi(FOP)beingamongstthemostimportantworldwide.RacesofFOParedifficulttodistinguishwiththecurrentmethodofpathogenicitytestsusingpeadifferentiallines.PathogenicityfactorsincludingeffectorgeneshavebeenreportedinotherpathogenicF.oxysporumwithSecretedinXylem(SIX)geneswidelystudied.ThemainaimsofthisPhDprojectaretoidentifytherangeofFusariumspeciesaffectingpeaintheUKandidentifyeffectorgenesassociatedwithpathogenicity.AsurveyofUKpeafieldsidentifiedfootrotcausingspeciesincludingF.oxysporum,F.solaniandF.redolens,withthemostcommonbeingF.oxysporum.TheseisolatescollectedfromUKfieldsandFOPrace-typedisolateswereusedintwopathogenicityteststodistinguishwiltcausingandfootrotcausingisolates.InitialresultsfromwholegenomesequencingandPCRhaverevealedthepresenceof7SIXgenesinFOPraces1,2and5,andfurthertestswillstudytheirexpressionlevelspostinoculation.*Finemapping of a major gene locus for resistance against Pyrenopeziza brassicae(lightleafspot)inBrassicanapusandidentificationofcandidateresistancegenesChinthaniS.KarandeniDewage,HenrikU.Stotz,BruceD.L.FittSchoolofLife&MedicalSciences,UniversityofHertfordshire,Hatfield,Herts,AL109AB,UKEmail:c.s.karandeni-dewage@herts.ac.ukPlantmajorgene-mediatedresistanceisimportantformanagementofdiseasethreatstocrops.Finemappingandcloningofthesegenescanprovideinsightsintohost-pathogeninteractionstoimprovediseasecontrolstrategies.ThisworkontheBrassicanapus–Pyrenopezizabrassicaepathosystemaimstoimproveunderstandingoftheoperationofmajorgene-mediatedresistanceagainsthemibiotrophicplantpathogens.PyrenopezizabrassicaecauseslightleafspotdiseaseonwinteroilseedrapeintheUKaswellasdiseaseonvegetablebrassicas,includingBrusselssprouts.AmajorgenelocusmapstothebottomendofBrassicanapuschromosomeA01.UsingnovelgenomicinformationandmolecularmarkerdataonB.napus,improvedmarkerresolutionandunderstandingofthegeneticbasisofthisresistancecanbeobtained.WeidentifiedthephysicallocationoftheclosestflankingmarkertotheresistancelocusonB.napuschrA01usingmarkersequenceandthesyntenybetweenB.rapaandB.napusgenomes.Singlenucleotidepolymorphism(SNP)markerinformationonthecorrespondinggenomicregionwasobtainedandc.400candidateSNPsspecifictochrA01wereidentified.Initially,38SNPswereselectedfordevelopmentofKASPmarkers,which

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weretestedonparentallinesoftheDHpopulationthatsegregateformajorgeneresistanceagainstP.brassicae.Polymorphicmarkersbetweenparentallineswereanalysedinthesegregatingpopulationtoimprovethegeneticmapresolution.WealsoanalysedthreedifferentBrassicanapusgenomesequencestoidentifygenecontentinthisgenomicregionandgenepredictionswillbeusedtoidentifycandidateresistancegenesagainstP.brassicae.*Fromthefieldtothelabandbackagain:exploringlegacyeffectsofrootgeneticsandfungicidefortake-allrootdiseasecontrolinwheatrotationsJosephMoughan1,VanessaMcMillan1,RodgerWhite2,JamesMelichar3,DavidRanner3andKimHammond-Kosack11Biointeractions and Crop Protection Department, Rothamsted Research, Harpenden,Hertfordshire, AL5 2JQ, UK; 2 Computational and Analytical Sciences Department,Rothamsted Research, Harpenden, Hertfordshire, AL5 2JQ, UK; 3SyngentaUK Limited,Fulbourn,Cambridge,CB215XE,UKEmail:joseph.moughan@rothamsted.ac.ukGaeumannomycestriticiisasoil-borne,rootpathogenofwheat,causingtake-alldisease.Somemodern,elite,winterwheatcultivarspossessagenetictraitthatpromoteslowtake-allinoculumbuild-up(LowTAB)infirstwheat(McMillanetal.,(2011),PlantPathology,60:200-206).Thisleadstoreduceddiseaseifwheatisgrowninthesamefieldthenextyearandhasgiventwotonnes/hayieldadvantagesinseverediseaseyears(McMillan,submitted).Usingfieldtrials,thisresearchinvestigatedifenhancedorsynergisticreductionoftake-allinoculumbuild-up,couldbeachievedwithfoliarapplicationsoftake-allactivefungicide,azoxystrobin,incombinationwithLowTABgenetics.Subsequentdiseasewasmeasuredinsecondwheatoversowexperiments.Tocomplementthis,laboratoryscreenswereperformedtocheckforcommontargetsitemutationstotheazoxystrobinfungicide,innewandhistoricG.triticiisolates.Reducedsecondwheattake-alldiseasewasfoundresultingfromlegacyeffectsinthesoilfromthepreviousyear’sfirstwheattreatments.Individualeffectsoffirstwheatrootgenetics(LowTAB)(P<0.001)andazoxystrobinapplication(P=0.009)wereobserved,butnosynergywasidentified.Firstandsecondwheatchemistryeffectsdidnotcorrelate.Mid-seasonfirstwheatspraysloweredinoculumbuild-up(P=0.014),butonlyearlyfirstwheatsprayseffectivelyreducedsecondwheatdisease.Theseinconsistentfungicideeffectscouldnotbelinkedtotargetsiteresistance,butmaybeduetolowlevelsoffungicideinsensitivityresultingfromactivityofanalternativerespirationpathwayinG.tritici.Weconcludethatinthefield,azoxystrobinalteredthenormaltake-alldiseaseprogressioncycle.

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ExploitingeffectorsinbreedinganddeploymentofRgenesinpotatoVivianne G.A.A. Vleeshouwers1 , Geert Kessel2, Theo van der Lee2, and Richard G.F.Visser11PlantBreeding,WageningenUniversity&Research,Wageningen,P.O.Box386,6700AJ,TheNetherlands;2Biointeractions,WageningenUniversity&Research,Wageningen,P.O.Box16,6700AA,TheNetherlandsEmail:vivianne.vleeshouwers@wur.nlTheIrishfaminepathogenPhytophthorainfestanscausesthedestructivelateblightdiseaseonpotato(Solanumtuberosum),amajorfoodcropworld-wide.Potatoplantsattainresistanceduetodiseaseresistance(R)genesthathaveco-evolvedwitheffectors,orso-calledavirulence(Avr)genesoftheoomycetepathogen.Sincethegenomicsera,sucheffectorshaveemergedastoolstoaccelerateandimprovetheidentification,functionalcharacterisationanddeploymentofRgenes.UpontheavailabilityofthereferencegenomeofP.infestans,wehavegeneratedagenome-wideinfection-readylibraryofcandidateAvrgenes.TheseareusedinfunctionalassaysfordetectingactivityofRgenes.ThiseffectoromicsstrategyhasacceleratedcloningandspecificityprofilingofRgenesfromwildpotatospeciesandproveneffectiveandcomplementarytoclassicalresistancebreeding.ForvariousR-AVRpairs,wearestudyingtheeffectordiversityandactivitytogetinsightinthemechanismsthatP.infestansemploysforevadingRproteinrecognition.WehavedesignedaPCR-basedmonitoringforcharacterizedAvrgenestodetectvirulentisolatesamonglargesetsofsamplesinfieldtrails.Theseeffectorprofilesareimplementedinanintegratedpestmanagement(IPM)strategy.Withtheseexperiments,weshowthatahigh-throughputspatio-temporalmonitoringofeffectorallelicdiversityinP.infestanspopulationscanenableamoreeducateddeploymentofRgenesinpotato.The RxLR Motif of the Host Targeting Effector AVR3a of Phytophthora infestans IsCleavedBeforeSecretionStephanWawra1,2,, Franziska Trusch1,, AnjaMatena3, Kostis Apostolakis1, Uwe Linne4,IgorZhukov5,6,JanStanek7,WiktorKoźmiński7,IanDavidson8,ChrisJ.Secombes9,PeterBayer3andPietervanWest11AberdeenOomyceteLaboratory,InstituteofMedicalSciences,UniversityofAberdeen,Aberdeen,AB252ZD,UK;2BotanicalInstitute,GeneticalInstitute,UniversityofCologne,50674,Cologne,Germany;3StructuralandMedicinalBiochemistry,CentreofMedicinalBiotechnology,University ofDuisburg-Essen, 45141Essen,Germany; 4Core Facility forMass Spectrometry, Chemistry, Philipps-Universität Marburg, D-35032, Marburg,Germany;5InstituteofBiochemistryandBiophysics,PolishAcademyofSciences,02-106Warsaw,Poland;6NanoBioMedicalCentre,AdamMickiewiczUniversity,61-614Poznań,Poland; 7Biological and Chemical Research Centre (CENT III), Faculty of Chemistry,UniversityofWarsaw,02-089Warsaw,Poland;8ProteomicsFacility,Institute

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ofMedical Sciences,University ofAberdeen,Aberdeen,AB252ZD,UK; 9 Scottish FishImmunology Research Centre, Institute of Biological and Environmental Sciences,UniversityofAberdeen,Aberdeen,AB242TZ,UKEmail:franziska.trusch@abdn.ac.ukWhenplant-pathogenicoomycetesinfecttheirhosts,theyemployalargearsenalofeffectorproteinstoestablishasuccessfulinfection.Someeffectorproteinsaresecretedandaredestinedtobetranslocatedandfunctioninsidehostcells.ThelargestgroupoftranslocatedproteinsfromoomycetesaretheRxLReffectors,definedbytheirconservedN-terminalArg-Xaa-Leu-Arg(RxLR)motif.However,thepreciseroleofthismotifinthehostcelltranslocationprocessisunclear.HeredetailedbiochemicalstudiesoftheRxLReffectorAVR3afromthepotatopathogenPhytophthorainfestansarepresented.MassspectrometricanalysisrevealedthattheRxLRsequenceofnativeAVR3aiscleavedoffpriortosecretionbythepathogenandtheN-terminusofthematureeffectorwasfoundlikelytobeacetylated.High-resolutionNMRstructureanalysisofAVR3aindicatesthattheRxLRmotifiswellaccessibletopotentialprocessingenzymes.ProcessingandmodificationofAVR3aistosomeextentsimilartoeventsoccurringwiththeexportelement(PEXEL)foundinmalariaeffectorproteinsfromPlasmodiumfalciparum.ThesefindingsimplyarolefortheRxLRmotifinthesecretionofAVR3abythepathogen,ratherthanadirectroleinthehostcellentryprocessitself.Knowyourenemy–engineeringresistanceagainstFusariumwiltinbananaLola Penadez-Gonzales, Rafal Zdrzalek, Christopher Stevens, Peter van Esse, SarahM.SchmidtTheSainsburyLaboratory,Norwich,NR47UH,UKEmail:sarah.schmidt@tsl.ac.ukBananasarenotjustdeliciousfruits,butarethe4thmostimportantfoodcropintheworld.Inmanypartsoftheworldbananasarestaplecropsthatareimportantforfoodsecurity.Currently,thebiggestthreattoworldwidebananaproductionisFusariumwiltdisease,avasculardiseasecausedbythesoil-bornefungusFusariumoxysporumf.sp.cubense(Foc).Inthe1990sanewFocracecalledTropicalRace4(TR4)emergedinSouth-EastAsiathatisspreadingaroundtheglobe.Fusariumwiltcannotbecontrolledchemicallyandbreedingbananasisdifficultbecauseediblecultivarsaresterileanddonotproduceseeds.Inthe2Bladesgroup,Iamexploringnewroutesofengineeringresistanceinbananasthroughabetterunderstandingofthepathogenfocusingonthefollowingquestions;Whichvirulencegenesdoesitneedtoinfectbananas?ArethereresistancegenesknownagainstFusariumwiltdiseaseinotherplantspeciesthatwecanintroduceintobanana?Andfinally,canweuseantagonisticmicrobestocontrolFusariumwiltdiseaseofbananas?

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TheagronomicpotentialofgenesilencingapplicationAlineKoch1,DagmarBiedenkopf1,ElkeStein1,LukasJelonek2,AlexanderGoesmann2andKarl-HeinzKogel11Department of Phytopathology, Justus Liebig University, D-35390 Giessen,Germany;2DepartmentforBioinformaticsandSystemsBiology,JustusLiebigUniversity,D-35392Giessen,GermanyEmail:karl-heinz.kogel@agrar.uni-giessen.deNon-codingRNAshaveemergedasapowerfulgenetictoolinresearchandpracticaluseinmedicineandagriculture.Thedemonstrationthatagriculturalpests/pathogens,suchasinsects,nematodes,andfungi,arekilledbyexogenouslysuppliedRNAtargetingtheiressentialgeneshasraisedthepossibilitythatnon-codingRNAcanbedevelopedintoanovelclassofpesticides.Recently,weprovidedproof-of-conceptthatdeliveryofdsRNAtargetingthethreecytochromeP450lanosterolC-14α-demethylases(CYP51)genesofthenecrotrophicfungalpathogenFusariumgraminearumeffectivelycontrolsFusariumHeadBlightandRootRotinbarley.1,2Here,wewilldiscussthemodeofactionofdsRNA-mediatedFusariumgrowthinhibitionindifferentexperimentalset-upsincludinginvitroliquidfungalculture,insituleafsprayapplication,andtransgenicdelivery.OtherlabsalsorecentlyshowedthatsprayapplicationsofRNAtoplants,includingArabidopsis,tomatoandtobacco,israthereffectiveincontrollingplantdiseasescausedbymicrobialpathogensandviruses.3,4Moreover,optimizationofRNAdeliverybycertainnanostructure-basedformulationswasdemonstrated.4Basedonthosefindingsandnewunpublisheddatafromourlab,wewillreportonadvancesandsetbacksincontrollingotherfungalpathogensbydsRNAapplication.Altogetherwewillprovideinformationonafundamentallynewplantprotectionstrategy,therebyopeningnovelavenuesforimprovingcropyieldsinanenvironmentallyfriendlyandsustainablemanner.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)NaturePlants,doi:10.1038/nplants.2016.207Combining proteomics of haustoria and gene silencing to discover key players ofvirulenceandsusceptibilityduringbarleypowderymildewinfection.Kate Orman, Sebastien Lambertucci-Bonnet, Shaoli Das Gupta, Laurence V.Bindschedler1SchoolofBiologicalSciences,RoyalHollowayUniversityofLondon,Egham,TW200EX,UKEmail:Laurence.bindschedler@RHUL.ac.uk

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Cerealpowderymildews,Blumeriagraminis,arerecognisedasmajorcontributorsaffectingyieldandgrainqualityofwheatandbarley.Usingaproteomicsapproach,Blumeriaeffectorcandidates(BECs),someofwhicharealsodefinedasCandidatesecretedeffectorproteins(CSEPs),werespecificallyassociatedwithhaustoria(Bindschedleretal2009,2011,Spanu2010,Pliegoetal,2013).Haustoriaareatthecentreofthebiotrophicinteraction,developingintheplantepidermiswithoutdestroyingtheinvadedcells.Haustoriaaresurroundedbyanextra-haustorialmembrane(EHM),whichisincontinuumbutdistinctfromthehostplasmamembrane.BecauselittleisknownabouttheEHMcomposition,weinvestigatedtheEHMproteometounravelputativekeyplayersofsusceptibilityandresistance.SinceBlumeriaisanobligatepathogen,wehadtodeviseanewconvenient,cloning-andGM-freeRNAialternativemethodthatdeliverssmall,modifiedoligodeoxynucleotidestowholeleaves,tovalidateeffectorfunctionbytransienthostinducedgenesilencing.Themethodwasequallysuitableforthevalidationofknownsusceptibilitygenes.Usingthissilencingapproach,wechallengedtheroleofaPR5thaumatin-likeproteinisoformwhichinteractswiththeRNAse-likeeffector,BEC1054(Penningtonetal2016)andwhichislikelyassociatedwiththeEHM.Tooursurprise,weestablishedthatthisPR5actsasasusceptibilityfactor,sinceitssilencingledtodecreasedBlumeriainfection.ThisincreaseinhostresistancebytargetingPR5wasassociatedwithadecreasedamountofthecognatemRNAandanincreasedpercentageofinfectedepidermalcellsproducinghydrogenperoxide.Directandindirecteffectsoffungalendophytesonplant-parasiticnematodesAlexanderSchoutenLaboratory of Nematology, Experimental Plant Sciences, Wageningen University,Droevendaalsesteeg1,6708PBWageningen,TheNetherlandsEmail:sander.schouten@wur.nlAlthoughnotorious for theirpathogenicity, it isbecomingevident thatmostFusariumoxysporum isolates encountered in nature are harmless to plants, someofwhich canevenplayabeneficial roleas trueendophytes,meaning that theycancoloniseplantswithouteverprovokingadiseasephenotype,byantagonizingplant-parasiticnematodes.The involved mechanisms in this quantitative antagonism are only superficiallyunderstood. There is evidence that the presence of the endophyte increases plantvigour and induces plant defence responses toward the nematode. Direct negativeeffectsoftheendophyteonnematodeperformanceandbehaviourcannotberuledoutaswell.Metabolites,producedbythefungalnematodecontrolendophyte,F.oxysporum162(Fo162),wereisolatedandfullycharacterized.Oftheelevenresolvedcompounds,sixwereisolatedfromaFusariumspp.forthefirsttime.Toxicityassaysshowedthat,4-hydroxybenzoicacid(4-HBA),indole-3-aceticacid(IAA)andgibepyroneDwerethemostpotenttowardnematodes,beingalmostasgoodasthebestcommercialnematocide.

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IAAisawell-knownphytohormoneandknowntoplayaroleinplantdevelopmentandtriggeringdefences.4-HBAisreportedtoincreaseabioticstresstolerance.Thisfindingthussuggeststhatindividualmetabolites,producedbyFo162,canhaveadualactivity,i.e.inducingdefencemechanismsand/ortolerancetowardnematodesinplantsanddirectlykillingorcompromisingnematodes.Suchcompoundsmayserveasimportantleadsinthedevelopmentofsustainableandenvironmentalfriendlynematocides.ImprovingPlantResistancetoFungalPathogensbyCalloseModificationTobiasHanak1andChristianA.Voigt21MolecularPhytopathology,BiocentreKleinFlottbek,UniversityofHamburg,Hamburg,22609,Germany;2DepartmentofAnimalandPlantSciences,P3CentreforPlantProductionandProtection,UniversityofSheffield,Sheffield,S102TN,UnitedKingdomEmail:c.a.voigt@sheffield.ac.ukTheplantcellwalliscomplexpolymernetworkthatformstheouterbarrierofplanttissuestothesurroundingenvironment.Itsarchitectureisofgreatimportancetoconferresistancetoinvadingpathogens,whichareamajorthreatforcropyieldandfoodsecurity.Plantdefenceresponsesincludecellwalladaptionatsitesofpathogeninteractionwithadecisiveroleofthecellwallpolymercallose,a(1,3)-β-glucan.Ourrecentresultshaveproventhatmodificationandoptimizationofcallosebiosynthesisprovidesresistancetoinvadingpathogensthroughnetworkformationwiththepreexistingcellulosiccellwall,notinthemodelplantArabidopsisthalianabutalsoinBrachypodiumdistachyonandwheat.Theapplicationoflocalizationmicroscopy,thefastestevolvingtechniqueinsuper-resolutionfluorescencemicroscopy,allowedvisualizationofpolymerrearrangementsinnanoscale3Dnetworksandfacilitatedinsitustructuralanalysisofcallosesynthasecomplexesinlivecellimaging.Withthis,insitulivecellsuper-resolutionmicroscopyrepresentsanewtoolinbridgingthegapbetweenthemolecularandsystemslevelinplantpathology.Understandingthesubcellularandmolecularmechanismsofplantcellwallbiosynthesisandarchitecturealsoonastructurallevelwillleadtotheidentificationofkeymodulatorsandfactorsforcellwallengineeringtosuccessfullydesigncropswithimprovedpathogenresistance.Understanding the genetic control of pathogenicity and resistance for FusariumoxysporuminonionA. Taylor1, A.C. Jackson1, G. Teakle1, A.D. Armitage2, R. de Heer3, N. Zhang3, W. vanLeeuwen3,S.Ott4,L.Baxter4,B.Greenfield2,R.J.Harrison2andJ.P.Clarkson11WarwickCropCentre,UniversityofWarwick,Wellesbourne,Warwick,CV359EF,UK;2 NIAB-EMR, New Road, East Malling, Kent, ME19 6BJ, UK; 3 Hazera Seeds B.V.,Schanseind 27, 4921 PMMADE, The Netherlands; 4University ofWarwick, Coventry,CV47AL,UK.

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Email:andrew.taylor@warwick.ac.ukBulbonion(AlliumcepaL.)isagloballyimportantcropbutthelackofagenomesequencehashamperedgeneticresearch.Fusariumoxysporumisanimportantsoilbornepathogenofmanycropsandcomprisesdifferentformaespeciales(f.spp.)thatinfectparticularhosts.Inonion,F.oxysporumf.sp.cepae(FOC)isamajorconstrainttoworldwideproduction,principallycausingabasalrotofbulbswithestimatedlossesof£11MperannumintheUK.AsFOCproduceschlamydosporesthatsurviveinsoilformanyyears,diseasemanagementischallengingandthereforedevelopingresistantcultivarsishighlydesirable.Initially,onionlineswithhighlevelsofbasalrotresistancewereidentifiedwithinauniquediversityset.NewonionpopulationssegregatingforFOCresistancewerethendeveloped,genotypedusingKASPmarkersandQTLsidentifiedlinkedtoresistance.Associatedgeneticmarkersarenowbeingdevelopedtoacceleratebreedingofbasalrotresistantvarieties.Inparallelresearch,thegeneticbasisforpathogenicityinFOCwasinvestigatedandthepresenceofputativeeffectorsidentifiedbygenomesequencingandPCRusingarangeofpathogenicandnon-pathogenicF.oxysporumisolatesfromonion.Thepresenceof‘secretedinxylem’(SIX)effectorgenes,whichhavehomologsinotherF.oxysporumf.spp.,wasclearlyassociatedwithpathogenicityandthesegeneswerestronglyupregulatedduringinfection.EffectorgenecomplementsandsequencesmayalsoprovidethebasisforadiagnostictestforFOC.Knock-outmutantstrainshavenowbeendevelopedforseveraleffectorgenesasafirststeptowardfunctionalanalysis.Understanding appressoriumdevelopment inMagnaporthe oryzaeusing analysis ofnetwork-extractedontologiesandpathogentranscriptomics.RyanMAmesWellcomeTrustCentreforBiomedicalModellingandAnalysis,LivingSystemsInstitute,UniversityofExeter,StockerRoad,Exeter,EX44QD,UKEmail:r.ames@exeter.ac.ukMagnaportheoryzaeisthecausalagentofriceblastdisease,themostimportantinfectionofriceworldwide.Halftheworld’spopulationdependsonriceforitsprimarycaloricintakeand,assuch,riceblastposesaseriousthreattofoodsecurity.ThestagesofM.oryzaeinfectionarewelldefined,withtheformationofanappressorium,acelltypethatallowspenetrationoftheplantcuticle,particularlywellstudied.However,manyofthekeypathwaysandgenesinvolvedinthisdiseasestageareyettobeidentified.Inthiswork,Ihaveusednetwork-extractedontologies(NeXOs),hierarchicalstructuresinferredfromRNA-Seqdata,toidentifypathwaysinvolvedinappressoriumdevelopment,whichinturnhighlightsnovelgeneswithpotentialrolesinthisprocess.ThisstudyillustratestheuseofNeXOsforpathwayidentificationfromlarge-scalegenomicsdataandalsoidentifiesnovelgenesandpathwayswithpotentialrolesindisease.Themethodspresentedherewillbeusefultostudydiseaseprocessesinother

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pathogenicspeciesandthesedatarepresentpredictionsofnoveltargetsforinterventioninM.oryzae.

Integrated deployment of host resistance and fungicides: managing evolution ofvirulenceandinsensitivityFrankvandenBoschDepartment of crop protection, Rothamsted Research, Harpenden, AL5 2JQ, UnitedKingdomEmail:Frank.vandenbosch@rothamsted.ac.ukSustainablecontrolofplantpathogensismaintainedbytwohighlydynamicinteractions:firstlybetweentheintroductionofnewhostresistancegenesandpathogenpopulationsovercomingthem;secondlybetweentheintroductionofnewfungicidemodesofactionandpathogenpopulationsbecominginsensitivetothem.Virtuallyallstudiesonsustainablediseasecontrolstudyoneofthecontroloptionsexclusively.Thisissurprising,giventhewidelyheldbeliefthatintegratedcontrolwillleadtomoredurablecontrolthanrelianceononecontroloption.Inpracticebothdiseasecontroloptionsareintegrated,buttheeffectsofintegrationonthesustainabilityofdiseasecontrolarenotsufficientlyunderstoodtopredictwhichintegrationstrategieswillbemosteffective.Weintroduceagoverningprinciplethatpredictsthedirectionofselectionforfungicideinsensitivityandthedirectionofselectionforpathogenvirulenceunderarangeofdiseasemanagementoptions.Thepredictionsarecomparedtoexitingevidence.Thisleadsustoformulatetwostrategiesimprovingdurablediseasecontrol:

1. Deploymentoffungicidesreducesselectionforvirulence.2. Deploymentofcropresistancereducestheselectionforfungicide

insensitivity.Usingthismechanisticunderstandingabouttheeffectofintegrationthenleadsustohypothesisethatitismoresustainabletointegrateandbalancechemicalandgeneticcropprotection,thantoattempttoreplacechemicalwithgeneticcontrolorrelyentirelyonchemicalcontrol.Resultsofmodellingstudiesontwocontrastingpatho-systemswillbeusedhere(andinthepaperbyPaveley)toexplorethepracticalimpactofexploitingthemutualprotectionmechanism.Integrated deployment of host resistance and fungicides: economics, efficacy andevolutionNeilD.PaveleyADAS,HighMowthorpe,Duggleby,Malton,NorthYorkshire,YO178BP,UKEmail:neil.paveley@adas.co.uk

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Theaimofdiseasecontrolistoprotectmarketableyieldinordertominimisetheunitcostofcropproduction.Butcontrolbyhostresistanceandfungicidescreatesselectionpressureforpathogenevolutiontowardsvirulenceandfungicideinsensitivity.Thecommercialandsocietalneedistobalancebetweenachievingcost-effectivecontrolintheshorttermandmanagingpathogenevolutiontoensurecontrolisdurableinthelongterm.TheabilitytocontroldiseasesinsomemajorEuropeancropsistenuouslybalancedcurrently.FrankvandenBosch(theseproceedings)describeswhyintegratinggeneticandchemicalcontrolislikelytobemoredurablethaneitheralone.Farmuptakeofintegratedcontrolinmajorcropsdependslargelyontheshort-termeconomics(yieldvalueminusfungicidecosts)ofgrowingdiseaseresistantorsusceptible,butsometimeshigheryielding,cultivars.Thechallengeforpathologists,cropphysiologistsandplantbreedersistominimisethe‘yielddrag’,whichcanbeassociatedwithbreedingfordiseaseresistance,sothatresistantvarietiesaremoreeconomicallyattractive.Datafromfieldexperimentsandanalysisbymathematicalmodellingshowthat:(i)thecombinedefficacyofhostresistanceandfungicidesispredictableusingasimplemultiplicativesurvivalapproach,enablingthedesignofoptimalintegratedcontrolprogrammes,(ii)diseaseforecastingprovidesgreatereconomicbenefitsonpartiallydiseaseresistant,thansusceptible,varieties,and(iii)thecombinationofvarietyresistanceandforecastingtoguidefungicidetreatment,canreducetheselectionpressureforevolutionoffungicideinsensitivestrains,thusincreasinglongtermdurabilityofcontrol.Fenpicoxamid(InatreqTMactivefungicide),anewfungicidewithutility incereals forcontrolofZymoseptoriatritici(Septorialeafblotch)andotherdiseasesAndrewJ.Leader1andGregoryMKemmitt21Dow AgroSciences, CPC2 Capital Park, Cambridge, United Kingdom; 2 DowAgroSciences,3BParkSquare,MiltonPark,Abingdon,Oxfordshire,UnitedKingdomEmail:aleader@dow.comFenpicoxamid(Inatreq™Active)isthefirstmoleculefromanewclassoffungicides(picolinamides).FenpicoxamidisderivedfromanaturalcompoundUK-2AwhichisproducedbyfermentationofanActinomycete(Streptomycesspp.).UK-2AissubjecttominoralterationpostfermentationtoproduceFenpicoxamidandimpartadditionalstabilitytothemolecule.InatreqinhibitsfungalrespirationinthemitochondriaattheQiubiquinonereductasebindingsiteincomplexIII,whichisanewtargetsitewithinthecerealfungicidesegment.Mostimportantly,Inatreqshowsnocross-resistancetoanyexistingcerealfungicidechemistries,includingtriazoles,strobilurinsorSDHIsandwillbeanimportantnewtoolinthearmouryofcerealgrowers.IntheEuropeancerealsmarket,Zymoseptoriatritici(synonymMycosphaerellagraminicola,Septorialeafblotch),thepathogenofgreatestconcernintermsofyieldlossesinwinterwheatproduction,hasdevelopedwidespreadresistancetostrobilurinsclassandsignificantsensitivitylosstothetriazoles.Recentlyisolateswithreduced

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sensitivitytoSDHI’shavebeendetectedintheUKandIreland.Anadditionalconcernisthepossibilitythatsomemoleculesthatarewidelyusedtodaywillbeunderregulatorypressureandmayexitthemarketthusreducingthenumberofsolutionsavailabletogrowers.FenpicoxamidwillbeaninnovativeandeffectivetoolinhelpingfarmerswhofaceincreasedresistancethreatsfromZ.triticiwithexistingtechnologies.However,asfenpicoxamidisasinglesiteinhibitor,theproductshouldonlybeusedincombinationwithotheractivestominimizetheriskofresistancedevelopment.ControllingStrawberryPowderyMildewrequiresallyearroundvigilanceHall,A.M.,Jin,X.,Liu,B.BiologicalandEnvironmentalSciences,UniversityofHertfordshire,Hatfield,AL109ABEmail:a.m.hall@herts.ac.ukStrawberryPowderyMildewcausedbyPodosphearaaphanisisthemostfeareddiseaseofprotectedstrawberriesintheUK,causinglossesof20%to70%ofthecropperannum,whichresultsinthelossof£56.8millionwithonly20%loss.WorkattheUniversityofHertfordshiresince2003/4hasresultedinthedevelopmentofanallyearroundcontrolstrategy,which,iffollowedbygrowers,canresultinsatisfactorydiseasecontrol.Commercially,therearemanydifferentgrowingsystemsforstrawberries;insomecasestheplantsare‘perennial’,plantedfor3wintersand4harvests,inothercasestheyareplantedinFebruary/MarchandgrownuntilSeptember/October.Inthecaseofoverwinteredcrops,thediseasecanoverwinteraschasmothecia,whereasplantsarrivingfromthepropagatorsarerarelyclean.Controllingthisinitialinoculumisvital.ThereisheavydiseasepressureinJuly,AugustandSeptember,buttheepidemicscanbereducedbya)theuseofasiliconnutrientandb)theuseofapredictionsystembasedondisease-conduciveconditionsoftemperatureandhumidity.ChasmotheciastartformingonleavesinAugustandSeptemberandgiverisetotheoverwinteringinoculumontheperennialcrops.Thesecanbecontrolledbyusingafungicideafterharvesthasfinished,andagaininthespring,asthecropiscoveredwithfleece.Theresearchreportedherehasresultedinanupdatedfactsheetforgrowers,whichincludesanintegratedprogrammeforallyearrounddiseasecontrol.

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ClosingPlenaryTalksTheScienceofQuornRobJohnsonScienceManager-FermentationSpecialist,Quorn,StationRoad,Stokesley,NorthYorks,TS97BR,UKEmail:rob.johnson@quornfoods.comQuornhasamere50yearhistoryandisthefirstnewfoodtypetoenterthemarketsincethepotato.ThistalkwillgiveanoverviewofthehistoryofQuorn,itsrelevancetothemodernworldandmoveontothesciencebehindtheproduct,includinghowitismade,wherethetexturecomesfrom,andwhatchallengesarebeingaddressedforthefuture.Coveringgenetics,fermentation,environmentalimpactandhealthbenefitsanoverviewofoneofthefastestgrowingfoodproductsintheworldwillbeoutlined.FungalweaponstomanipulateplantsNataliaRequena,MeikeHartmann,RubenBetz,SvenHeidtandCarolinHeckMolecularPhytopathologydepartment,KarlsruheInstituteofTechnology,Karlsruhe,76228,GermanyEmail:Natalia.requena@kit.eduThearbuscularmycorrhiza(AM)symbiosisbetweenGlomeromycotinafungiandmostplantrootsisoneofthemostancientandwidespreadsymbiosesonearth.AMfungiprovidephosphatetotheirhostsinexchangeforcarbonandtheestablishmentandmaintenanceofthesymbiosisrequiresacomplexcommunicationexchangebetweensymbiontsandwiththeenvironment,becausehighconcentrationsofPiinhibitthesymbiosis.Interestingly,AMfungiareabletocolonizethemajorityoflandplantsincontrasttomostpathogenicfungithathaveaquitenarrowhostrange.WehypothesizethatAMfungiemployconservedeffectorproteinstorewiretheplantphysiology.WepreviouslyidentifiedtheeffectorproteinSP7andagroupofrelatedproteins(SP2,SP5andSP31)fromRhizophagusirregularis.Allmembersofthisfamilyshowthesamebasicproteinstructurewithtandemhydrophilicandimperfectrepeatsandanuclearlocalizationsignal.Hereweshowthatallthreeeffectorsredundantlymodifytheplantphosphatestarvationresponse(PSR).Expressioninrootsofanyofthemleadstoaccumulationofpre-miRNA399sinleavesthatultimatelydownregulatesubiquitinE2conjugasePHO2inrootscausingincreasedPSR.Weproposethatthismechanismallowsthefungustocolonizetherootsystemfurtherdespiteanet-increaseinphosphate.InteractionandlocalizationassayssuggestthattheseeffectorsmodulatethePSRbyintersectingwiththeplantmRNAtranslationmachinery.Theseresultshighlightthefinesseofsymbioticarbuscularmycorrhizalfungitomanipulatetheplantcellprogram,

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guaranteeingtheirpermanenceintherootwhilecontributingtoplanthealthandnutrition.

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POSTERS

*JColhounCompetitionentrant**BMSStudentPosterCompetitionentrant

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Effects of Leaf Extracts ofOcimum gratissimum L. on Quality of Fresh Cut CucumissativusL.OluwagbengaO.Adeogun,AlfredMaroyiandAnthonyJ.AfolayanDepartmentofBotany,UniversityofFortHare,Alice,EasternCape,SouthAfricaEmail:adeogunoluwagbenga@gmail.com;oadeogun@ufh.ac.zaHighmoisturecontentandminimalprocessingoffruitsandvegetablesencouragemicrobialincursionandtheseraisedtheneedforpreservation.Ocimumgratissimumleaveswereextractedwithethanolandhexaneseparately.TheextractswereusedtoenhancethequalityoffreshcutCucumissativusstoredfor9daysusingdippingmethod.QualitativeandquantitativephytochemicalanalysesoftheextractsweredeterminedandessentialoilconstituentswereestablishedusingGC-MS.TheresultsshowedthatthemoisturecontentsofC.sativuswithextractswerelowercomparedtostoredC.sativuswithoutextracts.TheacidityofC.sativuswithextractswaslowerinrelationtothetestsamplewithoutextracts.TheincreaseintotalsolublesolidwashigherinC.sativuswithouttheextracts.Theascorbicacid,carotenoidsandphenoliccontentsofstoredC.sativuswithextractshadlowerrangeofreductioncomparedtoC.sativuswithouttheextracts.ThedifferencesinpotentialbrowningandturbidityofstoredC.sativuswithextractsduringstoragewerenotobvious.TherewerehigherloadsofmicrobesinC.sativuswithoutextractscomparedtoC.sativusstoredwithextracts.Theresultofthephytochemicalscreeningshowedthepresenceofflavonoid,alkaloid,cardiacglycoside,phenol,tannin,steroid,anthraquinoneandsaponinintheleaf.Someoftheessentialoilconstituentsareγ-Terpinene,Caryophyllene,Oleicacidand(E)-9-Octadecenoicacid.ThisstudywasabletoestablishthattheleafextractsofO.gratissimumhavebio-preservationpotentialthatcanbeusedtoenhancetheshelfofC.sativus.IntegratingfungicideswithvarietalresistanceagainstSeptoriatriticiblotchinwheatOlubukola.O.Ajigboye,DimitraJ.Angelopoulou,JohnFoulkes,Rumiana.V.RaySchool of Biosciences, University of Nottingham, Sutton Bonington Campus,Loughborough,Leicestershire,LE125RD,UnitedKingdomEmail:bukky.ajigboye@nottingham.ac.ukSeptoriatriticiblotch(STB),causedbytheascomycetefungusZymoseptoriatritici,isthemostimportantfoliardiseaseofwheat,reducinggrainyieldbyupto50%.Symptomsoftenoccurfollowingalatentphasebetween5-10daysbeyondwhichfungicideeffectivenessisimpaired.Varietalresistanceisnotyetfullyutilisedinfieldalthoughtherearenumberofcommerciallyavailablewheatcultivarswithmoderatetohighfieldresistance.IntegratingpreventativefungicideapplicationswithvarietalresistanceisessentialtooptimizecontrolofSTB.TheobjectivesofthepresentstudyweretodeterminethebesttimingandfungicideagainstSTBfortwodifferentwheatvarieties,onesusceptiblecv.Dickensandtheresistantcv.Cougar.Fieldexperimentswere

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conductedduringtwoseasons(2016/17)usingnaturalinfectionbyZ.tritici.Combinationsoffungicidesincludingmulti-site,demethylationandsuccinatedehydrogenaseinhibitorswereappliedatdifferentcropgrowthstages(GS)30,31/32,33,37/39or59/61.TheeffectoftimingandfungicidetreatmentontheprogressandseverityofSTBinthecanopywereassessedatGS31,61and75whilefungalbiomassofZ.triticiwasquantifiedinasymptomaticflagleavesatGS61.Theseparametersalongwiththecropphotosyntheticefficiency,biomassandfinalgrainyieldwereusedtoidentifythebeststrategiesforintegratingfungicidetiming,modeofactionandvarietalresistance.AcknowledgmentsThisstudyispartofcollaborationworkbetweentheUniversityofNottingham,AgriiandRAGTSeeds,co-fundedbyInnovateUK.*PowderyMildew:ATaleofTwoFungicidesCorinneJ.Arnold,DianeG.O.SaundersandJamesK.M.Brown.CropGenetics,JohnInnesCentre,Norwich,NR47UH,UKEmail:Corinne.Arnold@jic.ac.ukBlumeriagraminisisapowderymildewpathogenthatinfectsmanywildgrassesandcerealsacrosstheworld.Ifleftuncontrolled,itcancausesignificantyieldlossesincereals.OnemainmethodforcontrolistheapplicationoffungicidesbutresistancetostrobilurinandoldertriazolefungicidesalreadyoccursinB.graminis,andfewotherfungicidesareavailableformildewcontrol.IhavecharacterisedisolatesfromanoutbreakofB.graminisf.sp.tritici(Bgt)onwheatwheretwoofthemajorfungicidesusedagainstBgtwereineffective:cyflufenamid(Cyflamid),aneradicantfungicidewithanunknownmodeofaction,andfenpropimorph(Corbel),amorpholinefungicidetargetingasterolreductase.AllBgtisolatesfromthestudysitewerecompletelyresistanttoCyflamidsprayedattherecommendedfieldrate.However,overthefollowingtwoyears,theBgtpopulationrecoveredsensitivitytoCyflamid,suggestingthatresistancemayhaveasignificantfitnesspenalty.Additionally,followingtwospraysofCorbel,theseBgtisolateshadsignificantlylowersensitivitytofenpropimorphthanfieldisolates.Overuseoffenpropimorphwouldalmostcertainlyleadtostrongresistanceinmildew,soitshouldbeusedaspartofarotationoffungicideswithdifferentmodesofaction.Likewise,Cyflamidcancontinuetobeusedaspartofanintegrateddiseasecontrolprogramme.Geneticandgenomicanalysisisinprogress,withtheaimofidentifyingcandidategenesforresistancetocyflufenamidandtofenpropimorph.

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**Use of bulk segregant analysis to analyse azole drug resistance in AspergillusfumigatusGeorgeD.Ashton1,CélineM.O'Gorman1,DanielZadik2,MartinBlythe2,VictoriaWright2,SimonV.Avery1andPaulS.Dyer11School of Life Sciences, University of Nottingham, Nottingham, NG7 2UH, UnitedKingdom;2DEEPSeq facility, University of Nottingham, Nottingham, NG7 2UH, UnitedKingdomEmail:stxgdas@nottingham.ac.ukAspergillusfumigatusisanopportunistichumanfungalpathogencausingaspergilliosis.Infectionsarecontrolledthroughtheuseofazoleantifungaldrugs.Overthepast20yearstherehasbeenanincreaseinoccurrenceofazoleresistantstrains.Resistanceisduetoavarietyofdifferentgeneticmechanisms,someofwhichhavebeencharacterizedinclinicalandenvironmentalisolates.A.fumigatuswasoncethoughttoreproduceentirelyasexually.However,asexualcyclewasdiscoveredin2009.Thefungalsexualcycleisapowerfultoolandcanbeexploitedinanumberofdifferentareas.ItwashypothesizedthesexualcycleinA.fumigatusmightbeusedtoidentifyazoleresistancegenesusingthetechnique‘bulksegregantanalysis’(BSA)incaseswherethemechanismofresistanceisunknown.TheprocessofBSAinvolvesasexualcrossbeingperformedbetweenanazoleresistantandazolesensitivestrain.Followingthis,aseriesofbackcrossesbetweentheresistantprogenyandsensitiveparentareperformedtominimizegeneticdifferencesbetweenthem,withtheazoleresistancegene(s)andanylinkedgenesbeingtheonlydifferingregions.GenomesequencingisthenperformedontwopoolsofDNA:oneofprogenyexhibitingazoleresistance;andoneshowingazolesensitivity,aswellasgenomesequencingofthesensitiveparent.Fromthis,theaimistoidentifykeyareasandgenesthatmaybecausingazoleresistanceintheseisolates.Thismayleadtothedevelopmentofusefuldiagnosticstomonitorazoleresistanceandimprovethetreatmentofazole-resistantA.fumigatusinfections.*An investigation into silicon localization in strawberryplants linked to reduction indiseasesusceptibilitytoPodosphaeraaphanisC.I.Asiana,A.M.HallandK.DaviesDepartment of Biological and Environmental Sciences, School of Life and MedicalSciences,UniversityofHertfordshire,Hatfield,AL109AB,UKEmail:i.asiana@herts.ac.ukThemostimportantdiseaseofprotectedstrawberriesintheUKisstrawberrypowderymildewcausedbyPodosphaeraaphanis,whichhastobecontrolledbyfrequentuseoffungicides.Siliconisnotconsideredasessentialfortheplant;however,workcarriedoutattheUniversityofHertfordshirehasshownthatweeklyuseofasiliconnutrientinfertigationtubesatacommercialstrawberryfarmresultsinreducedsusceptibilitytothisdisease.Siliconcanonlybetakenupinabioavailableformandthenutrientusedin

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thisexperimentisabioavailableformofsilicon.Asiliconlocalizationexperimentwasconducted.Siliconwasappliedviathesoilintheglasshouse,thefertigationtubesatthecommercialfarmandwassprayedoninafieldplot.Inthisstudy,crosssectionsofleaves,petiolesandrootswerestainedwithafluorescencedye,“lysotrackeryellowHCK-123”.Sampleswereexaminedusingaconfocalmicroscopeatx400andawavelength450nm.Sofar,resultsshowedintheleaf,siliconwasfoundinthecuticle,epidermis,palisadelayer,stomataandvasculartissue.Inthepetiole,siliconwasfoundintheepidermisandxylemandintheroots,thesiliconwasfoundintheepidermis,endodermisandvasculartissue.Thefluorescenceintensityofthecrosssectionswasquantifiedandthisshowsthatthesiliconwas5timeshigherinthetreatedplantsthantheuntreated.Inaddition,thesiliconfertigationfieldtrialhasshownthatplantswithhigherlevelsofsiliconarelesssusceptibletothedisease.Chrysanthemum ChloroticMottle Viroidmediated Trafficking of ForeignmRNA intoChloroplastsEseulBaek1,MinjuPark1,Ju-YeonYoon2andPeterPalukaitis11Dept. of Horticultural Sciences, SeoulWomen’sUniversity, Seoul, 01797, Republic ofKorea; 2National Institute of Horticultural & Herbal Science, RDA, Wanju, 55365,RepublicofKoreaEmail:eseul@swu.ac.krChrysanthemumchloroticmottleviroid(CChMVd),amemberofthefamilyAvsunviroidae,wasidentifiedinchrysanthemumplantscausingchloroticleafsymptoms.ToshowthatCChMVdcantrafficintochloroplasts,whichisthoughttobethesiteofitsreplication,CChMVdwasfusedtotheleadersequenceofareportergeneencodingmonomerredfluorescenceprotein(mRFP)andexpressedtransientlyinagroinfiltratedNicotianabenthamiana.FluorescencefrommRFPwasobservedinchloroplastsbyconfocalmicroscopy,butonlyiftheviroidtranscriptionfusionswerepresent.Theviroidcomponentsofthefusionswerefull-length400-ntCChMVd,oroneoftwopartialfragments(nucleotides125to2and231to372).ThemRFPanditsmRNAweredetectedbywesternblottingandRT-PCR,respectively,inwholetissueextractsofplantsinfiltratedbyeachfusionconstruct.IsolatedchloroplastswereshownbyRT-PCRtocontaintheRNAsequencesofbothCChMVdandmRFP,ifbothwerepresent,butnotthemRFPsequenceintheabsenceoftheviroidsequences.Eggplantlatentviroid(ELVd),inthesamefamilyAvsunviroidae,hasbeenshowntotrafficmRNAintochloroplastsforexpressionofafluorescentmarkerproteininpreviousstudies(1-3).SincetherearedifferencesinsequencebetweenELVdandCChMVd,itsuggeststhattheabilityofbothRNAstotrafficRNAsintochloroplastswasprobablyduetoanRNAstructure,andnotaparticularsequence.1.Gómez,G.,Pallás,V.(2010).PLoSOne5(8):e12269.2.Gómez,G.,Pallás,V.(2010).PlantSignl.Behav.5(11):1517-1519.3.Gómez,G.,Pallás,V.(2012).PlantPhysiol.159:558-564.

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WorkwasfundedbytheRuralDevelopmentAgencyoftheRepublicofKorea.Generation ofwheat plasmamembrane reporter lines to further understand fungalinfectionmechanisms.Laura Baggaley1, Alison Huttly1, Caroline Sparks1, Smita Kurup1, Christine Faulkner2,MichaelDeeks3andKimHammond-Kosack11 Rothamsted Research, Harpenden, AL5 2JQ, UK; 2 John Innes Centre, Norwich, NR47UH,UK;3UniversityofExeter,Exeter,EX44QD,UKEmail:laura.baggaley@rothamsted.ac.ukThefungus,Fusariumgraminearum,istheprimarycauseoffloralcerealinfectionsworldwide,resultinginreductionsinwheatgrainquality,quantityandadverselyaffectinghuman,animalandecosystemhealth.HowF.graminearumestablishesinfectionwithinwheathasbeenwellstudied,howeverwhatcausesthetransitionbetweentheasymptomaticandsymptomaticphasesremainsunclear.Thistransitionininfectioncorrespondswithashiftfromprimarilyinter-tointracellularfungalgrowth,leadingtoextensivecelldeath.Ithasbeenhypothesisedthatthischangeinfungalgrowthisduetotheutilisationofplasmodesmata–thinstreamsofcytoplasmbetweenadjacentplantcellwallsvitalforcell-to-cellcommunication.IdentificationofthecellularandmolecularmechanismsinvolvedintheF.graminearum-wheatfloralinteractionhaspreviouslyrelieduponhistologicalstainingandfungalreporterstrains[1].Wehaveattemptedtocreatetwowheatplasmamembranereporterlines,usingthegenesROP7[2]andLTI6b,toallowforamoreefficient,detailed,spatio-temporalstudy,oftheinfectionprocess.Duetotheautofluorescentnatureofwheattissues,tissuespecificautofluorescentprofileshavealsobeenestablishedusingconfocalmicroscopy,toaidimagecaptureandanalysis.Thisnewinformationwillbecriticalfortheevaluationofthewheatplasmamembranereporterlinesandfungalreporterstrains.Togetherthesenewtoolsanddatasetswillallowfortheidentificationandcharacterisationofinfectionmechanismsinwheat,allowingforagreaterunderstandingofmultiplecrop-yieldlimitingpathogens.[1]Brownetal.(2010)FungalBiology114:555-571[2]Kirienkoetal.(2012)PlantPhysiology159:1309-1318

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Howtocontrolleafdiseasesoffababeans?BirutaBankina,GunitaBimšteineandArtursKatamadzeInstitute of Soil and Plant Sciences, Latvia University of Agriculture, Jelgava, LV 3001,LatviaEmail:Biruta.Bankina@llu.lvFababean(ViciafabaL.varminor)hasbecomeanimportantfieldcropinLatviaduringthelastfiveyears.FourBotrytisspeciesasthecausalagentsofbeandiseaseshavebeendetermined:B.fabae,B.fabiopsis,B.cinerea,andB.pseudocinerea.Also,severalspeciesofAlternariaandStemphyliumhavebeenfound.Theaimofthepresentstudywastoclarifytheefficacyoffungicideapplication.TrialswereconductedattheResearchandStudyfarm“Peterlauki”oftheLatviaUniversityofAgriculturein2015–2016.Athree-factorfour-replicatetrialwasestablished:factorA–variety(‘Laura’,‘Boxer’,‘Isabell’);factorB–sowingrate(30,40,50germinatingseedsperm2);andfactorC–fungicide(boskalid267.0gkg-1andpyraclostrobin67.0gkg-1)0.75kgha-1atthebeginningofflowering.Diseaseassessmentwasstartedwhenthefirstsymptomsoccurred:20randomplantswerechosenineachplot,anddiseaseseveritywasnoted(0–9pointsscale).TotalimpactofleafdiseasesduringvegetationwasevaluatedbycalculatingtheAUDPC(areaunderdiseaseprogresscurve).Fungicideapplicationsignificantly(P<0.0001)decreasedthediseaselevel:efficacyofchocolatespotcontrolwas21.3%in2015and31.7%in2016,andefficacyofAlternarialeafblotchcontrolwas40.7%and35.3%respectively.Furtherinvestigationsarenecessarytoessentiallyimprovetheefficacyoffungicideapplication.SupportedbytheStateResearchProgramme“AgriculturalResourcesforSustainableProductionofQualitativeandHealthyFoodsinLatvia”:project“Sustainableuseofsoilresourcesandabatementoffertilisationrisks”.HostandnonhostsinteractionsofpowderymildewswithvariouscropplantsBalázsBarna,CsillaJuhász,AdélNémethandGáborGullnerPlant Protection Institute, Centre for Agricultural Research, Hungarian Academy ofSciences,Budapest,P.O.box102,HungaryEmail:barna.balazs@agrar.mta.huPowderymildewsareoneofthemostdevastatingpathogensofcropsallovertheworld.Triticale,anartificialhybridofwheatandrye,wasresistanttopowderymildewuntil2001,butsincethenthisfungusbecomeaserioustriticalediseaseinEurope.RecentlyitwasfoundthatBlumeriagraminisf.sp.triticaleisahybridbetweenwheatandryepowderymildews.Noneof11testedtriticalegenotypesshowedsusceptibilitytoourwheatpowderymildewisolate.Inordertogetinformationontheroleofproteinkinasesandphosphatasesonmildewdevelopmenttheeffectofpre-treatmentofhostplantswiththeinhibitorsstaurosporineorokadaicacidwasevaluated.Furthermore,the

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electrophoreticribonuclease(RNase)enzymepatternsofbarleyandwheatwereverydifferentfromeachotherandfromBrachypodiumdistachyonplantextractsthatshowedhighRNaseactivity.Allinvestigatedhostandnon-hostplantspeciesreactedwithanincreaseofRNaseactivitytopowderymildeworrustinfections.Wefoundthatheatpre-pretreatmentincreasedboththesusceptibilityofbarleytopowderymildewanditsRNaseactivity,whichconfirmsthepossibleroleofRNasesinsusceptibility.InteractionsofGolovinomycesorontiiwithvarioustobaccolinesinducedastrongincreaseofRNaseenzymeactivity,andanewRNasebandappearedintheextractsfromallinfectedtobaccosfollowingPAGEandspecificstaining.Interestingly,powderymildewinfectionsup-regulatedtheexpressionofPR-1b,PR-1candWRKY12genesinallsusceptibletobaccolinestovariousextents.Thepossibleroleoftheabovechangesinplant-powderymildewinteractionisdiscussed.Phenotypic, comparative genomic and transcriptomic analysis of recently collectedEuropeanZymoseptoriatriticistrains.HongxinChen1,CarlosBayon1,RobertKing2, StefanoTorriani3, StephaneBieri3,KostyaKanyuka1,KimE.Hammond-Kosack1andJasonJ.Rudd11DepartmentofBiointeractionsandCropProtectionand2DepartmentofComputationaland Analytical Sciences, Rothamsted Research, Harpenden, Hertfordshire AL5 2JQ,United Kingdom; 3Syngenta Crop Protection AG, Schaffhauserstrasse, CH-4332 Stein,SwitzerlandEmail:hongxin.chen@rothamsted.ac.ukSeptorialeafblotch(STB),causedbytheascomycetefungusZymoseptoriatritici,isoneofthemostimportantfoliardiseasesofwheat,especiallyinEuropeandUSA.Duetothepoorgeneticbasedresistanceofwheattothispathogen,extensiveapplicationoffungicidesiscurrentlythemostcommonwaytocontrolSTB.ThishasledtorapidevolutionoffungicideresistanceinmanyZ.triticipopulations.ToascertainthevariabilityinvirulenceprofilesofacollectionofrecentisolatesofZ.triticicollectedfromEuropeanfieldsites,42strainswereinoculatedonto21wheatcultivars,alongwiththereferencestrainIPO323asacomparison.Diseaseprogresswasmonitoreddailyanddateswererecordedwhen1stdiseasesymptomappearedandfullnecrosisdeveloped.Fungalsporulationwasalsomeasuredforeachinteraction.Theresultsshowdiversevirulenceprofilesofdifferentisolatesandarangeofinteractionoutcomes.ManyofthetestedisolatesdisplayedenhancedvirulencerelativetoIPO323.Noobviouscorrelationwasfoundbetweenvirulenceandsporulation,indicatingthesetwotraitsmightbeunderthecontrolofdifferentgeneticloci.Tobetterunderstandthegeneticbasisofdifferentialvirulence,genomicDNAofthetop12mostvirulentisolateswillbesubjectedtofullgenomesequencing,andRNAseqbasedgeneexpressionanalysisofinvitrocultures,andinplantasamples,willbeperformed.

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Definingtheimpactofautophagyonthehost-pathogeninteractionofwheatandthefungusZymoseptoriatriticiHarryT.Child1,StevenBates1,JasonJ.Rudd2andKenHaynes11Biosciences, University of Exeter, Exeter, EX4 4QD, United Kingdom; 2Department ofBiointeractionsandCropProtection,RothamstedResearch,Harpenden,AL52JQ,UnitedKingdomEmail:hc515@exeter.ac.ukThefungalwheatpathogenZymoseptoriatriticicausesmajorcroplossesandcurrentlydemands€1billionworthoffungicidetocontrolinEuropeannually.TheinfectioncycleofZ.triticidisplaystwodistinctphases,beginningwithanextendedsymptomlessphaseofatleastaweek,beforethefungusinduceshostcelldeathandtissuecollapseintheleaf.Recentevidencesuggeststhatthefungususeslittlehost-derivednutritionduringtheearlystagesofinfection,insteadrelyingonmacromoleculeswithinthefungalspore.WehypothesisethatZ.triticiutilisestheself-degradativeprocessofautophagytorecyclestoredcellularresourcestosupportestablishmentofleafcolonisation.Usingmoleculartechniques,thisresearchwillassesstheimportanceoftheautophagypathwayduringZ.triticicellulardifferentiationandinfectionofwheat.Manipulatingthegenesinvolvedinautophagywillrevealitsimportancetofungalvirulence,whilefluorescentlytaggingkeyautophagyproteinswillvisualisethespatialandtemporaloccurrenceofautophagicprocessesinvivo.ThisresearchwillprovidefundamentalknowledgeofthemolecularmechanismsofZ.triticiinfectionaswellasidentifyingpotentialnewtargetsforcontrolofthisdevastatingdisease.*Resistance to Rhynchosporium: Characterisation and identification of candidategenesMaxCoulter1,MarkLooseley1,RobbieWaugh2andAnnaAvrova11TheJamesHuttonInstitute,Invergowrie,Dundee,UK;2UniversityofDundee,Dundee,UKEmail:Max.Coulter@hutton.ac.ukRhynchosporiumcommune,causingleafscald,isoneofthemostdestructivepathogensofbarleyworldwide,leadingtoyieldlossesofupto30-40%.Despiteadvancesinmolecularmarkertechnologyandthesequencingofthebarleygenome,understandingofbarleyresistancetoRhynchosporiumatthemolecularlevelislimited.TheSpanishlandraceCIho3515ishighlyresistanttoscald,containingresistancegenesRrs1andRrs18.Rrs1wasthefirstRgeneagainstRhynchosporiumtobeidentified,whileRrs18hasonlyrecentlybeencharacterised.Rrs18hasalsobeenidentifiedinaSteptoexMorexDHpopulation,andalargeSteptoexMorexBC1populationhasbeenusedtofinemapthegenetoa650Kbintervalon6HS,identifying17candidategenes.ToidentifyacandidateforRrs1,AssociationmappingusingelitespringbarleycultivarsandSyrianandJordanianlandraceshasnarrowedtheRrs1mappingintervaltoasinglegene

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on3H,aWallassociatedKinase(WAK).AnalysisofexomecapturedataformultiplecultivarscarryingRrs1showedevidenceofaWAKparaloguenotpresentinthesusceptibleMorexreferencesequence.PCRandSangersequencingwascarriedouttodecipherthesequenceoftheWAKparaloguewiththeaimofcloningRrs1.CloningofbothRrs18andRrs1willfurtherourunderstandingofRhynchosporiumresistanceandenablediagnosticmarkerstobedevelopedhelpingbreederstopyramidbothgenestocreatedurableresistancetoRhynchosporiuminbarley.*Isolation and characterisationof a chrysovirus fromDothistroma septosporum, thecausativeagentofpineneedleblightJohnDaudu1,UnnatiShah1,Kotta-LoizouIoly2andRobertH.A.Coutts11Department of Biological and Environmental Sciences, University of Hertfordshire,Hatfield,AL109AB,UK;2DepartmentofLifeSciences,ImperialCollegeLondon,London,SW72AZ,UKEmail:j.o.daudu@herts.ac.ukDothistromaneedleblightcausedbyDothistromaseptosporumhasemergedintheBritishIslesasamajorthreattoCorsicanpine,lodgepolepineandScotspine.Thereisincreasingevidencethatmycovirusescanreducethegrowthandpathogenicityoffungalplantpathogens.Forinstance,avirulence-attenuatingRNAmolecule,Cryphonectriahypovirus1(CHV1)hasbeendescribedandsuccessfullyusedasabiologicalcontrolagentofCryphonectriaparasitica.Theaimofthepresentstudyistoisolateandcharacterisedouble-stranded(ds)RNAvirusespresentinacollectionofD.septosporumisolates.Tothisend22D.septosporumisolateswerescreenedusingarapidsmall-scaledsRNAextractionprocessandonlyonewasfoundtoharbourdsRNAelements.Subsequentlytheseelementswereclonedandsequenced,usingtargetedsequencingandRNAligase-mediatedrapidamplificationofcDNAends(RLM-RACE).ComputationalanalysisofthesequencesobtainedrevealedtheirsimilaritytomembersoftheestablishedfamilyChrysoviridae,whichhave4genomicsegmentsencapsidatedinisometricparticlesandareknowntoelicithypovirulenceintheirhosts.Generationofisogeniclinesofvirus-freeandvirus-infectedisolatesisinprogresstodetermineanyeffectsofthemycovirusonfungalfitnessandpathogenicity;thepathogenicityofthevirus-infectedisolatewillbeassessedincomparisontothevirus-freeone,inordertodetectputativehypovirulence,acommonfeaturenotedformycoviruses,whichmightbeusedforbiologicalcontroltoinvasionbymoreaggressivestrainsofD.septosporum.Inconclusion,thisstudyreportsthefirstmycoviruseverfoundinD.septosporum.

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Anovelmodeofactioninoomyceteplantpathogencontrol:ORONDISTM

JudithSheldon,JillFoundling,MathiasBlum,SianDeller,DavidBeattieandMikeCsukaiSyngenta, Jealott’s Hill International Research Centre, Bracknell, Berkshire. RG42 6EY.UKEmail:sian.deller@syngenta.comOxathiapiprolin,discoveredbyDuPont,isthelatestoomycete-activecompoundwithanovelmodeofactiontoenterthemarket,andismarketedbySyngentasince2016asORONDISTM.DuPontinitiallyidentifiedthemoleculartargetofoxathiapiprolin.Thisreportconfirmsthatfindingbycharacterisingforwardgeneticsmutantswithanoxathiapiprolin-resistantphenotype.Non-synonymouspointmutationswerefoundinallresistantlabstrainsintheoxysterolbindingprotein-encodinggene(OSBP).OSBPisthoughttobeinvolvedinthemovementofsterolsandlipidsbetweenmembranes,maintaininglipidhomeostasis.Downstreamperturbationscausedbydisruptionofthisfunctionultimatelykillthecell.Toinvestigatethishypothesisintheoomycetepathogen,Phytophthorainfestans,wehaveusedmicroscopy-basedtechniquestoexaminetheultrastructureofgermlingstreatedwithsublethaldosesofoxathiapiprolinandcomparedthesetocontrolgermlings.ObservationsofdifferentialcytologywillprovideinsightstobetterunderstandtheroleofOSBPandthewayinwhichitsinhibitionbyoxathiapiprolincontrolsoomyceteplantpathogens.Anti-protozoal agents identified in the Pathogen Box can inhibit Aspergillus fumigatusgrowthThailaFernandadosReisandGustavoH.GoldmanFaculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo,RibeirãoPreto,Brazil.E-mail:thailaf@hotmail.com

Aspergillusfumigatusisanopportunistichumanfungalpathogenandallergenresponsibleforinvasiveaspergillosis,animportantinfectionwhichcausesahighlevelofmortalityinimmunocompromisedpatients.Althoughtheaspergillosisismainlytreatedbyadministrationoftriazoles,epidemiologicalstudieshaveindicatedthattheemergenceofazole-resistantA.fumigatusisolateshasincreasedsignificantlyoverthelastdecade.Despitethegrowingneeds,treatmentsforinvasivefungalinfectionsremainunsatisfactoryandthediscoveryofnovelantifungalsdrugsishighlydesirable.ThePathogenBox(https://www.pathogenbox.org/)isalibrarycomposedby400syntheticchemicalsmoleculeswithactivityagainstdifferentpathogensresponsibleforimportantdiseasessuchasmalaria,tuberculosis,toxoplasmosis,andothers.AimingtodiscovernewpotentialagentsagainstaspergillosiswescreenedthePathogenBoxchemicalcompoundsforantifungalactivityagainstA.fumigatus.Weidentifiedmiltefosine(MMV688990)asanantifungicidalagentagainstaspergillosis.Usinginvitroassays,wedemonstratedthatthiscompoundhasaMIC(MinimalInhibitoryConcentration)of10

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µM.Interestingly,anullmutantinthecAMPdependentproteinkinaseSchaSCH9hasaMICof20uM.Thisproteinkinaseinfluencessphingolipidsbiosynthesis,suggestingthatmiltefosinecouldactinhibitingthefungalgrowthviatargetingthelipids/sphingolipidssynthesis.Additionally,iodoquinol(MMV002817)showedanantifungistaticactionwithaMICof2µM.Basedonourresults,weproposethatthesetwocompoundscouldrepresentpotentialcandidatesforfurtherantifungaldrugdevelopmentagainstinvasiveaspergillosis.Financialsupport:FAPESPandCNPq,BrazilFunctional characterization of the Aspergillus nidulans non-essentialproteinkinasesinvolvedintheregulationofsorbicacidantifungalpropertiesThailaFernandadosReis1,IveyGeoghegan2,SimonAvery2andGustavoH.Goldman11Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo,RibeirãoPreto,Brazil;2SchoolofLifeSciences,UniversityofNottinghamUniversityPark,NottinghamNG72RD,UK.E-mail:thailaf@hotmail.comDuetoaconstantlyincreasingoftheworldpopulation,thedemandforfoodisalsoontherise.However,ahighpercentageofthisfoodislostdue,partially,tofoodstuffspoilagebymicroorganisms,includingfilamentousfungi.Weakorganicacidssuchassorbic,benzoic,propionicandaceticacidareusedasantimicrobialagentsinordertopreventfoodandbeveragespoilagecausedbymicroorganisms.Unfortunately,themicroorganismsshowagreatresistancetoweakacidpreservativesandtheemergenceofresistantpopulationisoftenreported.Theexactmechanismbehindtheantifungalpropertiesofthesecompoundsisstillunknown.Aimingtobetterunderstandthemechanismsthatcontrolthesorbicacidantifungalpropertiesandidentifypotentialtargetsofthisweakacid,wescreenedanA.nidulansnullmutantlibrarycomposedof103non-essentialproteinkinases.Atotalof16nullmutantshadalteredgrowthinpresenceofdifferentsorbicacidconcentrations.Interestingly,03ofthemthatcorrespondtogenesencodingproteinsinvolvedintheHighOsmolarityGlycerol(HOG)pathwayweresensitivetosorbicacid.WeobservedthatA.nidulansHogAphosphorylationwasmodulatedinthepresenceofsorbicacidsuggestingtheinvolvementofthispathwayinthisprocess.TheacquisitionofmoredetailsaboutthemechanismsinvolvedintheantifungalpropertiesofsorbicacidaswelltheinvolvementoftheHOGpathwayinthisprocesswillclearlyhelponthedevastatingimpactoffungalfoodspoilageonglobalfoodsecurity.Financialsupport:BBSRC(UnitedKingdom),FAPESPandCNPq(Brazil)

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FrontiersintheunderstandingandmanagementofArmillariainUKgardensJassyDrakulic,MatthewCromey,LizBeal1,GerardClover

PlantHealth,RoyalHorticulturalSociety,RHSGardenWisley,Woking,GU236QB,UK;Email:jassydrakulic@rhs.org.ukHoneyfungusrootrot,causedbyArmillariaspp.,isthediseasemostfrequentlyenquiredaboutbyUKgardenerstotheRoyalHorticulturalSociety(RHS).Armillariaepidemiologyhasbeenstudiedwithinforestryandagriculture,butdataislackingwithingardens,whichhavegreaterhostplantdiversitythanorchardsandvineyardsandgreaterdisturbancethanwoodlands.SurveysofmaterialsenttotheRHSbymembershashelpedanswerquestionsaboutwhichArmillariaspeciesareresponsibleforthediseaseingardens,andhowplantsfromthebroadrangeofsusceptibleornamentalsaredifferentiallyaffectedbythedisease.FurtherresearchdirectionswithintheRHSfocusonevaluationoftheprinciplesofmanagementpracticessuchasstudiesdeterminingthelifespanofrhizomorphsofdifferentsizewhenremovedfromafoodsource,screeningofTrichodermaendophytesforuseasbiocontrol,andelucidatingthenatureoftheinteractionsbetweendifferentArmillariaspeciesofdifferingaggressivenesstohostplants.Fullresultsfromthesurveyarepresentedalongsidepreliminaryfindingsfromthecurrentresearchprojectslistedabove.AphidinfestationincreasessusceptibilityofB.napustoRhizoctoniasolaniAG2-1FryniDrizou1,TobyJABruce2andNeilSGraham11Divisionof Plant andCrop Sciences, School of Biosciences,University ofNottingham,SuttonBoningtonCampus,Loughborough,Leicestershire,LE125RD,UK;2SchoolofLifeSciences,KeeleUniversity,Keele,Staffordshire,ST55BG,UKEmail:Fryni.Drizou@nottingham.ac.ukOilseedrape(OSR),Brassicanapus,istheprimaryhostofRhizoctoniasolaniAG2-1,asoil-bornepathogenthatinfectsyoungseedlingscausingdamping-offdiseaseandimpairedcropestablishment.Currentlythereisnoavailableresistantgermplasmanditisunknownhowthepathogenovercomesplantdefences.Additionally,OSRisoneofthesecondaryhostsofMyzuspersicae,anaphidknownforitsabilitytoadapthostplantdefences.TheaimofthepresentworkwastoidentifyifherbivorybyM.persicaeaffectsOSRresponsestoAG2-1.Weexaminedhowaphidinfestationaffectsresponseofcultivars‘Canard’and‘Temple’toAG2-1byvisuallyassessingdiseasesymptoms13dayspostinoculationonplantstemsandbyquantifyingthefungalDNA.AdditionallyweexaminediftherewasanalterationintheexpressionofthedefencerelatedgenesLOX3,MYC2,NPR1,PR1andWRKY38.Ourresultsshowthataphidinfestationresultedinincreasedsusceptibilityof‘Canard’plantstoAG2-1:plantswithaphidshadmoreseverediseasesymptomsbuttheamount

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offungalDNAwasthesamebetweentreatments.GeneexpressionanalysisrevealedthatM.persicaeinfestationdecreasedtheexpressionofLOX3andMYC2inplantsinfectedwithAG2-1,hencepossiblealternationsinthejasmonatesignallingpathwaycausedtheincreasedsusceptibilitytothepathogen.Also,theelevationofPR1inAG2-1inoculatedplantsandNPR1underdualattack,impliesthatsalicylicacidmayplayaroleindefenceagainstthenecrotrophicfungiR.solaniAG2-1.AntifungalandPhytochemicalEvaluationofAnnonamuricataplantMargaret1A.Ebabhi.,KanifeU.Claris2,JuliusS.Omogbai3,NnabugwuE.Chinalurum2andRaheemL.Omobolaji2.1Department of Education Science, Distance Learning Institute, University of Lagos,Akoka, Lagos Nigeria; 2Department of Biological Science, Yaba College of Technology,PMB2011,Lagos,Nigeria;3DepartmentofPharmacy,FacultyofPharmacy,UniversityofLagos,Lagos,NigeriaEmail:ebabhi.meg@gmail.comTheincidenceoffungalinfectionisontheincreaseaspathogenicfungiarebecomingresistanttotheconventionaldrugs.Thisresearchexaminedtheantifungalpropertiesoftheaqueous,N-hexaneandethanolextractsoftheleaves,fruits,stem,barkandrootsofAnnonamuricatausingthediscdiffusionagarassay.Thezoneofinhibitionvariedforthetesteddermatophyticorganisms-Candidaalbicans,Trichodermasp.andTrichophytonmentagrophytes,withrespecttotheplantpartsandtypeofextract.Theaqueousextractsofthetestplantpartsshowedmorepotencyonthetestfungalspecies.PreliminaryphytochemicalstudiesoftheextractsoftheplantpartsshowedthatAlkaloid,Cardiacglycosidesandphenolwerehighlypresentinallpartswhilesteroidandphobataninarelesspresent.ThequantitativeresultsofvariouspartsoftheplantreviewedthatPhenolfromleafextractsarehighlyquantifiablethanotherphytochemicalcomponents.Inall,Alkaloids,CardiacGlycosideandphenolwerehighlyquantifiable,whileTannin,SteroidandPhobataninwerelessquantifiable.ThequantitativeanalysiswasdoneusingANOVAmethodat0.05levelofsignificance.ThesefindingsgivecredencetothetraditionaluseofAnnonamuricataforthetreatmentoffungalinfections.CharacteristicsandcontrolofAlternariaalternataaffectingsweetpepperinEgyptAbdulmageedM. Kamara1, Ahmed. E. Elkorany2, Eman E. Ramadan2, andGhonemM.Amer21 Department of Plant Pathology, University of Alexandria, Alexandria 21561, Egypt;2DepartmentofPlantPathology,UniversityofDamanhour,Damanhour22511,Egypt.Email:drelkorany125@yahoo.com

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Thirty-nineisolatesofAlternariaalternatawererecoveredfromsweetpepperfruitsshowedblackmouldsymptomscollectedfromdifferentregionsinEl-Beheragovernorate,Egypt.Theisolateswerecharacterizedfortheircolonyphenotypesandvirulencewherenocorrelationwasdetected.TheRAPDanalysisrevealed100%polymorphismusingtheOPA-03primeramongsixA.alternateisolatesanalyzedandthedevelopeddendogramshowedpartialclusteringforregionsandvirulence.Alltestedcultivars,i.e.Emberu,Trezaa,Mazurka,andTarantoweresusceptibletotheartificialinfectionwithA.alternatatodifferentdegrees.Treatmentofpepperfruitswithresistanceinducers,i.e.salicylicacid(SA),abscisicacid(ABA),methyljasmonate(MJ),andcalciumchloride(CaCl2)significantlyinducedresistanceanddecreasedthedevelopedblackmouldwherecalciumchloride(1.5g/L)andSA(1mM)werethemosteffective.InductionofresistanceinpepperfruitswithCaCl2andSAaswellasthetolerancerevealedincv.Mazurkawasfoundtobeduetohighinductioninenzymeactivityofpolyphenoloxidase(PPO),peroxidase(POD),andphenylalanine(PAL)aswellasanincreaseinthetotalphenolicsinthetreatedpepperfruits.*PowderyMildewIdentification: ImprovingaccuracyusingpreviouslyuntappedDNAregionsOliverEllingham1,JohnDavid2,andAlastairCulham1

1School of Biological Sciences, University of Reading, Reading, RG6 6AS,UK;2DepartmentofPlantPathology,TheRoyalHorticulturalSociety,Wisley,GU236QB,UKEmail:o.h.ellingham@pgr.reading.ac.ukThepowderymildews(Ascomycota,Erysiphales)areagroupofobligatebiotrophicFungifoundonnearly10,000angiospermplanthostsglobally,manyofwhichareimportanthorticulturalandagriculturalplants.Infectioncanreduceattractivenessandyieldssignificantly.Areliablemethodisrequiredforunambiguousidentificationoftheseoftencrypticspeciessuchthatspreadtonewareasand/ornewhostscanbedetectedrapidlyandcontrolledearly.Thisprojecttakesauniqueapproachtosampling;usingacitizenscienceschemelaunchedin2014toassessthepresenceofthe876globalpowderymildewspeciesintheUKandtheefficacyoftechniquescurrentlyusedfortheiridentification–hostidentification,fungalmorphologicalanalysis,sequencingofthefungalrDNAITSregion.Thesurveywaspromotedviaanarrayofonlineandprintedmediaandover500infectedplantsampleswerereceived–mostwerepowderymildews.Ofthese94%wereidentifiedtogenusleveland79%tospecieslevel.ThisshowsthescopeforimprovementstothesetechniquesthroughanalysisofsupplementaryDNAregions.90differentpowderymildewspecieswerefoundoverthe2-yearperiod;60%ofthe200speciespreviouslyrecordedintheUK.DuringthesurveyperiodtwonewhostswerefoundforspeciesknowntooccurintheUK.ThesedataprovidethemostcomprehensivesurveyofpowderymildewspecieswithintheUK.This

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knowledgeiscriticalforearlydetectionofnewlyarrivedspeciesofpowderymildewandofhostshiftsornovelhosts.**PhenotypicheterogeneityasanadaptationoffungitoenvironmentalstressDavidS.Foster1,SteveP.McGrath2,PaulS.Dyer1andSimonV.Avery11School of Life Sciences, University of Nottingham, Nottingham, NG7 2RD, UK;2SustainableAgricultureSciences,RothamstedResearch,Harpenden,AL52JQ,UKEmail:mbxdf@nottingham.ac.ukPhenotypicheterogeneitydescribesvariationinagivencharacteristicthatisevidentbetweenindividualcellsofanisogenicpopulation.Theextentofheterogeneityinaparticularphenotypemaybedeterminedbyepigeneticorgeneticfactors,e.g.specificgene-promotersequencesproducingnoisiergeneexpression.Anumberofcharacteristicsdisplayingphenotypicheterogeneityareknowntoinfluencecompetitivefitnessunderstress,thereforeheterogeneitylikelyhasabearingonsurvivalunderfluctuating-stressconditions.Recentevidencefromourlaboratorysuggeststhatenvironmentalstressselectsforincreasedlevelsofheterogeneityinresistancetocertainstressors.Wildyeastisolatesfrompollutedsiteswerefoundtoshowelevatedheterogeneityinresistancetopollutants,incomparisontoisolatesfromnearbycontrolsites.However,theinfluenceofenvironmentalstabilityonselectionforheterogeneityisnotunderstood,noritslong-termstability.Yeastandfilamentousfungalisolatesfromlong-termstressedexperimentalsiteshavebeenstudiedtoprovideinsight.Dose-responsegradients,amongstothermeasuresofcell-cellvariation,indicatethatalong-term,stablelevelofstressmayselectforadecreaseinheterogeneityofresistance.Thedegreeofresistanceheterogeneitymayhavethepotentialtoevolveindifferentdirectionsdependingonthestabilityofthestressordoseintheenvironment.DevelopmentofCRISPR/Cas9geneeditingtoolsforthericeblastfungusAndrewJ.Foster1,MagdalenaMartinUrdiroz1,NicholasJ.Talbot11SchoolofBiosciences,UniversityofExeter,Exeter,Devon,UnitedKingdom.Email:A.J.Foster@exeter.ac.ukCRISPR/Cas9geneeditingisarapiddevelopingfieldwhichholdsgreatpromiseforacceleratingthepaceoftargetedmutagenesisinfungi.Currently,althoughverymanyfungalgenesarepredictedbasedongenomesequencing,theprocessofspecificallymutatingselectedgenes,eveninthebeststudiedfungi,requiresmonthsratherthanweeks.ThericeblastfungusMagnaportheoryzaeisthemostseriouspathogenofrice.ToacceleratetheprocessoftargetedmutationandspecificgeneeditsinthisimportantspeciesweareworkingtowardsthedevelopmentofarapidCrispr/Cas9basedgeneeditingtechnology.Technologiesallowingpreciseeditstocodingsequences,gene

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taggingaswellasdeletionsofsingleormultiplegenesarebeingoptimised.Thisresearchwillopenthedoortoanalysisofgenefamiliesorsubsetsoftargetgenes.Latestresultswillbediscussed.Tacklingresistancetofoodpreservatives:heterogeneityinfungalsporepopulationsIveyA.Geoghegan,MalcolmStratford,DavidB.ArcherandSimonV.AverySchoolofLifeSciences,UniversityofNottingham,Nottingham,NG72RD,UKEmail:mbzig@exmail.nottingham.ac.uk

Foodspoilagebyfungiisaglobalissuewithfar-reachingeconomic,environmentalandhealth-relatedconsequences.Spoilageisparticularlyseverewithinthesoft-drinksindustry,whereevenahandfulofspoiledbottlescanresultintherecallofanentirebatch,costingmillionsofpounds.Suchspoilageoccursdespitethepresenceoffoodpreservativesinsoftdrinks,whicharegenerallyeffectiveatrestrictinggrowthofyeastsandmoulds.However,certainspeciesoffungiarehighlyresistanttothesepreservatives,andareassociatedwithspoilageincidentsworldwide.Thus,effectivecontrolofthesefungiisanimportantavenueofresearch.Wearecharacterizingmechanismsofresistancetosorbicacid,awidelyusedpreservativeinsoftdrinks.ThesporesofthemouldAspergillusnigerarehighlyresistanttosorbicacid.Inaddition,A.nigersporepopulationsdisplayahighdegreeofheterogeneityinresistancetothisparticularstress.Bycombininganalysisofsinglesporephenotypeswithcutting-edgemolecularindexingtechnologyaswellasclassicalmoleculargeneticstechniques,wehavebeguntouncovertheinextricablelinksbetweenpopulationheterogeneityandpreservativeresistanceinthisspoilagefungus.*Xanthomonas citri pv. citri pathotypes: Lipopolysaccharide (LPS) structure andfunctionasmicrobe-associatedmolecularpatterns(MAMPs)LotteBettinaAndersenGersby1,OlivierPruvost2,AntonioMolinaro3andMari-AnneNewman11PLEN,UniversityofCopenhagen,Thorvaldsensvej40,1871Denmark;2CIRAD,PVBMT,SaintPierre,LaRéunion,France;3DepartmentofChemicalSciences,UniversityofNaplesFedericoII,Naples,ItalyEmail:lba@plen.ku.dkCitrusbacterialcanker(CBC)isadiseaseofcitrusplantswithremarkablesocioeconomicimpactonthecitrusindustry.ThecausalagentofAsiaticCBCistheGram-negativebacteriumXanthomonascitripv.citri(Xac).Xacstrainsareclassifiedintothreepathotypes.PathotypeAhasthebroadesthostrange,infectingalmostallCitrus,pathotypesA*andAWshowalimitedhostspecificity,onlyaffectingMexicanlime.

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LPSisknowntoactasaMAMPandelicitsimmuneresponsesineukaryoticorganisms.MAMP-inducedresponsesincludeproductionofreactiveoxygenandnitrogenspecies,alterationsintheplantcellwall,inductionofantimicrobialcompoundsandexpressionofpathogenesis-relatedgenes.LPSiscomposedofapolysaccharideregion(O-chain),acoreoligosaccharideandaglycolipidregiontermedlipidA.AgenomicregioninvolvedinLPSbiosynthesisthatishighlyvariableamongtheXacpathotypeswasrecentlyidentified.Furthermore,partialstructureelucidationofXacLPSrevealedthepresenceofarhamnosehomo-polymerastheO-chainmoiety.AfullelucidationofthestructuresofLPSfromseveralXacstrainsmightclarifythedifferencesindefenceresponsesindifferenthostplants,aswellasthehostspecializationprocessoftheXacpathotypes.Here,wepresentthestructuresofLPSfromtwoXacstrainsofpathotypesAandA*andtheabilityofthewholeLPSandlipidAofbothstrainstoactasMAMPsinthemodelplantArabidopsisthalianaandinthetwohosts,Mexicanlime(Citrusaurantifolia,pathotypeA*host)andlemon(Citruslemon,pathotypeAhost).**FungalandOomyceteBiodiversityintheFalklandIslandsandSouthGeorgiaJacobHargreaves1,JimI.Prosser2,PaulBrickle3andPietervanWest11Aberdeen Oomycete Laboratory, College of Life Sciences and Medicine, Institute ofMedical Sciences, University of Aberdeen, Aberdeen, UK; 2Institute of Biological andEnvironmental Sciences, University of Aberdeen, Cruickshank Building, Aberdeen, UK;3SouthAtlanticEnvironmentalResearchInstitute,Stanley,FalklandIslandsEmail:j.hargreaves@abdn.ac.ukSoilmicrobialcommunitiesareextremelycomplexandheterogeneousentities,whicharenotfullyunderstoodatmostlevels,fromsoilporestructuralmicroclimatestoglobaldiversityanddispersion.Thisisdespitetheircrucialinvolvementinbiogeochemicalcycles,whichaffectsagriculturalandnaturalecosystems.Amajorcomponentofthemicrobialcommunityarefungi,whichhavecriticalrolesinbreakingdownorganicmatter.Asubgroupcanalsoformsymbioticrelationshipswithplants,knownasmycorrhizae,whichcanincreaseplantproductivity.TheFalklandIslandsandSouthGeorgiarepresentanisolatednaturalecosystemwithintroducedherbivoressuchassheepandreindeer.Toinvestigatefungaldiversityinthesecircumstances,soilsamplesfromWestFalklandandSouthGeorgiaweretakeninthe2017southernsummertocarryouthighthroughputsequencingofbarcoderegions.Thesesampleswillgiveasnapshotofthenaturaleukaryoticmicrofloraoftheregions,withareasofrelativelylittleandareasofalotofgrazingpressure.Thedatawillallowfungalcommunitystudiestobecarriedout,lookingattheimpactofvegetationandreindeergrazing.Thesubsequentanalyseswillallowinferenceofthefungalcommunityfunctiontobedetermined,withapplicationstowardsagricultureandbioremediation.Herewepresentourlatestfindings.

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MappingtheAdaptiveLandscapeofZymoseptoriatriticiCYP51underAzoleFungicideSelectionNicholaJ.HawkinsandBartA.FraaijeBiointeractionsandCropProtection,RothamstedResearch,Harpenden,AL52JQ,UKEmail:nichola.hawkins@rothamsted.ac.ukFungicidesareakeycomponentofcontrolprogrammesformanyplantpathogens,buttheiruseexertsastrongselectivepressureforresistance.Forsomesite-specificfungicides,theevolutionofresistanceissimpleandrapid,withasinglemajortarget-sitemutationresultingineffectivelycompleteresistance.However,fortheazolefungicides,thepictureismorecomplex,withmultipletarget-sitemutationsandother,over-expressionbasedmechanismsaccumulatingtoproducequantitative,step-wisereductionsinsensitivity.Theeffectofmultiplemutationsonresistanceisoftenreferredtoas“additive”,butinfactthesituationisfurthercomplicatedbyepistaticinteractions,bothintermsoffungicideresistance,andofimpactsonenzymefunctionandstabilityofthetargetprotein.Thisresultsinaruggedadaptivelandscape.Consequently,adetailedunderstandingoftheeffectofeachmutation,individuallyandindifferentgeneticbackgrounds,onresistanceandfitness,isnecessaryinordertoattempttopredictevolutionarytrajectoriesunderdifferentfungicideselectionscenarios.WewillpresentanupdatedfitnesslandscapeofZ.triticiCYP51haplotypesreportedinfieldisolatestodate,discussingtheimplicationsforthepredictabilityoffutureevolutionandexperimentalapproachesfortestingthis.GeneticdiversityofFusariumspp.oncerealsinRussiaAleksandrIgnatov1,TatianaSuprunova2andNataliaZhemchuzhina31Russian University of People’s Friendship, Moscow, 117198, Russia and R&D Center“PhytoEngineering” LLC, Rogachevo, 141880, Russia; 2“Doka-Gene Technology” LLC,Rogachevo, 141880, Russia; 3 Russian Research Institute of Phytopathology, BolshieVyazemy,143050,Russia.Email:an.ignatov@gmail.comTherootrot,snowmold,andfusariumearblightcausedbyFusariumspp.andMicrodochiumspp.-relatedfungidamageawiderangeofcerealcropsinRussia.Thediseaseagentsarecharacteristictocertainecologicalandgeographicalareas.96strainsofFusariumspp.andMicrodochiumspp.fromtheStateCollectionofPlantPathogenicMicroorganismswereusedforMultiLocusSequenceTyping(MLST)analysisforgenesoftranslationelongationfactor-1alpha(TEF1),β-tubulin,ITSrRNA(with3differentprimersets),andRNApolymeraseII(RPB2)withprimersreportedelsewhere(O’Donnelletal.,2000;Debourgognetal.2010;CBS-KNAWFungalBiodiversityCentre’sFusariumMLSTwebsite).Resultsofpreviousphenotypicidentification,PCRFingerprint(IRAPandISSR),andMLSTanalysiswerecompared.StrainsofF.culmorumformedthemostpolymorphicgroupaccordingtoPCRfingerprintandMLST,whereasthemost

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homogeneousgroupincludedF.sporotrichioides.StrainsofM.nivale,F.avenaceum,F.gibbosumwerenotsignificantlydifferentiatedstatisticallyfromotherspecies.SomestrainsofM.nivale,F.sporotrichoides,andF.oxysporumcouldnotbecorrectlyidentifiedbyeitherphenotypeorPCRfingerprinting.ThespeciesofthepathogenswidespreadinRussiahadvariouspathogenicandtoxicproperties,indicatingtheirstrongintraspecificvariability.Rarefungalspeciespossessedtoxicitytotest-plantseedlings,butwerenotpathogenic.ThegeneticsimilaritybetweennewlysequencesMLSTlociandclosestGenebank(NCBI)accessionsrangedfrom81to99%indicatingpresenceofbothpreviouslycharacterizedanduniquegeneticlinagesamongstrainsofFusariumspp.andMicrodochiumspp.infectingcerealsinRussianFederation.BiologicalmanagementofsouthernblightofchilibyPenicilliumoxalicumandleavesofEucalyptuscitriodoraArshadJavaid,RabiaAfzalandAmnaShoaibInstitute of Agricultural Sciences, University of the Punjab, Quaid-e-Azam Campus,Lahore,Pakistan.Email:arshadjpk@yahoo.com,arshad.iags@pu.edu.pkInthepresentstudy,leafdrybiomassofEucalyptuscitriodoraandabiologicalcontrolagentPenicilliumoxalicumwereevaluatedformanagementofsouthernblightdiseaseofchilicausedbySclerotiumrolfsii.Inapottrial,soilwasmadesickwithinoculumofS.rolfsii.SoilwasamendedwithdryleafbiomassofE.citriodora(1,2,3%)andabiologicalcontrolagentPacicilliumoxalicumaloneorincombination.Soilamendmentwith3%leafbiomassofE.citriodorasignificantlyreducedplantmortalityby65%overpositivecontrol.Consequently,rootandshootgrowth,fruityieldaswellaschlorophyllcontentweresignificantlyenhanced.P.oxalicumincombinationwith3%leafbiomassalsosignificantlyreducedplantmortalityandenhancedcropgrowthandyield.ProteincontentinP.oxalicuminoculatedtreatmentswasalsosignificantlyhigherthancontrol.S.rolfsiiinoculationsignificantlyenhancedperoxidase(PO),phenylalanineammonia-lyase(PAL),catalaseandpolyphenoloxidase(PPO)activitiesinleavesofchiliwhiletheseparametersweredrasticallyreducedbyapplicationofP.oxalicumandleafbiomassofE.citriodora.Inlaboratorybioassay,differentconcentrationsofmethanolicleafextractofE.citriodoraviz.1,2,…,5%reducedfungalgrowthby3to51%.Inmethanolicleafextract,compoundsnamely2-Furancarboxaldehyde,5-(hydroxymethyl),Cyclohexanol,2-(2-hydroxy-2-propyl)-5-methyl,Pentadecanoicacid,14-methyl,methylester,and14,17-Octadecadienoicacid,methylesterwereidentifiedthroughGC-MSanalysis.Thepresentstudyconcludesthat3%leafbiomassofE.citriodoracansignificantlyreduceplantmortalityduetoS.rolfsiiandenhanceplantgrowthandyieldinchili.

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Tracing the transduction of a phenylpyrrole fungicide reveals its effect on cAMPsignallinginBotrytiscinereaJaafarKilani1,2,MonicaMiazzi3,ColetteAudéon1andSabineFillinger11UMRBIOGER, INRA,AgroParisTech,UniversitéParis-Saclay,Thiverval-Grignon,France;2Univ. Paris-Sud, Université Paris-Saclay, France; 3Department of Soil, Plant and FoodSciences,UniversityofBariAldoMoro,Bari,ItalyEmail:jaafar.kilani@inra.frFungirapidlyadapttotheirenvironmentinvolvingtosignallingpathwayslikethoseofmitogenactivatedproteinkinases(MAPKs).InBotrytiscinerea,thephenylpyrrolefungicidefludioxonilactivatestheSak1andBmp3MAPKs,whichareinvolvedinosmoregulation,cellwallintegrity,developmentandpathogenicity.Inordertotracethetransductionoffludioxonilandtoidentifythesignallingpathwaysinvolved,wehaveperformedcomparativephosphoproteomicsofthewild-typestrainsubjectedornottofludioxonil.Thisexperimentledtotheidentificationof“fludioxonil”candidatesignallingproteinssuchasthephosducin-likeproteinPhnA.Ineukaryotes,phosducinisinvolvedintheG-proteinsignallingandhencethecAMPpathway.InthephytopathogenicfungiCryphonectriaparasiticaandFusariumgraminearum,phosducinisinvolvedinpigmentation,sporulation,andpathogenicity.

WeperformedthefunctionalanalysisofphnAinB.cinerea.ItsdeletionrevealedthatphnAisinvolvedindevelopment,pathogenicityandadaptationtocellwallstress.TheroleofphsoducininG-proteinsignallingpushedustostudytheimpactoffludioxonilonthecAMPsignallingpathway.Inthewild-typeweobservedasteadydecreaseofintracellularcAMPaccumulationduringexposuretofludioxonil.WealsostudiedtheconnectionbetweenPhnAandSak1andweobservednoinfluenceofPhnAonSak1phosphorylation.

OuranalysesrevealedthatthephenylpyrrolefludioxonilactsoncAMPdependentsignaltransductioninadditiontoitsactivatingeffectontheosmosensingMAPKpathway.TheroleofPhnAandtheosmoticMAPKpathwayincAMPsynthesisunderfludioxoniltreatmentiscurrentlyunderinvestigation.*The impact of microbial communities on the incidence of foliar fungal diseasesymptomsinwheatGeorgeLund1,2,MatthewDickinson3,TimMauchline2andJasonRudd11Bio-interactionsandCropProtectionDepartment,RothamstedResearch,Harpenden,AL5 2JQ, UK; 2 Sustainable Agricultural Science Department, Rothamsted Research,Harpenden, AL5 2JQ, UK; 3 School of Biosciences, University of Nottingham,Loughborough,LE125RD,UKEmail:george.lund@rothamsted.ac.ukSeptoriatriticiblotchdisease(STB)causedbythefungusZymoseptoriatritici,isthemosteconomicallyimportantwheatdiseaseinWesternEurope.Withresistanceto

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conventionalfungicidesbecominganincreasingproblem,alternativesolutionsarerequired.Primarystagesofinfectionoccurintheextracellularspacesofthewheatleaf,wherebacteriasuchasPseudomonasfluorescenscanalsobefound.InourfirstexperimentaPseudomonadlibrarywasscreenedfordirectinhibitionofZ.triticionagarplates,todetermineifanyexhibitantifungalactivity.Usingahigh-throughputscreeningmethodologydevelopedinthisprojectwhichtested460bacterialendophytesandrhizosphereassociatedPseudomonads,40isolatesshowedanabilitytodirectlyinhibitthegrowthofZ.tritici.Ourresultsraisethepossibilitythatbacteriapresentonwheatleavesmayaffecttheincidenceand/orseverityofSTBviadirectinhibitionoffungalgrowth,throughsecretedfungalinhibitors.Inasecondexperiment,weplantogrowsurfacesterilisedwheatseedsinsterilegrowthmediumamendedwithsyntheticmicrobialcommunitiesbeforeinfectingtheplantswithZ.tritici.Thiswillallowustomonitortheimpactofthebelowgroundmicrobiomeontheplant’sresponsetofoliardisease,throughprimingorothermechanisms.Thesecombinedapproachesmayleadtonovelmechanisms,whichcouldbeexploitedforchemicaland/orbiologicalcontrolofZ.tritici.ExploringNGS,HIGSandsi-RNAtechnologiesforthecontrolofFusariumearblightinwheatAnaK.Machado1,RobertKing2,WingShamLee1,CarolineSparks1,NeilBrown1,KostyaKanyuka1,MartinUrban1,JonWest1,EleneYamazaki-Lau3,CasianeS.Tibola3,MariaI.P.M. Lima3, Roberto Togawa4, Natalia Martins4, Francisco Aragão4, Camila P. Nicolli5,EmersonM. Del Ponte5, Dauri J. Tessmann6, JoséMauricio C. Fernandes3 and Kim E.Hammond-Kosack11 Biointeractions and Crop Protection Department and 2 Computational and SystemsBiology Department, Rothamsted Research, Harpenden, UK, 3 Embrapa Trigo, PassoFundo, Rio Grande do Sul, Brazil 4 Embrapa Genetic Resources and Biotechnology,Brasilia,Brazil,5UniversidadeFederaldeViçosa,MG,Brazil,and6UniversidadeEstadualdeMaringá,PR,Brazil.Email:ana.machado@rothamsted.ac.ukFusariumearblight(FEB)isamajorprobleminmostsmall-graincerealgrowingregionsandnowthreatensglobalfoodsecurity.CurrentlystrategiestocontrolFEBarenotveryeffective,fungicidesgivepartialprotectionanddevelopmentofgenetic-basedresistantcultivarshasproventobedifficult.Therefore,newwaystocontrolFEBareurgentlyrequired.Here,wedescribetheframeworkandinitialresultsfromabilateralUK-Brazilprojectthatisusingabespokewholegenomesequencing,inplantatranscriptomeandreversegeneticsguidedapproachtounderstandandpinpointtheFusariumgenesandpathwaysrequiredtocausediseaseinwheatheads.OurintentionistoidentifyanewsuiteofFusariumgenesforinterventionthatcansimultaneouslybetargetedviahostinducedgenesilencing(HIGS).HIGSconstructsarebeingtestedforefficacyinArabidopsis,lettuceandwheat,andpromisingHIGSconstructshavebeenstably

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transformedintoacommercialmoderately-FEBresistantBrazilianwheatinpreparationforfieldtestinginsouthernBrazilduring2018and2019.BytakingaFusariumgenome/reversegeneticsguidedapproach,thisisenablingthedevelopmentofflexiblenewwaystocontrolFEBdiseaseinwheatcropsgrowninBrazil.Themainprerequisitesneededtoapplythisapproachinotherwheatgrowingregionswillbeaddressed,aswellasadditionalscientific,societalandindustrybenefitsthatcouldpotentiallyemergewhenusingHIGStechnologiesforplantdiseasecontrol.ThisresearchissponsoredbytheBBSRC,TheNewtonFundandEMBRAPA.HeterologousexpressionofferuloylesteraseofAspergillusterreusMiia R. Mäkelä1, Adiphol Dilokpimol2, Salla Koskela1, Ronald P. de Vries2, KristiinaHildén11Division ofMicrobiology and Biotechnology, Department of Food and EnvironmentalSciences, University of Helsinki, Helsinki, Finland; 2Westerdijk Fungal BiodiversityInstitute, Utrecht, The Netherlands; 3FungalMolecular Physiology, Utrecht University,Utrecht,TheNetherlandsEmail:miia.r.makela@helsinki.fiInthecellwallsofgramineousplants,hemicellulosesarecrosslinkedtothearomaticligninpolymerviahydroxycinnamicacids(ferulicacidandp-coumaricacid).Feruloylesterases(ferulicacidesterases,EC3.1.1.73),presentinCAZyfamilyCE1(www.cazy.org),areenzymesthatcatalysethecleavageofcovalentesterbondsbetweencarbohydrateandligninmoietiesinplantcellwalls.Duetotheabilitytospecificallycleaveesterlinkages,feruloylesterasesarepromisingbiocatalystsforabroadrangeofbiotechnologicalapplications.Theseincludee.g.pharmaceutical,agriculturalandfoodindustries,aswellastheproductionofbiofuel.AnalysisoftheCAZomesofeightAspergillusspeciesrevealedahighvariabilityinthegenesetsrelatedtoplantbiomassdegradation(1).OneofthesevariationswasintheputativeFAEsofCAZyfamilyCE1,whereallspeciescontainoneconservedfaegene,butsomespeciespossessadditionalcandidategenesforthisactivity.AspergillusterreuscontainstwoadditionalputativeFAEencodinggenes,oneofwhichhasorthologsinAspergillusoryzaeandAspergillusflavus,butnotinanyoftheothertestedspecies(Aspergillusniger,Aspergillusnidulans,Aspergillusclavatus,AspergillusfumigatusandAspergillusfisheri).OneoftheseputativeA.terreusFAEencodinggeneswaschosentobeheterologouslyexpressedinamethylotrophicyeastPichiapastoris.Therecombinantenzymewassuccessfullyproducedandfurtherstudiedforitsbiochemicalcharacteristics.(1)Closelyrelatedfungiemploydiverseenzymaticstrategiestodegradeplantbiomass.Biotechnol.Biofuels8,107.

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MeasuringresistancetoSeptoriatriticiblotchinelitewheatcultivarsusingautomatedimageanalysisAlexeyMikaberidzePlantPathology,InstituteofIntegrativeBiology,ETHZürich,Universitaetstr.2,ETHZentrum,LFWB28,CH-8092Zurich,SwitzerlandEmail:QuantitativeresistanceislikelytobemoredurablethanmajorgeneresistanceforcontrollingSeptoriatriticiblotch(STB)onwheat.Earlierstudieshypothesizedthatresistanceaffectingthedegreeofhostdamage,asmeasuredbythepercentageofleafareacoveredbySTBlesions,isdistinctfromresistancethataffectspathogenreproduction,asmeasuredbythedensityofpycnidiaproducedwithinlesions.Wetestedthishypothesisusingacollectionof335eliteEuropeanwinterwheatcultivarsthatwasnaturallyinfectedbyadiversepopulationofZymoseptoriatriticiinareplicatedfieldexperiment.Weusedautomatedanalysisof21214scannedwheatleavestoobtainquantitativemeasuresofSTBconditionalseveritythatwereprecise,objective,andreproducible.Thesemeasuresallowedustoexplicitlyseparateresistanceaffectinghostdamagefromresistanceaffectingpathogenreproduction,enablingustoconfirmthattheseresistancetraitsarelargelyindependent.Thecultivarrankingsbasedonhostdamageweredifferentfromtherankingsbasedonpathogenreproduction,indicatingthatthetwoformsofresistanceshouldbeconsideredseparatelyinbreedingprogramsaimingtoincreaseSTBresistance.Wehypothesizethatthesedifferentformsofresistanceareunderseparategeneticcontrol,enablingthemtoberecombinedtoformnewcultivarsthatarehighlyresistanttoSTB.ThesedatacanalreadybeusedbybreedingprogramsaimingtoincreaseSTBresistancetochoosewheatcultivarsthatarebroadlyresistanttonaturallydiverseZ.triticipopulationsaccordingtothedifferentclassesofresistance.*In-Vitro Antifungal Screening of Radish (Raphanus sativus L.) Extracts as SafeBotanicalFungicidesagainstAlternariaalternataandSclerotiumrolfsiiHassanB.MohamedandRashidM.MamunurDepartment of Plant Pathology, Hajee Mohammad Danesh Science and TechnologyUniversity,Dinajpur,5200,BangladeshEmail:medbarkad1991@gmail.com

Twolabexperimentswereconductedonradish(Raphanussativusvar.BARIMula-1)extractionstoquantitativelyinvestigatephytochemicalsinmethanolicradishrootextract(MRRE)andscreenantifungalactivityofmethanolicandaqueousradishextractsagainstAlternariaalternataandSclerotiumrolfsiiinAgriculturalChemistryandPlantPathologyLaboratories,HajeeMohammadDaneshScienceandTechnologyUniversityDinajpur,duringSeptember,2015–September,2016.Methanolicsolventwasused

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usingmacerationmethod,where15.12g(30.24%)brownpungentgreaselessjelly-likecrudephytochemicalsubstancewasextractedfrom50gradishrootpowderdissolvedwith200mlmethanol.TLCanalysisofMRREshowedat1:10ethylacetatehexane6spotsrecordedasRfvalues,wherehighestRfvaluewas0.91cmandlowestRfvalue0.29cm.In-vitroinhibitoryactivityoftheradishextractsagainstmycelialgrowthofthetestfungiwasscreenedusingpoisonedfoodtechnique.Threepoisonedmediaviz.methanolicradishrootextractmedia(MRREM,T1),aqueousradishrootextractmedia(ARREM,T2)andaqueousradishleafextractmedia(ARLEM,T3),andnon-poisonedPDAmedia(Tc)wereusedastreatments.Therewerehighlysignificancedifferencesamongthepoisonedmediainreductionofthemycelialgrowthoverthenon-poisonedcontrolPDAmedia.MRREMinhibitedthegrowthofA.alternataandS.rolfsii94%and58.1%,followedbyARREM93%and40%andbyARLEM90%and20%,respectively,overthecontrol.ItmaybeconcludedthatradishextractscanbeusedassafebotanicalfungicidestocontrolA.alternataandS.rolfsii.**ResistanceofSaccharomycescerevisiaetolignocellulose-derivedinhibitorsElenaMoreno-MartínezandSimonV.AverySchoolofLifeSciences,UniversityofNottingham,UniversityPark,NottinghamNG72RD,UKEmail:msxem3@nottingham.ac.ukProductionofbioethanolfromlignocellulosichydrolysatesisapromisingbiofueltechnologybutissubjecttoinhibitionbychemicalsreleasedduringthepretreatmentofbiomass.Weakacids,furansandphenolsimpairyeastgrowthandfermentation,thusreducingefficiencyoftheprocessandincreasingthecost.Someofthemolecularmechanismsofactionandresponsetothesestressorsarecharacterized,butlittleisknownabouthowuniformsucheffectsareacrossindividualcellswithinanisogeniccellpopulation.Non-genotypicheterogeneityreferstotheexpressionofdifferentphenotypeswithinaclonalcellpopulation,despiteallindividualsbeinggeneticallyidentical.Understandingindividual-celleffectsofhydrolysateinhibitorscouldhelpengineeringofpopulation-levelresistance.ThedegreeofheterogeneityinresistancetraitswasspecifictoparticularinhibitorsinthefermentingyeastS.cerevisiae.Inclusionofasecondinhibitorcouldalterheterogeneityrelativetoeitherinhibitoralone.Shortpretreatmentswithsub-inhibitoryconcentrationsoftheinhibitorsindicatedadaptiveresistanceandadaptiveheterogeneityresponses.RNAiassistedgenomeevolutionwasalsousedtorevealcombinationsofgeneknockdownsthatcansynergisticallyincreaseresistancetowardstheseinhibitors.Findingsderivedfromthisresearchwillprovidebetterunderstandingofthebehaviourofyeastinlignocellulosicethanolfermentations,andmighthelpimproveproduction.

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Carbohydrate arrays coupled with mass spectrometry to detect activity of plantpolysaccharidedegradativeenzymesfromAspergillusnigerJolandavanMunster1,2,BaptisteThomas1,MichelRiese1,AdrienneDavis3,ChristopherGray1,DavidArcher2,SabineFlitsch11Chemical Biology,Manchester Institute for Biotechnology,University ofManchester,Manchester, M1 7DN UK; 2 Fungal Biology and Genetics, School of Life Sciences,UniversityofNottingham,Nottingham,NG72RD,UK;3SchoolofChemistry,UniversityofNottingham,Nottingham,NG72RD,UKemail:Jolanda.vanMunster@Manchester.ac.ukRenewables-basedbiotechnologydependsonenzymestodegradeplantlignocellulosetosimplesugarsthatareconvertedtofuelsorhigh-valueproducts.Identificationandcharacterizationofsuchlignocellulosedegradativeenzymescouldbefast-trackedbyavailabilityofanenzymeactivitymeasurementmethodthatisfast,label-free,usesminimalresourcesandallowsdirectidentificationofgeneratedproducts.TheobjectiveofthisworkwastodevelopsuchamethodbyapplyingcarbohydratearrayscoupledwithMALDI-ToFmassspectrometrytoidentifyreactionproductsofcarbohydrateactiveenzymes(CAZymes)ofthefilamentousfungusAspergillusniger.Wedescribetheproductionandcharacterizationofplantpolysaccharide-derivedoligosaccharidesandtheirattachmenttohydrophobicself-assemblingmonolayersonagoldtarget.Weverifiedeffectivenessofthisarrayfordetectingexo-andendo-actingglycosidehydrolaseactivityusingcommercialenzymes,anddemonstratehowthisplatformissuitablefordetectionofenzymeactivityinrelevantbiologicalsamples,theculturefiltrateofA.nigergrownonwheatstraw.WewillnowapplythismethodtocharacterisethedegradativepotentialofthebroadsetofA.nigerCAZymesthataresecretedinresponsetoplantlignocellulose.Inconclusion,wedescribeaversatilemethodthatisbroadlyapplicableinscreeningandcharacterisationofactivityofCAZymes,suchasfungalenzymesforplantlignocellulosedegradationwithrelevancetobiotechnologicalapplicationsasbiofuelproduction,thefoodandanimalfeedindustry.*Detection and Distribution of Aphanomyces Root Rot in Peas within the UnitedKingdomBrianÓLoinsigh1,LeaHerold2,DebbieSparkes1andMatthewDickinson11PlantandCropSciencesDivision,UniversityofNottingham,LE125RD,UK;2ProcessorsandGrowersResearchOrganisation,Peterborough,PE86HJ,UKEmail:stxbb13@nottingham.ac.ukRootrot,causedbyAphanomyceseuteiches,astrictsoil-borneoomycete,isoneofthemostdamagingdiseasesofpea(Pisumsativum)andsignificantlyreducespeaproductionglobally.Sincetheinoculumpersistswithinthesoil,andnoresistantcultivarsormeansofchemicalcontrolexists,thediseaseisdifficulttomanage.

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Moreover,theUKviningpeaareasarefixedwithnoroomtomovefurtherafield.Forthesereasonsgrowersfinditusefultoobtaininformationonthedegreeofinfestationwithintheirfieldstoavoidthoseathighrisk.However,thespreadandseverityofA.euteichesacrosstheUKgrowingregionsiscurrentlyunknown.Inthisproject,soilsfrom38siteswerecollectedspanningtheviningpeaareasandtestedforthepresenceofAphanomycesrootrotthroughbaitingexperiments.ALoopMediatedIsothermalAmplification(LAMP)assayforthespecificdetectionofA.euteichesfrompeaplatebaitingexperimentswasdeveloped.TheresultssuggestthatthisLAMPassaycanbeusedtodetectA.euteichesDNAontherootsofinfectedseedlingsthathasbeenbaitedwithsoilwithindaysofreceivingthesamples.CombiningbothvisualdiseasescoresandLAMPresults,adistributionmapofthediseasewasdeveloped.WeshowthatA.euteichesisfoundacrossallareasoftheUKgrowingregionandismorewidespreadthanpreviouslyknown,withsomeareashavingahigherburdenthanothers.TheLAMPassayallowsforfastandspecificdetectionofthepathogen.ThisinformationcannowbeusedtofurtherunderstandthediseaseintheUKandhelpslowthespreadofthedisease.**Optimisation of sexual development in Pyrenopeziza brassicae to allow study ofhormonaltriggeredmorphogenesisTomPearson1,AlisonA.Ashby2,PaulS.Dyer11SchoolofLifeSciences,UniversityofNottingham,Nottingham,NG72RD,UK;2CrogganLtd,TheCoachHouse,LongLaneCambridgeshire,CB245LD,UKEmail:Stxtp4@nottingham.ac.ukFungicideresistanceisamajorproblemwithinagriculture,causingmillionsofpounds’worthofcroplossandthreateningfoodsecurity.Severaleconomicallyimportantfungalcroppathogenshavedevelopedsomelevelofresistancetocommonfungicides,oneexamplebeingPyrenopezizabrassicae,thecauseoflightleafspotonoil-seedrapeandotherBrassicaspecies.Itisclearthatnewmethodstocontroldiseaseneedtobedeveloped.ThereispreliminaryevidencethatP.brassicaeproducessexhormones(knownasSexFactor)duringmatingwhichalterdevelopment.Thesecouldbeutilisedasnovelfungicidestoinhibitasexualreproduction,thereforecontrollingthespreadofdisease.Toidentifymodelsystemsforhormonestudy,sexualcrossesarebeingsetupbetweenP.brassicaeisolatesofopposingmating(MAT)types.Currentworkisfocusedonoptimizingproductionoffertileapotheciainlaboratorycrosses,usingavarietyofcrossingmethodsandgrowthmedia.Ongoingresultswillbepresented.FutureworkwillfocusonfurtherpurificationandcharacterisationofSexFactor,whichpreviousstudiesindicateislipoidalinnatureandcanbeextractedusingethylacetateormethylchloroform.Bioassayswilllookforsuppressionofasexualsporulationandinductionofsex.

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*Virus-andsalicylicacid-inducedchangesinplant-insectandplant-virusinteractionsAnna Platoni1,2, Sanjie Jiang1, Alex M. Murphy1, Adrienne E. Pate1, Ju-Yeon Yoon1,3,BeverleyJ.Glover1,JohnP.Carr1andGerardR.G.Clover21PlantSciencesDepartment,UniversityofCambridge,Cambridge,CB23EA,UK;2PlantHealth,RoyalHorticulturalSociety,RHSGardenWisley,Woking,SurreyGU236QB;3VirologyUnit,DepartmentofHorticulturalEnvironment,NationalInstituteofHorticulturalandHerbalScience,RDA,Wanju,55365,SouthKoreaEmail:amp93@cam.ac.ukInteractionsbetweenplantsandinsectsarecomplexandmediatedbyplant-derivedsignalmolecules,includingvolatileorganiccompounds.Infectionbyvirusesaltersplantvolatileemissionandconsequentlyaffectsplantinteractionswithinsectsthatvectorviruses,aswellaswithbeneficialinsects.Forexample,infectionwithcucumbermosaicvirus(CMV)affectsinteractionswithaphids(1),whileinotherworkwefoundthatbumblebees(Bombusterrestris)areattractedtoCMV-infectedtomato(Solanumlycopersicum)plants(2).Salicylicacid(SA)andjasmonicacid(JA)aredefensivephytohormonesthatinfluenceinteractionsofplantswithviruses,microbesandinsects.Intomatoandpepper(Capsicumannuum)weareinvestigatingtherolesofSAandJAinregulatingresistancetoCMVinfection,andincontrollingemissionofvolatilesthatinfluenceinteractionsofplantswithaphids(Myzuspersicae)orwithbumblebees.Inparticular,weareinvestigatingtheeffectsofexogenousapplicationofSAandJAonemissionofinsect-perceivablevolatiles,andtheeffectsofSAonCMVreplicationandmovement.ThewideraimofthisworkistofurtherunderstandtheeffectsoftheSA-andJA-dependentdefensivesignallingpathwaysoninteractionsbetweenplants,virusesandharmfulaswellasbeneficialinsectstoprovideamateurandprofessionalhorticulturalistswithbetteroptionsforenvironmentallyfriendlyintegratedpestanddiseasemanagement.1.Westwoodetal.2013PLoSONE8(12):e83066.2.Groenetal.2016PLOSPathogens12(9):e1005906.WorkfundedbytheRoyalHorticulturalSociety,BBSRC,LeverhulmeTrust,andtheRuralDevelopmentAgencyoftheRepublicofKorea.MechanisticbasisofepithelialdamageinresponsetoAspergillusfumigatusinfection**NormanvanRhijn1,TakanoriFurukawa1,PanagiotisPapastamoulis2,SayemaKhan1,MikeBromley1,MagnusRattray2andElaineBignell11Manchester Fungal Infection Group, University of Manchester, Manchester, UnitedKingdom;2DivisionofInformatics,ImagingandDataSciences,UniversityofManchester,Manchester,UnitedKingdom.Email:norman.vanrhijn@manchester.ac.ukThe human lung is continually exposed to spores of the airborne mouldAspergillusfumigatus. Inhaled spores are small enough to bypass mucociliary clearance

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mechanisms and reach the alveoli of the lung where interaction between host andpathogen cells can lead to fungal clearance, or to development of inflammatory orinvasive fungal diseases. Fuelled by the results of extensive fungal transcriptomeresearchwehavedeveloped, inrecentyears,aprogrammeofresearchthatseeksthemechanistic basis of lung damage duringA. fumigatusinfection. In this study weintroduceanewhighthroughoutstudyaddressingtheA.fumigatusregulatorynetworkdriving epithelial damage in cultured human lung tissue and examine how thisknowledgemightleadtonoveltherapeuticinterventions.*InvestigatingeffectordeliverybythericeblastfungusMagnaportheoryzaeClaraRodriguez-Herrero,MagdalenaMartin-Urdiroz,GeorgeLittlejohn,MiriamOses-Ruiz,WasinSakulkoo,XiaYan,MichaelJ.Kershaw,DarrenM.SoanesandNicholasJ.TalbotSchoolofBiosciences,UniversityofExeter,EX44QD,UnitedKingdomEmail:cr425@exeter.ac.ukMagnaportheoryzaeisthecausalagentofriceblastdisease.ThelifecycleofM.oryzaestartswhenasporelandsonthesurfaceofaleaf.Thesporegerminatestoformagermtube,whichgeneratestheappressorium.Theappressoriumgeneratesenormousturgorpressurethatistranslatedintophysicalforcethatrupturesthericeleaf.Thefungusthendevelopsinvasivehyphae(IH),whichcolonizesplanttissue.M.oryzaesecreteseffectorstofacilitatehostcolonizationbysuppressingplantimmunityresponses.EffectorsinM.oryzaecanbeclassifiedascytoplasmic,orapoplastic,dependingonwheretheyaredelivered.Cytoplasmiceffectorscanbeseentoaccumulateinamembrane-richplantstructurecalledthebiotrophicinterfacialcomplex(BIC),whileapoplasticeffectorsareattheextra-invasivehyphalmembranesurroundingtheIH.TheBICisthoughttobeinvolvedintranslocationofeffectorsintothericecytoplasm.Cytoplasmicandapoplasticeffectorsfollowtwodifferentsecretionpathways.ApoplasticeffectorsaresecretedbytheconventionalER-Golgisecretorypathway,whilecytoplasmiceffectorsaresecretedviaanunconventionalroute.Littleisknownabouthowsecretedeffectorsaredeliveredtothecorrectdomains.Tounderstandthisprocess,wearetryingtoidentifyaspecifichosttargetingsequencebygeneratingalibraryofchimericeffector-encodinggenefusions.Weareanalysingthechimerasfortranslocationpotential.Inaddition,wearegeneratingstabletransgenicriceplantsinwhichwehaveGFP-taggedbothearlyandlateendosomalcompartmentstodeterminethepotentialroleofplantendocytosisineffectoruptakeintoplantcells.

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Azole sensitivity in Leptosphaeria pathogens of oilseed rape: the role of lanosterol14α-demethylase.ThomasRSewell1,2,NicholaJHawkins3,YongJuHuang2,HenrikUStotz2,StevenLKelly4,DianeEKelly4,BartFraaije3,BruceDLFitt21Department of Infectious Disease Epidemiology, Imperial College London, UK; 2CropProtection Group, Centre for Agriculture, Food and Environmental Management,University of Hertfordshire, Hatfield, UK; 3Biointeractions and Crop Protection,Rothamsted Research, Harpenden, Hertfordshire, UK; 4Centre for Cytochrome P450Biodiversity,InstituteofLifeScience,SchoolofMedicine,SwanseaUniversity,Swansea,UKEmail:t.sewell@imperial.ac.ukLanosterol14-αdemethylase(erg11,CYP51)isakeyenzymeinthefungalergosterolbiosyntheticpathway,andisthetargetofazolefungicides.PriorstudieshaveshownthatLeptosphaeriamaculansandL.biglobosa,thecausalpathogensofphomastemcankeronoilseedrape(Brassicanapus),differintheirsensitivitytosomeazoles.Hereweusedheterologousexpressionandproteinmodellingtodeterminewhetherthereareinter-specificsensitivitydifferencesatthetarget-sitelevel.Moreover,wedeterminedthepracticalsignificanceoffungicidesensitivityusinganewcollectionofisolatesfromthesoutheastoftheUK.HeterologousyeastexpressionofLmCYP51BandLbCYP51Bwithfungicidesensitivitytestingofthetransformantssuggeststhatthetwoproteinsaresimilarlysensitivetoazolefungicidesflusilazole,prothioconazole-desthioandtebuconazole.Thesefindingsaresupportedbyhomologyproteinmodelling,whichpredictsthatLmCYP51BandLbCYP51Barestructurallyverysimilar,specificallyattheazole-bindingsite.Fungicidesensitivitytestingonthenewcollectionofisolatesshowsthattheysometimeshaveaminordifferenceinsensitivityinvitroandinplantabutatpresentbothspeciesremainverysensitivetosomeazolefungicides.TheseresultsshowthatazolefungicidesremainausefulcomponentoftheintegratedmanagementofphomastemcankerintheUKduetotheireffectivenessatcontrollingLeptosphaeriaspp.growth,andsuggestthatotherfactors,suchasvarietalresistanceorclimate,maybedrivingfrequencychangesinthetwospecies.BioactivecompoundswithantimicrobialactivityextractedfrommushroomsgrowninLaRiojaVanessa Grifoll1, Ana Gonzalo1, Maria Luisa Tello1,Margarita Pérez1, Carmen Lozano2,BeatrizRojo2,YolandaSáenz21CentroTecnológicodeInvestigacióndelChampiñónenLaRioja(CTICH).Ctra.Calahorra,KM 4, 26560 Autol, La Rioja, Spain; 2Centro de Investigación Biomédica de La Rioja(CIBIR).CallePiqueras,983º,26006Logroño,LaRioja,Spain.Email:biotecnologia@ctich.com

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Theprolongeduseofantibiotics,inhumanmedicine,veterinaryandagriculture,hascausedanalarmingincreaseofantimicrobialresistantbacteriaworldwide.Concerningtothisseriousthreat,theWorldHealthOrganization(WHO)signedin2016anoverallagreementtoaddresstheproblemofantimicrobialresistance,andin2017publishedalistofprioritypathogensthatrequirethedevelopmentofnewantimicrobialagents.Inthatline,theaimofthisworkwastosearchnewantimicrobialalternativesobtainedfrommushrooms.Sixteenmushrooms(Pleurotuseryngii,Agaricusbisporus,Pleurotusostreatus,Pleurotussalmoneostramineus,Hypsizygusulmarius,Pleurotuscitrinopileatus,Agrocybeaegerita,Hericiumerinaceus,Lentinulaedodes,Agaricusbrunnescens,Agaricusbrazei,Hypsizygustessulatusgris,Hypsizygustessulatusblanca,Grifolafrondosa,GanodermalucidumandPholiotanameko)weregrownonsterilizedsubstratesinLaRioja(Spain)andsubsequentlyfreeze-driedtoobtainamushroompowder.Diversemethodologieswereusedtoobtainthelyophilisedpowdersandthedifferentaqueousandalcoholicextracts.AgardiffusionmethodswereusedtotesttheinvitroantimicrobialactivityagainstreferencestrainsofEscherichiacoli,Listeriamonocytogenes,Pseudomonasaeruginosa,Enterococcusfaecalis,MicrococcusluteusandStaphylococcusaureusspecies.PleurotuscitrinopileatusandLentinulaedodeslyophilisedpowderwereactiveagainstallstudiedbacteriaexcepttoP.aeruginosa.ThepowderofPleurotusostreatus,HypsizygustessulatusgrisandPholiotanamekosuppressedthegrowthofP.aeruginosaorS.aureus.Thefirstresultsareencouraging,andraiseapossibledevelopmentofnewextractswithantimicrobialpropertiesthatmaybeapplicableindifferentindustries.Methodologies development for the cultivation of new species of fungi in La Rioja(Spain)MariaLuisaTello,MargaritaPérezCentroTecnológicodeInvestigacióndelChampiñónenLaRioja(CTICH).Ctra.Calahorra,KM4,26560Autol,LaRioja,SpainEmail:biotecnologia@ctich.comThefungalkingdomischaracterizedbyitshighbiodiversity.Howeveronlyabout100speciesoffungihavebeencultivated.Inthelastyearsthenumberofcultivatedmushroomshasbeenincreasedgradually,optimizingtechniquesusedforgrowingtraditionalspeciestocultivatenewvarieties.Inaddition,themushroommarketinEuropehasgrownandconsumerssearchfornewhealthyproductswithhighqualityandthebestprice.Thus,themainpurposeofthisworkistodevelopthecultivationofnewmushroomspeciesendemictoLaRioja.Toachievethis,CTICHhaveselected12strainsfromthespeciesconservedintheMushroomsBiobankatCTICH.ThisbiobankisafungalgeneticresourcecollectionthatenablestheconservationofwildfungalbiodiversityintheregionofLaRioja(Spain).Thespeciesselectedwerethosewhosecultivationseemed

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morefeasiblebearinginmindseveralfactors,suchas:costofrawmaterialsandproduction,simplicityofcultivationandcollection,marketingoracceptancebypotentialconsumers.AmongtheselectedspeciestherearespecieswithmedicinalpropertieslikeGanodermalucidum,protectedspeciessuchasRhodocybegeminaandediblemushroomslikeCalocybegambosa.Forthecultivationoftheselectedspeciesitwasnecessarytodeveloptechniquesforconservationofthemyceliumandspawnproliferationandnewsubstrateformulae,inadditiontodesigningandoptimizingtheconditionsforincubation,cultureandcropmanagement.Doesnitricoxideregulatecroppathogenresistance?AnnaM.M.TileyandAngelaFeechan

School of Agriculture and Food Science, University College Dublin, Dublin, Dublin 4,IrelandEmail:anna.tiley@ucd.ieZymoseptoriatriticiisapathogenicfungusthatcausesoneoftheworstdiseasesofwheatworldwide,SeptoriaTriticiBlotch.Theaimofourprojectistoinvestigatetheimportanceofnitricoxide(NO)duringZ.triticiinfection.NOisproducedasadefenceresponsebytheplant,andcanactdirectlyasanantimicrobialorasasignallingmolecule.WefirstlyaimtoassesswhetherNOmightregulatetheeffectorproteinsreleasedbyZ.triticiduringinfection.CandidateeffectorshavebeenidentifiedinZ.tritici,andarebeingstudiedtoassesswhethertheyareNO-regulatedduringplantinfection.ThisprojectalsoaimstoidentifywhetherZ.triticiemploysmechanismstocountertheplant’sNOburst.TheenzymeGSNOreductase(GSNOR)controlstheNOpoolinotherspecies,andsoweareinvestigatingtheeffectofdisruptingthegsnorgeneinZ.tritici.BioinformaticsstudieshaveidentifiedapotentialgsnorgenehomologinZ.tritici,ztgsnor.Knock-outplasmidshavebeenconstructedandtransformedintoZ.triticitocreateztgsnorknock-outmutants.Theresultsfromtheseexperimentsareongoing,andthemostrecentfindingsfromthesestudieswillbepresented.EvidenceforinvolvementofanRLXReffectorproteininPhytophthorainfestanssexualreproductionGeorgios Tzelepis, Anna Åsman, Ramesh Vetukuri, Johan Fogelqvist, and ChristinaDixeliusDepartment of Plant Biology, Uppsala Biocenter, Swedish University of AgriculturalSciences,LinneanCenterforPlantBiology,P.O.Box7080,S-75007,Uppsala,SwedenEmail:Georgios.tzelepis@slu.se

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Phytophthorainfestansisanotoriousfungal-likeplantpathogen,responsibleformajoreconomiclossesinpotatoandtomatocultivationsworldwide.Inordertoestablishasuccessfulinfection,thispathogendeploysanarsenalofsmall-secretedproteins,termedeffectors.Amongthem,effectorsharbouringtheRXLRmotifarethebest-studiedones.Theyarehighlyinducedduringearlybiotrophicstageofinfectionandareabletomanipulateplantdefencemechanismsoperatinginsidethehostcell.InthisstudyweinvestigatedthetranscriptionprofilesofdifferentP.infestansisolates(originatedfromdifferentpartsinEurope)duringthematingprocess,sincematingfrequenciesareknowntodifferamongEuropeancountries.OurresultsrevealedthatanRXLReffector-encodinggeneishighlyinducedduringmatinginmostofourcrossings.Furtheranalysisshowedthatoosporesproductionissignificantlyreducedwhenthiseffectorhasbeensilenced.Moreover,weobservedthatthisproteinisaccumulatedinhyphaltipsduringmating,whileinfectionofN.bethamianaplantswiththeGFP-taggedstrainandtransientexpressioninpotatoleavesconfirmedthecytoplasmiclocalizationofthiseffectoruponinfection.Inconclusion,thisstudyrevealedapotentialmultifunctionalroleofanRXLReffector.Interestingly,thepresenceofthiseffectoronlyinheterothalliccloselyrelatedtoP.infestansspecies,supportsitspotentialroleinsexualreproduction.WecanspeculatethatcertaineffectorsmightbeinvolvedinotheraspectsofP.infestansbiologythaninoomycete-plantinteractions.Ifthisfunctionreflectsevolutionaryadaptationremainstobeclarified.PHI-base:HarnessingcommunityexpertisetofightplantdiseaseMartinUrban1,AlayneCuzick1,KimRutherford2,HelderPedro3,NishadiDeSilva3,PaulKersey3andKimE.Hammond-Kosack11DepartmentofBiointeractionsandCropProtection,RothamstedResearch,Harpenden,Herts, AL5 2JQ, UK; 2 Cambridge Systems Biology and Department of Biochemistry,University of Cambridge, Sanger Building, 80 Tennis Court Road, Cambridge,Cambridgeshire CB2 1GA, UK; 3 The European Molecular Biology Laboratory, TheEuropeanBioinformaticsInstitute,Hinxton,Cambridgeshire,CB101SD,UKEmail:Martin.Urban@rothamsted.ac.ukThepathogen-hostinteractionsdatabasePHI-base(www.phi-base.org1)isagold-standarddatabasestoringphenotypesongenesimplicatedinvirulence.Itisaprimaryinformationsourceforresearchersstudyingplant-pathogeninteractionsaswellashumanfungalpathogens.Morethan2300researcharticlesaremadeaccessibleforresearcherstoeasilyfamiliarisethemselveswithrelevantinformationontheirtargetgenesofinterest.PHI-baseprovidesbothmanuallycuratedmolecularandbiologicalphenotypeinformationonpathogengenes.Themulti-speciesdatabasesummarisesinformationfrom264pathogenstestedon173hoststoallowcomparativegenomicsapproachestofacilitatenovelfungicidetargetdiscoveryandidentifycriticallyimportantgenesforbiotech/breedingcontrolstrategies.Allmajorplantpathogensfromthebacterial,fungalandoomycetekingdomsareincluded.

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HerewedescribeournewPHI-baseVersion4.3release(May2017).Ourcurrentdataplatformincludesimprovedsearch,filteringandextendeddatadisplayfunctions.Aprotein-to-phenotypeBLASTsearchfunctionisincluded.Developmentofacommonlanguage(ontology)todescribecomparablephenotypesinawiderangeofpathogen-hostinteractionsisdiscussed.ExemplarusecasesexploringthevastinformationinPHI-baseforfungaldiseasecontrolarepresented.1Urban,M.,Cuzick,A.,Rutherford,K.,Irvine,A.,Pedro,H.,Pant,R.,Sadanadan,V.,Khamari,L.,Billal,S.,Mohanty,S.,etal.(2017).PHI-base:anewinterfaceandfurtheradditionsforthemulti-speciespathogen-hostinteractionsdatabase.NucleicAcidsRes,doi:10.1093/nar/gkw1089.ThisworkissupportedbytheUKBiotechnologyandBiologicalSciencesResearchCouncil(BBSRC)(BB/I/001077/1,BB/K020056/1).PHI-basereceivesadditionalsupportfromtheBBSRCasaNationalCapability(BB/J/004383/1).*Effector discovery and characterisation in the Fusarium graminearum-wheatinteractionCatherineWalker1,2,Wing-ShamLee1,NeilBrown1,MartinUrban1andKim-HammondKosack11BiointeractionsandCropProtection,RothamstedResearch,Harpenden,AL52JQ,UK;2SchoolofBiosciences,UniversityofNottingham,SuttonBonington,LE125RD,UKEmail:catherine.walker@rothamsted.ac.ukFusariumgraminearum(Fg),thecausalpathogenofFusariumHeadBlight,exhibitsanunusualhemibiotrophiclifestylecharacterisedbyspatiallyandtemporallydistinctsymptomlessandsymptomaticphasesofinfection.Thesedistinctphasesarefacilitatedbythebiphasicsecretionofanarsenalofproteins,includingsmall,cysteine-richeffectors,whichmodifylivinghostplants,aidingcolonisationandinfection.Itishypothesisedthatthesecretionofeffectorsactstosubvertrecognitionbythehostand/orsuppresshostdefencesduringthesymptomlessphaseofinfection.Toidentifycandidateeffectors,aspecies-specificmicroarrayanalysiswasusedtointerrogatetheinvitroandinplantaexpressionpatternsofgenesbelongingtotheFgsecretome.TwentyputativeFusariumeffectorswereidentifiedandhavebeguntobecharacterisedusingtheBarley-StripeMosaicVirus-mediatedoverexpressionsystem(BSMV-VOX),stableArabidopsistransformationandviathegenerationofFggenedeletionmutants.InitialdatafromtheBSMV-VOXexperimentssuggeststhatseveralcandidateeffectorproteinsarebiologicallyactiveintheFg-wheatinteraction.DatafromtheseexperimentsindicatesthatviraltitreiscriticaltoproteinexpressionintheVOXexperimentsandthataninteractionbetweenthevirusandsomeproteinsofinterestmaybeoccurring.Preliminaryexperimentssuggestthatoverexpressionofthese

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proteinsinthenon-hostNicotianabenthamianaresultsinthedifferentialexpressionofplantdefence-relatedgenes;ifso,thiscouldofferahigh-throughputmethodforeffectorscreening.Furtherworkisrequiredtodeterminewhetherthisisaviablerouteforeffectorcharacterisationandtomakeeffortstodecipherthefungal-viral-plantinteractionoccurringduringBSMV-VOXexperiments.AnalysisofpathogenicityshiftofCucumbermosaicvirusisolatesinCapsicumannuuminSouthKoreaJu-YeonYoon1,3,BongNamChung1,Seung-KookChoi2andJohnP.Carr31VirologyUnit,HorticulturalandHerbalCropEnvironmentDivision,NIHHS,RDA,Wanju55365 South Korea; 2 Vegetable Research Division, Department of Horticultural CropResearch, NIHHS, RDA, Wanju 55365, South Korea; 3 Department of Plant Science,UniversityofCambridge,Cambridge,CB23EA,UnitedKingdomEmail:juyeon74@gmail.comCucumbermosaicvirus(CMV),isanaphid-bornevirusthatinfectsawiderangeofplantspeciesincludingvegetablecropsandweeds.CMVisoneofthemostimportantviraldiseasesinpepper(Capsicumannuum)inSouthKorea.Since1990s,commercialchillipeppercultivarshavebeenbredforresistanceagainstCMVisolates(pathotype-0,CMV-P0)thatbelongtosubgroupIA(S-IA)becausetheS-IAisolatesweredominant.Recently,twoCMVisolates(CMV-CaP1andCMV-GTN),whichsystemicallyinfectthechillipeppercultivarsresistanttoCMV-P0,werereportedandbelongtoCMVsubgroupIB;theseweredesignatedaspathotype-P1(CMV-P1).Since2009,nationwideannualsurveysofpeppersfieldsshowedthemostdominantCMVpopulation(ca.90%)inpepperandweedswassubgroupIB.Inpathogenicityassays,CMV-GTNinducedseveremosaicandyellowingonchillipepper(cultivarCheong-Yang)21dayspost-inoculation(dpi),whileCMV-CaP1inducedmildoroccasionalmosaic.DetachedleafandDAS-ELISAassaysclearlyshowedthatCMV-GTNandCMV-Fny(S-IA)spreadfasterthanCMV-CaP1inthecultivarCheong-Yang,resultinginmoreseveresystemicsymptoms.PeppercultivarsManitta,BaerottaandLAM32containaresistancegene(s)againstCMV-P1.SystemicinfectionbyCMV-CaP1andCMV-GTNwasverifiedonupperleavesofboth‘Manitta’and‘Baerotta’butnoton‘LAM32’usingELISAanalysis.Interestingly,CMV-GTNinducedmoreseveresymptomsonupperleavesandhadfastersystemicmovementthanCMV-CaP1inBaerotta.TheseresultssuggestapathogenicityshiftinCMVisolatesinpepperplantsinSouthKorea.FundedbyRuralDevelopmentAgency(PJ012426)oftheRepublicofKorea.

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Identification of QTLs underlying resistance to Cucumber mosaic virus in CapsicumannuumJu-YeonYoon1,3, SuKim2,Seung-KookChoi2,EunYoungYang2,BongNamChung1,andJohnP.Carr31VirologyUnit,HorticulturalandHerbalCropEnvironmentDivision,NIHHS,RDA,Wanju55365, South Korea; 2 Vegetable Research Division, Department of Horticultural CropResearch, NIHHS, RDA, Wanju 55365, South Korea; 3 Plant Sciences Department,UniversityofCambridge,Cambridge,CB23EA,UKEmail:juyeon74@gmail.comCucumbermosaicvirus(CMV)isoneofthemostimportantvirusesinfectingchilipepper(Capsicumannuum).RecentlyCMVisolatesthatbelongtosubgroupIB(namedCMV-P1pathotype)overcomeCMVresistanceofpepperinKorea.ResistanceevaluationofpeppergeneticresourcesidentifiedthatCapsicumannuum‘LAM32’showednoveltolerance(hereafterresistance)againstCMVisolatesCMV-P1pathotype.ToidentifyacandidateresistancegeneofLAM32forCMVresistanceQTL,C.annuum‘Subi’asasusceptibleparentwerecrossedandprogeniesofF1,F2andbackcross1populationswereproduced.ParentsandtheirprogenieswereinoculatedwithCMV-GTNisolatethatbelongstosubgroupIBandwereevaluatedforresistanceusinglateralflowimmunoassayandDAS-ELISA.TheresultsshowedthattheresistanceofC.annuum‘LAM32’wascontrolledbyasinglerecessivegene.Asubsetof119putativelymolecularmarkers,welldistributedoverthepeppergenome,wasselectedforQTLmappingusingthecompositeintervalmappingmethod.Sixgenomicregions,includingtwomajoreffectsandfourminoreffectQTLs,maybeassociatedwithresistanceagainstCMV.Subsequentmappinganalysis,basedonapepperlinkagemap,revealedthatgenescontrolledCMVresistancemightbelocatedatchromosome4&5.FurtherfinegeneticmappingforCMVresistanceisinprogress.FundedbyRuralDevelopmentAgency(PJ012426)oftheRepublicofKorea.

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BSPPPresidentialMeeting2018

10–11thDecember2018

@UniversityofWarwick

Prof.MurrayGrant

BSPPPresidentElect