! 1"

The$16th$International$Aspergillus$Meeting$

Asperfest"16"

March"11/12,"2019"

Asilomar"Conference"Center,"Pacific"Grove,"CA"

$$ $

! 2"

Aspergillus$Genomes$Research$Policy$Group$(AGRPG)$

An"Aspergillus"Genomics"workshop"was"held"at"the"March"2003"Asilomar"Fungal"Genetics"meeting."From"discussions"in"that"workshop"it"was"obvious"that"our"community"needed"to"organize"to"fully"exploit"genomics"resources."A"provisional"Aspergillus"Genomes"Research"Policy"Committee"(AGRPC)"was"conscripted"and"charged"with"creating"a"structure"for"community/wide"coordination"and"organizing"an"annual"meeting."The"First"Aspergillus"Meeting"was"held"in"Copenhagen,"April"21,"2004,"as"a"satellite"meeting"of"the"European"Congress"on"Fungal"Genetics/7."In"addition"to"scientific"presentations,"bylaws"were"approved,"community" research"directions"were"discussed"and" the"2004"AGPRC"was"elected."The" name" Aspergillus" Genomes" Research" Policy" Group" was" adopted" for" the" community." The"objectives"of"the"AGRPG"are:"(1)"Provision"of"an"educational"and"discussion"forum"for"issues"pertaining"to"Aspergillus"genomics,"in"its"widest"sense,"and"for"the"various"Aspergillus"research"communitiesS"(2)"Influencing"grant"making"bodies"and"other" institutions"on"behalf"of" the"various"Aspergillus" research"communitiesS"(3)"Coordinating"research"activities"internationally,"as"and"when"required,"to"further"the"science"base"of"the"Aspergillus"genus."For"more"information"on"the"activities"of"the"AGRPG"and"other"Aspergillus"news"see"our"homepage"at"FGSC"(http://www.fgsc.net/Aspergillus/asperghome.html).""

2018$AGRPC$

Richard"Todd,"2016/2018,"Chair,"Local"Organizer,"Kansas"State"University,"USAS"rbtodd@ksu.edu""Mikael"R."Andersen,"2016/2018,"Technical"University"of"Denmark"(DTU),"DenmarkS"mr@bio.dtu.dk""Robert"Cramer,"2016/2018,"Dartmouth,"USAS"robert.a.cramer.jr@dartmouth.edu""Ling"Lu,"2016/2018,"Nanjing"Normal"University,"ChinaS"linglu@njnu.edu.cn"Gerhard"Braus,"2017/2019,"Georg/August/University"Goettingen,"GermanyS"gbraus@gwdg.de"Elaine"Bignell,"2017/2019,"University"of"Manchester,"UKS"elaine.bignell@manchester.ac.uk""David"Cánovas,"2017/2019,"University"of"Seville,"SpainS"davidc@us.es"Vera"Meyer,"2017/2019,"Berlin"University"of"Technology,"GermanyS"vera.meyer@tu/berlin.de""Chris"Koon"Ho"Wong,"2017/2019,"University"of"Macau,"Macau"SARS"koonhowong@umac.mo""Isabelle"Benoit/Gelber,"2018/2020,"Concordia"University,"CanadaS"isabelle.benoit@concordia.ca""Nancy"Keller,"2018/2020,"University"of"Wisconsin"Madison,"USAS"npkeller@wisc.edu""Nir"Osherov,"2018/2020,"Tel"Aviv"University,"IsraelS"nosherov@post.tau.ac.il""Norio"Takeshita,"2018/2020,"University"of"Tsukuba,"JapanS"takeshita.norio.gf@u.tsukuba.ac.jp""Michelle"Momany,"2018,"Past"Chair,"University"of"Georgia,"USAS"mmomany@uga.edu""Kevin"McCluskey"(Ex"officio),"Curator,"Fungal"Genetics"Stock"Center.""

THANKS$TO$OUR$MEETING$SPONSORS!$

!

!!!!! !!!!!!! !

!

!

PROGRAM'

The'Sixteenth'International'Aspergillus'Meeting''Asperfest,16,

Monday!March*11*–*Tuesday*March*12,*2019*Merrill*Hall,*Asilomar*Conference*Center,*Pacific*Grove,*CA.*

*Monday'March'11''

'4:00PM'' ' Poster'hang'up''

5:30PM'B'8:00PM' Registration''

7:00PM'B'10:00PM' Poster'and'Welcome'Reception'(sponsored*by*Novozymes,*Inc.)*'

' 7:00*J*8:30* EvenJnumbered*posters** 8:30*J10:00* OddJnumbered*posters**Judging*for*Novozymes*Student*Poster*Prize,*coordinated*by*Mikael*Andersen,*DTU,*Denmark** * * 'Tuesday'March'12''

'8:30AM' ' Welcome,'introductions'and'announcements* ****** Richard*Todd*

* * ** * * * * * * Kansas*State*University,*USA** *

8:40'' ' Session'I' * * * * * Vera*Meyer*Berlin*University*of*Technology,*Germany*

*Expanding*the*Aspergillus!fumigatus!CRISPR/Cas9*toolbox*with!in!vitro*assembled*ribonucleoproteins.'* Jarrod*Fortwendel,*University!of!Tennessee!Health!Science!Center,!Memphis,!TN,!USA.!**

** Spliceosomal*twin*introns:*diversity,*evolution*and*function.*

* Erzabet*Fekete,*University!of!Debrecen,!Debrecen,!Hungary.***' Surface*analysis*tools*identify*how*Aspergillus!niger*and*its*enzymes*modify*lignocellulose.'

* Jolanda*van*Munster,*University!of!Manchester,!Manchester,!UK.*!** How*do*fungi*get*along*with*other*microbes*in*the*ecosystem?** ** Daisuke*Hagiwara,*University!of!Tsukuba,!Ibaraki,!Japan.*

*10:00B10:25' Coffee'Break'''

10:25''' Session'II'''' * ' ' ' '''''''''''' ''''''''David*Canovas'* * * * * * * * * Universidad*de*Sevilla,*Spain*

* Filamentous*fungi*as*cell*factories*and*microbial*products*for*industrial*uses.* *Debbie*Yaver,*Novozymes**

'High*throughput*genetic*engineering*of*filamentous*fungi*using*automated*liquid*handling*robotics

Ken*Bruno,*Zymergen*** *

* A*manually*curated*resource*for*the*completely*finished*Aspergillus!nidulans*A4*genome*Adrian*Tsang,*Concordia!University,!Montreal,!Canada.!**

**

* '11:20''Community'directions:'FungiDBO'DiscussionO'Elections.'''' ' Richard*Todd'

* * * * * * * * * * Kansas*State*University,*USA*

FungiDB: integrating genomic data for pathogens and model organisms and providing advanced search capabilities and large-scale data analysis

Omar Harb, FungiDB '' Discussiona*Elections**'12:00PM'B'1:15PM'''Lunch'''

Lunch!with!industry!representatives!–!to!participate!please!meet!in!Merrill!Hall!and!enter!Crocker!Dining!Hall!as!a!group.**

'1:15PM' Session'III:'Talks'from'Abstracts' * * * Isabelle*Benoit*Gelber,*

*Concordia*University,*Canada*!! How*do*Microbes*Choose*their*Neighbors?**

* Kurt*Dahlstrom,*California!Institute!of!Technology,!Pasadena,!CA,!USA.**

Molecular*mass*of*αJ1,3Jglucan*affects*the*degree*of*hyphal*aggregation*and*its*localization*in*Aspergillus*nidulans.*** Ken*Miyazawa,*Tohoku!Univ.,!Sendai,!Japan.*

** Investigation*of*proteins*associated*with*hyphal*growth*and*endocytosis*** * Blake*Commer,*Texas!A&M!University,!College!Station,!TX,!USA.***

A*LowJOxygen*Responsive*Alanine*Aminotransferase*is*Important*for*Aspergillus!fumigatus!Biofilm*Formation,*Germination,*and*Growth*During*Infection.*** Joshua*Kerkaert,*Dartmouth!College,!Hanover,!NH,!USA.' ' ' ' '

' '' '' ' ' * * * * * * '

2:15' ' Pontecorvo'Lecture' (sponsored*by*Zymergen,*Inc.)'' ' Robb*Cramer*'Dartmouth*College,*USA*

Ken*Bruno*' ' ' ' Zymergen,*Inc.,*USA*

** Baskerville*fumigatus:*Opportunistic*Pathogen*

* Judy*Rhodes,*Professor*Emerita,**University!of!Cincinnati,!Cincinnati,!OH,!USA.'

*2:45PM* Election'resultsO'other'discussion'items** * * * ******Richard*Todd*

Kansas*State*University,*USA**

' Novozymes'student'poster'prize'presentation** * * Mikael*Andersen,**DTU,*Denmark*

* * * * Novozymes*Inc.,*USA*'3:00PM** Dismiss**(Remove*posters)*

! 5"

List$of$Posters$$Presenter'indicated'in'bold'type,'*"denotes'a'student'poster'presenter'

1.$Importance$of$the$NLterminal$region$of$the$Aspergillus*oryzae$hydrophobin$RolA$in$the$ionic$interaction$with$cutinase$CutL1.""H."Sato,"T."Tanaka,"Y."Terauchi,"K."Nanatani,"A."Yoshimi,"K.$Abe."""*2.$Tracing$monosaccharide$degradation$in$the$Aspergillus*niger$metabolic$network""M.V.$Aguilar$Pontes,"T."Choumpri,"A."Wiebenga,"D."Fulton,"A."Lipzen,"V."Ng,"I."Grigoriev,"A."Tsang,"R.P."de"Vries."""3.$The$Identification$of$novel$mechanisms$for$cell$wall$biosynthesis$in$Aspergillus*fumigatus""Q.$Al$Abdallah,"A."C."O."Souza,"A."Martin/Vicente,"A."Nywening,"W."Ge,"J."R."Fortwendel.""$4.$Sequencing$a$fungal$genus$and$applying$hundreds$of$genomes$for$biotechnology$and$basic$research""M.R.$Andersen,"T.C."Vesth,"J.L."Nybo,"S."Theobald,"J.C."Frisvad,"T.O."Larsen,"J.B."Hoof,"A."Salamov,"I."Grigoriev,"U."Mortensen,"R."Riley,"M."Hainaut,"B."Henrissat,"R.P."de"Vries,"M."Mäkelä,"J."Magnusson,"B."Simmons,"A."Tsang,"M."Machida,"S.E."Baker."""5.$Influence$of$conventional$vs.$organic$agricultural$practices$on$the$ecology,$population$genomics,$and$azole$resistance$status$of$Aspergillus*fumigatus""A.$E.$Barber,"J."Born,"T."Sae/Ong,"I."Schliebner,"K."Kang,"G."Panagiotou,"H."B."Deising,"O."Kurzai."""*6.$Investigation$of$enhanced$virulence$mechanisms$in$Aspergillus*fumigatus*isolates$from$the$International$Space$Station""A.$Blachowicz,"J."Romsdahl,"N."Raffa,"N."P."Keller,"K."Venkateswaran,"C."C."C."Wang."""7.$Inducible$cell$fusion$permits$use$of$competitive$fitness$profiling$in$the$human$pathogenic$fungus$Aspergillus*fumigatus.""Mike$Bromley.""$8.$High$throughput$genetic$engineering$of$filamentous$fungi$using$automated$liquid$handling$robotics""K.S.$Bruno,"E."Szewczyk,"J."Fredlund,"H."Abdulla,"S."Jain,"J."vanDijk,"B.P."Knox,"M."Siddiqui,"A."Fong,"D."Vaughn,"C."Cizauskas,"Z."Serber."""*9.$Characterisation$of$fungal$enzymes$responsible$for$xylan$degradation$in$plant$lignocellulose""G.$Bulmer,"J.M."van"Munster.""$10.$The$central$regulatory$model$of$Aspergillus$conidiation$didn't$work$in$Monascus*ruber""L."Jia,"Z."Huang,"J."Yu,"F."Chen,"W.$Chen.""'*11.$Unraveling$the$genetics$of$pentose$catabolism$in$Aspergillus*niger""Tania$Chroumpi,"Ronald"P."de"Vries.""$*12.$Investigation$of$proteins$associated$with$hyphal$growth$and$endocytosis""B.$Commer,"Z."Schultzhaus,"B.D."Shaw.""$13.$How$do$Microbes$Choose$their$Neighbors?""K.$Martin$Dahlstrom,"Marko"Kojic,"Dianne"Newman."""$$

! 6"

14.$An$Aspergillus*niger$colony$differentiates$its$molecular$responses$to$spatially$separated$substrates"Paul$Daly,"Mao"Peng,"Ad"Wiebenga,"Hugh"D."Mitchell,"Lye"Meng"Markillie,"Charles"Ansong,"Heather"M."Brewer,"Young/Mo"Kim,"Galya"Orr,"Scott"E."Baker,"Peter"de"Gijsel,"Mirjam"A."Kabel,"Kristiina"S."Hilden,"Miia"R."Mäkelä,"Ronald"P."de"Vries""*15."“Expanding$the$fungal$color$chart:$a$novel$PKLNRP$conidial$pigment$acts$together$with$AspLmelanin$to$give$Aspergillus*aurantiobrunneus$its$characteristic$orangeLbrown$color”""P.E.$de$Reus,"H.E."Frederiksen,"S."Theobald,"K.T."Kongsted,"R.J.N."Frandsen."""16.$Population$subdivision$and$the$frequency$of$aflatoxigenic$isolates$in$Aspergillus*flavus$in$the$United$States.""Milton"Drott,"Lauren"Fessler,"Michael"Milgroom.""$*17.$New$unexplored$functions$of$a$C2H2$zinc$finger$transcription$factor,$SltA$for$drug$responses$and$virulence$in$Aspergillus*fumigatus""W.$Du,"L."Lu."""18.$Development$of$a$high$protein$titer$Aspergillus*niger*expression$host$which$is$available$for$licensing""Nigel$Dunn$Coleman,"Wim"de"Laat,"Joyce"Arnouts,"Lucie"Parenicova"and"Wim"de"Laat""*19.$Analysis$of$sclerotia$formation$and$morphology$in$Aspergillus*niger""Valeria$Ellena,"Elsa"Arcalis,"Michael"Sauer,"Matthias"Steiger."""*20.$The$AGC$kinase$YpkA$regulates$sphingolipids$biosynthesis$and$physically$interacts$with$SakA$MAP$kinase$in*Aspergillus*fumigatus""J.$H.$T.$M."Fabri,"N."L."Godoy,"M."C."Rocha,"M."Munshi,"T."P."Fill,"T."A."Coccio,"M."R."V."Z."Kress,"A."F."da"Cunha,"M."Del"Poeta,"I."Malavazi.""'21.$A$spliceosomal$twin$intron$conserved$across$all$classes$of$the$Pezizomycotina$subphylum""N."Kavalecz,"N."Ág,"L."Karaffa,"C."Scazzocchio,"M."Flipphi,"E."Fekete."""22.$Distribution$of$secondary$metabolites$in$Aspergillus$and$Penicillium""J.C.$Frisvad.""$*23.$Dissecting$novel$secondary$metabolites$pathways$in$Aspergillus*homomorphus""M.E.$Futyma,"U."H."Mortensen,"C."Hoeck,"L."M."Petersen,"Y."Guo,"T."O."Larsen,"C."H."Gotfredsen,"J."B."Hoof.""$24.$Morphological,$Molecular$and$antimicrobial$activity$of$endophytic$fungi$isolated$from$forest$plants$at$Taif$region$in$Saudi$Arabia""R.$Gashgari,"A."Alsini.""$*25.$Genomic$analysis$provides$insight$into$secondary$metabolism$of$two$novel,$biosynthetically$talented$of$Aspergillus$from$Australia""C.L.M."Gilchrist,"A.M."Piggott,"E."Lacey,"J.I."Pitt,"Y.H."Chooi.""$26.$Interaction$between$CreD$and$HulA$involved$in$endocytic$degradation$of$the$maltose$transporter$MalP$in*Aspergillus*oryzae""K.$Gomi,"H."Tada,"T."Hiramoto,"Y."Matsuura,"M."Tanaka,"T."Shintani."""*27.$Hybrid$Transcription$Factor$Engineering$Activates$Expression$and$Helps$to$Elucidate$the$Biosynthesis$of$the$Silent$(+)LAsperlin$Gene$Cluster$in$Aspergillus*nidulans""Michelle$F."Grau,"Ruth"Entwistle,"Yi/Ming"Chiang,"Manmeet"Ahuja,"C."Elizabeth"Oakley,"Tomohiro"Akashi,"Clay"C."C."Wang,"Richard"B."Todd,"Berl"R."Oakley.""

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'28.$The$reader$protein$SntB$is$involved$in$regulating$secondary$metabolism$in$Aspergillus*flavus""C."Greco,"B."T."Pfannenstiel,"N."P."Keller."""29.$Diversified$transcriptional$regulation$of$secondary$metabolic$gene$clusters$in$closely$related$Aspergillus$species""H."Takahashi,"M."Umemura,"A."Ninomiya,"M."Shimizu,"Y."Kusuya,"S."Urayama,"A."Watanabe,"K."Kamei,"T."Yaguchi,"D."Hagiwara."""30."MpkB$MAP$kinase$pathway$is$required$for$sexual$development$but$not$for$mycotoxin$production$in$Aspergillus*nidulans$and$Aspergillus*flavus""Sang/Cheol"Jun,"Bohan"Zhu,"Jong/Hwa"Kim,"KapLHoon$Han.""$31.$NsdD$GATA$factorLdependent$regulation$of$fungal$development$and$sporeLspecific$gene$expression$of$Aspergillus*nidulans""Dong/Soon"Oh,"Mi/Kyung"Lee,"Hee/Soo"Park,"Heungyun"Moon,"Xiaoxiao"Huang,"Dong/Min"Han,"Jae/Hyuk"Yu,"KapLHoon"Han."""32.$FungiDB:$integrating$genomic$data$for$pathogens$and$model$organisms$and$providing$advanced$search$capabilities$and$largeLscale$data$analysis""Evelina"Basenko,"Omar"Harb,"Achchuthan"Shanmugasundram,"David"Roos,"..."on"behalf"of"the"EuPathDB"project.""$33.$Fungal$strategy$of$virulence$by$counteracting$lipidLraft$microdomain$formation$of$phagolysosomes""F."Schmidt,"M."Roecker,"A."Thywissen,"H."Schmidt,"Z."Cseresnyés,"S."Galiani,"M"Strassburger,"M.T."Figge,"C."Eggeling,"T.$Heinekamp,"S.G."Filler,"A.A."Brakhage."""34.$The$sepD5$mutation$in$Aspergillus*nidulans$is$located$in$the$gene$encoding$PaxB""T."Hill,"L."Jackson/Hayes,"S."Harris,"A."Virag,"M."Williamson.""$*35.$MultiLtiered$regulation$of$nuclear$accumulation$of$the$GATA$transcription$factor$AreA$in$Aspergillus*nidulans.""Cameron$C."Hunter,"Koon"Ho"Wong3,"Michael"J."Hynes,"Meryl"A."Davis,"Richard"B."Todd."""*36.$The$early$asexual$development$regulator$fluG$codes$for$two$putative$enzymes""M.$IradiLSerrano,"L."Tola/García,"M."S."Cortese,"U."Ugalde.""$37.$CRISPRLmediated$expression$platform$for$multiLspecies$Aspergilli""Z.D.$Jarczynska,"M.E."Futyma,"J.K."Rendsvig,"F.H."Kirchner,"C.S."Nødvig,"U.H."Mortensen."""*38."Mitochondrial$citrate$transporters$CtpA$and$YhmA$are$involved$in$cytosolic$acetylLCoA$biosynthesis$in$the$white$koji$fungus,$Aspergillus*luchuensis*mut.*kawachii.""C."Kadooka,"K."Izumitsu,"M."Onoue,"K."Okutsu,"Y."Yoshizaki,"K."Takamine,"M."Goto,"H."Tamaki,"T."Futagami."""*39.$Sporulation$environment$alters$conidial$transcriptome$and$germination$potential.""S.$Earl$Kang,"Michelle"Momany."""40.$Effects$of$manganese$(II)$ion$transporter$mutations$in$citric$acid$accumulation$in$Aspergillus*niger""B."Fejes,"J.P."Ouedraogo,"E."Fekete,"Á."Soós,"B."Kovács,"E."Sándor,"Á.P."Molnár,"A."Tsang,"L.$Karaffa."""41.$An$AoFus3Linteracting$protein$FipC$is$a$novel$regulator$of$cell$fusion$in$Aspergillus*oryzae""T."Katayama,"T."Mo,"Ö."Bayram,"D."Takemoto,"G."Braus,"K."Kitamoto,"J."Maruyama."""

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*42.$A$LowLOxygen$Responsive$Alanine$Aminotransferase$is$Important$for$Aspergillus*fumigatus*Biofilm$Formation,$Germination,$and$Growth$During$Infection.""Joshua"Kerkaert,"Carey"Nadell,"Robert"Cramer."""43."ProteomeLwide$analysis$of$cysteine$oxidation$in$Aspergillus*fumigatus$reveals$increased$levels$of$oxidatively$modified$thiol$groups$by$hypoxiaLinduced$reactive$oxygen$species""E."Shekhova,"T."Krüger,"M."Stroe,"J."Macheleidt,"L."Ivanova,"O."Kniemeyer,"A.A."Brakhage."""*44.$Fungal$Collagen$Dictates$Colony$and$Biofilm$Morphology$through$Filament$Interactions$to$impact$A.*fumigatus*Disease$Progression""Caitlin$H."Kowalski,"Josh"D."Kerkaert,"Jason"E."Stajich,"Robert"A."Cramer."""*45.$Genetic$engineering$of$the$Aspergillus*niger*regulatory$system$using$CRISPR/Cas9$technology""Roland$Sándor$Kun,"Ronald"P."de"Vries."""46.$Carbon$catabolite$repression$of$cellulase$genes$via$protein$kinase$A$pathway$in$Aspergillus*nidulans""E."Kunitake,"Y."Li,"R."Uchida,"T."Nohara,"K."Asano,"A."Hattori,"T."Kimura,"K."Kanamaru,"M."Kimura,"T."Kobayashi."""47.$The$VosA/VelBLactivated$McrA$governs$cellular$and$metabolic$integrity$in$Aspergillus*nidulans""Mi$Kyung$LEE,"Ye"Eun"Son,"Jae"Hyuk"Yu."""*48."Reeingineering$of$Asperfuranone$pathway$to$yield$Sclerotiorin""P.W.$Lehman,"T.L."Lin,"C.W."Wang."""49.$GenomeLwide$analyses$of$cadmium$stress$response$in$Aspergillus*nidulans""Tamás"Emri,"Erzsébet"Orosz,"Bianka"Huszti,"Éva$Leiter,"Károly"Antal,"Jae/Hyuk"Yu,"István"Pócsi."""*50.$Development$of$a$CRISPRLCas9Lbased$genetic$system$for$Chernobyl$isolated$Cladosporium*cladosporioides""Su$Jeung"Lim,"Adriana"Blachowicz,"Ngan"Pham,"Jingyi"Wang,"Tamas"Torok,"Kasthuri"Venkateswaran,"Clay"C.C."Wang."""*51.$Flexible$assembling$platform$built$up$for$heterologous$expression$of$secondary$metabolites$in$filamentous$fungi""Jun"Lin,"Sandra"Hoefgen,"Janis"Fricke,"Maria"C."Stroe,"Derek"J."Mattern,"Johann"E."Kufs,"Peter"Hortschansky,"Axel"A."Brakhage,"Dirk"Hoffmeister,"Vito"Valiante."""52.$Demystifying$the$NIH$grants$process$and$current$funding$opportunities$for$researchers.$Dona$Love.$$"53.$A$integrated$view$of$calcium$signaling$in$response$to$azole$antifungal$stress$in$Aspergillus*fumigatus""L.$Lu,"J."Ye,"W.L."Du."""*54.$Unravelling$the$aromatic$metabolic$pathways$of$Aspergillus*niger""R.$Lubbers,"Adiphol"Dilokpimol,"Isabelle"Benoit,"Jaap"Visser,"Ronald"de"Vries."""55.$Alkaline$thermostable$recombinant$laccase$from$Coprinopsis*cinerea$uses$syringylLtype$phenols$as$mediators$and$depolymerizes$lignin""C."Lyra,"M."Kähkönen,"P."Nousiainen,"J."Kontro,"J."Sipilä,"M.R."Mäkelä,"K."Hildén."""56.$Fungal$proteins$with$antiLbacterial$properties$secreted$during$infection""Silke$Machata,"Roland"Lehmann,"Hortense"Slevogt,"Axel"Brakhage,"Ilse"Jacobsen.""

! 9"

"57.$Contribution$of$maize$polyamine$and$amino$acid$metabolism$towards$resistance$against$Aspergillus*flavus$infection$and$aflatoxin$production.""R."Majumdar,"R."Minocha,"M."Lebar,"K."Rajasekaran,"S."Long,"C."Carter/Wientjes,"S."Minocha,"J."Cary."""*58.$A$novel$gelsolinLlike$protein$required$for$proper$septum$formation$by$regulating$contractile$actin$ring$formation""M."Mamun,"T."Katayama,"J."Maruyama."""59.$Aspergillus*fumigatus$RasGEFLmediated$invasive$growth""A.$MartinLVicente,"Q."Al"Abdallah,"A."Oliveira"Souza,"J."Fortwendel."""60.$Am$I$my$brother’s$keeper?$Revealing$the$pathogenomic$differences$between$Aspergillus*fumigatus*and$a$(mostly)$innocent$relative""M.E."Mead,"S.L."Knowles,"H.A."Raja,"S.R."Beattie,"C.H."Kowalski,"J.L."Steenwyk,"L.P."Silva,"J."Chiaratto,"L.N.A."Ries,"G.H."Goldman,"R.A."Cramer,"N.H."Oberlies,"A."Rokas."""*61.$Septins$and$stress$response$signaling$pathways$in$A.*nidulans""Alexander$Mela,"Michelle"Momany."""*62.$Molecular$mass$of$αL1,3Lglucan$affects$the$degree$of$hyphal$aggregation$and$its$localization$in$Aspergillus$nidulans""K."Miyazawa,"A."Yoshimi,"T."Yamashita,"A."Koizumi,"S."Yano,"K."Abe."""63."Dissecting$the$role$of$catalytic$residues$of$Human$peroxiredoxins$using$yeast$model""A.$Mohammad,"A.K."Saini.""$64.$Conserved$septin$residues$are$located$in$interfaces$between$monomers""M."Momany,"B."Auxier,"J."Dee,"M."Berbee."""65.$AtrR$is$a$critical$determinant$of$azole$resistance$in$Aspergillus*fumigatus""W.$Scott$MoyeLRowley.$$"66."Enhanced$production$of$antimicrobial$polyketides,$violaceols$in$Aspergillus$nidulans$by$coLculture$with$Aspergillus$fumigatus""A.$Ninomiya,"S."Urayama,"D."Hagiwara."""67."SCF*Complex$Components$Suppress$Septation$Near$the$Hyphal$Tip,$Regulate$Cell$Cycle$Progression$and$Interact$Genetically$with$γLtubulin$in$Aspergillus*nidulans""Vitoria"Paolillo,"James"Dohn,"Berl$Oakley."""68.$Defining$the$importance$of$protein$prenylation$in$A.*fumigatus$growth$and$virulence:$characterization$of$the$αL$(RamB)$and$βL$(Cdc43)$subunits$of$the$GeranylgeranyltransferaseLI$complex""A.$C.$Oliveira$Souza,"A."Martin/Vicente,"Q."Al"Abdallah,"K."DeJarnette,"A."Nywening,"W."Ge,"J."Fortwendel."""*69.$Delta3(E)Ldesaturation$of$glycosylceramides$protects$fungi$against$the$antifungal$protein$AFP$from$Aspergillus*giganteus""N.$Paege,"S."Jung,"V."Meyer."""*70.$Isolation$and$characterization$of$secondary$metabolites$from$Cladosporium*sphaerospermum""Ngan$Pham,"Adriana"Blachowicz,"Jingyi"Wang,"Su"Jeung"Lim,"Tamas"Torok,"Kasthuri"Venkateswaran,"Clay"C."C."Wang.""$

! 10"

71.$The$potential$application$of$Aspergillus*oryzae$in$the$biosorption$of$rare$earth$element$ions$present$in$seepage$waters$from$a$postLuraniumLmining$area""Imre"Boczonádi,"Ágnes"Jakab,"Edina"Baranyai,"Csilla"Noémi"Tóth,"Lajos"Daróczi,"Gréta"Kiss,"Miklós"Antal,"Tamás"Emri,"Tünde"Pusztahelyi,"István"Fábián,"Erika"Kothe,"István$Pócsi."""*72.$Activation$of$Silent$Gene$Clusters$in$Aspergillus*nidulans$using$Hybrid$Transcription$Factors""Christian$A.$Rabot,"Michelle"F."Grau,"Ruth"Enwistle,"Yi/Ming"Chiang,"Manmeet"Ahuja,"C."Elizabeth"Oakley,"Clay"C."C."Wang,"Richard"B."Todd,"Berl"R."Oakley."$*73.$A$Systems$Biology$Approach$to$Signal$Transduction$Gene$Regulation$Response$to$an$Antifungal$Drug$in$Aspergillus*nidulans""S.$Reese,"C."Chelius,"M."Marten,"R."Srivastava,"W."Riekhof,"S."Harris."""*74.$Quantitative$gene$expression$of$Aspergillus*bidirectional$histone$promoters$and$application$as$gene$expression$platform""J.K.$Rendsvig,"J.B."Hoof."""75.$Cell$wall$integrity$pathway$controls$the$asexual$development$of$Aspergillus*fumigatus""M.C.$Rocha,"J.H.T.M"Fabri,"R."Silva/Rocha,"A.F"Cunha,"D."Canovas,"I."Malavazi.""'*76.$Increased$Hypoxia$fitness$of$clinical$Aspergillus*fumigatus$isolates$from$chronic$Aspergillosis$patients""B."Ross,"J."Stajich,"R."Cramer."""*77.$What’s$in$a$spore?$Comparative$proteomics$of$the$mycelia$to$the$spore""K.$RothschildLMancinelli,"L."Bundgaard,"T."Vesth,"U."auf"dem"Keller,"M.R."Andersen."""*78.$Unexpected$production$of$rhizobium$signaling$molecules$across$the$Fungal$Kingdom""T.A."Rush,"A."Bascaules,"P."Jargeat,"F."Maillet,"K.R."Cope,"C."Johnson,"B."Kleven,"A."QuyManh/Maës,"Q.J."Choudhury,"J."Labbé,"C."Swift,"M.A."O’Malley,"J.W."Bok,"S."Cottaz,"S."Fort,"J."Nett,"N.P."Keller,"V."Puech/Pagès,"G."Bécard,"J.M."Ané."""*79.$The$monothiol$glutaredoxin$GrxD$interacts$with$the$iron$regulator$HapX$and$CmtA,$a$protein$with$yet$unknown$function,$in$Aspergillus*fumigatus""M.T.$MLL."Scheven,"M."Misslinger,"P."Hortschansky,"T."Krüger,"J."Weber,"H."Haas,"A."A."Brakhage.""'*80."Examination$of$ploidy$in$Aspergillus*fumigatus""B.A.$Shuman,"E."Scopel,"S.E."Kang,"D."Bensasson,"M."Momany."""*81."Extensive$loss$of$cell$cycle$and$DNA$repair$genes$in$an$ancient$lineage$of$bipolar$budding$yeasts""J.L."Steenwyk,"D.A."Opulente,"J."Kominek,"X./X."Shen,"X."Zhou,"A.L."Labella,"N.P."Bradley,"B.F."Eichman,"N."Cadez,"D."Libkind,"J."DeVirgilio,"A.B."Hulfachor,"C.P."Kurtzman,"C.T."Hittinger,"A."Rokas."""*82.$A$robust$phylogenomic$timetree$for$biotechnologically$and$medically$important$fungi$in$the$genera$Aspergillus*and$Penicillium$Jacob$L.$Steenwyk,"Xing/Xing"Shen,"Abigail"L."Lind,"Gustavo"H."Goldman,"and"Antonis"Rokas.$$*83.$Regulation$of$Leucine$Biosynthesis$and$a$Key$Nitrogen$Assimilation$Gene$by$the$Transcription$Factor$LeuR$in$Aspergillus*nidulans""Joel$T.$Steyer,"Damien"J."Downes,"Cameron"C."Hunter,"Richard"B."Todd."""$$$

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*84.$FungalLbacterial$coLculture$induces$biosynthesis$of$cryptic$natural$products$in$the$genus$Aspergillus*and$leads$to$identification$of$BasR$as$key$regulatory$node$for$translating$bacterial$signals""Maria$C."Stroe,"Tina"Netzker,"Kirstin"Scherlach,"Vito"Valiante,"Nils"Jaeger,"Juliane"Fischer,"Volker"Schroeckh,"Christian"Hertweck,"Thorsten"Heinzel,"Axel"Brakhage."""*85.$Comparative$genomics$of$endoparasitic$fungi,$Esteya*vermicola""Jongoh"Shin,"Jaemin$Sung,"Byungkwan"Cho."""*86."IntraLspecific$aflatoxin$inhibition$profile$by$atoxigenic$Aspergillus*flavus$reveals$touchLindependent$inhibition""R.R.$Sweany,"C.D."DeRobertis,"K.E."Damann"Jr.."""87.$Live$cell$imaging$analysis$of$fungalLbacterial$interaction""N.$Takeshita,"M."Kuchira,"G."Abeysinghe,"D."Hagiwara,"N."Nomura,"N."Takaya,"N."Obana."""88."Heterologous$production$and$the$productivity$enhancement$of$polyunsaturated$free$fatty$acids$by$metabolic$engineering$of$Aspergillus*oryzae""K."Tamano,"A."Itoh,"Y."Yasunaka,"A."Miura,"E."Kan,"Y."Koyama,"T."Tamura."""89.$Identification$and$distinct$regulation$of$oligopeptide$transporter$genes$in$Aspergillus*oryzae$"M.$Tanaka,"K."Ito,"Y."Kawarasaki,"K."Gomi."'90."The$secondary$metabolism$regulator$LaeA$in$Penicillium*sp.$influences$the$cheese$rind$community$assembly""J."Tannous,"C."Cosetta,"T."Rush,"J."Jiler,"N."Keller,"B."Wolfe.""$91.$Peptidyl$compound$synthetic$factors$widely$conserved$and$highly$diverse$in$the$Fungi$kingdom""Maiko"Umemura,"Linh"Dao,"Yuki"Nakashima."""92.$Surface$analysis$tools$identify$how$Aspergillus*niger*and$its$enzymes$modify$lignocellulose""J.M.$van"Munster,"T.V."Vuong,"S.E."Marcus,"J.P."Knox,"E.R."Master.""$93."Discovery$and$elucidation$of$the$biosynthesis$of$aspernidgulenes,$novel$polyenes$from$Aspergillus*nidulans,$using$serial$promoter$replacement""Tzu/Shyang"Lin,"Bethany"Chen,"Yi/Ming"Chiang,"Clay$C.$C.$Wang.""$*94."Elucidating$biosynthetic$pathway$of$a$compound$isolated$from$Aureobasidiumpullulans""Jingyi"Wang,"Adriana"Blachowicz,"Ngan"Pham,"Su"Jeung"Lim,"Tamas"Torok,"Kasthuri"Venkateswaran,"Clay"C."C."Wang.""$*95.$The$function$of$long$nonLcoding$RNA$lncR6731$in$a$plant$pathogenic$fungus$Fusarium*graminearum""J."Wang1,2,"W"Zeng,"Y"Fu,"D"Jiang,"J"Cheng."""96."FineLtuning$gene$expression:$Pantothenic$acid$inducible$promoters$in$Trichoderma*reesei""Franziska"Wanka,"Robert"H."Bischof,"Benjamin"Metz,"Christian"Gamauf,"Georg"Schirrmacher,"Christian"P."Kubicek,"Bernhard"Seiboth.""$97.$The$unfolded$protein$response$is$linked$with$Ca2+$homeostasis$to$mediate$stress$adaptation$and$cell$wall$integrity$in$Aspergillus*fumigatus""M."Weichert,"J.P."Guirao/Abad,"K."Krishnan,"V."Aimanianda,"C."Grisham,"R."Abbu,"A."Sheehan,"D.S."Askew.""$$

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*98.%Discovery%and%activation%of%a%tetracycline5like%producing%gene%cluster%in%Aspergillus*sydowii##P.%B.%Wolff,#K.#Subko,#S.#Theobald,#J.#B.#Hoof,#J.#C.#Frisvad,#C.#H.#Gotfredsen,#M.#R.#Andersen,#U.#H.#Mortensen,#T.#O.#Larsen.###*99.%The%Zn2Cys65type%transcription%factor%LeuB5involved%interconnection%between%BCAA%biosynthesis%and%the%Ca2+%signaling%pathways%in%Aspergillus*fumigatus##J.%Ye,#N.#Long,#L.#Gao,#L.#Lu.###100.%In*silico%screened%compound%Z5705%specifically%inhibits%protein%kinase%C%of%filamentous%fungi.##A.#Yoshimi,#A.#Sugahara,#F.#Shoji,#T.#Fujioka,#K.#Kawai,#H.#Umeyama,#K.#Komatsu,#K.#Miyazawa,#K.#Abe.##%*101.%Genome%Mining%of%the%Biosynthetic%Gene%Cluster%of%Citrinalin%A%in%Penicillium*citrinum*using%CRISPR5Cas9##B.#Yuan,#J.#van#Dijk,#Y.#Ye,#C.#Wang,#D.#Sherman.##%102.%Towards%minimal%Aspergillus%genomes:%editing%Aspergillus*niger%mycotoxin%clusters%using%an%optimized%CRISPR/Cas%system##X.#Zheng,#P.#Zheng,#T.#Cairns,#V.#Meyer,#Y.#Ma.###103.%A%manually%curated%resource%for%the%completely%finished%Aspergillus*nidulans%A4%genome#Erin#McDonnell,#Kimchi#Strasser,#Ian#Reid,#Alexandre#Beaudoin,#Nicholas#Geoffrion,#Debra#Fulton,#Sylvester#Palys,#Gregory#Butler,#Isabelle#Benoit#Gelber,#Thi#Truc#Minh#Nguyen,#Justin#Powlowski,#Kenneth#Dewar,#Jessica#Wasserscheid,#Elizabeth#Oakley,#Damien#Downes,#Chris#Wong,#Tomohiro#Akashi,#Arthur#Ram,#Xin#Xiang,#Gustavo#Goldman,#Ronald#de#Vries,#Richard#Todd,#Berl#Oakley,#Adrian%Tsang.##104.#Pharmacologic%inhibition%of%the%UPR%sensor%IreA%has%antifungal%effects%in%Aspergillus*fumigatus.%#Jose%P.%Guirao%Abad,%Aaron#Albee,#Martin#Weichert,#David#S.#Askew%#!

% %

! 13"

Presenting$Authors$$(Alphabetical6'Student'presenters'in'bold'type'with'asterisk)'Presenter$$ $ $ Poster$number$Abe,"K.""" " " " 1"Aguilar$Pontes,$M.V.$$ $ $ 2*"Al"Abdallah,"Q.""" " " 3"Andersen,"M.R."" " " 4"Barber,"A."E."" " " " 5"Basenko,"Evelina"" " " 32"Blachowicz,"A.$$ $ $ 6*"Bromley,"Mike"" " " " 7"Bruno,"K.S.""" " " " 8"Bulmer,"G.$"" " " " 9*"Chen,"W."" " " " 10"Chroumpi,$Tania"" " " 11*"Commer,"B.$ $ $ $ 12*"Dahlstrom,"K."Martin" " " 13"""Daly,"Paul" " " " 14"de$Reus,"P.E."" " " " 15*"Drott,"Milton"" " " " 16"Du,$W.""" " " " 17*"Dunn"Coleman,"Nigel" " " 18"Ellena,$Valeria""" " " 19*"Fabri,$J.$H.$T.$M."" " " 20*"Fekete,"E.""" " " " 21"Frisvad,"J.C."" " " " 22"Futyma,$M.E.$$ $ $ $ 23*"Gashgari,"R."" " " " 24"Gilchrist,$C.L.M."$$ $ $ 25*""Gomi,"K."" " " " 26"Grau,$Michelle$F."" " " 27*"Greco,"C."" " " " 28"Guirao"Abad,"Jose"P."" " " 104""Hagiwara,"D."" " " " 29"Han,"Kap/Hoon""" " " 30,"31"Harb,"Omar" " "" " 32"Heinekamp,"T."""" " " 33" "Hill,"T."" " " " " 34"Hunter,$Cameron$C." " " 35*"IradiLSerrano,$M."" " " 36*"Jarczynska,"Z.D."" " " 37"Kadooka,$C.$ $ $ $ 38*"Kang,$S.$Earl"" " " " 39*"Karaffa,"L."" " " " 40"Katayama,"T."" " " " 41"Kerkaert,$Joshua"" " "" 42*"Kniemeyer,"O."" " " " 43"Kowalski,$Caitlin$H."" " " 44*"Kun,$Roland$Sándor"" " " 45*"Kunitake,"E.""" " " " 46"LEE,"Mi"Kyung""" " " "47"Lehman,$P.W."" " " " 48*"Leiter,"Éva"" " " " 49"Lim,"Su$Jeung$$$ $ $ 50*"Lin,"Jun$$ $ $ $ 51*"

Presenter$$ $ $ Poster$number$Love,"Dona"" " " " 52"Lu,"L."" " " " " 53"Lubbers,$R."" " " " 54*"Lyra,"C."" " " " 55"Machata,"Silke.""" " " 56"Majumdar,"R."" " " " 57"Mamun,$M."" " " " 58*""Martin/Vicente,"A."" " " 59""M.E."Mead"" " " " 60"Mela,$Alexander"" " " 61*"Miyazawa,$K."$$ $ $ $ 62*"Mohammad,"A.""" " " 63"Momany,"M."" " " " 64"Moye/Rowley,"W."Scott""" " 65"Ninomiya,"A."" " " " 66""Oakley,"Berl"" " " " 67"Oliveira"Souza,"A."C.""" " "" 68"N.$Paege,$N.$" " " " 69*""Pham,$Ngan"" " " " 70*"Pócsi,"István"" " " " 71"Rabot,$Christian$A."" " " 72*"Reese,$S."" " " " 73*"Rendsvig,$J.K.""" " " 74*"Rocha,$M.C."" " " " 75"Ross,$B."" " " " 76*"RothschildLMancinelli,$K." " 77*"Rush,"T.A.$$ $ $ $ 78*"Scheven,$M.T.$MLL."" " " 79*"Shuman,$B.A."" " " " 80*"Steenwyk,$J.L.""" " " 81*"Steenwyk,$Jacob$L."" " " 82*$Steyer,$Joel$T.""" " " 83*"Stroe,$Maria$C."" " " 84*"Sung,$Jaemin"" " " " 85*"Sweany,$R.R."" " " " 86*"Takeshita,"N."" " " " 87"Tamano,"K."" " " " 88"Tanaka,"M."" " " " 89"Tannous,"J."" " " " 90"Tsang,"A."" " " " 103""Umemura,"Maiko"" " " 91"van"Munster,"J.M.""" " " 92"Wang,"Clay"C."C."" " " 93"Wang,$Jingyi"" " " " 94*"Wang,$J."" " " " 95*"Wanka,"Franziska"" " " 96"Weichert,"M."" "" " " 97"Wolff,$P.$B."" " " " 98*"Ye,$J."" " " " " 99*"Yoshimi,"A."" " " " 100"Yuan,$B."" " " " 101*"Zheng,"X."" " " " 102

! 14"

Student$Presenting$Authors$$(Alphabetical)'

Presenter$$ $ $ Poster$number$Aguilar$Pontes,$M.V.$$ $ $ 2*"Blachowicz,"A.$$ $ $ 6*"Bulmer,"G.$"" " " " 9*"Chroumpi,$Tania"" " " 11*"Commer,"B.$ $ $ $ 12*"de$Reus,"P.E."" " " " 15*"Du,$W.""" " " " 17*"Ellena,$Valeria""" " " 19*"Fabri,$J.$H.$T.$M."" " " 20*"Futyma,$M.E.$$ $ $ $ 23*"Gilchrist,$C.L.M."$$ $ $ 25*""Grau,$Michelle$F."" " " 27*"Hunter,$Cameron$C." " " 35*"IradiLSerrano,$M."" " " 36*"Kadooka,$C.$ $ $ $ 38*"Kang,$S.$Earl"" " " " 39*"Kerkaert,$Joshua"" " "" 42*"Kowalski,$Caitlin$H."" " " 44*"Kun,$Roland$Sándor"" " " 45*"Lehman,$P.W."" " " " 48*"Lim,"Su$Jeung$$$ $ $ 50*"Lin,"Jun$$ $ $ $ 51*"Lubbers,$R."" " " " 54*"Mamun,$M."" " " " 58*""Mela,$Alexander"" " " 61*"Miyazawa,$K."$$ $ $ $ 62*"N.$Paege,$N.$" " " " 69*""Pham,$Ngan"" " " " 70*"Rabot,$Christian$A."" " " 72*"Reese,$S."" " " " 73*"Rendsvig,$J.K.""" " " 74*"Rocha,$M.C."" " " " 75"Ross,$B."" " " " 76*"RothschildLMancinelli,$K." " 77*"Rush,"T.A.$$ $ $ $ 78*"Scheven,$M.T.$MLL."" " " 79*"Shuman,$B.A."" " " " 80*"Steenwyk,$J.L.""" " " 81*"Steenwyk,$Jacob$L."" " " 82*$Steyer,$Joel$T.""" " " 83*"Stroe,$Maria$C."" " " 84*"Sung,$Jaemin"" " " " 85*"Sweany,$R.R."" " " " 86*"Wang,$Jingyi"" " " " 94*"Wang,$J."" " " " 95*"Wolff,$P.$B."" " " " 98*"Ye,$J."" " " " " 99*"Yuan,$B."" " " " 101*"!

$ $

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Abstracts$$Presenter'indicated'in'bold'type'*"denotes'a'student'poster'presenter'$1.$Importance$of$the$NLterminal$region$of$the$Aspergillus*oryzae$hydrophobin$RolA$in$the$ionic$interaction$with$cutinase$CutL1.""H."Sato1,"T."Tanaka1,"Y."Terauchi1,"K."Nanatani1,"A."Yoshimi2,"K.$Abe1,2.""1)'Dept'Microbial'biotechnology,'Grad.'Sch.'Agricult.'Sci.,'Tohoku'Univ.,'Sendai,,'Miyagi,'JP6'2)'New'Industry'Creation'Hatchery'Center,'Tohoku'Univ.,'Sendai,,'Miyagi,'JP"""When"the"industrial"fungus'Aspergillus'oryzae"is"grown"with"the"biodegradable"plastic"polybutylene"succinate/co/adipate"(PBSA)"as"the"sole"carbon"source,"hydrophobin"RolA"and"PBSA"degrading"enzyme"cutinase"CutL1"are"co/expressed"and"consequently"hydrolyze"PBSA."1)"Hydrophobins"are"amphipathic"proteins"unique"to"filamentous"fungi."Hydrophobin"RolA"pre/attached"to"PBSA"particles"recruits"and"concentrates"CutL1"on"the"surface"of"the"particles"to"promote"decomposition"of"PBSA."In"previous"studies,"we"found"that"these"two"proteins"interact"via"ionic"interaction"and"positively"charged"amino"acid"residues"in"the"N/terminal"region"of"RolA"and"negatively"charged"residues"on"the"molecular"surface"of"CutL1"are"directly"involved"in"the"ionic"interaction.2)"It"was"also"found"that"the"hydrophobin"RodA"of"the"model"filamentous"fungus"Aspergillus'nidulans"also"ionically"interacts"with"CutL1."2)"However,"the"dissociation"constant�KD�between"RodA/CutL1"was"about"40"times"larger"than"that"between"RolA/CutL1."We"wonder"whether"the"core"structures"of"the"two"hydrophobins"cause"the"large"difference"in"the"two"KD"values"besides"the"secondary"structures"and"amino"acid"sequences"of"the"N/terminal"regions"of"RolA"and"RodA."In"the"present"study,"we"constructed"chimeric"mutants"in"which"the"N/terminal"region"was"exchanged"between"RolA"and"RodA,"and"determined"KD"values"of"the"chimeric"hydrophobins"for"CutL1"by"using"a"Quartz"Crystal"Microbalance"with"purified"chimeric"mutants."The"chimeric"mutants"having"the"N/terminus"derived"from"RolA"wild/type"and"RodA"core/structure"showed"significantly"lower"KD"values"compared"to"RodA"wild/type."On"the"contrary,"the"chimeric"mutants"with"the"N/terminal"sequence"of"RodA"wild/type"and"RolA"core/structure"had"significantly"higher"KD"values"than"RolA"wild/type."These"results"suggest"that"the"N/terminal"region"of"RolA"mainly"contributes"to"the"RolA/CutL1"interaction,"and"its"affinity"depends"on"the"density"of"positive/charged"residues"and"primary"sequence"in"the"N/terminal"region"of"RolA."1)"Takahashi"T.,"et"al."Mol.'Microbiol."57:1780–1798"(2005)"2)"Takahashi"T.,"et"al."Mol.'Microbiol."96:14/27"(2015)""*2.$Tracing$monosaccharide$degradation$in$the$Aspergillus*niger$metabolic$network""M.V.$Aguilar$Pontes1,"T."Choumpri1,"A."Wiebenga1,"D."Fulton2,"A."Lipzen3,"V."Ng3,"I."Grigoriev3,"A."Tsang2,"R.P."de"Vries1.""1)'Fungal'Physiology,'Westerdijk'Fungal'Biodiversity'Institute,'Utrecht,'NL6'2)'CSFGQConcordia'University,'Canada6'3)'Department'of'Energy'Joint'Genome'Institute,'Walnut'Creek'CA,'USA"""Depolymerization"of"plant"biomass"results"in"a"large"variety"of"soluble"sugars"ready"to"be"taken"up"by"the"degrading"organisms."Fungi"are"able"to"utilize"them"in"a"highly"efficient"and"controlled"manner."One"of"the"best"studied"species"is"Aspergillus'niger,"an"industrial"workhorse"used"for"enzyme"and"metabolite"production."In"order"for"A.'niger"to"achieve"its"full"potential"as"a"cell"factory,"deeper"knowledge"of"its"metabolism"is"needed."In"this"project,"we"focused"on"identifying"the"pathways"and"corresponding"enzymes"involved"in"the"conversion"of"plant"biomass"derived"monosaccharides,"and"its"regulatory"system."Based"on"the"manually"curated"annotation"of"the"A.'niger'NRRL3"genome,"we"automatically"reconstructed"the"genome/scale"metabolic"network"and"manually"curated"reactions"involved"in"plant"cell"wall"degradation"and"carbon"catabolism"based"on"literature"references,"as"well"as"protein,"metabolite"and"reaction"databases"and"expression"data."In"total,"the"carbon"metabolic"network"contains"329"enzymes"and"274"enzymatic"reactions,"linked"to"149"literature"references."RNA/seq"data"from"transfer"experiments"after"2"hours"grown"on"single"monosaccharides"was"used"to"identify"changes"in"gene"expression"distinguishing"between"constant"and"specific"expression"under"different"conditions."Additionally,"we"were"able"to"validate"expression"for"

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alternative"genes"and"predict"new"enzymes"assigned"to"orphan"reactions."During"plant"biomass"degradation,"activation"of"gene"expression"via"specific"inducers"is"balanced"with"gene"repression"via"carbon"catabolite"repression"mediated"by"CreA."RNA/seq"data"of"a"ΔcreA"strain"grown"on"monosaccharides"showed"that"the"strongest"influence"of"CreA"is"on"genes"encoding"plant"polysaccharides"degrading"enzymes"and"specific"sugar"transporters.""3.$The$Identification$of$novel$mechanisms$for$cell$wall$biosynthesis$in$Aspergillus*fumigatus""Q.$Al$Abdallah,"A."C."O."Souza,"A."Martin/Vicente,"A."Nywening,"W."Ge,"J."R."Fortwendel.""College'of'Pharmacy,'University'of'Tennessee'Q'Health'Science'Center,'MEMPHIS,'TN"""Despite"recent"advances,"the"mechanisms"that"regulate"cell"wall"biosynthesis"are"still"poorly"defined"in"the"human"pathogenic"fungus"Aspergillus'fumigatus."We"performed"a"forward"genetics"screen"to"evolve"cell"wall"stress"resistant"strains"under"increasing"concentrations"of"Calcofluor"White"(CFW)"and"Congo"Red"(CR)."Analysis"of"these"evolved"strains"is"expected"to"reveal"novel"mechanisms"regulating"cell"wall"biosynthesis"and"stress"response"in"A.'fumigatus."Whole/genome"sequencing"analysis"of"nine"resistant"strains"identified"86"potential"resistance/associated"mutations"in"the"coding"regions"of"over"60"genes."Interestingly,"mutations"in"the"tscC"gene"were"identified"in"all"resistant"strains."Previous"studies"reported"that"deletion"of"A.'fumigatus"tcsC,"a"hybrid"histidine"kinase"(HHK)"of"the"Two/component"system"(TCS),"results"in"hyper/"and"hypo/susceptibility"to"osmotic"and"cell"wall"stresses,"respectively."One"of"the"mutations"among"our"evolved"strains,"tcsCT813I,"occurs"within"the"conserved"kinase"domain"of"the"TcsC"protein."To"examine"the"functional"consequence"of"this"mutation,"we"performed"an"allele"swap"replacing"the"tcsC"locus"within"a"wild/type"strain"with"the"tcsCT813I"mutant"allele."The"resulting"strain"(Af293/tcsCT813I)"grew"normally"under"non/stress"conditions,"yet"was"highly"resistant"to"cell"wall"stress"induced"by"CFW"and"CR."In"contrast,"Af293/tcsCT813I"exhibited"no"changes"in"response"to"osmotic"stress"when"compared"to"the"wild"type"control."Together,"these"data"suggest"that"the"tcsCT813I"point"mutation"impacts"TcsC"activity"in"a"manner"that"uncouples"the"roles"of"TcsC"in"osmotic"and"cell"wall"stress"responses."Therefore,"this"mutant"will"be"a"valuable"tool"for"probing"regulatory"mechanisms"modulated"through"the"TcsC"two/component"system."To"further"TcsC"pathway"characterization"in"A.'fumigatus,"we"next"performed"a"promoter"exchange"of"the"downstream"histidine"phosphotransferase,"ypdA,"replacing"the"native"promoter"with"a"tetracycline/inducible"promoter"(tetON)."Loss"of"YpdA"expression"in"this"strain"resulted"in"severe"growth"reduction,"suggesting"an"essential"role."Future"studies"will"focus"on"how"the"TcsCT813I"mutation"regulates"cell"wall"biosynthesis"and"further"identifying"cell"wall"biosynthesis"mechanisms"via"our"forward"genetics"analyses."$4.$Sequencing$a$fungal$genus$and$applying$hundreds$of$genomes$for$biotechnology$and$basic$research""M.R.$Andersen1,"T.C."Vesth1,"J.L."Nybo1,"S."Theobald1,"J.C."Frisvad1,"T.O."Larsen1,"J.B."Hoof1,"A."Salamov2,"I."Grigoriev2,14,"U."Mortensen1,"R."Riley2,"M."Hainaut6,7,"B."Henrissat6,7,8,"R.P."de"Vries11,"M."Mäkelä13,"J."Magnusson3,4,"B."Simmons3,12,"A."Tsang5,"M."Machida10,"S.E."Baker3,9.""1)'Department'of'Biotechnology'and'Biomedicine,'Technical'University'of'Denmark,'Kgs.'Lyngby,'DK6'2)'US'Department'of'Energy'Joint'Genome'Institute,'Walnut'Creek,'CA,'USA.6'3)'US'Department'of'Energy'Joint'BioEnergy'Institute,'Emeryville,'CA,'USA.6'4)'Chemical'and'Biological'Process'Development'Group,'Energy'and'Environment'Directorate,'Pacific'Northwest'National'Laboratory,'Richland,'WA,'USA.6'5)'Centre'for'Structural'and'Functional'Genomics,'Concordia'University,'Montreal,'Quebec,'Canada.6'6)'Architecture'et'Fonction'des'Macromolécules'Biologiques,'CNRS'UMR'7257,'AixQMarseille'University,'Marseille,'France.6'7)'Institut'National'de'la'Recherche'Agronomique,'USC'1408'Architecture'et'Fonction'des'Macromolécules'Biologiques,'Marseille,'France.6'8)'Department'of'Biological'Sciences,'King'Abdulaziz'University,'Jeddah,'Saudi'Arabia.6'9)'Molecular'Sciences'Division,'Earth'and'Biological'Sciences'Directorate,'Pacific'Northwest'National'Laboratory,'Richland,'WA,'USA.'6'10)'Kanazawa'Institute'of'Technology,'IshikawaQken,'Japan6'11)'Fungal'Physiology,'Westerdijk'Fungal'Biodiversity'Institute'and'Fungal'Molecular'Physiology,'Utrecht'University,'Utrecht,'The'Netherlands.6'12)'Lawrence'Berkeley'National'Laboratory,'Berkeley,'CA,'USA.6'13)'Department'of'Microbiology,'University'of'Helsinki,'Helsinki,'Finland.6'14)'Department'of'Plant'and'Microbial'Biology,'University'of'California,'Berkeley,'CA,'USA.""

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"In"the"Aspergillus"whole/genus"sequencing"project,"we"are"sequencing"at"least"one"member"of"each"of"~300"species"in"the"genus,"focusing"on"type"strains"and/or"the"most"representative"isolates"of"the"species."The"goals"of"this"project"include"the"study"of"fungal"speciation,"diversity"of"primary"metabolism,"mapping"of"biofuel/relevant"CAZymes,"and"elucidation"of"bioactive"secondary"metabolites."Currently,"we"have"sequenced"the"genomes"of"197"species,"and"98"more"are"currently"being"sequenced."In"this"talk,"I"will"give"an"overview"of"the"status"and"goals"of"the"project."Furthermore,"I"will"present"how"we"are"analysing"the"genomes,"and"show"the"most"interesting"results"from"our"analyses."In"particular,"I"will"present"how"we"use"the"principles"of"association"analysis"–"as"known"from"human"genetics"–"at"a"genus"level"to"identify"the"genes"responsible"for"specific"phenotypes."I"will"show"examples"of"cases"from"a"wide"range"of"species,"spanning"A.'nidulans,'A.'flavus,'A.'oryzae,'A.'niger,'A.'fumigatus,"and"a"host"of"other"new"and"interesting"species,"while"covering"both"basic"research"and"direct"applications."The"talk"will"show"the"discovery"of"thousands"of"new"secondary"metabolite"gene"clusters,"and"tens"of"thousands"of"carbohydrate/acting"enzymes"(CAZymes),"and"showcase"some"of"the"unique"features"of"the"Aspergillus'genus."Of"particular"interest"is"the"linking"of"compounds"to"clusters""such"as"for"the"potential"anti/cancer"agent"malformin"and"novofumigatonin,"mapping"of"the"diversity"of"CAZymes,"and"identification"of"variation"of"genome"content"between"fungal"species.""5.$Influence$of$conventional$vs.$organic$agricultural$practices$on$the$ecology,$population$genomics,$and$azole$resistance$status$of$Aspergillus*fumigatus""A.$E.$Barber1,"J."Born2,"T."Sae/Ong1,"I."Schliebner2,"K."Kang1,"G."Panagiotou1,"H."B."Deising2,"O."Kurzai1,3.""1)'Leibniz'Institute'of'Natural'Product'Research'and'Infection'Biology'–'Hans'Knöll'Institute,'Jena,'DE6'2)'Martin'Luther'University'HaaleQWittenberg.'Halle'(Saale),'DE6'3)'University'of'Würzburg.'Würzburg,'DE""'Aspergillus'fumigatus"is"a"ubiquitous"saprophyte"capable"of"causing"invasive"infections"and"current"management"strategies"rely"on"the"azoles."Unfortunately,"over"the"last"decade"there"has"been"a"rapid"and"global"emergence"resistance"to"the"azoles"in"A.'fumigatus"and"the"dominant"resistance"mechanism"is"of"environmental"origin,"suggesting"that"resistance"is"emerging"through"a"selective"pressure"applied"by"the"widespread"usage"of"azoles"in"agriculture."To"examine"the"link"between"the"use"of"azoles"in"agriculture"and"the"emergence"of"clinical"resistance,"systematic"soil"sampling"was"performed"over"a"three"year"period"on"seven"conventional"agricultural"sites"utilizing"azole"fungicides"and"on"six"organic"sites"withholding"these"compounds."Conventional"sites"were"evaluated"both"before"and"after"azole"application,"and"in"total,"2875"soil"samples"were"analyzed"between"2016"and"2018."We"observed"a"high"degree"of"variation"in"the"abundance"of"A.'fumigatus'across"sitesS"however,"fields"with"high"A.'fumigatus"density"tended"to"be"consistently"so"from"year"to"year."Strikingly,"we"observed"a"reduction"in"the"abundance"of"A.'fumigatus"on"conventional"fields"following"azole"treatment"–"a"finding"that"was"not"repeated"on"an"organic"control"field"–"indicating"that"the"application"of"azoles"is"imposing"a"bottleneck"on"A.'fumigatus.""Resistance"to"medical"azoles"was"assessed"for"≈800"isolates"and"6%"of"isolates"collected"in"2016"and"1%"of"isolates"from"2017"displayed"resistance"to"one"or"more"medical"azole."Susceptibility"to"agricultural"azoles"was"also"assessed"and"isolates"resistant"to"medical"azoles"typically"had"elevated"MICs"to"agricultural"azoles,"suggesting"cross"resistance."Importantly,"we"observed"an"increased"tolerance"to"both"agricultural"and"medical"azoles"in"the"samples"taken"after"the"growing"season"and"application"of"azoles.'47%"of"resistant"isolates"carry"the"environmentally/derived"TR34/L98H"mutation,"while"the"mechanism"of"resistance"in"the"remaining"isolates"is"cyp51a/independent"and"remains"unknown.""To"quantify"the"impact"of"azole"application"on"A.'fumigatus'at"the"genome"level,"we"performed"whole"genome"sequencing"on"250"isolates."When"compared"to"Af293,"we"detected"an"average"of"77,000"SNVs"per"isolate."Isolates"from"different"regions"and"types"of"agriculture"did"not"cluster"separately,"however"we"observed"reduced"genetic"diversity"in"the"isolates"collected"from"conventional"fields"after"azole"application,"compared"to"before"application."Ongoing"work"is"focused"on"comparing"this"cohort"of"environmentally/derived"A.'fumigatus"isolates"with"sequenced"clinical"isolates"to"identity"fungal"

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determinants"critical"for"human"infection"as"well"as"genetic"markers"associated"with"increased"pathogenicity"and/or"cyp51a/independent"drug"resistance.""*6.$Investigation$of$enhanced$virulence$mechanisms$in$Aspergillus*fumigatus*isolates$from$the$International$Space$Station""A.$Blachowicz1,"2,"J."Romsdahl1,"N."Raffa3,"N."P."Keller3,"K."Venkateswaran2,"C."C."C."Wang1,"4.""1)'Department'of'Pharmacology'and'Pharmaceutical'Sciences,'School'of'Pharmacy,'University'of'Southern'California,'Los'Angeles,'CA6'2)'Biotechnology'and'Planetary'Protection'Group,'Jet'Propulsion'Laboratory,'California'Institute'of'Technology,'Pasadena,'CA6'3)'Department'of'Medical'Microbiology'and'Immunology,'University'of'WisconsinQMadison,'Madison,'WI'6'4)'Department'of'Chemistry,'Dornsife'College'of'Letters,'Arts'and'Sciences,'University'of'Southern'California,'Los'Angeles,'CA.""'Aspergillus'fumigatus,"one"of"many"fungal"isolates"identified"in"a"recent"Microbial"Observatory"Study"on"the"International"Space"Station"(ISS/MO"study),"is"a"ubiquitous"saprophytic"fungus."Its"vast"adaptation"capacity"enables"it"to"be"omnipresent"in"the"environment"and"to"be"a"successful"opportunistic"pathogen."A.'fumigatus"causes"variety"of"health"conditions"ranging"from"allergies"to"potentially"life/threatening"invasive"aspergillosis"(IA)"in"immunocompromised"individuals."Initial"characterization"of"two"A.'fumigatus'ISS/isolates,"ISSFT/021"and"IF1SW/F4,"showed"no"outstanding"differences"in"their"genomes"and"secondary"metabolite"(SM)"profiles"when"compared"to"clinical"isolates"CEA10"and"Af293,"however"both"isolates"were"significantly"more"lethal"in"a"larval"zebra"fish"model"of"IA."Considering"that"A.'fumigatus"becomes"more"virulent"in"space"posing"a"potential"threat"to"the"astronauts’"health,"we"investigated"molecular"changes"in"the"proteome"of"ISS/isolated"strains."Furthermore,"we"attempted"to"elucidate"the"mechanisms"of"observed"enhanced"virulence"in"“space”"strains."We"created"Af293"mutants"deficient"in"the"five"spore"associated"SMs"(DHN/melanin,"endocrocin,"trypacidin,"fumigaclavine"and"fumiquinazoline)"that"are"also"known"to"contribute"to"virulence"or"toxicity."We"tested"these"mutants"for"UV/C"sensitivity"and"virulence"in"in"two"larval"zebra"fish"models"characterized"by"neutrophil"and"macrophage"deficiency."Further,"an"evolution"study"using"clinical"isolate"Af293"was"conducted"to"examine"its"adaptation"to"sub/lethal"doses"of"UVC."The"first"and"last"generations"of"the"controlled"evolution"study"will"be"tested"for"virulence"and"analyzed"for"any"molecular"changes."Such"in"depth"analyses"are"imperative"for"understanding"the"mechanisms"of"enhanced"virulence"in"A.'fumigatus"and"may"result"in"the"development"of"preventive"measures"against"IA."Understanding"the"possible"molecular"alterations"triggered"by"irradiation"remains"crucial"in"the"light"of"future"long/term"manned"space"flights"to"both"ensure"the"astronaut’s"health"and"maintain"the"closed"habitat.""7.$Inducible$cell$fusion$permits$use$of$competitive$fitness$profiling$in$the$human$pathogenic$fungus$Aspergillus*fumigatus.""Mike$Bromley.""Manchester'Fungal'Infection'Group,'The'University'of'Manchester,'Manchester,'GB""'Aspergillus'fumigatus"is"the"most"important"airborne"mould"pathogen"of"man."Estimates"suggest"that"over"3"million"people"have"invasive"or"chronic"infections"that"lead"to"in"excess"of"600,000"deaths"every"year."Very"few"drugs"are"available"to"treat"the"various"forms"of"aspergillosis"and"we"rely"predominantly"on"the"azole"class"of"agents."Without"treatment,"the"disease"is"almost"always"fatal."Resistance"to"these"compounds"is"emerging"and"in"some"areas"is"reaching"levels"that"prevent"their"use"as"sole"first"line"therapeutics."There"is"a"significant"need"to"address"the"lack"of"available"antifungals"with"novel"mechanisms"of"action."High"throughput"functional"genomic"studies"that"employ"competitive"fitness"profiling"have"been"used"effectively"in"yeasts"to"address"gaps"in"functional"annotation,"determine"drug"mechanism"of"action,"and"identify"genes"associated"with"drug"resistance."Competitive"fitness"profiling"assumes"that"each"cell"in"a"pool"is"a"distinct"entity"and"no"exchange"of"genetic"material"takes"place"between"cells."Unfortunately,"all"filamentous"fungi"studied"thus"far"undergo"a"process"of"constitutive"and"rapid"self/fusion"(anastomosis)"with"nuclei"being"readily"transferred"between"cells."This"strongly"suggested"that"use"of"competitive"fitness"profiling,"of"the"nature"so"powerfully"used"in"yeasts,"would"not"be"possible"in"filamentous"fungi"such"as"A.'fumigatus."Here"we"present"compelling"data"showing"that,"unlike"the"situation"in"other"filamentous"fungi"such"as"Neurospora'crassa"and"Fusarium'oxysporum,"anastomosis"occurs"rarely"in"standard"culture"conditions"for"A.'fumigatus."Furthermore,"we"show"that"cell"fusion"can"be"induced"by"various"stressors,"

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including"antifungals,"but"only"in"specific"culture"settings."Using"this"information,"we"have"been"able"to"develop"a"competitive"fitness"profiling"strategy"to"assess"strain"fitness"in"pooled"cultures."I"will"also"present"evidence"to"show"that"we"can"use"competitive"fitness"profiling"to"identify"defects"in"pathogenicity"using"in'vitro"and"in'vivo"(murine)"models."$8.$High$throughput$genetic$engineering$of$filamentous$fungi$using$automated$liquid$handling$robotics""K.S.$Bruno,"E."Szewczyk,"J."Fredlund,"H."Abdulla,"S."Jain,"J."vanDijk,"B.P."Knox,"M."Siddiqui,"A."Fong,"D."Vaughn,"C."Cizauskas,"Z."Serber.""Zymergen,'Inc.,'Emeryville,'CA"""Industrial"fermentation"utilizing"filamentous"fungi"is"a"useful"method""for"large/scale"manufacturing"of"enzymes,"pharmaceuticals,"organic"acids,"and"other"valuable"commercial"products."While"fungi"are"excellent"producers"of"many"of"these"molecules,"strain"improvement"is"still"vital"for"increasing"production"efficiency"at"industrial"scales."Here,"we"demonstrate"the"use"of"automated"liquid"handling"robotics"(LHR)"for"the"purpose"of"genetic"manipulation"of"filamentous"fungi."This"process"at"Zymergen"begins"when"we"upload"a"genetic"sequence"into"our"Laboratory"Information"Management"System"(LIMS)S"this"system"is"capable"of"automated"design"of"genetic"constructs,"including"PCR"primers"for"thousands"of"genetic"edits"such"as"introduction"of"targeted"Single"Nucleotide"Polymorphisms"(SNPs),"gene"deletions,"and"promoter"exchange"constructs."We"then"construct"individual"genetic"cassettes"in"96"well"format"with"help"from"LHR."Transformation"of""the"constructs"into"target"fungal"strains"is"also"carried"out"using"LHR"and"96"well"microtiter"plates"allowing"for"HTP"screening"for"the"desired"strains"and"activities."Because"filamentous"fungi"can"form"heterokaryons,"we"employ"single/cell/sorting"technology"to"print"individual"spores"onto"desired"media"so"we"can"select"only"homokaryons."We"screen"these"homokaryons"for"desired"transformations"with"a"rapid"genomic"DNA"prep,"followed"by"PCR"screening,"Next"Generation"Sequencing"of"amplicons,"and"then"phenotypic"screening."The"efficacy"of"this"approach"is"demonstrated"using"Aspergillus"niger"but"our"platform"is"designed"to"accommodate"almost"any"fungal"host."With"our"platform,"we"can"rapidly"build"and"screen"thousands"of"engineered"fungal"strains"and"determine"which"of"these"are"improved"for"production.""Furthermore,"we"can"continue"to"reprocess"and"refine"these"strains,"combining"desired"traits"within"an"improved"lineage"that"is"based"on"defined"genetic"edits."The"platform"is"currently"being"deployed"for"multiple"industrial"partnerships.""*9.$Characterisation$of$fungal$enzymes$responsible$for$xylan$degradation$in$plant$lignocellulose""G.$Bulmer,"J.M."van"Munster."Manchester'Institute'of'Biotechnology,'University'of'Manchester,'Greater'Manchester,'GB"""Fungi"produce"a"diverse"repertoire"of"carbohydrate"active"enzymes"(CAZymes)"which"enable"them"to"degrade"the"lignocellulosic"biomass"of"dead"plants."This"economically"relevant"ability"is"exploited"in"biotechnology"to"release"sugars"from"renewable"resources"which"are"subsequently"used"to"produce"high/value"chemicals"and"second/generation"biofuels."However,"in"order"to"fully"harness"the"degradative"ability"of"fungi,"the"biochemical"processes"involved"must"be"characterised"in"greater"detail."For"example,"for"the"well/known"and"industrially"important"fungus"Aspergillus'niger,"transcriptional"regulation"and"protein"secretion"during"carbohydrate"degradation"is"relatively"well"understood."However,"over"50%"of"the"CAZymes"predicted"in"its"genome"have"not"been"biochemically"characterised"meaning"we"have"an"incomplete"understanding"of"the"changes"these"enzymes"could"cause"to"lignocellulose.""This"project"focuses"on"the"characterisation"of"CAZymes"involved"in"the"degradation"of"arabinoxylan,"the"main"hemicellulose"found"in"wheat"straw:"one"of"Europe's"main"agricultural"by/products."Glycan"arrays"containing"defined"branched"and"linear"plant"glycan"substrates"are"generated"and"used"to"determine"CAZyme"substrate"and"product"specificity"via"a"high"throughput"screening"method"involving"MALDI/ToF"MS."Enzyme"characterisation"will"further"the"understanding"of"fungal"degradation"and"allow"the"development"of"novel"enzymes"for"the"engineering"of"complex"plant/derived"carbohydrates."$$ $

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10.$The$central$regulatory$model$of$Aspergillus$conidiation$didn't$work$in$Monascus*ruber""L."Jia1,"Z."Huang1,"J."Yu2,"F."Chen1,"W.$Chen1.""1)'College'of'Food'Science'and'Technology,'Huazhong'Agricultural'University,'China6'2)'Department'of'Bacteriology,'University'of'WisconsinQMadison,'USA""'Monascus'spp."are"widely"used"in"the"production"of"lovastatin"and"food"grade"pigments"in"East"Asia."Unlike"their"close"relative"Aspergillus'spp.,"the"asexual"spores"of"Monascus"spp."are"produced"either"direct"from"hyphae"or"produced"laterally"on"short"pedicels"either"singly"or"in"short"chains."The"bioinformatics"analyses"supported"the"difference"in"conidiation"form"between"Aspergillus"spp."and"Monascus"spp."that"Monascus"genomes"lack"the"abaA"homolog,"responsible"for"the"differentiation"of"the"phialides"in"Aspergillus'nidulans."Therefore,"unlike"the"well/known"central"regulator"cascade"BrlA–AbaA–WetA"in"Aspergillus"spp."and"Penicillium'spp.,"Monascus"spp."have"a"novel"central"regulatory"mechanism"and"worth"further"analysis."Firstly,"we"heterologous"expressed"the"A.'nidulans"abaA"gene"in"Monascus'ruber."It"failed"to"induce"development"of"complete"conidiophores"as"seen"in"the"aspergilli,"but"did"result"in"increased"conidial"production"and"viability."Then,"we"respectively"generated"the"abaA'and"wetA"knockouts"of"Monascus'ruber"by"Agrobacterium/mediated"transformation"method."The"knockouts"were"confirmed"by"PCR,"cDNA"sequencing"and"southern"blotting."The"results"showed"that"deletion"of"brlA"and"wetA"had"no"apparent"change"in"the"conidia"morphology"and"formation"compared"to"the"wide"type."Moreover,"no"significant"change"was"observed"in"the"conidial"production"number"and"germination"rates"between"the"knockouts"and"the"wild"type."Altogether,"our"results"suggested"that"the"well/established"central"regulatory"model"of"asexual"sporulation"in"Aspergillus'was"not"applicable"in"their"close"relative"Monascus."$*11.$Unraveling$the$genetics$of$pentose$catabolism$in$Aspergillus*niger""Tania$Chroumpi,"Ronald"P."de"Vries.""Fungal'Physiology,'Westerdijk'Fungal'Biodiversity'Institute'&'Fungal'Molecular'Physiology,'Utrecht'University,'Utrecht,'The'Netherlands"""The"filamentous"ascomycete"Aspergillus'niger"is"able"to"efficiently"degrade"lignocellulosic"biomass"to"obtain"monomeric"sugars"that"can"use"as"a"carbon"source."In"nature,"D/xylose"and"L/arabinose"are"the"most"abundant"monosaccharides"after"D/glucose,"being"major"components"of"the"plant"cell"wall"polysaccharides"xylan,"pectin"and"xyloglucan."These"sugars"are"converted"through"the"pentose"catabolic"pathway"(PCP),"of"which"the"final"step"links"to"the"pentose"phosphate"pathway."Only"some"of"the"PCP"genes"and"enzymes"of"A.'niger"have"been"characterized"in"detail,"and"the"induction"of"the"PCP"genes"has"not"been"thoroughly"studied."Thus,"the"aim"of"this"study"was"to"obtain"a"better"understanding"of"the"pentose"catabolism,"and"the"induction"mechanism"of"the"PCP"genes"and"the"arabinanolytic"and"xylanolytic"regulatory"systems"of"A.'niger."To"this"end,"phenotypic"and"transcriptomic"analysis,"intracellular"accumulation"of"metabolites"and"enzymatic"activities"of"single/"and"multi/gene"deletion"mutants"of"PCP"genes"were"conducted."This"not"only"resulted"in"a"better"understanding"of"the"induction"and"role"of"the"PCP"genes,"but"also"identified"additional"genes"involved"in"this"pathway"in"A.'niger."The"results"of"this"study"will"therefore"also"facilitate"strain"engineering"of"A.'niger"as"a"cell"factory"for"the"production"of"metabolites"related"to"this"pathway,"such"as"xylitol."$*12.$Investigation$of$proteins$associated$with$hyphal$growth$and$endocytosis""B.$Commer,"Z."Schultzhaus,"B.D."Shaw.""Plant'Pathology'&'Microbiology,'Texas'A&M'University,'College'Station,'TX"""Filamentous"fungi"grow"by"producing"polarized"tubular"cells"called"hyphae."Each"hypha"elongates"by"synthesizing"and"adding"new"cell"wall"and"membrane"exclusively"at"the"apex."Growth"is"maintained"by"an"apical"recycling"mechanism"that"balances"endocytosis"and"exocytosis"at"the"forefront"of"growth."This"equilibrium"requires"both"an"apical"formation"of"secretory"vesicles"deemed"the"Spitzenkörper"(SPK),"and"an"endocytic"sub/apical"collar"located"approximately"1/5μm"from"the"SPK."As"extension"occurs"at"the"tip,"cargo"is"also"internalized"in"the"area"immediately"distal"to"the"apex."Evidence"of"this"relationship"is"observed"through"the"investigation"of"landmark"proteins"along"the"plasma"membrane"that"mark"areas"of"impending"growth."Here,"a"bioinformatics"approach"was"utilized"to"identify"39"proteins"in"Aspergillus'nidulans"that"are"predicted"to"be"cargo"of"endocytosis"based"on"the"presence"of"an"NPFxD"peptide"motif,"

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which"is"a"necessary"endocytic"signal"sequence"in"yeast."I"hypothesize"that"NPFxD"motif/containing"proteins"that"are"cargo"for"endocytosis"will"localize"to"at"least"one"of"three"apical"regions"(the"SPK,"sub/apical"collar,"and"apical"dome)"where"cargo"are"typically"observed.$In"order"to"test"this"hypothesis,"the"expression"and"dynamic"localization"of"motif/containing"proteins"in"A.'nidulans"is"evaluated"by"endogenously"inserting"GFP"or"mCherry."The"localization"patterns"studied"thus"far"support"the"hypothesis"that"endocytic"cargo"proteins"which"contain"this"motif"will"localize"to"one"or"more"of"the"three"predicted"regions"near"the"growing"fronts"of"hyphae."Those"that"are"cargo"for"endocytosis"are"also"likely"to"be"associated"with"the"establishment"or"maintenance"of"polarized"growth.$Mutants"from"initial"deletion"experiments"demonstrate"an"inability"to"maintain"polarization"and"many"display"atypical"development"in"various"cell"types,"which"also"suggests"that"the"genes"in"question"are"involved"with"membrane"turnover."$13.$How$do$Microbes$Choose$their$Neighbors?""K.$Martin$Dahlstrom1,"Marko"Kojic2,"Dianne"Newman1.""1)'Biology'and'Biological'Engineering,'California'Institute'of'Technology,'Pasadena,'CA6'2)'Faculty'of'Biology,'University'of'Belgrade""Rhizosphere"diversity"is"critical"to"the"yield"of"agricultural"crops.""The"rhizosphere"contains"several"species"of"biocontrol"bacteria"that"produce"phenazines,"toxic"molecules"which"help"keep"phytopathogenic"fungi"at"bay."However,"there"are"biocontrol"strains"of"fungi"which"are"also"susceptible"to"phenazine"assault"and"must"live"directly"next"to"phenazine"producers.""We"have"isolated"a"plant/promoting"biocontrol"fungus,"Aspergillus'ustus,"that"was"found"with"an"adherent"bacterial"partner,"Paraburkholderia'S0S3.""P.'SOS3'is"able"to"detoxify"and"sequester"phenazines,"including"the"environmentally"relevant"phenazine/1/carboxylic"acid"(PCA)."Intriguingly,"P.'SOS3"only"does"so"in"the"presence"of"A.'ustus,"allowing"the"fungus"to"grow"even"in"the"presence"of"phenazines"and"phenazine/producing"bacteria."This"is"accomplished"by"a"structural"rearrangements"P.'SOS3'within"the"fungal"colony.""In"the"absence"of"PCA,"P.'SOS3"is"dispersed"throughout"the"fungal"colony.""However,"in"the"presence"of"PCA"this"bacterium"forms"spheres"within"the"fungal"colony"that"reduce"and"sequester"PCA,"permitting"A.'ustus"growth.""These"bacterial"aggregates"appear"to"be"anoxic,"keeping"PCA"in"the"reduced,"less"toxic,"state.""This"behavior"appears"to"be"species/specific"as"other"biocontrol"bacteria,"including"other"species"of"Paraburkholderia"cannot"effectively"protect"A.'ustus.""Given"that"the"relative"abundance"of"phenazine"producers"in"the"soil"rises"with"temperature,"understanding"how"biocontrol"fungi"interact"with"partnered"bacteria"to"withstand"these"kinds"of"stresses"is"important"to"the"future"of"agriculture"in"a"warmer"climate.""14.$An$Aspergillus*niger$colony$differentiates$its$molecular$responses$to$spatially$separated$substrates"Paul$Daly1,"Mao"Peng1,"Ad"Wiebenga1,"Hugh"D."Mitchell2,"Lye"Meng"Markillie2,"Charles"Ansong2,"Heather"M."Brewer2,"Young/Mo"Kim2,"Galya"Orr2,"Scott"E."Baker2,"Peter"de"Gijsel3,"Mirjam"A."Kabel3,"Kristiina"S."Hilden4,"Miia"R."Mäkelä4,"Ronald"P."de"Vries1,"4"1Westerdijk"Fungal"Biodiversity"Institute"and"Utrecht"University,"The"Netherlands"2Pacific"Northwest"National"Laboratory,"United"States"3Wageningen"University"&"Research,"The"Netherlands"4University"of"Helsinki,"Finland"Email:"p.daly@westerdijkinstitute.nl"$

Saprotrophic"fungi,"such"as"Aspergillus'niger,"grow"as"mycelial"colonies"that"are"often"considered"uniform"entities."To"test"this"uniformity,"we"analysed"pie/slice"sections"of"a"colony"grown"on"spatially"separated"substrates"(glucose,"wheat"bran,"sugar"beet"pulp)"using"transcriptomics,"proteomics"and"metabolomics."

The"colony"tuned"its"response"to"the"local"carbon"source"composition."Plant"biomass"degrading"CAZymes"and"intracellular"carbon"catabolic"enzymes"were"more"abundant"in"parts"of"the"colony"containing"the"corresponding"substratesS"e.g."a"stronger"pectinolytic"response"was"observed"in"the"part"of"the"colony"grown"on"pectin/rich"sugar"beet"pulp"accompanied"by"higher"expression"of"the"genes"of"D/galacturonic"acid"catabolism,"the"main"component"of"pectin."Similarly,"highest"expression"of"xylanolytic"genes"was"observed"on"parts"exposed"to"wheat"bran.""

Our"results"argue"against"a"situation"in"which"small"molecules"are"transported"efficiently"throughout"the"colony"and"indicate"a"high"ability"within"the"fungal"colony"to"respond"to"local"environmental"conditions."This"fits"well"with"a"natural"biotope"in"which"the"substrate"is"typically"

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heterogeneous"and"where"colonies"can"reach"significant"sizes."In"such"a"situation"it"would"likely"not"be"energetically"efficient"to"move"carbon"sources"all"across"the"colony"and"maintain"a"uniform"physiology"throughout"the"colony.""*15."Expanding$the$fungal$color$chart:$a$novel$PKLNRP$conidial$pigment$acts$together$with$AspLmelanin$to$give$Aspergillus*aurantiobrunneus$its$characteristic$orangeLbrown$color""P.E.$de$Reus1,"H.E."Frederiksen1,"S."Theobald2,"K.T."Kongsted3,"R.J.N."Frandsen1.""1)'DTU'Bioengineering,'Technical'University'Denmark,'Copenhagen,'DK6'2)'Novo'Nordisk'Foundation'Center'for'Biosustainability,'Technical'University'Denmark,'Copenhagen,'DK6'3)'Department'of'Drug'Design'and'Pharmacology,'Copenhagen'University,'Copenhagen,'DK"""Conidial"pigmentation"plays"a"key"role"in"stress"resistance,"survival,"and"pathogenicity"of"filamentous"fungi."Ascomycetes"commonly"produce"pigments"that"originate"from"polyketide"monomers"YWA1"(pksP'in"A.'fumigatus)"or"dixhydroxynaphtalene/"(DHN)"melanin"(albA"in"A.'niger)."However,"certain"species"lack"these"PKSs"and"have"evolved"alternative"pigmentation"strategies."A.'terreus,'section"Terrei,"compensates"for"the"lack"of"a"pksP/albA'homologue"by"relying"on"the"NRPS/like"MelA"and"tyrosinase"TyrP"to"produce"the"cinnamon/brown"pigment"Asp/melanin"in"its"conidia."The"orange/brown"A.'aurantiobrunneus,'section"Nidulantes,"displays"a"similar"genetic"composition"with"MelA"and"TyrP'but"no"pksP/albA"homologue,"suggesting"that"this"species"also"relies"on"Asp/melanin"for"conidial"pigmentation."We"show,"however,"that"the"dominant"conidial"pigment"in"A.'aurantiobrunneus"is"a"tangerine/orange"compound"produced"by"a"novel"PKS/NRPS"encoding"gene"cluster,"which"we"name"Tng"after"its"tangerine"colored"product.""Deleting"the"PKS/NRPS"encoding"gene"tngS"in"A.'aurantiobrunneus'resulted"in"beige"spores,"while"heterologous"expression"of"the"entire"Tng"gene"cluster"in"a"white"A.'nidulans'strain'(∆yA,"∆wA)"resulted"in"bright"orange"conidia."HR/HPLC/MS"analysis"linked"the"Tng"gene"cluster"to"the"production"of"two"novel"compounds,"currently"undergoing"HR/NMR"structure"elucidation."Deletion"of"MelA"in"A.'aurantiobrunneus"wild"type"and"∆tngS'backgrounds"confirmed"the"contribution"of"Asp/melanin"as"spore"pigment."While"Asp/melanin"confers"resistance"to"UV"irradiation"in"A.'terreus,"preliminary"data"show"that"the"UV/protective"effect"of"Tng"pigments"is"limited,"suggesting"their"biological"functions"do"not"overlap.""This"study"describes"a"novel"combination"of"spore"pigments,"emphasizing"the"diversity"of"conidial"pigmentation"of"the"Aspergilli."Elucidating"the"biological"function"of"the"various"pigments"will"be"important"in"understanding"their"evolutionary"history"and"identifying"which"sequence"of"gene"gains/losses"or"horizontal"transfers"best"explain"the"present"combination"of"pigment"synthases.""16.$Population$subdivision$and$the$frequency$of$aflatoxigenic$isolates$in$Aspergillus*flavus$in$the$United$States.""Milton"Drott1,"Lauren"Fessler2,"Michael"Milgroom2.""1)'Molecular'and'Environmental'Toxicology,'University'of'Wisconsin'Madison,'Madison,'WI6'2)'Plant'Pathology,'Cornell'University,'Ithaca,'NY"""Consumption"of"food"contaminated"with"aflatoxin,"from"crops"infected"by"Aspergillus'flavus,"is"associated"with"acute"toxicosis,"cancer,"and"stunted"growth."While"such"contamination"is"more"common"in"the"lower"latitudes"of"the"United"States,"it"is"unclear"if"this"pattern"is"associated"with"differences"in"the"relative"frequencies"of"aflatoxigenic"individuals"of"A.'flavus."To"determine"if"the"frequency"of"aflatoxin/producing"ability"of"A.'flavus"increases"as"latitude"decreases,"we"sampled"281"isolates"from"field"soils"in"two"north/south"transects"in"the"U.S."and"tested"them"for"aflatoxin"production."We"also"genotyped"161"isolates"using"10"microsatellite"markers"to"assess"population"structure."While"the"population"density"of"A.'flavus"was"highest"at"lower"latitudes,"there"was"no"difference"in"the"frequency"of"aflatoxigenic"A.'flavus"isolates"in"relation"to"latitude."We"found"that"the"U.S."population"of"A.'flavus"is"subdivided"into"two"genetically"differentiated"subpopulations"that"are"not"associated"with"chemotype"or"geographic"origin"of"isolates."The"two"populations"differ"markedly"in"allelic"and"genotypic"diversity."The"less"diverse"population"is"more"abundant"and"may"represent"a"clonal"lineage"derived"from"the"more"diverse"population."Overall,"increased"aflatoxin"contamination"in"lower"latitudes"may"be"explained"partially"by"differences"in"population"density"of"A.'flavus,"not"genetic"population"structure."

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$*17.$New$unexplored$functions$of$a$C2H2$zinc$finger$transcription$factor,$SltA$for$drug$responses$and$virulence$in$Aspergillus*fumigatus""W.$Du,"L."Lu.""Nanjing'Normal'University,'Nanjing,'Jiangsu,'CN"""SltA,"a"C2H2"zinc"finger"transcription"factor,"is"highly"conserved"in"Aspergilii'with"no"identifiable"homolog"in"yeast/like"fungi."Previous"studies"verified"that"SltA"prevents"toxicity"by"the"cations"Na+,"K+,"Li+,"Cs+"and"Mg2+"but"shows"growth"defects"in"the"limited"extracellular"Ca2+"addition"in"model"filamentous"fungus"Aspergillus'nidulans."Here,"we"demonstrated"that"SltA"homolog"in"human"fungal"pathogen"A.'fumigatus"has"consistent"functions"with"that"of"A.'nidulans"for"cation"detoxification"and"calcium"auxotrophy"such"that"deletion"of"AfsltA"causes"hypersensitive"to"calcium"chelator"EGTA"while"addition"of"calcium"is"able"to"rescue"growth"defects."The"amplitudes"of"the"[Ca2+]c"transients"are"reduced"by"33%"in"AfsltA"mutant"compared"to"the"parental"strain"which"may"result"from"overexpression"of"putative"vacuolar"Ca2+/ATPases"PmcA,"PmcB"and"PmcC."Interestingly,"deletion"of"AfsltA'has"the"reduced"ergosterol"content"under"itraconazole"treatment"and"shows"hypersensitive"to"antifungals"itraconazole,"voriconazole,"bifonazole"and"terbinafine"in"a"calcium"supplementation"independent"way"but"displays"no"detectable"phenotypes"to"amphotericin"B"and"caspofungin."Furthermore,"adding"hemin"to"media"or"over/expressed"cyp51A"is"able"to"rescue"azoles"susceptibilities,"suggesting"AfsltA"may"affect"ergosterol/biosynthesis"functions."Most"importantly,"Galleria'mellonella"larvae"virulence"test"shows"a"significantly"attenuated"pathogenicity"in"the"AfsltA"deletion"strain,"indicating"that"AfSltA"has"unexplored"important"functions"in"human"pathogens.""18.$Development$of$a$high$protein$titer$Aspergillus*niger*expression$host$which$is$available$for$licensing""Nigel$Dunn$Coleman2,"Wim"de"Laat1.""1)'FunGeneX.USA,'Los'Gatos,'CA6'2)'FunGeneX'BV,'Breda'The'Netherlands""Nigel$Dunn$Coleman*,$Joyce"Arnouts,"Lucie"Parenicova"and"Wim"de"Laat"FunGeneX"BV,"Breda,"The"Netherlands,"*FunGeneX"USA,"Los"Gatos,"CA,"USA.""Aspergillus'niger'has"a"long"history"of"commercial"use"as"a"host"for"the"production"of"both"homologous"and"heterologous"proteins,"products"which"have"received"GRAS"status.""At"present"there"is"an"unmet"need"for"a"licensable"A.niger'platform"for"companies"to"evaluate"for"the"production"of"their"propriety"proteins."FunGeneX"BV"has"isolated"stains"which"have"optimized"by"classical"strain"improvement"techniques,"developed"have"preferred"non/viscous"morphology,"produce"high"amounts"of"enzyme"protein,"do"not"produce"any"mycotoxins,"genome"is"sequenced"and"annotated,"and"transformation"systems"have"been"developed."Key"features"High"level"transformation"frequency."Microtiter"screening"system."Scaleable"shake"culture"screening."Robust"42l"fermentation"protocol"using"low"cost"media"components."Prior"experience"in"technology"transfer"to"large"scale"fermentations."Regulatory"experience"with"EPA"MCAN"and"FDA"GRAS"filings."This"platform"is"available"on"a"protein"by"protein"basis"with"an"exclusive"or"non"exclusive"license"option."Nigel"Dunn"Coleman,"is"available"at"the"meeting"to"discuss"the"platform."ndunncoleman.fungenex@gmail.com.""$ $

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*19.$Analysis$of$sclerotia$formation$and$morphology$in$Aspergillus*niger""Valeria$Ellena1,2,"Elsa"Arcalis3,"Michael"Sauer1,2,"Matthias"Steiger1,2.""1)'ACIB'GmbH,'Austrian'Centre'of'Industrial'Biotechnology,'Vienna,'Austria'6'2)'Department'of'Biotechnology,'University'of'Natural'Resources'and'Life'Sciences,'Vienna,'Austria6'3)'Department'of'Applied'Genetics'and'Cell'Biology'(DAGZ),'University'of'Natural'Resources'and'Life'Sciences,'Vienna,'Austria""'Aspergillus'niger"is"a"relevant"industrial"microorganism"used"as"a"versatile"cell"factory"for"the"production"of"organic"acids"and"proteins."This"fungus"is"considered"asexual"and"able"to"reproduce"only"through"the"mitotic"formation"of"asexual"conidia."The"discovery"of"a"sexual"cycle"in"this"organism"would"not"only"broaden"its"current"biological"knowledge"but"also"allow"to"unravel"new"strategies"for"strain"improvement."An"important"step"in"the"induction"of"a"sexual"cycle"in"filamentous"fungi"is"the"formation"of"sexual"structures,"generally"named"fruiting"bodies,"where"the"fertile"ascospores"derived"by"meiosis"can"form."In"this"study,"we"were"able"to"induce"the"formation"of"pre/mature"sexual"structures,"namely"sclerotia,"by"the"A.'niger"strain"ATCC1015"in"different"conditions."In"particular,"sclerotia"can"be"stably"formed"by"the"wild/type"strain"on"malt"extract"agar"medium"supplemented"with"raisins."Also"a"pyrG"auxotroph"mutant"is"able"to"form"abundant"sclerotia"on"malt"extract"agar"medium"and"oatmeal"agar"medium."The"formation"of"sclerotia"is"often"accompanied"by"the"presence"of"liquid"droplets"on"top"of"them"and"by"a"reduction"of"the"asexual"conidiation,"suggesting"a"potential"implication"of"the"sexual"cycle"in"this"process."A"detailed"analysis"of"the"sclerotia"was"performed"by"both"light"and"electron"microscopy,"revealing"the"internal"morphology"of"the"A.'niger"sclerotia."The"results"show"that"these"structures"are"composed"of"three"layers"with"different"cell"types"and"are"comparable"to"the"sclerotia"formed"by"other"fungi.""*20.$The$AGC$kinase$YpkA$regulates$sphingolipids$biosynthesis$and$physically$interacts$with$SakA$MAP$kinase$in*Aspergillus*fumigatus""J.$H.$T.$M."Fabri1,"N."L."Godoy1,"M."C."Rocha1,"M."Munshi2,"T."P."Fill3,"T."A."Coccio4,"M."R."V."Z."Kress5,"A."F."da"Cunha1,"M."Del"Poeta2,"I."Malavazi1.""1)'Department'of'Genetics'and'Evolution,'Federal'University'of'São'Carlos,'São'Carlos,'BR6'2)'Department'of'Molecular'Genetics'and'Microbiology,'Stony'Brook'University,'Stony'Brook,'NY,'USA6'3)'Institute'of'Chemistry,'State'University'of'Campinas,'Campinas,'BR6'4)'Medical'School'of'Ribeirão'Preto,'University'of'São'Paulo,'Ribeirão'Preto,'BR6'5)'Department'of'Toxicological'and'Bromatological'Clinical'Analysis,'University'of'São'Paulo,'Ribeirão'Preto,'BR""'Aspergillus'fumigatus"is"the"main"causative"pathogen"of"invasive"pulmonary"aspergillosis"(IPA)"in"immunocompromised"individuals."Sphingolipids"(SL)"are"components"of"the"plasma"membrane"which"are"important"for"numerous"cellular"processes"and"virulence"of"different"fungal"pathogens."In"yeast,"SL"biosynthesis"is"regulated"by"the"“AGC"kinases”"Ypk1"and"Ypk2,"which"also"seem"to"connect"the"SL"biosynthesis"with"the"cell"wall"integrity"(CWI)"and"the"High"Osmolarity"Glycerol"(HOG)"pathways."Here,"we"investigate"the"role"of"ypkAYPK1"in"SL"biosynthesis"and"its"relationship"with"the"CWI"and"the"HOG"pathways"in"Aspergillus'fumigatus."We"found"that"ypkA"is"important"for"fungal"viability,"since"the"ΔypkA"strain"presented"a"terminal"phenotype"and"complete"absence"of"conidiation."We"observed"that"under"repressive"condition,"the"conditional"mutant"niiA::ypkA'exhibited"vegetative"growth"defects,"impaired"germination"and"thermosensitivity."In"addition,"the"ypkA"loss"of"function"caused"a"decrease"in"glycosphingolipids"(GSL)"levels,"especially"the"metabolic"intermediates"belonging"to"the"neutral"GSL"branch"including"dihydroceramine"(DHC),"ceramide"(Cer)"and"glucosylceramide"(GlcCer),"but"interestingly"a"small"increase"in"ergosterol"content."Genetic"analyzes"showed"that"ypkA"genetically"interacts"with"the"MAP"kinases"of"CWI"and"HOG"pathways,"mpkA"and"sakA,"respectively,"while"only"SakA"physically"interacts"with"YpkA."Our"results"suggest"that"YpkA"is"important"for"fungal"survival"through"the"regulation"of"GSL"biosynthesis"and"cross"talks"with"A.'fumigatus"MAP"kinase"pathways."Financial$support:"FAPESP"and"CNPq."$$ $

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21.$A$spliceosomal$twin$intron$conserved$across$all$classes$of$the$Pezizomycotina$subphylum""N."Kavalecz1,"N."Ág1,"L."Karaffa1,"C."Scazzocchio2,3,"M."Flipphi1,"E."Fekete1."1)'Dept.'of'Biochemical'Engineering,'University'of'Debrecen,'Debrecen,'HU6'2)'Dept.'of'Microbiology,'Imperial'College'London,'UK6'3)'I2BC'UMR'9198,'GifQsurQYvette,'FR"""In"the"primary"transcript"of"nuclear"genes,"coding"“exons”"usually"alternate"with"non/coding"“introns”."These"latter"have"to"be"precisely"excised"to"form"the"functional"mRNA,"in"the"large"majority"of"cases,"by"the"major"U2"spliceosome."Spliceosomal"twin"introns"(“stwintrons”)"are"complex"intervening"sequences"(IS)"where"a"standard"“internal”"U2"intron"interrupts"one"of"the"canonical"splicing"motifs"of"an"“external”"U2"intron"(viz."5’"donorS"3’"acceptorS"motif"around"branch"point"A),"i.e.,"the"external"intron"can"only"be"removed"after"excision"of"the"internal"intron."To"study"spliceosomal"introns,"we"can"use"fungal"stwintrons"as"model"systems."Thereto,"we"created"an"informatics"tool"to"search"for"stwintrons"in"Aspergillus'nidulans'in'silico."Here,"we"describe"a"[D5,6]"stwintron"in"the"gene"at"locus"AN5404"(encoding"a"reticulon/like"protein)"where"the"internal"intron"is"situated"within"the"donor"element"of"the"external"intron,"between"the"5th"and"the"6th"nt"[5’/GUAAG|U]."This"stwintron"is"old:"it"is"present"in"9"classes"of"Pezizomycotina"as"the"first"IS"after"the"AUG,"including"in"Pezizomycetes"and"Orbiliomycetes."In"certain"taxa,"the"second"IS"is"also"a"[D5,6]."In"the"specific"context"of"this"gene,"the"ubiquitous"stwintron"could"be"involved"in"alternative"splicing"by"“skipping”"the"exon"behind"it"in"certain"species,"like"Aspergillus'niger,"where"it"never"results"in"frameshift."In"most"species"of"Onygenales,"however,"inclusion"of"that"exon"II"does"not"lead"to"a"full/length"ORF."We"have"shown"in"Malbranchea'cinnamomea,"that"the"stwintron"and"the"next"U2"intron"need"to"be"excised"first"to"create"a"new"functional"U2"intron"in"the"transcript."It"thus"takes"4"consecutive"splicing"reactions"to"remove"the"whole"complex"IS."Research"was"supported"by"the"EU"and"co/financed"by"the"European"Regional"Development"Fund"under"the"project"GINOP/2.3.2/15/2016/00008S"by"the"EFOP/3.6.1/16/2016/00022"project"co/financed"by"the"EU"and"the"European"Social"Fund."EF"and"NK"are"supported"by"ÚNKP/18/4/DE/3"and"ÚNKP/18/3/II/DE/140"“New"National"Excelence"Program"of"the"Ministry"of"Human"Capacities”,"respectively."$22.$Distribution$of$secondary$metabolites$in$Aspergillus$and$Penicillium""J.C.$Frisvad."Department'of'Biotechnology'and'Biomedicine,'Technical'University'of'denmark,'Kongens'Lyngby,'DK"""Profiles"of"secondary"metabolites"are"species"specific"in"Aspergillus"(427"spp.)"and"Penicillium"(443"spp.),"but"interestingly"a"large"number"of"secondary"metabolites"(SMs)"are"common"in"both"genera,"but"pretty"rare"outside"these"major"genera."Most"species"in"both"genera"can"produce"a"large"number"of"secondary"metabolites."Genome"sequencing"has"shown"that"only"approximately"20%"of"these"secondary"metabolite"families"are"expressed"on"laboratory"media,"while"the"80%"need"special"xenoextrolite,"physiological"or"nutritional"stimuli"to"be"actually"produced."By"examining"many"isolates"in"each"species,"some"additional"secondary"metabolites"can"be"expressed"in"few"of"them,"while"the"core"secondary"metabolome"is"expressed"in"nearly"all"isolates."Aspergillus"and"Penicillium"species"were"compared"and"they"share"important"secondary"metabolites,"including"antibiotics,"mycotoxins"and"antioxidants."Among"the"shared"metabolites"are"3/nitropropionic"acid,"citric"acid,"oxalic"acid,"kojic"acid,"terrestric"acid,"patulin,"penicillic"acid,"xanthomegnins,"citreoviridins,"secalonic"acids,"melleins,"asperentins,"griseofulvins,"terrein,"compactins"/"mevinolins,"citrinin,"cyclopaldic"acids,"sorbicillins,"flavoglaucins,"viridicatumtoxins,"territrems"aflavinins,"penitrems,"fumigaclavines,"fumitremorgins,"gliotoxins,"brasiliamides,"chrysogines,"roquefortines,"ochratoxins,"pseurotins,"bicyclo[2.2.2]diazaoctane"core"containing"metabolites,"penicillins,"tryptoquivalines,"cytochalasins,"viridicatins,"asperphenamate,"mycophenolic"acids,"fumagillin,"austins"and"many"others,"while"few"known"secondary"metabolites"are"only"present"in"Aspergillus"or"Penicillium"species."Several"rather"different"Aspergillus"species"can"produce"aflatoxins"and"sterigmatocystins,"while"these"have"never"been"found"in"any"Penicillium."Other"SMs"found"in"Aspergillus,"but"never"Penicillium,"include"trichothecenes"(only"in"one"Aspergillus"hancockii"however),"fumonisins,"cordycepin,"echinocandins,"malformins,"echinulins,"tensidols,"pyranonigrins,"terphenyllin,"atromentins,"chlorflavonin,"while"SMs"such"as"curvulic"acid,"pulvilloric"acid,"xanthoepocin,"haenamindole,"psychrophilins,"and"PR/toxin"has"until"now"only"been"found"in"certain"Penicillium"spp."Examples"will"be"given"on"SMs"that"are"co/produced"in"certain"species"in"both"genera,"such"as"the"pair""fumagillin"and"pseurotin"(coded"by"an"intertwined"gene"cluster),"and"the"pair"kojic"acid"and"griseofulvin,"indicating"they"may"have"a"synergistic"effect."

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$*23.$Dissecting$novel$secondary$metabolites$pathways$in$Aspergillus*homomorphus""M.E.$Futyma1,"U."H."Mortensen1,"C."Hoeck2,"L."M."Petersen1,"Y."Guo1,"T."O."Larsen1,"C."H."Gotfredsen2,"J."B."Hoof1.""1)'DTU'Bioengineering,'Technical'University'of'Denmark,'Lyngby,'DK6'2)'DTU'Chemistry,'Technical'University'of'Denmark,'Lyngby,'DK""Aspergillus'homomorphus"is"a"filamentous"fungus"belonging"to"section"Nigri"of"Aspergillus'genus."It"is"a"genus"known"to"produce"harmful"mycotoxins"or"industrially"and"pharmaceutically"useful"secondary"metabolites,"making"it"relevant"to"identify"biosynthetic"gene"clusters"(BGC)"responsible"for"their"biosynthesis."Recently,"we"discovered"that"A.'homomorphus"is"particularly"rich"in"interesting"metabolites."Two"examples"are"the"family"of"homomorphosins"(A/F)"and"a"yellow"compound"named"CHO_015404."Here"we"present"our"efforts"towards"elucidation"of"the"genetic"requirements"for"these"two"biosynthetic"pathways."As"a"first"step"we"determined"the"putative"BGCs"of"the"discovered"SMs"by"FungiSMASH"analysis"and"predicted"SM"gene"clusters"available"on"JGI"genome"portal."In"case"of"the"homomorphosins,"we"based"our"search"for"a"BGC"on"clusters"containing"a"non/ribosomal"peptide"synthetase"with"two"adenylation"(AA)"domains"accompanied"by"two"dimethylallyl"tryptophan"synthetases."For"CHO_015404"our"top"BGC"candidate"contained"type"I"Polyketide"Synthase"and"Phenylalanine"Amonnia"Lyase."Performed"bioinformatics"analysis"pointed"into"limited"number"of"candidates"that"we"chose"to"validate"experimentally."With"the"advancing"gene/editing"toolbox"for"filamentous"fungi"developed"in"our"group,"we"used"CRISPR/Cas9"based"deletion"method"expressing"multiple"protospacers"targeting"the"gene"of"interest."At"the"current"stage"we"have"successfully"identified"the"homomorphosins"responsible"BGC."We"assigned"the"enzymatic"functions"encoded"by"the"gene"cluster"by"generating"deletion"mutants"and"metabolites"analysis"by"UHPLC/DAD/MS."The"potential"BGC"of"the"yellow"compound"CHO_015404"is"currently"in"the"validation"phase."$24.$Morphological,$Molecular$and$antimicrobial$activity$of$endophytic$fungi$isolated$from$forest$plants$at$Taif$region$in$Saudi$Arabia""R.$Gashgari1,2,"A."Alsini2.""1)'Biology'Department','King'Abdulaziz'university,'Jeddah,'SA6'2)'Biology'Department','Jeddah'university,'Jeddah,'SA"""Endophytic"fungi,"which"have"been"reported"in"numerous"plant"species,"are"important"components"of"the"forest"community"and"contribute"significantly"to"the"diversity"of"natural"ecosystems."Fungi"growing"on"plant"segments"were"isolated"and"identified"based"on"morphological"and"molecular"characteristics."The"isolates"were"grouped"into"35"distinct"operational"taxonomic"units,"based"on"the"sequence"of"the"internal"transcribed"spacer"regions"in"the"rRNA"gene."The"colonization"frequency"and"the"dominant"fungi"percentage"of"these"endophytic"fungi"were"calculated."A"dual"culture"technique"was"adopted"to"investigate"the"antifungal"activity"of"these"endophytes."Tamarix"nilotica"showed"the"highest"endophytic"diversity"with"a"relative"frequency"of"27.27%,"followed"by'Cressa'cretica"with"a"relative"frequency"of"19.27%."The"most"frequently"isolated"species"was'Penicillium'chrysogenum"with"an"overall"colonization"rate"of"98.57%."Seven"out"of"35"endophytic"fungi"exhibited"strong"antifungal"activity"to"all"plant"fungal"pathogens"tested."P.'chrysogenum,'Fusarium'oxysporum,"and"F.'nygamai"exhibited"the"highest"inhibition"against"the"human"pathogenic"bacteria"Staphylococcus'aureus,'Escherichia'coli,'Pseudomonas'aeruginosa,"and'Klebsiella'pneumoniae.'Aspergillus'sydowii,'P.'chrysogenum,'and'Eupenicillium'crustaceum'showed"strong"antimicrobial"activity"against"Enterococcus'faecalis."$$ $

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*25.$Genomic$analysis$provides$insight$into$secondary$metabolism$of$two$novel,$biosynthetically$talented$of$Aspergillus$from$Australia""C.L.M."Gilchrist1,"A.M."Piggott2,"E."Lacey3,"J.I."Pitt4,"Y.H."Chooi1.""1)'School'of'Molecular'Sciences,'The'University'of'Western'Australia,'Crawley,'AU6'2)'Department'of'Molecular'Sciences,'Macquarie'University,'Sydney,'NSW'2109,'Australia.6'3)'Microbial'Screening'Technologies,'Smithfield,'Sydney,'NSW'2164,'Australia.6'4)'Commonwealth'Scientific'and'Industrial'Research'Organisation,'North'Ryde,'Australia"""Fungi"from"the"genus"Aspergillus"are"prolific"producers"of"bioactive"secondary"metabolites"(SMs)."SMs"produced"by"members"of"the"Aspergillus"genus"include"clinical"drugs"such"as"the"cholesterol/lowering"lovastatin"and"the"antifungal"echinocandin,"and"pharmaceutical"leads"such"as"the"anti/angiogenic"fumagillin."Genomic"analysis"of"the"SM"biosynthetic"capabilities"of"Aspergillus"fungi"has"further"strengthened"the"notion"of"Aspergillus"as"a"talented"genus"for"production"of"structurally"diverse"bioactive"metabolites."Therefore,"the"search"for"rare"or"novel'Aspergillus"species"guided"by"chemotaxonomy"could"be"a"fruitful"strategy"for"discovering"novel"bioactive"metabolites.'In"this"manner,"our"group"sequenced"two"novel"Aspergillus"species"from"Australia,"A.'burnettii'FRR"5400"and"A.'hancockii'FRR"3425."Phylogenetic"analysis"placed"both"species"within"section"Flavi,"which"includes"A.'alliaceus,'A.'flavus'and"A.'oryzae.'We"then"analysed"the"SM"biosynthetic"gene"clusters"(BGCs)"in"each"genome,"and"their"conservation"across"the"broader"Aspergillus'genus."Both"species"are"biosynthetically"talented:"A.'burnettii'encodes"175"putative"BGCs,"of"which"only"19"show"any"similarity"to"previously"characterized"BGCsS"A.'hancockii"encodes"185"putative"BGCs,"with"34"showing"similarity"to"previously"characterized"BGCs."The"A.'burnettii'genome"contains"52"polyketide"synthases"(PKS),"27"nonribosomal"peptide"synthases"(NRPS)"and"20"genes"predicted"to"be"involved"in"terpene"biosynthesisS"A.'hancockii"contains"37"PKS,"26"NRPS"and"13"terpene"biosynthetic"genes."Metabolites"were"isolated"from"both"species"grown"on"7"different"media."All"7"isolated"metabolites"in"A.'burnettii,"and"8"in"A.'hancockii,"were"able"to"be"matched"back"to"candidate"BGCs"via"comparative"genomics"approaches."However,"even"with"many"culture"conditions"tested,"the"amount"of"metabolites"we"observe"is"dwarfed"by"how"much"biosynthetic"machinery"is"encoded"by"their"genomes,"suggesting"that"there"is"still"a"lot"of"biosynthetic"potential"yet"to"be"harnessed."$26.$Interaction$between$CreD$and$HulA$involved$in$endocytic$degradation$of$the$maltose$transporter$MalP$in*Aspergillus*oryzae""K.$Gomi,"H."Tada,"T."Hiramoto,"Y."Matsuura,"M."Tanaka,"T."Shintani.""Tohoku'University,'Sendai,'JP""'Aspergillus'oryzae"produces"copious"amount"of"amylolytic"enzymes,"and"MalP,"a"major"maltose"transporter,"is"required"for"the"expression"of"amylolytic"genes."The"expression"of"these"genes"is"strongly"repressed"by"carbon"catabolite"repression"(CCR)"in"the"presence"of"glucose."MalP"is"transported"from"the"plasma"membrane"to"the"vacuole"by"endocytosis,"which"requires"the"HECT"ubiquitin"ligase"HulA,"an"ortholog"of"yeast"Rsp5."In"the"yeast,"arrestin/like"proteins"mediate"endocytosis"as"adaptors"of"Rsp5"and"transporters."In"this"study,"we"have"examined"the"involvement"of"CreD,"an"arrestin/like"protein,"in"glucose/induced"MalP"endocytosis."Deletion"of"creD"inhibited"the"glucose/induced"endocytosis"of"MalP,"and"CreD"showed"physical"interaction"with"HulA."One"PPXY"and"three"PXY"motifs"exist"in"the"C/terminal"region"of"CreD"and"the"strains"harboring"the"PXY"motif"mutants"of"CreD"showed"the"retardation"of"the"MalP"endocytosis"induced"by"glucose."Futhermore,"pull/down"assay"using"the"PXY"motif"mutants"of"CreD"and"WW"domains"of"HulA"showed"that"the"sequence"PDY"located"at"the"proximal"C/terminus"of"CreD"would"be"most"involved"in"the"interaction"between"HulA"and"CreD.""$ $

! 28"

*27.$Hybrid$Transcription$Factor$Engineering$Activates$Expression$and$Helps$to$Elucidate$the$Biosynthesis$of$the$Silent$(+)LAsperlin$Gene$Cluster$in$Aspergillus*nidulans""Michelle$F."Grau1,"Ruth"Entwistle2,"Yi/Ming"Chiang1,3,"Manmeet"Ahuja2,"C."Elizabeth"Oakley2,"Tomohiro"Akashi4,"Clay"C."C."Wang1,5,"Richard"B."Todd6,"Berl"R."Oakley2.""1)'Department'of'Pharmacology'and'Pharmaceutical'Sciences,'School'of'Pharmacy,'University'of'Southern'California,'Los'Angeles,'CA'6'2)'Department'of'Molecular'Biosciences,'University'of'Kansas,'Lawrence,'KS'6'3)'Department'of'Pharmacy,'Chia'Nan'University'of'Pharmacy'and'Science,'Tainan'City,'Taiwan6'4)'Division'of'OMICS'analysis,'Center'for'Neurological'Diseases'and'Cancer,'Nagoya'University'Graduate'School'of'Medicine,'Nagoya,'Japan6'5)'Department'of'Chemistry,'University'of'Southern'California,'Dornsife'College'of'Letters,'Arts,'and'Sciences,'Los'Angeles,'CA6'6)'Department'of'Plant'Pathology,'Kansas'State'University,'Throckmorton'Plant'Sciences'Center,'Manhattan,'KS'""Some"of"the"most"valuable"medicinal"and"agricultural"compounds"are"secondary"metabolites"(SMs)"produced"by"fungi."As"the"number"of"sequenced"fungal"genomes"continues"to"grow,"it"has"become"evident"that"most"SM"gene"clusters"are"inaccessible"under"common"laboratory"growth"conditions."This"finding"highlights"the"untapped"potential"that"exists"in"discovering"new"and"potentially"valuable"fungal"SMs."Activation"of"pathway/specific"transcription"factors"has"helped"to"express"several"dormant"gene"clustersS"however,"this"approach"is"often"unsuccessful."We"sought"to"design"a"new"approach"to"activate"silent"gene"clusters"that"exploits"the"modularity"of"transcription"factor"domains."The"DNA"binding"domain"of"a"transcription"factor"associated"with"a"silent"SM"gene"cluster"was"fused"to"the"activation"domain"of"a"potent"transcription"factor,"AfoA,"and"the"construct"was"placed"under"the"control"of"the"alcA"promoter"in"Aspergillus'nidulans."Cultivation"of"the"hybrid"transcription"factor"expressing"strain"in"inducing"conditions"activated"expression"of"all"genes"in"the"target"cluster"and"produced"the"antibiotic"(+)/asperlin."Hybrid"transcription"factor"expressing"deletion"strains"were"generated"to"confirm"the"boundaries"of"the"(+)/asperlin"gene"cluster"and"help"elucidate"the"biosynthesis"of"(+)/asperlin."Our"novel"hybrid"transcription"factor"strategy"has"the"potential"to"uncover"the"products"of"other"silent"SM"gene"clusters.""28.$The$reader$protein$SntB$is$involved$in$regulating$secondary$metabolism$in$Aspergillus*flavus""C."Greco1,"B."T."Pfannenstiel2,"N."P."Keller1,3.""1)'Department'of'Medical'Microbiology'and'Immunology,'University'of'WisconsinQMadison,'Madison,'WI,'USA'6'2)'Department'of'Genetics,'University'of'WisconsinQMadison,'Madison,'WI,'USA6'3)'Department'of'Bacteriology,'University'of'WisconsinQMadison,'Madison,'WI,'USA"""Filamentous"fungi"are"an"incredible"source"of"bioactive"secondary"metabolites"(SMs)."Most"SM"biosynthetic"gene"clusters"(BGCs)"are"silent"under"laboratory"conditions"but"can"be"‘turned/on’"by"manipulation"of"histone"post/translational"modifications"(PTMs).""Proteins"containing"reading"domains"have"been"shown"to"facilitate"the"interaction"of"enzymes"that"perform"PTMs."We"discovered"that"the"reader"protein"SntB"in"Aspergillus"spp."globally"regulates"histone"PTMs"and"BGC"activity"and"we"investigate"the"impact"on"secondary"metabolite"production."SntB"was"deleted"in"A.'flavus"which"resulted"in"an"increase"of"global"levels"of"H3K9K14"acetylation."Comparative"analysis"of"secondary"metabolite"production"was"performed"using"UPLC/HRMS"and"HPLC/UV"between"the"wild/type"and"∆sntB"strains."It"was"found"that"SntB"positively"regulates"the"production"of"the"mycotoxin"aflatoxin"B1"as"well"as"the"metabolites"aflavarin"and"asparasone"A."The"contrast"higher"production"titers"of"the"bioactive"leporin"B"and"ditryptophenaline"was"observed."In"addition,"two"additional"metabolites"were"produced"in"the"∆sntB"strain,"upon"isolation,"HRMS"and"NMR"analysis"they"were"characterized"to"be"the"depsipeptide"aspergillicin"A"and"E,"previously"isolated"from"other"Aspergillus'spp.,"but"never"in"A.'flavus."Bioinformatic"analysis"of"the"A.'flavus"genomes"identified"a"putative"non/ribosomal"peptide"synthetase"(NRPS)"in"close"proximity"to"the"different"siderophore"genes."Deletion"of"this"NRPS"and"neighboring"gene"encoding"a"O/methyltransferase"were"required"for"depsipeptide"synthesis."The"NRPS"features"an"unusual"starting"condensation"domain"which"has"been"linked"by"phylogenetic"analysis"to"the"N/acetyl"capped"peptide."Disruption"of"the"neighboring"siderophore"genes"did"not"affect"the"production"of"the"two"aspergillicins.""$ $

! 29"

29.$Diversified$transcriptional$regulation$of$secondary$metabolic$gene$clusters$in$closely$related$Aspergillus$species""H."Takahashi1,"M."Umemura2,"A."Ninomiya3,"M."Shimizu4,"Y."Kusuya1,"S."Urayama3,"A."Watanabe1,"K."Kamei1,"T."Yaguchi1,"D."Hagiwara1,3.""1)'Medical'Mycology'Research'Center'(MMRC),'Chiba'University,'Chiba,'Japan6'2)'Bioproduction'Research'Institute,'National'Institute'of'Advanced'Industrial'Science'and'Technology'(AIST),'Ibaraki,'Japan6'3)'Faculty'of'Life'and'Environmental'Sciences,'University'of'Tsukuba,'Ibaraki,'Japan6'4)'Department'of'Biology,'Faculty'of'Science,'Chiba'University,'Chiba,'Japan"""Filamentous"fungi"produce"a"variety"of"bioactive"compounds"affecting"other"organisms"in"their"surroundings."In"general,"the"secondary"metabolites"(SMs)"are"biosynthesized"by"a"set"of"proteins"encoded"from"contiguous"gene"cluster."Although"fungal"genome"sequences"are"accumulated,"transcriptomic"view"for"evolution"of"SM"was"lacking."Here,"we"conducted"a"series"of"comparative"genomics"and"transcriptomics"analyses"using"closely"related"5"species"of"Aspergillus"section"Fumigati,"namely"medically"important"pathogenic"fungi"Aspergillus'fumigatus,"Aspergillus'lentulus,"Aspergillus'udagawae,"Aspergillus'pseudoviridinutans,"and"Neosartrya'fischeri."A"total"of"263"SM"core"genes"and"21"ribosomal"peptide"synthetic"(RiPS)"genes"was"identified."14"and"2"types"of"the"SM"and"RiPS"genes,"respectively,"are"conserved"in"all"5"species,"whilst"the"rate"of"SM"core"genes"unique"to"each"species"ranged"from"22.2"to"49.3"%."The"fungi"were"then"cultured"under"different"four"conditions"(Potato"dextrose"broth,"Sabouraud"dextrose"broth,"Czapek/Dox,"and"Potato"dextrose"agar),"where"37%"of"the"lineage/conserved"SM"core"genes"(26/70"genes)"were"expressed"in"either"of"cultures."In"contrast,"the"expressed"unique"ones"were"12%"(12/98"genes)."Of"note,"expression"profile"of"the"lineage/conserved"SM"core"genes"was"pronouncedly"diverse."These"results"indicated"an"evolutionary"diversification"of"fungal"SM"gene"cluster"at"transcriptional"level"besides"genomic"rearrangement"or"cluster"gain"and"loss."As"a"good"example"for"this"transcriptional"diversification,"gliotoxin"(gli)"biosynthetic"gene"cluster"exists"in"the"5"species"of"section"Fumigati."In"A.'fumigatus"and"A.'udagawae,"the"gli"cluster"was"active"with"most"genes"expressed"under"either"of"conditions"tested,"whereas"the"expression"level"was"intermediate"in"A.'lentulus,"and"almost"silent"in"A.'pseudoviridinutans"and"N.'fischeri."The"production"of"gliotoxin"was"only"detected"in"A.'fumigatus."Finally,"our"transcriptome"analysis"illuminated"media/dependent"expression"of"terrein"cluster"in"A.'lentulus"and"confirmed"extraordinary"high"level"of"terrein"production,"which"altogether"highlighted"the"potential"of"transcriptomic"approach"to"further"access"unknown"fungal"SMs.""30."MpkB$MAP$kinase$pathway$is$required$for$sexual$development$but$not$for$mycotoxin$production$in$Aspergillus*nidulans$and$Aspergillus*flavus""Sang/Cheol"Jun,"Bohan"Zhu,"Jong/Hwa"Kim,"KapLHoon$Han.""Dept.'of'Pharmaceutical'Engineering,'Woosuk'University,'Wanju,'Jeonbuk,'KR"""MAP"kinase"pathways"regulate"stress"responses"as"well"as"growth"and"development"in"most"eukaryotic"systems."MpkB"MAP"kinase"in"a"model"fungus"Aspergillus'nidulans"has"been"known"to"coordinate"sexual"development"and"secondary"metabolism,"including"sterigmatocystin"(ST)"production."In"this"study,"however,"the"results"of"the"ST"production"analysis"of"wild"type"and"mpkB"deletion"mutants"showed"that"the"mpkB"gene"did"not"affect"the"ST"production"and"ST"related"gene"expression.""Furthermore,"ST"production"of"ΔmpkB,"ΔmkkB,"and"ΔmpkBΔmkkB"mutants"in"the"veA+"background"were"similar"with"wild"type."Also,"MpkB"constitutive"activation"or"inactivation"mutants"showed"no"significant"effect"on"the"ST"production."Interestingly,"ST"production"of"mpkB"and"mkkB"mutants"was"remarkably"delayed"in"the"veA1"background,"suggesting"that"the"ST"production"is"affected"primarily"by"the"veA"gene."Similarly,"in"Aspergillus'flavus,"MpkB"ortholog"AflmpkB"mutant"couldn’t"produce"any"sclerotia,"but"it"produced"aflatoxin"B1"normally."Taken"together,"the"mpkB"gene"alone"does"not"affect"mycotoxin"production"such"as"ST"in"A.'nidulans"or"aflatoxin"B1"in"A.'flavus,"indicating"that"the"signaling"of"MpkB"MAP"kinase"and"mycotoxin"production"were"governed"by"independent"pathways."$$ $

! 30"

31.$NsdD$GATA$factorLdependent$regulation$of$fungal$development$and$sporeLspecific$gene$expression$of$Aspergillus*nidulans""Dong/Soon"Oh1,"Mi/Kyung"Lee2,"Hee/Soo"Park2,"Heungyun"Moon2,"Xiaoxiao"Huang1,"Dong/Min"Han3,"Jae/Hyuk"Yu2,"KapLHoon"Han1.""1)'Dept.'of'Pharmaceutical'Engineering,'Woosuk'University,'Wanju,'Jeonbuk,'KR6'2)'Dept.'of'Bacteriology,'University'of'WisconsinQMadison,'Madison,'WI,'USA6'3)'Dept.'of'Life'Sciences,'Wonkwang'University,'Iksan,'Jeonbuk,'KR"""Many"fungi"produce"sexual"spores"as"a"result"of"stress"responses"to"cope"unfavorable"conditions"or"for"propagating"to"their"habitats"appropriately."A"genetic"model"fungus,"Aspergillus'nidulans,"is"a"homothallic"ascomycete"that"has"been"studied"sexual"and"asexual"developmental"processes"in"molecular"level"for"many"years."Previous"studies"showed"that"a"GATA/type"transcription"factor,"NsdD,"plays"a"pivotal"role"in"activating"sexual"development"of"A.'nidulans."The"nsdD'deletion"mutants"failed"to"form"the"sexual"structures,"cleistothecia,"even"under"conditions"where"sexual"development"preferentially"occurs"in"wild"type."Over/expressors"of"the"gene"formed"larger"numbers"of"cleistothecia"than"wild"type,"even"under"conditions"where"wild"type"strains"form"little"sexual"structure,"indicating"that"the"nsdD"gene"is"positive"regulator"of"sexual"development."Recently,"we"also"found"that"the"deletion"of"nsdD"bypassed"the"need"for"fluG"and"flbA~flbE,"but"not"brlA"or"abaA,"in"conidiation,"and"partially"restored"production"of"the"mycotoxin"sterigmatocystin"(ST)"in"the"ΔfluG,"ΔflbA,"and"ΔflbB"mutants,"suggesting"that"NsdD"plays"an"important"negative"role"in"conidiation"and"is"positioned"between"FLBs"and"BrlA"in"A.'nidulans."We"have"further"revealed"that"NsdD"is"a"key"repressor"affecting"the"quantity"of"asexual"spores"in"Aspergillus."To"identify"the"direct"and/or"indirect"target"genes"of"NsdD,"ChIP/seq"and"RNA/seq"analyses"have"been"performed."As"a"result,"NsdD"Responsible"Element"or"NRE"has"been"identified"as"CMGATCT"and"many"possible"target"promoters"have"been"elucidated."Moreover,"since"physiological"and"genetic"studies"of"ascospores"are"remained"to"be"characterized,"we"also"performed"RNA/seq"analysis"from"conidia"and"ascospores"RNA"samples"in"A.'nidulans'for"obtaining"more"information"about"ascospore"biology."Comparative"analysis"of"transcription"profiles"revealed"many"genes"that"are"expressed"specifically"in"ascospores."Detailed"investigation"of"the"differentially expressed genes will be discussed. $32.$FungiDB:$integrating$genomic$data$for$pathogens$and$model$organisms$and$providing$advanced$search$capabilities$and$largeLscale$data$analysis""Evelina"Basenko1,"Omar"Harb2,"Achchuthan"Shanmugasundram1,"David"Roos2,"..."on"behalf"of"the"EuPathDB"project.""1)'Centre'for'Genomic'Research,'Functional'&'Comparative'Genomics,'University'of'Liverpool,'UK6'2)'Department'of'Biology,'University'of'Pennsylvania,'Philadelphia'PA,'US""$FungiDB.org"is"a"free,"online"data"mining"resource"supporting"fungal"and"oomycete"genera"and"providing"functional"analysis"of"genomic/scale"datasets."FungiDB"is"a"component"of"the"EuPathDB"(http://EuPathDB.org)"family"of"databases"that"integrate"diverse"information,"including:"//"genome"sequences"and"population/level"diversity"data"for"both"pathogenic"and"non/pathogenic"species"//"diverse"functional"genomics"information,"generated"on"multiple"platforms:"chromatin"marks,"transcriptomes,"proteomes,"genome/wide"phenotypic"information,"etc."//"information"on"structure,"molecular"interactions,"metabolic"pathways"and"signaling"networks,"publications,"etc."//"automated"analyses"(InterPro"scan,"orthology"predictions,"etc.)"'User"Workspaces'"permit"researchers"to"analyze"their"own"data"using"Galaxy"workflows"and"examine"the"results"in"the"context"of"publicly/accessible"datasets"(e.g."RNAseq"analysis"results"can"be"exported"into"private"EuPathDB"user"workspace"and"visualized"in"GBrowse"along"with"publicly"available"RNA/Seq"tracks)."Analytical"tools"(ClutalOmega,"MitoProt,"WoLF"PSORT,"etc.)"and"sophisticated"'Search"Strategies'"support"in"silico"queries"against"a"wealth"of"integrated"and"automatically"generated"data,"encouraging"scientists"to"ask"their"own"questions."FungiDB"supports"exploration"of"genomes"from"Agaricomycetes,"Blastocladiomycetes,"Chytridiomycetes,"Dothideomycetes,"Eurotiomycetes,"Leotiomycetes,"Pneumocystidomycetes,"Pucciniomycetes,"Saccharomycetes,"Schizosaccharomycetes,"Sordariomycetes,"Tremellomycetes,"Ustilaginomycetes,"and"Zygomycetes"genera."It"integrates"a"wealth"of"datasets"including"functional"genomic"and"phenotypic"information"for"Neurospora"(the"Neurospora"genome"project"and"knockout"collection),"Aspergilli"(AspGD"

! 31"

curated"data),"and"Fusarium."FungiDB"also"accommodates"community/driven"submission"of"images,"phenotype"descriptions,"supporting"files,"PubMed"records,"etc."as"'User"Comments',"which"become"immediately"visible"and"searchable."Interested"in"joining"FungiDB"advisory"calls"or"nominating"a"dataset"for"integration?"Have"questions"or"comments?"Email"help@fungidb.org"FungiDB"is"supported"in"part"by"NIH"#HHSN272201400030C"and"Wellcome"Trust"#WT108443MA"and"Wellcome"Biomedical"Resources"Grant"#212929/Z/18/Z."$33.$Fungal$strategy$of$virulence$by$counteracting$lipidLraft$microdomain$formation$of$phagolysosomes""F."Schmidt1,2,"M."Roecker1,2,"A."Thywissen1,2,"H."Schmidt1,2,"Z."Cseresnyés3,"S."Galiani4,"M"Strassburger5,"M.T."Figge3,"C."Eggeling4,"T.$Heinekamp1,2,"S.G."Filler6,"A.A."Brakhage1,2.""1)'Department'of'Molecular'and'Applied'Microbiology,'Leibniz'Institute'for'Natural'Product'Research'and'Infection'Biology'(HKI),'Jena,'DE6'2)'Department'of'Microbiology'and'Molecular'Biology,'Institute'of'Microbiology,'Friedrich'Schiller'University'Jena,'Jena,'DE6'3)'Research'Group'Applied'Systems'Biology,'HKI,'Jena,'DE6'4)'MRC'Human'Immunology'Unit,'Weatherall'Institute'of'Molecular'Medicine,'University'of'Oxford,'UK6'5)'Transfer'Group'AntiQInfectives,'HKI,'Jena,'DE6'6)'Division'of'Infectious'Diseases,'Los'Angeles'Biomedical'Research'Institute'at'HarborQUCLA'Medical'Center,'Torrance,'CA,'David'Geffen'School'of'Medicine'at'University'of'California,'Los'Angeles,'CA""'Aspergillus'fumigatus"represents"the"most"important"air/borne"fungal"pathogen."In"lung"alveoli,"resident"alveolar"macrophages"belong"to"the"first"line"of"defense"against"inhaled"conidia."The"presence"of"dihydroxynaphthalene"(DHN)"melanin"on"the"conidial"surface"is"crucial"to"inhibit"phagolysosomal"acidification"and"to"cause"increased"damage"to"macrophages"compared"to"non/pigmented"pksP'mutant"conidia."Here,"we"discovered"the"importance"of"lipid"rafts"for"immune"cells"to"counteract"the"attack"of"A.'fumigatus."DHN"melanin"leads"to"disturbance"of"lipid"raft"microdomains"in"the"membrane"of"phagolysosomes,"thereby"preventing"the"assembly"and"activity"of"vATPase."As"a"consequence,"phagolysosomal"acidification"is"inhibited."The"conidia/containing"phagolysosomal"membrane"is"characterized"by"a"high"content"of"cholesterol"and"sphingolipids."Further"classification"of"the"lipid"raft"microdomains"showed"that"flotillin/enriched"lipid"rafts"co/localized"with"sphingolipids"on"phagolysosomal"membranes"containing"pksP"conidia"but"not"wild/type"conidia."In"human"and"mouse,"two"flotillins,"Flot/1"and"2,"are"present."As"chaperon,"flotillins"stabilize"lipid/raft"microdomains"and"thus"represent"a"marker"protein."To"further"classify"the"lipid/raft"microdomains"we"improved"a"protocol"for"isolation"of"phagolysosomes"from"macrophages."This"allowed"to"monitor"the"dependence"of"the"recruitment"of"receptors"and"enzyme"complexes"on"lipid/raft"microdomains"and"also"to"co/localize"microdomains"and"receptor/enzyme"complexes"by"high"resolution"fluorescence"microscopy."In"addition,"bone"marrow/derived"macrophages"(BMDMs)"of"Flot/1/2"knockout"mice"were"compared"with"C57BL7/6"wild/type"macrophages."BMDMs"were"co/incubated"with"melanized"wild/type"conidia"as"well"as"pigmentless"pksP"conidia."Consistently,"acidification"of"phagolysosomes"containing"pksP'conidia"was"drastically"reduced"in"Flot/1/2"knockout"BMDMs"as"well"as"vATPase"assembly"and"phagocytosis."Infection"of"Flot/1/2"knockout"mice"with"pksP"conidia"resulted"in"increased"cell"damage"and"killing"compared"to"wild/type"mice."Taken"together,"these"data"indicate"that"lipid"raft"microdomains"provide"important"platforms"for"signaling"and"defense"of"immune"cells"and"can"be"manipulated"by"A.'fumigatus.""34.$The$sepD5$mutation$in$Aspergillus*nidulans$is$located$in$the$gene$encoding$PaxB""T."Hill1,"L."Jackson/Hayes1,"S."Harris2,"A."Virag3,"M."Williamson1.""1)'Biochemistry'&'Molecular'Biology'Program,'Rhodes'College,'Memphis,'TN6'2)'Department'of'Biological'Sciences,'University'of'Manitoba,'Winnipeg,'MB6'3)'Dupont'Industrial'Biosciences,'Palo'Alto,'CA"""The"study"of"septation"in"filamentous"fungi"received"a"significant"boost"through"the"identification"and"characterization"of"several"temperature/sensitive"septation"(sep)"mutants"in"Aspergillus'nidulans"(Morris,"1975,"Genet."Res."26:"237/254S"Harris"et"al.,"1994,"Genetics"136:"517/532)."""One"continuing"loose"end"from"the"latter"study"has"been"the"identity"of"the"mutation"designated"sepD5,"which"is"completely"aseptate"at"restrictive"temperatures""Meiotic"mapping"places"sepD5"on"Chromosome"II"in"the"vicinity"of"

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trpA"(AN3695)"and"adD"(AN3626)"with"recombination"frequencies"of"approximately"1%"and"8%,"respectively.""Using"next/generation"whole"genome"sequencing"of"gDNA"from"a"sepD5"strain,"we"have"identified"a"point"mutation"in"the"coding"region"of"AN3659,"predicted"to"encode"a"homologue"of"S.'cerevisiae'and"S.'pombe"paxillin"Pxl1.""This"locus"has"been"designated"paxB.""The"sepD5"phenotype"is"complemented"by"the"cloned"wild"type"paxB"allele"and"is"replicated"in"a"paxB"deletion"strain.""The"mutation"in"sepD5"is"a"G/to/A"transition"at"position"92"of"the"2513"base"pair"open"reading"frame,"predicted"to"cause"a"glutamine/to/arginine"substitution"at"residue"31"of"the"predicted"776/amino"acid"product.""This"residue"is"in"a"region"of"predicted"intrinsic"disorder"(MobiDB"database)"having"no"counterpart"in"the"yeast"homologues.""The"Q31"residue"is,"however,"a"perfectly"conserved"feature"of"the"paxillin"homologues"of"filamentous"ascomycetes"(based"on"the"top"50"homologues"identified"in"a"UniProtKB"database"search)"in"a"highly"conserved"ca."38/residue"region,"suggesting"a"functionality"of"filamentous"fungus"paxillins"not"found"in"paxillins"of"yeasts."""GFP/tagged"PaxB"is"located"apically"in"a"point"consistent"with"the"core"components"of"the"Spitzenkörper,"and"in"a"strain"co/expressing"the"Lifeact"actin"marker"PaxB::GFP"colocalizes"with"actin"at"the"final"stages"of"compaction"of"the"contractile"actin"ring"(CAR).""It"was"not"observed"in"association"with"actin"during"the"preceding"“string”"stage"of"ring"assembly.""PaxB"remains"associated"with"the"CAR"as"it"constricts"during"septum"formation."""In"a"paxB"deletion"strain"CARs"form"abundantly,"but"they"fail"to"constrict,"and"after"a"prolonged"existence"they"dissipate"into"the"cytoplasm.""If"CAR"formation"is"blocked"in"a"strain"in"which"the"gene"for"profilin"is"deleted,"PaxB"fails"to"localize"at"sites"consistent"with"septation"landmarks,"indicating"that"PaxB"localization"at"septation"sites"is"dependent"upon"actin."""$*35.$MultiLtiered$regulation$of$nuclear$accumulation$of$the$GATA$transcription$factor$AreA$in$Aspergillus*nidulans.""Cameron$C."Hunter1,"Koon"Ho"Wong3,"Michael"J."Hynes2,"Meryl"A."Davis2,"Richard"B."Todd1.""1)'Kansas'State'University,'Manhattan,'KS6'2)'The'University'of'Melbourne,'Parkville,'Australia6'3)'The'University'of'Macau,'Taipa,'China"""The"primary"regulator"of"nitrogen"metabolic"genes"in"fungi"is"the"ortholog"of"the"GATA"transcription"factor"AreA."In"Aspergillus'nidulans,"the"activity"of"AreA"is"regulated"by"multiple"mechanisms"including"regulated"nuclear"accumulation."When"A.'nidulans"is"grown"in"the"presence"of"a"good"nitrogen"source,"AreA"is"found"predominantly"throughout"the"cytoplasm,"but"when"transferred"from"a"good"nitrogen"source"to"an"environment"depleted"of"nitrogen"AreA"accumulates"in"the"nucleus."In"a"previous"study"we"demonstrated"that"AreA"has"multiple,"conserved,"nuclear"localization"sequences"which"mediate"AreA"nuclear"import."The"mechanisms"underlying"AreA"nuclear"import"and"accumulation"are"largely"unknown."We"have"attempted"to"elucidate"both"the"causes"for"and"the"mechanisms"behind"AreA"nuclear"import"and"accumulation."Using"strains"with"HA/epitope"tagged"versions"of"wild/type"and"mutant"AreA"we"have"shown"that"DNA"binding"is"not"required"for"AreA"nuclear"accumulation,"that"nuclear"import"of"AreA"is"dependent"on"the"α/importin"KapA,"and"that"the"deletion"of"some"individual"Target"of"Rapamycin"pathway"genes"prevents"AreA"nuclear"accumulation."Collectively"our"data"reveals"complex"and"multi/tiered"regulation"of"the"subcellular"localization"of"the"GATA"transcription"factor"AreA.""*36.$The$early$asexual$development$regulator$fluG$codes$for$two$putative$enzymes""M.$IradiLSerrano,"L."Tola/García,"M."S."Cortese,"U."Ugalde."Microbial'Biochemistry,'Applied'Chemistry,'University'of'the'Basque'Country,'Donostia'Q'San'Sebastian,'Gipuzkoa,'ES"""FluG"is"a"long"recognized"early"regulator"of"asexual"development"in"Aspergillus'nidulans."fluG"null"mutants"(ΔfluG)"show"profuse"aerial"growth"but"no"conidial"production."Initial"studies"reported"sequence"homology"of"FluG"with"a"prokaryotic"type"I"glutamine"synthetase,"but"catalytic"activity"was"not"demonstrated."FluG"signaling"has"been"reported"to"involve"extracellular"low"molecular"weight"factors"(Lee"and"Adams,"1994)."Their"search"by"Rodriguez/Urra"et"al."(2012)"identified"an"adduct"of"two"excreted"secondary"metabolites"(diorcinol"and"dehydroaustinol"(DHO))"that"was"capable"of"inducing"partial"sporulation"when"applied"onto"a"ΔfluG"colony."However,"a"report"by"Nielsen"et"al."(2011)"described"the"phenotype"of"a"null"mutant"of"the"gene"coding"for"the"synthesis"of"the"austinol'(ausA),"the"precursor"in"the"DHO"biosynthesis,"as"conidial."In"this"study"we"have"confirmed"that"an"ausA"null"mutant"displayed"a"conidiating"phenotype"and"was"able"to"extracellularly"complement"the"sporulation"defect"of"an"adjacent'ΔfluG"strain,"thus"showing"that"FluG/related"signaling"is"not"directly"connected"to"DHO/induced"

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conidiation."In"addition,"we"conducted"an"in/depth"analysis"of"the"FluG"sequence,"which"revealed"a"putative"N/terminal"amidohydrolase"region"linked"to"a"C/terminal"γ/glutamyl"ligase"region."Separate"expression"of"these"domains"confirmed"that"the"C/terminal"region"is"essential"for"asexual"development."The"N/terminal"region,"in"turn,"was"required"to"maintain"the"balance"between"development"and"vegetative"growth"under"high"nutrient"availability."The"analysis"of"a"strain"expressing"both"N/"and"C/terminal"regions"as"two"separate"polypeptides"not"only"proved"that"both"regions"are"functional"when"separately"expressed,"but"that"the"N/terminal"region"performs"an"important"supportive"role"in"conidia"production,"especially"when"both"regions"are"jointly"expressed"and"under"nutrient/rich"conditions."Point"mutations"directed"at"respective"key"catalytic"residues"in"each"region"demonstrated"that"they"were"essential"for"biological"function."Moreover,"the"substitution"of"the"N/and"C/terminal"regions"with"homologs"from"Lactobacillus'paracasei"and"Pseudomonas'aeruginosa"respectively,"resulted"in"conidiating"phenotypes."In"line"with"the"aforementioned"results,"the"phenotype"observed"in"a"chimera"expressing"a"fusion"protein"of"both"bacterial"genes,"confirmed"that"both"bacterial"enzymes"can"work"jointly"as"a"FluG"substitute"in'vivo."Taken"together,"the"results"lead"us"to"conclude"that"FluG"is"a"bifunctional"enzyme"that"participates"in"an"as"yet"unidentified"metabolic"or"signaling"pathway"involving"a"γ/glutamylated"intermediate"required"for"asexual"development."$37.$CRISPRLmediated$expression$platform$for$multiLspecies$Aspergilli""Z.D.$Jarczynska,"M.E."Futyma,"J.K."Rendsvig,"F.H."Kirchner,"C.S."Nødvig,"U.H."Mortensen."DTU'Bioengineering,'Technical'University'of'Denmark,'Lyngby,'DK"""The"recent"sequencing"survey"on"many"fungal"species"revealed"a"large"repertoire"of"industrially"or"medically"relevant"enzymes"and"secondary"metabolites."However,"the"industrial"potential"of"these"species"is"often"hindered"by"the"fact"that"it"is"difficult"to"predict"whether"a"given"host"possesses"a"physiology"and"metabolism"compatible"for"production"of"a"given"productS"and"by"poorly"developed"genetic"toolboxes."Hence,"most"heterologous"production"is"performed"in"a"few"different"hosts"where"toolboxes"are"in"place."To"increase"the"chance"of"successful"heterologous"production,"we"have"created"a"flexible"expression"platform"in"various"Aspergillus"species."The"system"is"based"on"a"reporter"gene,"which"has"been"inserted"into"a"defined"locus"in"each"of"the"different"expression"hosts."The"reporter"serves"two"purposes."Firstly,"it"allows"to"assess"the"strength"of"expression"from"the"defined"locus."Secondly,"the"reporter"gene"can"be"replaced"by"a"gene/expression"cassette"containing"your"favorite"gene"or"pathway"via"marker/free"homologous"recombination"mediated"by"CRISPR/Cas9"technology."Importantly,"to"minimize"DNA"construction"work,"our"setup"allows"different"species"to"be"transformed"by"the"same"gene/expression"cassette,"which"can"be"assembled"by"in'vivo"recombination"using"bio/bricks"that"overlap"with"40"bp."As"a"proof/of/concept,"we"chose"three"representatives"of"Aspergillus:"the"model"fungus"A.'nidulans,"and"the"fungal"industrial"workhorses"A.'niger"and"A.'aculeatus."We"have"used"β/glucuronidase/encoding"gene"(gus)"as"a"reporter"gene"and"inserted"it"into"a"defined"integration"site"in"all"three"species."These"strains"develop"blue"color"on"media"supplemented"with"GUS"susbtrate"(X/gluc)"and"constitute"our"versatile"Aspergillus"expression"platform."Strains"where"new"genes"have"been"correctly"inserted"into"the"platform"via"CRISPR"mediated"recombination"can"easily"be"identified"by"using"simple"blue/white"screening"as"the"new"genes"replace"gus"in"the"process."Using"our"platform"we"have"tested"and"compared"production"of"relevant"enzymes"and"secondary"metabolites"in"all"three"species.""*38."Mitochondrial$citrate$transporters$CtpA$and$YhmA$are$involved$in$cytosolic$acetylLCoA$biosynthesis$in$the$white$koji$fungus,$Aspergillus*luchuensis*mut.*kawachii.""C."Kadooka1,"K."Izumitsu2,"M."Onoue1,"K."Okutsu1,"Y."Yoshizaki1,"K."Takamine1,"M."Goto3,"H."Tamaki1,"T."Futagami1.""1)'Kagoshima'Univ.,'Kagoshima,'JP6'2)'Univ.'of'Shiga'Pref.,'Shiga,'JP6'3)'Saga'Univ.,'Saga,'JP""'Aspergillus'luchuensis'mut.'kawachii'produces"a"large"amount"of"citric"acid"during"the"process"of"fermenting"shochu,"a"traditional"Japanese"distilled"spirit."In"this"study,"we"characterized"A."kawachii"CtpA"and"YhmA,"which"are"homologous"to"the"yeast"Saccharomyces'cerevisiae"mitochondrial"citrate"transporters"Ctp1"and"Yhm2,"respectively."CtpA"and"YhmA"were"purified"from"A.'kawachii"and"reconstituted"into"liposomes."The"proteoliposomes"exhibited"only"counter/exchange"transport"activityS"CtpA"transported"citrate"using"counter"substrates"especially"for"cis/aconitate"and"malate,"whereas"YhmA"

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transported"citrate"using"a"wider"variety"of"counter"substrates,"including"citrate,"2/oxoglutarate,"malate,"cis/aconitate,"and"succinate."Disruption"of"ctpA"and"yhmA"caused"deficient"hyphal"growth"and"conidia"formation"with"reduced"mycelial"weight–normalized"citrate"production."Because"we"could"not"obtain"a"ΔctpA"ΔyhmA"strain,"we"constructed"a"ctpAQS"conditional"expression"strain"in"the"ΔyhmA'background"using"the"Tet/On"promoter"system."Knockdown"of"ctpA/S"in"ΔyhmA"resulted"in"a"severe"growth"defect"on"minimal"medium"with"a"significantly"reduced"acetyl/CoA"level,"indicating"that"double"disruption"of"ctpA"and"yhmA"leads"to"synthetic"lethalityS"however,"we"subsequently"found"that"the"severe"growth"defect"was"relieved"by"addition"of"acetate,"which"can"be"converted"to"acetyl/CoA."Our"results"indicate"that"CtpA"and"YhmA"are"mitochondrial"citrate"transporters"involved"in"citric"acid"production"and"that"transport"of"citrate"from"mitochondria"to"the"cytosol"plays"an"important"role"in"acetyl/CoA"biogenesis"in"A.'kawachii.""*39.$Sporulation$environment$alters$conidial$transcriptome$and$germination$potential.""S.$Earl$Kang,"Michelle"Momany."Plant'Biology,'University'of'Georgia,'Athens,'GA""''''''Aspergillus'fumigatus,"a"thermotolerant"saprophytic"fungus,"causes"over"3"million"global"cases"of"acute"and"chronic"pulmonary"infections.""A.'fumigatus"is"also"responsible"for"food"spoilage"on"cured/processed"meats"and"stored"commodities"such"as"cereal"grains,"spices,"and"cocoa."The"infectious"propagules"of"A.'fumigatus"are"small,"airborne"conidia"(asexual"spores)"which"are"produced"in"a"wide"range"of"environments"and"distributed"ubiquitously"throughout"the"globe."""""""To"determine"the"impact"of"environment"on"conidial"development,"dormancy,"and"germination,"A.'fumigatus"isolate"CEA10"was"sporulated"and"germinated"under"various"environmentally"and"medically"relevant"conditions.""We"found"that"variation"in"sporulation"environment"leads"to"phenotypic"variation"in"genetically"identical"conidia"and"impacts"germination"rate"and"early"post"germination"growth."""""""To"identify"genes"responsible"for"the"phenotypic"variation,"transcriptome"analysis"was"performed"on"conidia"from"various"sporulation"environments."We"identified"genes"that"are"always"present"in"conidia"regardless"of"sporulation"environment."We"also"found"that"conidia"contain"transcripts"specific"to"individual"sporulation"environments"suggesting"conidia"might"be"primed"by"sporulation"conditions"to"respond"to"specific"germination"environments.""Comparison"of"conidia"from"sporulation"environments"resulting"in"high"and"low"germination"phenotypes"showed"differential"expression"of"secondary"metabolite"clusters"suggesting"secondary"metabolites"might"play"roles"in"conidial"development,"dormancy,"and"germination."""""""Taken"together,"our"results"suggest"that"sporulation"environment"generates"heterogeneity"in"a"population"of"genetically"identical"conidia"which"contributes"to"the"fitness"of"the"conidia"and"survival"of"A.'fumigatus."""40.$Effects$of$manganese$(II)$ion$transporter$mutations$in$citric$acid$accumulation$in$Aspergillus*niger""B."Fejes1,"J.P."Ouedraogo2,"E."Fekete1,"Á."Soós3,"B."Kovács3,"E."Sándor3,"Á.P."Molnár1,"A."Tsang2,"L.$Karaffa1.""1)'Dept.'of'Biochemical'Engineering,'University'of'Debrecen,'Debrecen,'HU6'2)'Centre'for'Structural'and'Functional'Genomics,'Concordia'University,'Montreal,'CA6'3)'Institute'of'Food'Science,'University'of'Debrecen,'HU"""Citric"acid"(2/hydroxy/propane/1,2,3/tricarboxylic"acid)"is"a"well/known"example"of"fungal"“overflow"metabolism”,"a"term"used"to"describe"the"apparently"wasteful"strategy"of"certain"fungi"to"incompletely"oxidize"their"carbon"source."The"estimated"2.1"million"tons"of"citric"acid"manufactured"annually"is"mostly"performed"by"fermentation"using"the"filamentous"fungus'Aspergillus'niger."One"critical"prerequisite"for"high/yield"citric"acid"overflow"is"to"adjust"the"concentration"of"Mn(II)"ions"in"the"medium."Mn(II)"concentrations">4"ppb"reduce"citric"acid"accumulation"by"about"20%."Understanding"manganese"metabolism"in"A.'niger"is"critical"to"improve"citric"acid"production."The"A.'niger"gene"NRRL3_07789"is"predicted"to"encode"a"manganese"transporter."We"constructed"a"∆NRRL3_07789"strain"as"well"as"a"strain"overexpressing"NRRL3_07789."Mn(II)"uptake"rate"of"the"deletion"strain"dropped"to"zero,"while"that"of"the"overexpression"strain"doubled."Under"citric"acid"producing"conditions"both"the"reference"and"the"deletion"mutant"strains"displayed"molar"yields"over"80%,"but"maximal"biomass"of"the"mutant"decreased"by"a"third."In"contrast,"molar"yield"of"the"OE"cultures"dropped"to"17%"of"that"of"the"reference"strain,"while"biomass"formation"doubled."Notably,"the"overexpression"strain"displayed"an"overwhelmingly"filamentous"morphology"characterized"by"branched"

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mycelial"clumps"as"opposed"to"the"pellet/dominated"cultures"of"the"reference"and"deletion"strains."Our"results"demonstrate"that"NRRL3_07789"is"involved"in"the"transport,"and"hence"metabolism"of"Mn(II)"ionsS"and"is"highly"relevant"to"citric"acid"overflow'A.'niger."Research"was"supported"by"the"EU"and"co/financed"by"the"European"Regional"Development"Fund"under"the"project"GINOP/2.3.2/15/2016/00008S"by"the"EFOP/3.6.1/16/2016/00022"project"co/financed"by"the"European"Union"and"the"European"Social"FundS"and"the"Industrial"Biocatalysis"Strategic"Network"of"the"Natural"and"Engineering"Research"Council"of"Canada."EF"is"supported"by"the"ÚNKP/18/4"New"National"Excellence"Program"of"the"Ministry"of"Human"Capacities.""41.$An$AoFus3Linteracting$protein$FipC$is$a$novel$regulator$of$cell$fusion$in$Aspergillus*oryzae""T."Katayama1,2,"T."Mo1,"Ö."Bayram3,"D."Takemoto4,"G."Braus5,"K."Kitamoto6,"J."Maruyama1,2.""1)'Department'of'Biotechnology,'The'University'of'Tokyo,'Tokyo,'JP6'2)'CRIIM,'UTokyo,'Tokyo,'JP6'3)'Department'of'Biology,'Maynooth'University,'Maynooth,'IE6'4)'Graduate'School'of'Bioagriculutural'Sciences,'Nagoya'University,'Aichi,'JP6'5)'Department'of'Molecular'Microbiology'and'Genetics,'GeorgQAugustQUniversity'Göttingen,'Göttingen,'DE6'6)'Nihon'Pharmaceutical'University,'Saitama,'JP"""Cell"fusion"is"crucial"process"in"sexual"development"such"as"mating"of"yeasts"and"also"occurs"during"vegetative"growth"in"filamentous"fungi"but"not"yeasts."The"components"of"signaling"pathway"regulating"mating"of"yeasts"such"as"a"mitogen/activated"protein"kinase"Fus3"and"a"transcription"factor"Ste12"are"essential"for"cell"fusion"in"filamentous"fungus"Neurospora'crassa."Previously,"we"also"showed"an"Fus3"orthologue"AoFus3"is"required"for"cell"fusion"in"Aspergillus'oryzae1."In"the"present"study,"we"identified"FipC"and"an"Ste12"orthologue"AoSte12"as"AoFus3/interacting"proteins"by"tandem"affinity"purification."FipC"also"interacts"with"AoSte12,"and"localizes"in"nuclei."In"a"fipC'deletion"mutant,"the"cell"fusion"efficiency"and"the"mRNA"levels"of"cell"fusion/related"genes"greatly"decreased."By"contrast,"the"mRNA"levels"of"cell"fusion/related"genes"were"increased"by"overexpression"of"fipC."These"results"indicate"that"FipC"is"a"novel"positive"regulator"of"the"cell"fusion/related"genes."On"the"other"hand,"the"cell"fusion"efficiency"and"the"mRNA"levels"of"cell"fusion/related"genes"moderately"decreased"in"an"Aoste12'deletion"mutant,"indicating"that"AoSte12"positively"regulates"cell"fusion"but"is"not"essential"for"cell"fusion"in"A.'oryzae'unlike"N.'crassa."Interestingly,"the"decreased"cell"fusion"efficiency"and"mRNA"levels"of"cell"fusion/related"genes"in"the"fipC'deletion"mutant"were"partially"recovered"by"deletion"of"Aoste12,"suggesting"that"AoSte12"also"negatively"regulates"cell"fusion."The"cell"fusion"efficiency"was"indeed"decreased"by"overexpression"of"Aoste12."Our"results"suggest"that"a"novel"protein"FipC"regulates"filamentous"fungi/specific"cell"fusion"by"controlling"AoSte12,"which"has"both"positive"and"negative"functions"in"cell"fusion."1Tsukasaki"et'al.,"Biosci.'Biotechnol.'Biochem.,"78:"1254/1262"(2014)""*42.$A$LowLOxygen$Responsive$Alanine$Aminotransferase$is$Important$for$Aspergillus*fumigatus*Biofilm$Formation,$Germination,$and$Growth$During$Infection.""Joshua"Kerkaert1,"Carey"Nadell2,"Robert"Cramer1."1)'Department'of'Microbiology'and'Immunology,'Geisel'School'of'Medicine'at'Dartmouth,'Hanover,'NH,'USA6'2)'Department'of'Biological'Sciences,'Dartmouth'College,'Hanover,'NH,'USA""'Aspergillus'fumigatus"encounters"low"oxygen"microenvironments"during"infection."Adaptation"to"low"oxygen"microenvironments"is"essential"for"virulence"and"is"transcriptionally"regulated"by"the"sterol"regulatory"binding"proteins,"SrbA"and"SrbB."Additionally,"A.'fumigatus'grows"in"a"biofilm/like"structure"in'vivo'and"this"phenotype"requires"SrbA"for"full"biofilm"formation."To"identify"metabolic"genes"important"within"the"infection"environment"and"for"biofilm"formation,"we"identified"genes"with"increased"expression"in"low"oxygen"conditions."An"alanine"aminotransferase,"alaA,"was"found"to"be"one"of"the"few"amino"acid"metabolism"genes"that"was"both"directly"regulated"by"SrbA,"as"determined"by"ChiP/seq,"and"had"increased"mRNA"levels"in"low"oxygen."To"determine"how"alanine"metabolism"via"alaA"affects"fungal"biology"and"virulence,"alaA'null"mutants"were"generated"in"2"different"A.'fumigatus"strains,"CEA10"and"Af293."alaA'null"mutants"are"not"auxotrophic"and"were"capable"of"catabolizing"alanine"as"a"sole"carbon"or"nitrogen"source."Loss"of"alaA'resulted"in"a"moderate"decrease"in"growth"under"biofilm/forming"conditions"that"was"independent"of"oxygen"levels."This"growth"decrease"was"found"to"be"largely"attributable"to"a"germination"delay"that"is"rescued"by"supplementation"with"yeast"extract,"but"not"alanine"or"any"other"nitrogen"sources"tested."Intriguingly,"loss"of"alaA"results"in"decreased"expression"of"galactosaminogalactan"biosynthetic"genes"and"a"concomitant"decrease"in"adherence"to"abiotic"surfaces."

! 36"

Histological'data"from"a"murine"invasive"aspergillosis"model"suggests"that"alaA"is"involved"in"germination"and"growth"within"the"murine"lung,"and"qPCR"analysis"indicate"that"loss"of"alaA"results"in"a"reduced"fungal"burden."Taken"together,"these"data"reveal"that"alanine"metabolism"via"alaA'plays"a"significant"role"in"A.'fumigatus’"ability"to'germinate,"adhere"to"surfaces,"and"grow"within"the"murine"lung.""43."ProteomeLwide$analysis$of$cysteine$oxidation$in$Aspergillus*fumigatus$reveals$increased$levels$of$oxidatively$modified$thiol$groups$by$hypoxiaLinduced$reactive$oxygen$species""E."Shekhova1,"3,"T."Krüger1,"M."Stroe1,"J."Macheleidt1,"L."Ivanova1,"O."Kniemeyer1,2,"A.A."Brakhage1,2.""1)'Molecular'and'Applied'Microbiology,'Leibniz'Institute'for'Natural'Product'Research'and'Infection'Biology'(HKI),'Jena,'DE6'2)'Department'of'Microbiology'and'Molecular'Biology,'Institute'of'Microbiology,'Friedrich'Schiller'University,'Jena,'DE6'3)'Medical'Research'Council'Centre'for'Medical'Mycology'at'the'University'of'Aberdeen,'Institute'of'Medical'Sciences,'Foresterhill,'Aberdeen,'UK"""The"filamentous"fungus"Aspergillus'fumigatus"is"a"ubiquitously"distributed,"opportunistic"pathogen,"which"causes"a"variety"of"diseases"ranging"from"allergic"responses,"to"chronic,"but"also"life/threatening,"invasive"infections."At"the"site"of"infection,"the"lungs,"A.'fumigatus"is"often"subjected"to"declines"in"molecular"oxygen"concentrations."An"increasing"number"of"studies"have"demonstrated"that"an"exposure"of"fungal"cells"to"hypoxia"induces"increased"levels"of"ROS,"which"are"able"to"introduce"protein"thiol"modifications."These"may"either"impair"protein"functions"or"modulate"hypoxia"adaptation"by"activating"hypoxic"genes."To"learn"more"about"the"role"of"hypoxic"ROS,"we"determined"intracellular"ROS"production"in"A.'fumigatus"exposed"lo"low"oxygen"levels."Indeed,"the"oxidant/sensing"probe"DCFH/DA"revealed"increased"amounts"of"ROS"in"A.'fumigatus"mycelium"exposed"to"hypoxia."Moreover,"nuclear"accumulation"of"the"oxidative"stress"response"regulator"AfYap1"was"observed"under"hypoxia."The"iodo/TMT"labeling"of"redox/sensitive"cysteine"residues"of"A.'fumigatus"proteins"followed"by"LC/MS/MS"analysis"identified"protein"that"undergo"substantial"thiol"modifications"under"hypoxia."These"included"proteins"with"important"roles"in"maintaining"redox"balance"and"protein"folding,"such"as"the"thioredoxin"Asp"f"29"and"the"protein"disulphide"isomerase"PdiA."Our"data"also"showed"that"the"mitochondrial"respiratory"complex"IV"assembly"protein"Coa6"was"significantly"oxidized"by"hypoxic"ROS."Deletion"of"the"corresponding"gene"resulted"in"a"complete"absence"of"hypoxic"growth,"indicating"the"importance"of"complex"IV"during"adaptation"of"A.'fumigatus"to"low"oxygen"levels.""*44.$Fungal$Collagen$Dictates$Colony$and$Biofilm$Morphology$through$Filament$Interactions$to$impact$A.*fumigatus*Disease$Progression""Caitlin$H."Kowalski1,"Josh"D."Kerkaert1,"Jason"E."Stajich2,"Robert"A."Cramer1.""1)'Department'of'Microbiology'and'Immunology,'Geisel'School'of'Medicine'at'Dartmouth,'Hanover,'NH,'USA6'2)'Department'of'Plant'Pathology'&'Microbiology'and'Center'for'Plant'Cell'Biology,'University'of'CaliforniaQRiverside,'Riverside,'California,'USA"""Microbial"populations"arrange"in"intricately"organized"colonies"with"morphological"features"that"have"intrigued"and"perplexed"scientists"for"decades."Within"a"single"microbial"species,"heterogeneity"in"colony"morphotypes"is"appreciated,"but"in"only"several"cases"are"the"genetic"determinates"of"macroscopic"morphological"variation"identified."For"the"human"pathogen"Aspergillus'fumigatus"recent"work"has"highlighted"intra/species"heterogeneity"in"colony"morphotypes"during"low"oxygen"growth."For"A.'fumigatus,"the"ability"to"grow"in"low"oxygen"is"required"for"virulence,"suggesting"a"relationship"between"fungal"colony"morphology,"hypoxic"stress,"and"fungal"disease"progression."To"identify"the"genetic"factors"that"connect"these"aspects"of"A.'fumigatus"biology"we"utilized"an"experimental"evolution"approach"in"low"oxygen"that"generated"a"hyper/virulent"strain"that"is"more"fit"in"low"oxygen"and"that"has"an"altered"colony"morphology."Through"genome"sequencing,"we"have"characterized"a"hypothetical"protein,"hrmA'(hypoxia/responsive/morphology"factor"A),"that"is"necessary"for"the"morphology,"virulence,"and"hypoxia"fitness"of"the"evolved"strain."In"the"parent,"the"mutant"allele,"or"overexpression"of"the"parent"allele"generates"the"morphological"change,"and"this"change"requires"hrmA"to"localize"to"the"nucleus."RNA/sequencing"identified"a"cluster"of"genes"surrounding"the"hrmA"locus"that"are"regulated"by"hrmA,"one"of"which"is"a"fungal"collagen"like"protein,"cgnA,"a"class"of"proteins"completely"unstudied"in"human"pathogenic"fungi."cgnA"is"necessary"for"the"colony"morphology"and"virulence"of"the"evolved"strain."Interestingly,"cgnA"also"alters"the"interactions"of"fungal"filaments"in"biofilms,"where"its"expression"reduces"the"interconnectivity"of"the"mycelia."This"biofilm"phenotype"translates"in'vivo,"where"fungal"lesions"of"the"evolved"strain,"with"

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elevated"cgnA'mRNA"levels,"are"more"diffuse"and"result"in"increased"inflammation"and"pulmonary"damage,"contributing"to"altered"and"more"severe"disease"progression.""*45.$Genetic$engineering$of$the$Aspergillus*niger*regulatory$system$using$CRISPR/Cas9$technology""Roland$Sándor$Kun,"Ronald"P."de"Vries."Fungal'Physiology,'Westerdijk'Fungal'Biodiversity'Institute,'Utrecht,'Utrecht,'NL"""CRISPR/Cas9"technology"has"been"well/established"in"the"industrial"workhorse"Aspergillus'niger,"allowing"for"the"efficient"construction"of"strains"with"gene"deletions,"point"mutations"or"swapped"domains."We"applied"the"CRISPR/Cas9"system"in"Aspergillus'niger'to"study"the"regulatory"system"of"this"fungus"that"controls"plant"biomass"conversion.""Here"we"studied"the"hierarchy"of"four"transcriptional"activators"involved"in"this"process:"XlnR,"AraR,"ClrA"and"ClrB."We"constructed,"single,"double,"triple"and"quadruple"deletion"mutants"of"these"regulators"and"compared"them"to"the"wild"type"during"growth"on"various"plant"biomass"related"substrates."Protein"profiles,"enzyme"activities"and"gene"expression"was"analyzed"to"determine"the"relative"importance"of"each"regulator,"and"how"this"was"affected"by"the"composition"of"the"substrate"the"fungus"was"growing"on.""46.$Carbon$catabolite$repression$of$cellulase$genes$via$protein$kinase$A$pathway$in$Aspergillus*nidulans""E."Kunitake1,"Y."Li2,"R."Uchida2,"T."Nohara2,"K."Asano2,"A."Hattori2,"T."Kimura1,"K."Kanamaru2,"M."Kimura2,"T."Kobayashi2."1)'Graduate'School'of'Bioresources,'Mie'University,'Tsu,'Mie,'JP6'2)'Graduate'School'of'Bioagricultural'Sciences,'Nagoya'University,'Nagoya,'Aichi,'JP"""""CreA"orthologs"are"the"central"transcription"factors"that"mediate"carbon"catabolite"repression"(CCR)"of"genes"involved"in"carbon"utilization."However,"deletion"of"creA"in"A.'nidulans"did"not"lead"to"de/repression"of"cellulase"production"on"an"agar"plate"containing"carboxymethylcellulose"plus"glucose,"although"amylase"production"was"substantially"de/repressed"on"a"starch"plus"glucose"plate."Conversely,"deletion"of"pkaA"(protein"kinase"A),"which"was"reported"to"be"partially"involved"in"CCR,"led"to"significant"de/repression"of"cellulase"production,"while"amylase"production"was"not"de/repressed."As"to"the"three"G/protein"α"subunit"genes"(ganA,"ganB,"and"fadA),"the"ganB"deletant"behaved"similarly"to"the"pkaA"deletantS"production"of"cellulase"was"de/repressed"but"that"of"amylases"was"not."These"results"indicated"that"molecular"mechanisms"underlining"CCR"of"cellulase"genes"are"different"from"those"of"amylase"genes."In"submerged"culture"with"cellobiose"as"the"inducer"and"glucose"or"2/deoxyglucose"as"the"repressing"carbon"source,"both"creA"and"pkaA"single"deletions"led"to"partial"de/repression"of"cellulase"genes"based"on"RT/qPCR"analysis,"and"the"creA/pkaA"double"deletion"and"the"ganB"single"deletion"caused"the"highest"level"of"de/repression."""As"cellulase"gene"expression"in"A.'nidulans"is"more"or"less"repressed"by"any"carbon"sources"but"inducers"according"to"our"preliminary"results,"de/repression"from"CCR"by"the"above"deletions"were"examined"using"repressing"monosaccharides"other"than"glucose."Plate"assay"and"transcriptional"analysis"revealed"that"CCR"by"fructose"or"mannose"were"similar"to"that"by"glucoseS"CreA,"PkaA,"and"GanB"were"all"participated."By"contrast,"CCR"by"xylose"and"arabinose"was"predominantly"regulated"by"CreA"with"the"participation"of"GanB"but"not"PkaA."In"concluding"this"study,"CreA,"PkaA,"and"GanB"differently"function"in"CCR"depending"on"culture"conditions.""47.$The$VosA/VelBLactivated$McrA$governs$cellular$and$metabolic$integrity$in$Aspergillus*nidulans""Mi$Kyung$LEE1,"Ye"Eun"Son2,"Jae"Hyuk"Yu3.""1)'Biological'Resource'Center(BRC),'Korea'Research'Institute'of'Bioscience'and'Biotechnology,'Republic'of'Korea6'2)'School'of'Food'Science'and'Biotechnology,'Kyungpook'National'University,Republic'of'Korea6'3)'Department'of'Bacteriology,'University'of'Wisconsin,'Madison,'WI,'USA"""The"multicluster"regulator"A"(McrA)"C6"factor"acts"as"a"global"repressor"of"secondary"metabolism"in"diverse"filamentous"fungi."Here,"we"report"that"the"mcrA"gene"is"positively"regulated"by"the"velvet"regulators"VosA/VelB"during"conidiogenesis"and"McrA"governs"cellular"and"metabolic"integrity"of"conidia"in"Aspergillus'nidulans."An"mcrA"promoter"region"contains"a"VosA"responsive"element,"TGGCTTGGGCTGG,"and"its"mRNA"accumulation"in"conidia"requires"both"VosA"and"VelB."The"deletion"

! 38"

(Δ)"of"mcrA'results"in"a"reduced"number"of"conidia"and"decreased"mRNA"levels"of"the"key"asexual"developmental"activator"brlA."In"conidia,"ΔmcrA'causes"a"rapid"loss"of"conidia"viability,"which"is"likely"associated"with"the"lack"of"trehalose"accumulation"and"increased"the"ß/(1,3)/glucan"levels."In"supporting"its"repressive"role,"ΔmcrA"results"in"about"4/fold"increased"production"of"the"mycotoxin"sterigmatocystin"(ST)"and"an"unknown"metabolite"in"conidia."While"overexpression"(OE)"of"mcrA"causes"fluffy/autolytic"and"accelerated"cell"death"phenotypes,"ΔmcrA"does"not"suppress"the"developmental"defects"caused"by"the"lack"of"the"RGS"protein"FlbA."However,"ST"production"is"restored"in"the"ΔflbA"ΔmcrA"double"mutant,"and"OEmcrA"completely"blocked"the"production"of"ST"and"many"secondary"metabolites,"corroborating"the"global"repressive"role"of"McrA"in"secondary"metabolism."A"genetic"model"depicting"the"roles"of"McrA"in"governing"cellular"and"metabolic"integrity"is"presented.""*48."Reeingineering$of$Asperfuranone$pathway$to$yield$Sclerotiorin""P.W.$Lehman,"T.L."Lin,"C.W."Wang.""School'of'Pharmacy,'University'of'Southern'California,'Los'Angeles,'CA"""Sclerotiorin,"first"isolated"from"Penicillium'sclerotiorium'in"1940,"belongs"to"a"family"of"fungal"polyketides"called"Azaphilones."Azaphilones"are"characterized"by"their"diverse"biological"activity,"highly"oxygenated"pyranoquinone"bicyclic"core,"and"reactivity"with"nitrogen"to"form"vinylogous"γ/pyridones."Sclerotiorin"has"been"found"to"have"weak"antimicrobial,"anticancer,"aldose"reductase"inhibitory,"protein"kinase"G"inhibitory,"and"lipoxygenase"inhibitory"activity."Previously,"our"lab"elucidated"the"biosynthetic"pathway"for"the"azaphilone"asperfuranone."From"these"findings,"mutants"were"discovered"to"make"an"intermediate"of"asperfuranone"which"was"utilized"in"a"semi/synthetic"effort"to"yield"a"racemic"mixture"of"sclerotiorin."In"this"study,"we"reengineered"these"mutants’"asperfuranone"pathway"to"modify"the"intermediate"to"the"natural"product"sclerotiorin"via"transferases."The"new"mutants"produce"the"enantiomerically"pure"sclerotiorin"natural"product"along"with"several"intermediates.""49.$GenomeLwide$analyses$of$cadmium$stress$response$in$Aspergillus*nidulans""Tamás"Emri1,"Erzsébet"Orosz1,"Bianka"Huszti1,"Éva$Leiter1,"Károly"Antal2,"Jae/Hyuk"Yu3,"István"Pócsi1.""1)'Department'of'Biotechnology'and'Microbiology,'Faculty'of'Science'and'Technology,'University'of'Debrecen,'Debrecen,'Hungary6'2)'Department'of'Zoology,'Eszterházy'Károly'University,'Eger,'Hungary6'3)'Departments'of'Bacteriology'and'Genetics,'University'of'Wisconsin,'Madison,'WI,'USA"""The"b/Zip"transcription"factor"AtfA"is"a"principal"regulator"governing"oxidative"stress"responses"in"the"filamentous"fungus"Aspergillus'nidulans."To"understand"the"role"of"AtfA"in"response"to"the"heavy"metal"cadmium"(Cd),"we"carried"out"genome/wide"expression"analyses"using"the"60/mer"oligonucleotide/based"whole/genome"DNA"microarrays"and"exposing"wild/type"(WT)"and"aftA"null"mutant"strains"to"0.2"mM'of'CdCl2"(0.2"mM)."In"the"control"strain,"cadmium"stress"up/regulated"eight"genes"predicted"to"encode"FeS"cluster"proteins,"those"involved"in"biosynthesis"of"biotin,"NAD+"and"Lys,"and"in"the"formation"of"the"nitrate"reductase"complex,"as"well"as"antioxidant"enzymes,"including"CatB"catalase"and"GpxA"glutathione"peroxidase."These"suggest"cadmium"exposure"likely"inactivated"FeS"clusters"and"induced"oxidative"stress."As"cadmium"replaces"zinc"in"various"metalloproteins"including"DNA"repair"enzymes,"the"observed"down/regulation"of"DNA"repair"enzyme"genes"(altogether"21"genes)"may"protect"A.'nidulans'cells"from"the"deleterious"effects"of"caused"by"Cd2+/substituted"and,"hence,"improperly"functioning"enzymes"under"the"cadmium"stress."Furthermore,"the"extensive"down/regulation"of"DNA"repair"enzymes"in"WT"may"explain"the"widely"known"genotoxicity"of"this"acutely"harmful"heavy"metal."Among"the"potential"cadmium"detoxification"processes,"up/regulations"of"genes"associated"with"glutathione"synthesis"and"oxalate"formation"in"WT"were"recorded"meanwhile"genes"putatively"involved"in"CdS"formation"were"not"affected."Interestingly,"up/regulation"of"four"out"of"the"six"hydrophobin"genes"was"also"observed."The"absence"of"atfA"in"A.'nidulans"prevented"the"down/regulation"of"DNA"repair"genes"as"well"as"the"up/regulation"of"hydrophobin"genes,"which"were"in"line"with"the"increased"cadmium"sensitivity"of"this"mutant.""Acknowledgements:"The"research"was"financed"by"the"European"Union"and"the"European"Social"Fund"through"the"project"EFOP/3.6.1–16–2016/00022,"by"the"National"Research,"Development"and"Innovation"Office"(Hungary)"projects"K"112181"and"NN"125671,"and"by"the"Higher"Education"Institutional"Excellence"Program"of"the"Ministry"of"Human"Capacities"in"Hungary,"within"the"framework"of"the"Biotechnology"thematic"program"of"

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the"University"of"Debrecen."The"work"at"UW/Madison"was"support"by"the"National"Institute"of"Food"and"Agriculture,"United"States"Department"of"Agriculture,"Hatch"project"1009695.""*50.$Development$of$a$CRISPRLCas9Lbased$genetic$system$for$Chernobyl$isolated$Cladosporium*cladosporioides""Su$Jeung"Lim1,"Adriana"Blachowicz1,2,"Ngan"Pham1,"Jingyi"Wang1,"Tamas"Torok3,"Kasthuri"Venkateswaran2,"Clay"C.C."Wang1,4.""1)'Pharmacology'and'Pharmaceutical'Sciences,'University'of'Southern'California,'CA6'2)'Biotechnology'and'Planetary'Protection'Group,'Jet'Propulsion'Laboratory,'California'Institute'of'Technology,'Pasadena,'CA6'3)'Ecology'Department,'Lawrence'Berkeley'National'Laboratory,'Berkeley,'CA6'4)'Department'of'Chemistry,'Dornsife'College'of'Letters,'Arts'and'Sciences,'University'of'Southern'California,'Los'Angeles,'CA"""Fungi"are"a"major"source"of"secondary"metabolites"(SMs),"which"are"structurally"heterogeneous"low/molecular/mass,"bioactive"compounds."They"are"not"directly"involved"in"the"growth"of"the"organisms,"but"SMs"often"contribute"to"the"adaptation"of"fungi"to"different"environmental"stimuli"and"enable"the"survival"of"the"species."Cladosporium'cladosporioides'is"ubiquitous"saprophytic"fungus"isolated"from"wide"range"of"sources,"including"air,"soil"and"textiles."This"plant"pathogen"is"also"commonly"observed"in"indoor"mycobiome."C.'cladosporioides,'IMV"00236,'is"Chernobyl/isolated"strain"that"survived"and"adapted"to"high"irradiation"environment."Due"to"its"unique"isolation"site,"the"strain"was"selected"for"initial"SM"characterization.""We"started"with"testing"varying"growth"condition"to"optimize"production"of"SM"of"interest."LC/MS"analysis"revealed"production"of"four"major"compounds."Three"of"the"compounds"were"identified"via"HPLC"and"NMR"as"aromatic"polyketides,"the"remaining"fourth"compound"is"to"be"identified."Further,"we"will"use"CRISPR/Cas9"to"conduct"a"RNA/guided"mutagenesis"to"create"mutant"strains,"which"will"help"us"to"propose"the"biosynthetic"pathway"of"unidentified"compound."For"the"fungal"transformation,"we"will"use"the"plasmid"with"hygromycin"B"as"a"fungal"marker"kindly"provided"by"professor"Uffe"Mortensen’s"Lab."Upon"successful"gene"targeting"we"will"use"HPLC"to"purify"the"unknown"compound"and"its"intermediates,"and"NMR"to"verify"the"structure"and"further"elucidate"the"biosynthetic"pathway"of"an"unknown"compound."Discovering"of"the"unknown"pathway"may"lead"to"optimizing"production"of"this"compound.""*51.$Flexible$assembling$platform$built$up$for$heterologous$expression$of$secondary$metabolites$in$filamentous$fungi""Jun"Lin1,"Sandra"Hoefgen1,"Janis"Fricke2,"Maria"C."Stroe3,4,"Derek"J."Mattern3,"Johann"E."Kufs1,"Peter"Hortschansky3,"Axel"A."Brakhage3,4,"Dirk"Hoffmeister2,"Vito"Valiante1,5.""1)'Leibniz'research'cluster'–'Biobricks'of'Microbial'Natural'Product'Syntheses,'Leibniz'Institute'for'Natural'Product'Research'and'Infection'Biology'Q'Hans'Knöll'Institute,'Jena,'DE6'2)'Department'Pharmaceutical'Microbiology'at'the'HansQKnöllQInstitute,'Friedrich'Schiller'University,'Jena,'DE6'3)'Department'of'Molecular'and'Applied'Microbiology,'Leibniz'Institute'for'Natural'Product'Research'and'Infection'Biology'Q'Hans'Knöll'Institute,'Jena,'DE6'4)'Friedrich'Schiller'University,'Jena,'Germany6'5)'Department'of'General'Microbiology'and'Microbial'Genetics,'Institute'of'Microbiology,'Faculty'of'Biology'and'Pharmacy,'Friedrich'Schiller'University,'Jena,'DE"""In"natural"product"discovery,"the"heterologous"expression"of"entire"biosynthetic"pathways"can"be"used"to"uncover"compounds"produced"by"organisms"normally"recalcitrant"to"lab"conditions."Nevertheless,"laborious"and"time/consuming"routine"in"both"expression"vector"construction"and"mutant"strain"selection"inspired"scientists"to"develop"new"platforms"to"speed/up"the"discovery"process."We"designed"the"pV2A/T"vector"system"using"2A"peptide"sequences"from"picornaviruses"for"parallel"translation"of"multiple"genes"(transcribed"as"one"large"mRNA),"the"TEV"protease"to"cleave"off"2A"peptides"from"translated"proteins,"and"a"split"YFP"for"easy"screening"of"positive"transformants."This"system"enabled"a"facile"assembly"and"expression"of"gene"clusters"in"eukaryotic"hosts"as"polycistrons."After"ectopic"integration"into"the"genome"of"the"host"organism,"in"our"case"the"model"fungus"Aspergillus'nidulans,"positive"colonies"with"correct"and"complete"gene"expression"could"be"screened"by"nuclear"fluorescence."First,"we"proved"the"efficiency"of"this"methodology"by"expressing"four"genes"isolated"from"Aspergillus'

! 40"

calidoustus,"including"a"large"gene"coding"for"a"non/iterative"polyketide"synthase"([1]),"to"rewire"the"biosynthesis"of"austinoids"in"the"host"A.'nidulans."Second,"our"system"was"applied"for"the"production"of"psilocybin,"a"basidiomycete"psychotropic"compound"currently"in"phase"III"clinical"trials."By"transferring"the"psi"pathway"into"A.'nidulans,"we"could"show"the"production"of"110"mg"psilocybin"per"liter"culture."([2,"3])"The"developed"pipeline"allows"for"easier"identification"and"isolation"of"compounds"of"interest."We"are"currently"applying"this"method"to"explore"unknown"secondary"metabolic"gene"clusters"predicted"from"fungal"genomic"databases."[1]"Valiante,"V.,"Mattern,"D."J.,"Schuffler,"A.,"Horn,"F.,"Walther,"G.,"Scherlach,"K.,"Petzke,"L.,"Dickhaut,"J.,"Guthke,"R.,"Hertweck,"C.,"Nett,"M.,"Thines,"E."&"Brakhage,"A."A."ACS'Chem'Biol"2017,"12,"1227/1234."[2]"Fricke"J,"Blei"F,"Hoffmeister"D."Angew'Chem'Int'Ed"2017,"56,"12352/12355."[3]"Hoefgen"S,"Lin"J,"Fricke"J,"Stroe"MC,"Mattern"DJ,"Kufs"JE,"Hortschansky"P,"Brakhage"AA,"Hoffmeister"D,"Valiante"V."Metab'Eng"2018,"48,"44/51.""52.$Demystifying$the$NIH$grants$process$and$current$funding$opportunities$for$researchers.$Dona$Love.$$Mycology'Program'Officer,'BMB/DMID/NIAID,'National'Institutes'of'Health,'Bethesda'MD,'USA.''"Information"will"be"presented"by"the"NIAID"mycology"program"officer"on"grant"submission,"peer"review,"and"the"current"funding"opportunities"available"for"researchers.$"53.$A$integrated$view$of$calcium$signaling$in$response$to$azole$antifungal$stress$in$Aspergillus*fumigatus""L.$Lu,"J."Ye,"W.L."Du.""Nanjing'Normal'University,'Nanjing,'210023,'CN""'Aspergillus'fumigatus"has"evolved"numerous"signal/transduction"systems"to"sense"and"respond"to"stresses"to"survive"and"proliferate"in"harsh"environmental"conditions."Among"these"signal/transduction"systems,"the"Ca2+"signaling"pathway"is"one"of"most"important"response"systems,"which"has"been"verified"to"be"involved"in"stress"adaptation."Fungal"cytosolic"Ca2+"concentrations"are"maintained"at"extreme"low"level"to"function"as"the"second"messenger"while"high"concentrations"of"Ca2+"are"immediately"available"for"influx"from"outside"of"the"cell"into"the"cytoplasm"and"the"intracellular"Ca2+"stores."The"Ca2+"channels,"pumps,"exchangers,"and"antiporters"localized"in"the"membrane"as"well"as"Ca2+"sensors"with"their"signal"effectors"in"the"cytoplasm"are"highly"networked"to"regulate,"conduct"and"translate"various"signals."In"this"study,"we"demonstrate"that"the"activation"of"the"calcium"signaling"pathway"induced"by"azole"stress"contributes"to"the"survival"of"fungal"cells"during"drug"exposure."In"contrast,"the"blocking"of"the"calcium"signaling"pathway"or"the"loss"of"the"functional"calcium"signaling"pathway"components"in"A.'fumigatus"cause"enhanced"drug"susceptibilities."Thus,"effectively"utilizing"the"inhibitors"or"blockers"of"the"calcium"signaling"pathway"might""enhance"the"pharmacology"of"limited"antifungal"agents"for"azole"resistance"isolates"in"clinical"A.'fumigatus.""*54.$Unravelling$the$aromatic$metabolic$pathways$of$Aspergillus*niger""R.$Lubbers,"Adiphol"Dilokpimol,"Isabelle"Benoit,"Jaap"Visser,"Ronald"de"Vries.""Fungal'phsyiology,'Westerdijk'fungal'biodiversity'institute,'Utrecht,'Utrecht,'NL"""Aromatic"compounds"derived"from"lignin"are"of"great"interest"for"renewable"biotechnical"applications."These"compounds"can"serve"in"many"industries"e.g."as"biochemical"building"blocks"for"bioplastics"or"(bio)chemicals"or"as"antioxidants,"flavor"agents"or"preservatives"in"the"food"industry."For"most"microorganisms,"aromatic"compounds"have"a"toxic"effect"already"at"low"concentration"hence"these"compounds"are"quickly"degraded"to"non/"or"less"toxic"compounds."Many"possible"pathways"have"been"suggested"for"fungi,"however"these"observations"are"scattered"over"different"species"and"only"a"few"fungal"aromatic"metabolic"enzymes"are"characterized."In"order"to"better"explore"these"fungal"aromatic"metabolic"pathways"and"to"identify"novel"fungal"aromatic"metabolic"enzymes,"we"created"an"Aspergillus'niger"ferulic"acid"adaptive"evolution"mutant"(FA6)"by"growing"A.'niger"N402"on"incrementally"increasing"concentrations"of"ferulic"acid,"which"led"to"the"identification"of"several"pathways"and"their"genes,"which"were"confirmed"by"the"construction"of"deletion"strains"and"biochemical"characterization"of"the"pathway"enzymes."Highlights"from"this"study"will"be"presented."

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"55.$Alkaline$thermostable$recombinant$laccase$from$Coprinopsis*cinerea$uses$syringylLtype$phenols$as$mediators$and$depolymerizes$lignin""C."Lyra1,"M."Kähkönen1,"P."Nousiainen2,"J."Kontro2,"J."Sipilä2,"M.R."Mäkelä1,"K."Hildén1.""1)'Department'of'Microbiology,'University'of'Helsinki,'Helsinki,'FI6'2)'Department'of'Chemistry,'University'of'Helsinki,'Helsinki,'FI"""Laccases"are"multi/copper"oxidases"that"use"molecular"oxygen"as"the"electron"acceptor"to"oxidize"phenolic"and"indirectly"also"non/phenolic"substrates"by"radicals."Due"to"their"eco/friendliness"and"broad"substrate"specificity,"laccases"span"a"wide"range"of"biotechnological"applications."We"have"heterologously"expressed"laccase"from"the"litter/degrading"basidiomycete"Coprinopsis'cinerea"(CcLcc9)"in"methylotrophic"Pichia'pastoris"yeast."The"recombinant"CcLcc9"oxidized"2,6/dimethoxyphenol"in"the"neutral"pH"range"and"showed"thermostability."The"oxidation"of"non/phenolic"compounds"by"CcLcc9"was"tested"with"low"molecular"weight"mediators,"such"as"syringyl/type"plant"phenolics,"which"have"shown"potential"as"natural"co/oxidants"for"lignocellulosic"materials."The"results"show"that"syringyl"nitrile"and"methyl"syringate"efficiently"mediate"the"oxidation"of"veratryl"alcohol"to"veratraldehyde."In"addition,"CcLcc9"is"able"to"depolymerize"biorefinery"hardwood"lignin"in"the"presence"of"these"mediators"analyzed"by"Gel"Permeation"Chromatography."The"depolymerization"of"lignin"by"Cclcc9"enzyme"harbor"a"great"potential"for"the"sustainable"processes"of"circular"economy"and"modern"biorefineries.""56.$Fungal$proteins$with$antiLbacterial$properties$secreted$during$infection""Silke$Machata1,"Roland"Lehmann2,"Hortense"Slevogt2,"Axel"Brakhage3,"Ilse"Jacobsen1,4.""1)'Microbial'Immunology,'Leibniz'Institute'for'Natural'Product'Research'and'Infection'BiologyQ'Hans'Knöll'Institute,'Jena,'Germany6'2)'Septomics'Research'Center,'Jena'University'Hospital,'Jena,'Germany6'3)'Molecular'and'Applied'Microbiology,'Leibniz'Institute'for'Natural'Product'Research'and'Infection'BiologyQ'Hans'Knöll'Institute,'Jena,'Germany6'4)'FriedrichQSchillerQUniversity,'Jena,'Germany""'Aspergillus'fumigatus"is"a"saprophytic"mold"that"is"responsible"for"various"severe"pulmonary"fungal"diseases"including"invasive"aspergillosis"(IA)."Various"studies"have"focused"their"efforts"on"finding"fungal"secreted"antigens"to"identify"novel"biomarkers"and"vaccine"candidates"using"human"patient"serum"against"fungal"protein"extracts."However,"these"attempts"are"limited"to"the"detection"of"fungal"antigens"that"are"produced"in"liquid"growth"media"and"are"not"suitable"for"the"identification"of"proteins"that"are"induced"only"during"infection."In"the"present"study,"we"took"advantage"of"a"mouse"infection"model"for"IA"to"analyze"the"secretome"of'A.'fumigatus"in'vivo"by"mass"spectrometry"allowing"the"detection"of"infection/specific"fungal"proteins"in"bronchoalveolar"lavage"samples."Among"the"ten"most"abundant"secreted"fungal"proteins"were"three"proteins"that"have"not"been"previously"described"and"share"motifs"commonly"known"in"bacteria."They"contain"NlpC/p60"domains"and/or"bacterial"SH3"domains"that"are"often"found"in"bacterial"murein"peptidases."Recombinant"His/fusion"proteins"were"generated"for"two"of"the"proteins"and"their"lytic"activity"was"determined"against"Gram"positive"and"Gram"negative"bacteria.""We"found"that"both"proteins"are"able"to"cleave"Gram"positive"peptidoglycan"with"distinct"substrate"specificity."For"at"least"one"of"the"proteins""we"also"demonstrate"bacteriolytic"activity"against"various"Gram"positive"bacteria."The"regulatory"factors"leading"to"protein"expression"and"secretion"still"remain"to"be"elucidated."The"proteins"characterized"in"this"study"are"not"commonly"produced"under"in'vitro"conditions."However,"hypoxia"seems"to"play"a"role"for"the"induction"of"gene"expression"of"two"of"the"proteins."Furthermore,"the"role"of"common"transcriptional"regulators"in"the"expression"of"these"proteins"is"currently"investigated"by"analyzing"various'A.'fumigatus"mutant"strains."Our"current"study"presents"a"new"group"of"proteins"of"A.'fumigatus"with"anti/bacterial"activity."While"the"production"of"secondary"metabolites"in"Aspergillus'acting"against"the"host"and"other"organisms"has"been"investigated"in"great"detail,"there"have"been"no"descriptions"on"fungal"proteins"acting"against"bacteria."The"detailed"analysis"of"newly"generated"fungal"mutants"overexpressing"our"genes"of"interest"will"help"us"to"further"understand"the"biological"role"of"these"proteins"for"the"lifestyle"of"A.'fumigatus"and"its"interaction"with"bacteria"in"microbial"communities.""$ $

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57.$Contribution$of$maize$polyamine$and$amino$acid$metabolism$towards$resistance$against$Aspergillus*flavus$infection$and$aflatoxin$production.""R."Majumdar1,"R."Minocha2,"M."Lebar1,"K."Rajasekaran1,"S."Long2,"C."Carter/Wientjes1,"S."Minocha3,"J."Cary1.""1)'USDAQARSQSRRC,'New'Orleans,'LA6'2)'USDA'Forest'Service,'Northern'Research'Station,'Durham,'NH6'3)'Department'of'Biological'Sciences,'University'of'New'Hampshire,'Durham,'NH"""Polyamines"(PAs)"are"ubiquitous"polycations"found"in"plants"and"other"organisms"that"are"essential"for"growth,"development,"and"resistance"against"abiotic"and"biotic"stresses."The"role"of"PAs"in"plant"disease"resistance"depends"on"the"relative"abundance"of"higher"PAs"[spermidine"(Spd),"spermine"(Spm)]"vs."the"diamine"putrescine"(Put)"and"PA"catabolism."With"respect"to"the"pathogen,"PAs"are"required"to"achieve"successful"pathogenesis"of"the"host."Maize"is"an"important"food"and"feed"crop,"which"is"highly"susceptible"to"A.'flavus"infection."Upon"infection,"the"fungus"produces"carcinogenic"aflatoxins"and"numerous"other"toxic"secondary"metabolites"that"adversely"affect"human"health"and"crop"value"worldwide."To"evaluate"the"role"of"PAs"in"aflatoxin"resistance"in"maize,"in'vitro"kernel"infection"assays"were"performed"using"maize"lines"that"are"susceptible"(SC212)"or"resistant"(TZAR102,"MI82)"to"aflatoxin"contamination."Results"indicated"significant"induction"of"both"PA"biosynthetic"and"catabolic"genes"upon"A.'flavus"infection."As"compared"to"the"susceptible"line,"the"resistant"maize"lines"showed"higher"basal"expression"of"PA"metabolism"genes"in"mock/inoculated"kernels"that"increased"upon"fungal"infection."In"general,"increased"biosynthesis"and"conversion"of"Put"to"Spd"and"Spm"along"with"their"increased"catabolism"was"evident"in"the"resistant"lines"vs."the"susceptible"line"SC212."Higher"accumulation"of"amino"acids"such"as"glutamate"(Glu),"glutamine"(Gln),"and"γ/aminobutyric"acid"(GABA)"correlated"with"a"significant"increase"in"aflatoxin"production"in"SC212."The"resistant"lines"showed"a"significant"reduction"in"fungal"load"and"aflatoxin"production"compared"to"the"susceptible"line."The"data"presented"here"demonstrate"an"important"role"of"PA"metabolism"in"the"resistance"of"maize"to"A.'flavus"colonization"and"aflatoxin"contamination."These"results"provide"future"directions"for"the"manipulation"of"PA"metabolism"and/or"the"use"of"Spd/Spm"as"biomarkers"to"screen"maize"genotypes"for"improved"aflatoxin"resistance"and"overall"stress"tolerance.""*58.$A$novel$gelsolinLlike$protein$required$for$proper$septum$formation$by$regulating$contractile$actin$ring$formation""M."Mamun1,"T."Katayama1,2,"J."Maruyama1,2.""1)'Department'of'Biotechnology,'The'University'of'Tokyo,'Tokyo,'JP6'2)'CRIIM,'The'University'of'Tokyo"""Septum"divides"the"hyphae"into"individual"compartments"in"filamentous"fungi,"and"cell/to/cell"communication"depends"on"the"septal"pore,"which"allows"the"exchange"of"cytoplasmic"constituents"between"adjacent"cells."Septum"is"formed"by"constriction"of"contractile"actin"ring"(CAR),"and"CAR"formation"involves"regulatory"components"such"as"formin"conserved"both"in"yeast"and"filamentous"fungi."However,"the"specific"machineries"regulating"actin"dynamics"in"septum"formation"is"unknown"in"filamentous"fungi."In"this"study"using"BLAST"and"gene"ontology/based"selection"for"Pezizomycotina/specific"actin"regulators,"we"found"an"uncharacterized"gelsolin/like"protein,"GelA."Gelsolin"is"known"as"an"actin/modulating"protein"with"severing"activity."GelA"has"three"gelsolin"domains"and"a"domain"of"unknown"function"(Pfam:"DUF4045)."Phylogenetic"analysis"with"gelsolin"superfamily"members"exhibited"that"GelA"forms"an"independent"clade"consisting"of"Pezizomycotina"orthologs."We"generated"a"gelA"deletion"strain"in"Aspergillus'oryzae,"one"of"the"filamentous"fungi"extensively"characterized"with"fungal"septal"pore"regulation,"resulting"in"moderate"growth"defect."In"particular,"gelA"deletion"caused"unusually"larger"interval"of"septa"in"all"part"of"the"hypahe"and"abnormal"septum"morphology."To"evaluate"the"septal"pore"function,"intercellular"communication"via"the"septal"pore"was"analyzed"by"tracking"cell/to/cell"translocation"of"Dendra2,"a"green/to/red"photoconvertible"fluorescent"protein."Upon"cold"stress"in"which"Dendra2"translocation"via"the"septal"pore"was"ceased"in"wild"type"due"to"septal"pore"closure,"the"protein"was"still"translocated"to"the"next"cell"in"ΔgelA."This"result"indicated"that"the"regulation"of"septal"pore"closure"was"defective"in"ΔgelA."When"observing"actin"dynamics"with"actin"marker"Lifeact/GFP"during"septum"formation,"ΔgelA"showed"improper"CAR"formation"with"actin"accumulation"only"at"one"cortical"site"or"abnormal"orientation"of"CAR,"finally"not"forming"a"septum."Localization"of"GelA"was"found"at"the"site"of"ongoing"septum"formation"

! 43"

similarly"to"CAR"formation."Finally,"in"this"study"we"elucidated"a"novel"actin"modulating"component,"which"specifically"participates"in"septum"formation"by"regulating"CAR"formation"in"filamentous"fungi.""59.$Aspergillus*fumigatus$RasGEFLmediated$invasive$growth""A.$MartinLVicente,"Q."Al"Abdallah,"A."Oliveira"Souza,"J."Fortwendel.""Clinical'Pharmacy'and'Translational'Science,'University'of'Tennessee'Health'Science'Center,'Memphis,'TN"""The"Aspergillus'fumigatus"genome"encodes"four"Ras"Guanine"Exchange"Factors"(RasGEFs)"to"regulate"Ras"protein"activity,"two"of"which"belong"to"the"fungus/specific"SH3/class."These"two"proteins,"GefA"and"GefB,"harbor"putative"SH3"domains"at"their"N/termini"that"likely"contribute"to"protein/protein"interactions."Whereas"GefA"is"homologous"to"the"Saccharomyces'cerevisiae"Cdc25p"RasGEF,"GefB"is"homologous"to"Bud5p,"the"RasGEF"for"Rsr1p,"a"Ras/like"GTPase"essential"for"bud"site"selection."Recently,"we"reported"that"loss"of"both"A.'fumigatus'gefA"and"gefB"(ΔgefA/ΔgefB)"resulted"in"inviability."Therefore,"we"hypothesized"that"coordinated"regulation"of"Ras/"and"Rsr1/signaling"is"essential"to"viability"in"A."fumigatus."Previous"studies"in"our"lab"revealed"that"RasA"alone"is"not"essential"for"A.'fumigatus"viability,"though"its"deletion"causes"impaired"hyphal"morphogenesis"and"reduced"virulence."To"test"the"requirement"for"Rsr1"in"A.'fumigatus"growth"and"development,"we"first"generated"a"single/deletion"mutant"lacking"the"Rsr1"homolog."Phenotypic"characterization"of"the"Δrsr1"strain"revealed"only"a"moderate"reduction"in"radial"growth"rates"with"no"apparent"abnormalities"in"morphogenesis,"suggesting"that"Rsr1/mediated"signaling"alone"was"not"essential."To"examine"the"importance"of"coordinated"regulation"of"both"pathways"for"viability,"we"next"generated"a"double"mutant"by"replacing"the"rasA"endogenous"promoter"with"the"tetracycline/inducible"promoter"(pTetOn)"in"the"Δrsr1"background"(Δrsr1/pTetOn/rasA)."Under"inducing"conditions,"this"mutant"displayed"growth"reductions"similar"to"the"Δrsr1"strain."In"contrast,"under"non/inducing"conditions,"the"double"mutant"phenotype"mimicked"that"of"ΔrasA,"suggesting"that"SH3/class"RasGEF"morphogenetic"signaling"is"at"least"partially"RasA/"and"Rsr1/independent."We"next"sought"to"determine"if"the"SH3"protein"domain"was"required"for"GefA"function"and"potentially"contribute"to"the"synthetic"lethality"of"the"ΔgefA/ΔgefB"mutant."Truncation"of"the"GefA"SH3"domain"resulted"in"a"strain"that"phenocopied"the"ΔgefA"single"mutant,"suggesting"that"SH3"domain/dependent"interactions"are"important"for"GefA"function"in"A.'fumigatus."Future"work"will"define"the"importance"of"the"GefB"SH3"domain"and"identify"SH3/dependent"RasGEF"interactions"in"A.'fumigatus.""60.$Am$I$my$brother’s$keeper?$Revealing$the$pathogenomic$differences$between$Aspergillus*fumigatus*and$a$(mostly)$innocent$relative""M.E."Mead1,"S.L."Knowles2,"H.A."Raja2,"S.R."Beattie3,"C.H."Kowalski3,"J.L."Steenwyk1,"L.P."Silva4,"J."Chiaratto4,"L.N.A."Ries4,"G.H."Goldman4,"R.A."Cramer3,"N.H."Oberlies2,"A."Rokas1.""1)'Biological'Sciences,'Vanderbilt'University,'Nashville,'Tennessee6'2)'Department'of'Chemistry'and'Biochemistry,'University'of'North'Carolina'at'Greensboro,'Greensboro,'North'Carolina6'3)'Department'of'Microbiology'and'Immunology,'Geisel'School'of'Medicine'at'Dartmouth,'Hanover,'New'Hampshire6'4)'Faculdade'de'Ciencias'Farmacêuticas'de'Ribeirão'Preto,'Universidade'de'São'Paulo,'Brazil""'Aspergillus'fischeri"is"a"close"evolutionary"relative"of"the"major"cause"of"invasive"mold"infections,"Aspergillus'fumigatus."In"contrast"to"A.'fumigatus,"A.'fischeri"rarely"causes"invasive"disease"even"though"it"is"commonly"found"in"diverse"environments,"including"hospitals."Thus,"it"remains"unclear"why"A.'fischeri"causes"less"human"disease."To"begin"addressing"this"question,"we"characterized"the"pathogenic,"genomic,"and"secondary"metabolic"similarities"and"differences"between"A.'fischeri"and"A.'fumigatus."We"observed"multiple"differences"between"the"two"species"for"phenotypes"related"to"pathogenesis,"including"that"A.'fischeri"is"less"virulent"than"A.'fumigatus"in"multiple"animal"models"of"invasive"disease,"exhibits"slower"growth"in"low"oxygen"environments,"and"is"more"sensitive"to"oxidative"stress."In"contrast,"the"genomic"similarity"between"the"two"species"is"highS"~90%"of"the"A.'fumigatus"proteome"is"conserved"in"A.'fischeri,"including"48/49"genes"known"to"be"involved"in"A.'fumigatus"virulence."However,"only"10/33"A.'fumigatus'biosynthetic"gene"clusters"(BGCs)"likely"involved"in"the"production"of"secondary"metabolites"are"conserved"in"A.'fischeri'and"only"13/48"A.'fischeri'BGCs"are"conserved"in"A.'fumigatus."Detailed"chemical"characterization"of"A.'fischeri"cultures"grown"on"multiple"substrates"identified"multiple"secondary"metabolites,"including"two"new"compounds"and"one"never"before"isolated"as"a"natural"product."Additionally,"an"A.'fischeri"deletion"mutant"of"laeA,"a"master"regulator"of"secondary"metabolism,"produced"

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fewer"secondary"metabolites"and"in"lower"quantities,"suggesting"that"regulation"of"secondary"metabolism"is"at"least"partially"conserved"between"the"two"species."These"results"suggest"that"the"non/pathogenic"A.'fischeri"possesses"many"of"the"genes"important"for"A.'fumigatus"pathogenicity"but"is"divergent"with"respect"to"its"ability"to"thrive"under"host/relevant"conditions"and"its"secondary"metabolism."Ongoing"work"is"expanding"these"studies"to"other"species"closely"related"to"A.'fumigatus"as"well"as"other"aspects"of"genome"evolution"that"influence"the"severity"of"human"disease"caused"by"some"fungi."Leveraging"the"diversity"of"species"closely"related"to"A.'fumigatus"is"giving"us"a"better"understanding"of"the"nature"and"evolution"of"human"fungal"pathogenesis"as"the"appreciation"for"the"health"burden"caused"by"fungi"continues"to"grow.""*61.$Septins$and$stress$response$signaling$pathways$in$A.*nidulans""Alexander$Mela,"Michelle"Momany.""Plant'Biology,'University'of'Georgia,'Athens,'GA"""The"Cell"Wall"Integrity"(CWI)"and"High"Osmolarity"Glycerol"(HOG)"MAPK"signaling"pathways"are"integral"for"remediation"of"extracellular"stress"in"fungi.""These"stress"response"signaling"pathways"have"been"shown"to"be"responsible"for"upregulation"of"environmental"stress"response"genes"and"consequently"compensatory"alterations"to"the"cell"wall"and"plasma"membrane"architecture"of"fungi.""Septins"are"small"GTPases"in"the"cytoskeleton,"which"among"many"roles"in"the"cell,"act"as"molecular"scaffolds"to"facilitate"recruitment"of"chitin"and"beta/glucan"synthases"at"the"mother"bud"neck"and"septum"during"growth"and"cell"division"in"yeast"and"filamentous"fungi."""Septins"have"also"been"shown"to"colocalize"with"sterol/rich"regions"and"axes"of"polarity,"which"have"been"implicated"in"organizing"specific"plasma"membrane"domains,"such"as"sterol/rich"lipid"rafts.""Here"we"provide"evidence"supporting"the"hypothesis"that"septins"serve"a"role(s)"in"coordinating"one"or"more"MAPK"signaling"pathways"involved"in"sensing"the"extracellular"environments"and"modulating"the"cell"wall"and"plasma"membrane"in"response"to"external"stimuli"within"the"filamentous"fungus"Aspergillus'nidulans.""*62.$Molecular$mass$of$αL1,3Lglucan$affects$the$degree$of$hyphal$aggregation$and$its$localization$in$Aspergillus$nidulans""K."Miyazawa1,"A."Yoshimi2,"T."Yamashita1,"A."Koizumi1,"S."Yano3,"K."Abe1,2.""1)'Grad.'Sch.'Agric.'Sci.,'Tohoku'Univ.,'Sendai,'JP6'2)'NICHe,'Tohoku'Univ.,'Sendai,'JP6'3)'Grad.'Sch.'Eng.,'Yamagata'Univ.,'Yonezawa,'JP"""α/1,3/Glucan"is"one"of"the"main"polysaccharide"in"the"cell"wall"of"filamentous"fungi,"and"is"known"as"an"aggregation"factor"of"hyphae."The"model"filamentous"fungus"Aspergillus'nidulans"has"two"α/1,3/glucan"synthases"genes,"agsA"and"agsB."We"previously"reported"that"AgsB"is"a"primary"α/1,3/glucan"synthase"in"vegetative"hyphae,"but"the"function"of"AgsA"remains"unclear."Here"we"constructed"strains"overexpressing"agsA"(agsAOE)"or"agsB"(agsBOE)"for"clarifying"the"role"of"α/1,3/glucan"in"hyphal"aggregation."In"liquid"culture,"the"hyphae"of"the"wild/type"and"agsBOE"strain"formed"tightly"aggregated"pellet,"whereas"the"agsAOE"aggregated"loosely."To"reveal"these"phenotypic"difference"between"the"overexpression"strains,"we"analyzed"the"chemical"structure"of"α/1,3/glucan"from"both"strains."Interestingly"peak"molecular"mass"of"α/1,3/glucan"from"the"agsAOE"strain"(1,480"±"80"kDa)"was"four"times"than"that"from"agsBOE"strains"(372"±"47"kDa)."We"also"analyzed"localization"of"α/1,3/glucan"in"the"cell"wall"by"fluorescent"labelling"of"α/1,3/glucan"with"α/1,3/glucan/binding"domain/fused"GFP"(AGBD/GFP)."α/1,3/Glucan"of"the"agsBOE"cells"was"clearly"labelled"with"AGBD/GFP,"whereas"only"weakly"labelled"in"the"agsAOE"cells."In"addition,"β/1,3/glucanase"treated"agsAOE"cells"were"clearly"labelled"with"AGBD/GFP."These"results"suggest"that"α/1,3/glucan"was"localized"outermost"layer"in"the"agsBOE"strain,"whereas"β/1,3/glucan"covered"most"of"α/1,3/glucan"in"the"agsAOE"strain."Together"the"degree"of"hyphal"aggregation"seemed"to"be"controlled"not"only"by"the"amount"of"α/1,3/glucan"but"also"by"the"molecular"mass"and"localization"of"α/1,3/glucan"in"the"cell"wall."The"different"distribution"of"α/1,3/glucan"in"the"cell"wall"of"the"agsAOE"and"agsBOE"strains"may"be"caused"by"the"difference"in"the"physicochemical"properties"of"α/1,3/glucans"synthesized"by"AgsA"and"AgsB."In"general,"polysaccharides"of"which"molecular"mass"is"lager"are"usually"less"soluble"than"the"smaller"one."Therefore,"the"α/1,3/glucan"having"larger"molecular"mass"might"be"localized"in"the"inner"layer,"whereas"smaller"one"in"the"outer"layer"of"the"cell"wall."Here,"we"discuss"the"relationship"between"molecular"mass"and"distribution"of"α/1,3/glucan"in"the"cell"wall.""

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63."Dissecting$the$role$of$catalytic$residues$of$Human$peroxiredoxins$using$yeast$model""A.$Mohammad,"A.K."Saini.""Faculty'of'biotechnology'and'applied'sciences,'Shoolini'university'of'biotechnology'and'management'sciences,'Solan,'IN""$Statement$of$purpose:"Peroxiredoxin"class"of"proteins"play"a"critical"role"in"scavenging"the"harmful"free"radicals"via"their"peroxidase"activity"mediated"by"residues"which"are"proposed"to"stabilize"the"radicals"in"catalytic"pocket."The"direct"role"of"these"residues"is"still"not"clear."Methods:"We"used"yeast"model"to"understand"the"role"of"catalytic"residues"Arg128,"Pro45"and"Thr49"of"human"peroxiredoxin"I"(hPrxI)."For"this"we"created"their"Ala"substitution"and"expressed"the"mutant"alleles"through"TSA1"promoter"in"yeast"strain"lacking"the"hPrxI"homologues,"tsa1"and"tsa2"genes"(tsa1tsa2del)."Growth"in"the"presence"of"redox"stressors"was"analyzed."Dimerization"of"hPrx1"due"to"disulfide"bond"formation,"an"indicator"of"peroxidase"activity,"was"also"analyzed"in"vivo"using"immunoblotting."Results:$As"expected"hPrx1"supported"the"growth"of"tsa1tsa2del"strain"in"the"presence"of"SNP"(sodium"nitroprusside)"and"H2O2."We"found"that"Ala"substitution"of"Arg128,"Pro45"and"Thr49"compromised"the"hPrx1"function"to"support"growth"under"nitrosative"stress."The"cells"expressing"these"mutants"were"unable"to"grow"in"the"presence"of"SNP"as"compared"to"WT/hPrxI."Using"immunoblotting,"we"found"that"these"mutants"were"unable"to"form"dimer"as"compared"to"WT/hPrxI"strengthening"the"canonical"pathway"where"dimerization"is"required"for"quenching"the"stressors."We"also"checked"the"growth"of"these"mutants"in"the"presence"of"H2O2"stress."Surprisingly,"under"oxidative"stress"these"mutants"were"able"to"support"the"growth"to"the"level"of"WT/hPrxI,"indicating"their"redundancy"in"supporting"the"growth"even"after"losing"the"peroxidase"function."We"reasoned"this"might"be"due"to"the"ability"of"these"mutants"to"boost"the"expression"of"other"antioxidant"genes"like"SOD"or"GPx"in"the"absence"of"dimeric"hPrxI."Conclusion:$Catalytic"residue"other"than"peroxidatic"and"resolving"cystines"are"essential"to"overcome"nitrosative"stress"but"are"redundant"to"support"growth"under"oxidative"stress."This"indicates"quenching"mechanism"for"both"the"stressors"could"be"different.""64.$Conserved$septin$residues$are$located$in$interfaces$between$monomers""M."Momany1,"B."Auxier2,3,"J."Dee2,"M."Berbee2.""1)'Fungal'Biology'Group'and'Plant'Biology'Department,'University'of'Georgia,'Athens,'GA,'USA'6'2)'Department'of'Botany,'University'of'British'Columbia,'Vancouver,'Canada6'3)'Laboratory'of'Genetics,'Wageningen'University,'The'Netherlands"""Septins"are"cytoskeletal"elements"that"play"many"roles"in"the"cell,"including"determining"division"planes"and"diffusion"barriers."Most"animals"and"fungi"contain"multiple"septin"paralogs"that"fall"into"seven"previously"defined"clades."Individual"septin"paralogs"interact"to"form"nonpolar"heteropolymeric"rods"that"assemble"into"higher"order"structures."The"rules"governing"septin"paralog"polymerization"and"assembly"are"not"well/understood."We"analyzed"septins"from"ophisthokonts,"the"clade"that"unites"fungi"and"animals,"with"a"focus"on"newly"available"genome"sequences."We"found"that"septins"diverged"earlier"than"previously"realized,"with"early/diverging"lineages"containing"septins"representing"at"least"two,"and"possibly"four,"of"the"seven"clades."Using"protein"crystal"structures"to"model"septin"interactions,"we"found"that"previously"identified"conserved"sequence"motifs"(S1,"S2,"S3,"S4,"and"SUE)"were"all"located"in"interface"regions"between"monomers."This"localization"along"interacting"surfaces"suggests"that"these"amino"acids"are"conserved"because"they"are"important"for"the"monomer/monomer"interactions"that"drive"assembly"of"septin"heteropolymers"and"higher"order"structures.""65.$AtrR$is$a$critical$determinant$of$azole$resistance$in$Aspergillus*fumigatus""W.$Scott$MoyeLRowley.$$Dept'Molec'Physiology/Biophy,'Univ'Iowa,'Iowa'City,'IA"""Fungal"infections"caused"by"azole"resistant"isolates"of"Aspergillus"fumigatus"have"an"alarmingly"high"mortality.""Azole"resistance"in"A.'fumigatus"is"most"typically"associated"with"a"compound"mutation"in"the"cyp51A"gene"that"encodes"the"enzymatic"target"of"this"class"of"drugs.""This"unusual"mutant"allele"consists"of"a"34"bp"duplication"in"the"promoter"regions"(TR34)"linked"with"a"single"amino"acid"substitution"mutation"in"the"coding"sequence"(L98H).""We"are"studying"a"Zn2Cys6"DNA/binding"domain/containing"

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transcription"factor"called"AtrR"that"is"a"key"regulator"of"azole"resistance"in"general"and"cyp51A"in"particular.""Previous"work"demonstrated"that"AtrR"also"controlled"expression"of"an"ATP/binding"cassette"transporter/encoding"gene"called"abcG1S"with"expression"of"this"gene"also"required"for"normal"azole"resistance.""We"have"carried"out"chromatin"immunoprecipitation"coupled"with"Next"Generation"Sequencing"(ChIP/seq)"to"map"the"genomic"target"sites"for"AtrR"that"we"have"designated"AtrR"response"elements"or"ATREs.""GO"term"analysis"of"ATRE/containing"genes"indicated"that"the"main"enriched"class"of"gene"encoded"other"sequence/specific"transcription"factors.""We"have"mapped"ATREs"upstream"of"abcG1"as"well"as"cyp51A.""Importantly,"the"ATRE"in"cyp51A"lies"within"the"34"bp"region"that"is"repeated"in"azole"resistant"isolates.""Gene"disruption"experiments"have"also"confirmed"that"loss"of"the"atrR"gene"from"clinical"isolates"containing"the"TR34"L98H"allele"of"cyp51A"eliminates"azole"resistance.""We"also"describe"two"different"hyperactive"alleles"of"atrR"that"provide"insight"into"the"likely"regulatory"mechanisms"controlling"the"activity"of"this"transcription"factor.""Our"data"implicate"AtrR"as"an"important"central"coordinator"of"azole"resistance"and"other"phenotypes"likely"through"its"regulation"of"expression"of"a"range"of"different"transcription"factors.""66."Enhanced$production$of$antimicrobial$polyketides,$violaceols$in$Aspergillus$nidulans$by$coLculture$with$Aspergillus$fumigatus""A.$Ninomiya,"S."Urayama,"D."Hagiwara.""Faculty'of'Life'and'Environmental'Sciences,'University'of'Tsukuba,'Ibaraki,'JP""""""""Fungal"genome"contains"a"large"number"of"cryptic"biosynthetic"gene"clusters"for"new"compounds"which"are"not"expressed"under"standard"laboratory"conditions."For"expression"of"cryptic"gene"clusters,"several"strategies"have"been"employed"by"using"chemical"elicitors"or"heterologous"gene"expression."Recently,"co/cultivation"has"been"the"method"of"choice"to"obtain"the"biosynthetic"products"of"cryptic"genes."Thus,"a"close"study"on"the"molecular"mechanisms"underlying"the"activation"of"biosynthetic"gene"clusters"by"co/cultivation"is"necessary"for"development"of"technology"to"exploit"potential"of"fungal"secondary"metabolism.""""""By"using"several"Aspergillus"species,"we"discovered"a"fungal/fungal"interaction"that"stimulates"biosynthesis"of"antimicrobial"polyketides,"violaceols"in"Aspergillus"nidulans."The"production"of"violaceols"was"promoted"by"co/cultivation"with"a"pathogenic"fungus,"Aspergillus"fumigatus."Violaceols"are"known"to"act"against"several"microorganisms"including"Aspergillus"niger,"Candida"stellatoides,"and"Bacillus"subtilis.1"The"promotion"of"violaceols"production"was"not"observed"in"co/culture"of"A."nidulans"and"A."fumigatus"separated"by"a"dialysis"membrane,"which"suggested"that"the"contact"with"A."fumigatus"is"necessary"for"the"stimulation"of"violaceols"biosynthesis"in"A."nidulans.""""""We"also"found"the"activation"of"several"biosynthetic"gene"clusters"encoded"in"the"genome"of"the"two"fungi"for"secondary"metabolites"including"the"orsellinic"acid"gene"cluster"(ors"cluster)"by"the"fungal/fungal"interaction"mentioned"above"by"RNA/seq"analysis."The"ors"cluster"is"involved"in"several"bioactive"compounds"including"violaceols"in"A."nidulans.2,"3"Thus,"we"concluded"that"interaction"between"A."nidulans"and"A."fumigatus'promoted"biosynthesis"of"violaceols"in"A."nidulans"through"activation"of"the"ors"cluster."Interestingly,"the"interaction"between"A."nidulans"and"a"streptomycete"also"up/regulates"the"ors"cluster"of"A."nidulans"to"trigger"biosynthesis"of"four"polyketides,2"each"of"which"is"different"from"the"main"products"of"our"co/culture,"violaceols.""""""Using"the"co/culture"system"of"A."nidulans"and"A."fumigatus"as"a"model"system,"we"will"elucidate"the"molecular"mechanisms"underlying"activation"of"biosynthetic"gene"clusters."1."Yamazaki,"M."et"al."Chem.*Pharm.*Bull."30:"514/518,"1982."2."Schroeckh,"V."et"al."PNAS"106:"14558/14563,"2009."3."Nielsen,"M."L."et"al."FEMS"Microbiol.*Lett."321:"157/166,"2011.""$ $

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67."SCF*Complex$Components$Suppress$Septation$Near$the$Hyphal$Tip,$Regulate$Cell$Cycle$Progression$and$Interact$Genetically$with$γLtubulin$in$Aspergillus*nidulans""Vitoria"Paolillo,"James"Dohn,"Berl$Oakley.""Dept'Molec'Biosci,'Univ'Kansas,'Lawrence,'KS"""We"previously"demonstrated"that"mipAD159,"a"cold/sensitive"γ/tubulin"allele"of"Aspergillus'nidulans,"causes"a"nuclear"autonomous"failure"of"the"Cdh1"homolog,"CdhA,"to"dissociate"from"spindle"pole"bodies"at"the"G1/S"transition,"resulting"in"continuous"activation"of"the"anaphase"promoting"complex/cyclosome"(APC/C)"in"affected"nuclei."Failure"of"inactivation"of"the"APC/C"results"in"continuous"destruction"of"NimECyclin"B"and"failure"of"affected"nuclei"to"progress"through"the"cell"cycle."These"data"reveal"that"γ/tubulin"plays"a"role"in"regulation"of"CdhA,"but"the"underlying"molecular"mechanisms"are"unknown."One"known"regulator"of"Cdh1"in"other"organisms"is"the"Skp1/Cullin1/F/box"(SCF)"complex."This"complex"is"an"E3"ubiquitin"ligase"that"regulates"Cdh1"ubiquitination"and"subsequent"destruction."We"were"consequently"interested"in"investigating"the"function"of"the"SCF"complex"in"A.'nidulans.'Homologs"of"skp1"and"cul1"(skpA'and"culA)"have"been"identified"in"A.'nidulans'and"both"were"shown"to"be"essential."Localization"and"functional"data"of"SkpA"and"CulA"were"lacking,"however."We"report"the"successful"N/terminal"GFP/tagging"and"localization"of"SkpA"and"CulA."Both"localize"to"the"nucleus"during"interphase"and"exit"at"mitotic"onset."Interestingly,"both"also"localize"to"small"cytoplasmic"bodies"in"the"tip"region."Surprisingly"SkpA,"but"not"CulA,"localizes"to"the"Spitzenköper"in"the"majority"of"tip"cells."Using"heterokaryon"rescue"to"determine"the"phenotypes"of"SkpA"and"CulA"deletions,"we"have"found"that"the"SCF"complex"plays"a"role"in"regulating"the"abundance"of"CdhA"and,"surprisingly,"it"also"plays"a"critical"role"in"suppressing"septation"near"the"growing"hyphal"tip."We"have"also"found"that"deletion"of"culA'recapitulates"a"mipAD159"phenotype."Finally,"we"found"that"mipAD159"exhibits"a'strong,"genetic"interaction"with"a"partially"functional"GFP/tagged"culA'allele,"indicating"that"the"SCF"complex"and"γ/tubulin"are"both"involved"in"a"common"function"required"for"growth"in"A.'nidulans.""68.$Defining$the$importance$of$protein$prenylation$in$A.*fumigatus$growth$and$virulence:$characterization$of$the$αL$(RamB)$and$βL$(Cdc43)$subunits$of$the$GeranylgeranyltransferaseLI$complex""A.$C.$Oliveira$Souza,"A."Martin/Vicente,"Q."Al"Abdallah,"K."DeJarnette,"A."Nywening,"W."Ge,"J."Fortwendel.""Department'of'Clinical'Pharmacy'and'Translational'Science,'University'of'Tennessee'Health'Science'Center,'Memphis,'TN"""Protein"prenylation"is"a"post/translational"modification"that"controls"protein"localization"and"consists"of"the"attachment"of"an"isoprenoid"lipid"to"a"C/terminus"consensus"motif"referred"to"as"CaaX"box."This"pathway"is"mediated"by"two"heterodimeric"enzyme"complexes,"farnesyltransferase"and"Geranylgeranyltransferase"type/I"(GGTase/I),"each"composed"of"a"shared"α/subunit"and"a"unique"β/subunit."In"this"study"we"investigated"the"importance"of"the"shared"α/subunit,"RamB,"and"the"GGTase/I"β/subunit,"Cdc43,"during"A.'fumigatus"hyphal"growth"and"virulence."We"found"that"deletion"of"cdc43"resulted"in"impaired"hyphal"morphogenesis"and"thermo/sensitivity."The"Δcdc43"mutant"also"displayed"hypersensitivity"to"cell"wall"stress"agents"and"to"cell"wall"synthesis"inhibitors."Analyses"of"cell"wall"content"revealed"decreased"amounts"of"β/glucan"in"the"Dcdc43"strain."Despite"strong"in'vitro"phenotypes,"the"Δcdc43"mutant"was"fully"virulent"in"two"models"of"murine"invasive"aspergillosis."In"contrast,"a"strain"expressing"the"α/subunit"gene,"ramB,"from"a"tetracycline/inducible"promoter"was"inviable"under"non/inducing"in'vitro"growth"conditions"and"was"virtually"avirulent"in"both"mouse"models."These"findings,"together"with"previously"published"work,"indicate"a"differential"requirement"for"protein"geranylgeranylation"during"fungal"virulence"and"further"support"that"simultaneous"inhibition"of"both"farnesyltransferase"and"GGTase/I"may"be"a"promising"strategy"for"the"therapy"of"A.'fumigatus"infections.""$ $

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*69.$Delta3(E)Ldesaturation$of$glycosylceramides$protects$fungi$against$the$antifungal$protein$AFP$from$Aspergillus*giganteus""N.$Paege,"S."Jung,"V."Meyer.""Applied'and'Molecular'Microbiology,'TU'Berlin,'Berlin,'Berlin,'DE"""The"emergence"and"spread"of"pathogenic"fungi"that"are"resistant"to"virtually"all"available"antimicrobials"represents"a"serious"challenge."Smarter"antimicrobials,"which"affect"multiple"cellular"targets,"are"one"discussed"strategy"to"prevent"the"development"of"resistance"mechanisms."Of"special"interest"is"the"exploitation"of"the"AFP"family"of"antimicrobial"peptides"including"its"founding"member"AFP."It"is"a"highly"potent"inhibitor"of"chitin"synthesis"and"plasma"membrane"integrity"in"many"human"and"plant"pathogenic"fungi"without"adverse"effects"on"bacteria,"plants"or"mammalian"cells."AFP"is"a"cysteine"stabilized,"small,"cationic"and"amphipathic"γ/core"motif"peptide"secreted"by"the"filamentous"fungus"Aspergillus'giganteus."It"induces"cell"wall"stress"in"fungal"cells"which"will"only"survive"if"they"respond"by"fortification"of"the"cell"wall"with"chitin"via"the"calcineurin"signaling"pathway."Recent"studies"suggested"a"relation"between"membrane"glycosylceramides"(GlyCer)"and"the"antifungal"activity"of"antimicrobial"peptides,"which"were"found"to"specifically"recognize"and"bind"to"GlyCer"of"fungal"but"not"of"mammalian"origin."To"investigate"this"further,"we"analyzed"the"importance"of"the"fungal/specific"Δ3/desaturation"of"GlyCer"fatty"acid"on"the"susceptibility"of"fungi"towards"AFP."Here"we"show"that"filamentous"fungi"deleted"for"the"gene"encoding"the"Δ3/desaturatase"became"more"resistant"towards"AFP,"whereas"knock/in"strains"of"naturally"less"susceptible"yeast"strains"became"less"resistant"towards"AFP."Using"MS"analysis"we"could"prove"that"the"desaturation"of"fatty"acid"chain"indeed"determines"susceptibility."To"the"best"of"our"knowledge,"this"is"the"first"study"demonstrating"how"resistance/susceptibility"towards"an"antifungal"peptide"in"filamentous"fungi"and"yeast,"respectively,"is"controlled"by"specific"desaturation"of"a"glycosylceramide.""*70.$Isolation$and$characterization$of$secondary$metabolites$from$Cladosporium*sphaerospermum""Ngan$Pham1,"Adriana"Blachowicz1,2,"Jingyi"Wang1,"Su"Jeung"Lim1,"Tamas"Torok3,"Kasthuri"Venkateswaran2,"Clay"C."C."Wang1,4.""1)'Department'of'Pharmacology'and'Pharmaceutical'Sciences,'School'of'Pharmacy,'University'of'Southern'California,'Los'Angeles,'CA6'2)'Biotechnology'and'Planetary'Protection'Group,'Jet'Propulsion'Laboratory,'California'Institute'of'Technology,'Pasadena,'CA6'3)'Ecology'Department,'Lawrence'Berkeley'National'Laboratory,'Berkeley,'CA6'4)'Department'of'Chemistry,'Dornsife'College'of'Letters,'Arts'and'Sciences,'University'of'Southern'California,'Los'Angeles,'CA"""Fungi"are"producers"of"unique"natural"products"with"different"biological"activities"and"often"important"pharmaceutical"potential."Secondary"metabolites"(SMs)"are"not"directly"involved"in"the"normal"growth,"reproduction"or"development"of"the"organism."Fungi"produce"myriad"of"SMs"to"gain"environmental"advantage"by"inhibiting"the"growth"of"other"microorganisms"to"protect"their"habitat."SM"biosynthesis"depends"on"various"environmental"factors,"including"the"carbon"and"nitrogen"source,"temperature,"light,"salinity,"and"pH."Cladosporium'sphaerospermum'is"a'ubiquitous,"allergenic"mold"found"in"man/made"and"natural"environments."C.'sphaerospermum"was"predominant"isolate"from"the"Chernobyl"Nuclear"Power"Plant"accident"site,"including"the"wall"of"the"exploded"reactor"and"the"surrounding"Exclusion"Zone"soil."This"highly"melanized"fungus"exhibited"radiotropism,"a"phenomenon"of"growing"towards"the"radiation"source."C.'sphaerospermum'IMV"00045"is"unique"extremophilic"strain"that"was"selected"for"further"characterization"and"genetic"modification."Initial"SM"characterization"via"LCMS"revealed"production"of"four"compounds"with"uncharacterized"biosynthetic"pathways."Our"goal"is"to"establish"a"genetic"system"for"this"organism"using"CRISPR/Cas9"plasmid"provided"by"Professor"Uffe"Mortensen."We"will"disrupt"the"backbone"genes"of"targeted"SM"clusters"and"select"the"correct"transformants"by"using"hygromycin"B"as"a"selection"marker."We"have"optimized"the"protoplasting"protocol"and"determined"the"effective"hygromycin"B"concentration."The"next"step"is"to"target"and"disrupt"the"IMV"00045"melanin"PKS"gene"to"visualize"the"successful"gene"transformation."Upon"establishing"the"effective"gene"transformation"protocol,"we"will"target"the"backbone"and"tailoring"enzyme"genes"of"the"produced"compounds"to"discover"unknown"biosynthetic"pathways."Implementation"of"the"genetic"system"in"IMV"00045"will"enable"in/depth"characterization"of"the"SM"produced"by"this"unique"isolate.""

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71.$The$potential$application$of$Aspergillus*oryzae$in$the$biosorption$of$rare$earth$element$ions$present$in$seepage$waters$from$a$postLuraniumLmining$area""Imre"Boczonádi1,"Ágnes"Jakab1,"Edina"Baranyai2,"Csilla"Noémi"Tóth2,"Lajos"Daróczi3,"Gréta"Kiss4,"Miklós"Antal4,"Tamás"Emri1,"Tünde"Pusztahelyi5,"István"Fábián2,"Erika"Kothe6,"István$Pócsi1.""1)'Department'of'Biotechnology'and'Microbiology,'Faculty'of'Science'and'Technology,'University'of'Debrecen,'Debrecen,'Hungary6'2)'Department'of'Inorganic'and'Analytical'Chemistry,'Faculty'of'Science'and'Technology,'University'of'Debrecen,'Debrecen,'Hungary6'3)'Department'of'Solid'State'Physics,'Faculty'of'Science'and'Technology,'University'of'Debrecen,'Debrecen,'Hungary6'4)'Department'of'Anatomy,'Faculty'of'Medicine,'University'of'Debrecen,'Debrecen,'Hungary6'5)'Central'Laboratory'of'Agricultural'and'Food'Products,'Faculty'of'Agricultural'and'Food'Sciences'and'Environmental'Management,'University'of'Debrecen,'Debrecen,'Hungary6'6)'Microbial'Communication,'Institute'of'Microbiology,'Friedrich'Schiller'University'Jena,'Jena,'Germany"""Biosorption"of"heavy"metal"ions"by"microbial"biomass"is"a"well/known"and"effective"tool"to"remove"these"metals"from"wastewaters,"e.g."seepage"waters"flowing"out"from"abandoned"mine"sites."Biosorption/based"metal"binding"is"also"an"interesting"and"cost"effective"option"to"separate"high/value"rare"earth"elements"(REEs)"or"other"precious"metals"from"wastewaters."In"our"studies,"we"used"Aspergillus'oryzae"biomass"to"capture"REE"ions"under"conditions"mimicking"those"typically"observed"in"seepage"waters,"e.g."at"the"former"uranium"mining"site"close"to"Ronneburg,"Germany."The"strain"A.'oryzae"RB40"was"pre/grown"and"mycelium"re/suspended"in"metal"ion"solution"(16.5"ºC,"pH"3.85±0.07S"NH4

+"concentration"0.69±0.05"mg"l/1)."After"incubation"with"shaking"for"24"h,"no"autolysis"was"observed"and"the"average"diameter"of"the"pellets"was"not"affected"by"the"metal"ions"while"the"pH"and"NH4

+"concentration"of"the"solution"increased"up"to"4.78±0.06"and"43.5±4.7"mg"l/1,"respectively."The"biosorption"efficiency"reached"100"%"for"the"REE"ions"Y3+"(7"μM),"Ce4+"(2"μM)"and"Nd3+"(0.9"μM)"even"in"the"presence"of"high"concentrations"of"Li+"(0.06"mM),"Mg2+"(30"mM),"Al3+"(1.4"mM),"Fe3+"(0.6"mM),"Ni2+"(0.2"mM)"and"Zn2+"(0.04"mM)"ions."It"is"noteworthy"that"the"great"majority"of"Fe3+"and"Al3+"ions"was"absorbed"by"12"h"incubation,"while"the"removal"of"REE"ions"was"shifted"clearly"to"between"6/24"h."When"A.'oryzae'biomass"was"added"to"100"ml"aliquots"of"a"solution"containing"only"Y3+,"Ce4+"and"Nd3+"ions,"the"biomass"absorbed"all"REE"ions"completely"within"1"h"of"incubation."To"shed"light"on"the"localization"and"mechanism"of"the"observed"biosorption"processes,"Energy"Dispersive"X/Ray"Spectroscopy"and"Transmission"Electron"Microscopy"analyses"were"carried"out"on"the"metal"ion"solution/exposed"hyphae"of"A.'oryzae."These"experiments"clearly"indicated"that"Fe3+"ions"were"retained"on"the"surface"of"the"cell"wall"in"the"form"of"complex"aggregates."""The"research"was"financed"by"the"European"Union"and"the"European"Social"Fund"through"the"project"EFOP/3.6.1–16–2016/00022,"by"the"National"Research,"Development"and"Innovation"Office"(Hungary)"projects"K112181"and"K119494,"and"by"the"Higher"Education"Institutional"Excellence"Program"of"the"Ministry"of"Human"Capacities"in"Hungary,"within"the"framework"of"the"Biotechnology"thematic"program"of"the"University"of"Debrecen.""*72.$Activation$of$Silent$Gene$Clusters$in$Aspergillus*nidulans$using$Hybrid$Transcription$Factors""Christian$A.$Rabot1,"Michelle"F."Grau1,"Ruth"Enwistle2,"Yi/Ming"Chiang1,"Manmeet"Ahuja2,"C."Elizabeth"Oakley2,"Clay"C."C."Wang1,3,"Richard"B."Todd4,"Berl"R."Oakley2."1)'Dept.'of'Pharmacology'and'Pharmaceutical'Sciences,'University'of'Southern'California,'Los'Angeles,'CA,'USA6'2)'Dept.'of'Molecular'Biosciences,'University'of'Kansas,'Lawrence,'KS,'USA6'3)'Dept.'of'Chemistry,'University'of'Southern'California,'Los'Angeles,'CA,'USA6'4)'Dept.'of'Plant'Pathology,'Kansas'State'University,'Manhattan,'KS,'USA.''"Fungal"secondary"metabolites"(SMs)"represent"a"largely"underexploited"source"of"clinically"and"agriculturally"useful"compounds."Bioinformatic"analyses"of"fungal"genomes"suggest"that"the"number"of"putative"biosynthetic"gene"clusters"far"exceeds"the"number"of"known"SMs."However,"around"70%"of"these"putative"gene"clusters"are"silent"under"standard"laboratory"conditions."Over"the"past"few"decades,"many"groups"have"employed"a"number"of"different"strategies"to"activate"these"silent"gene"clusters,"such"as"varying"growth"conditions,"promoter"replacement,"co/culturing"with"other"species,"or"epigenetic"modifications."One"additional"strategy"to"activate"these"gene"clusters"is"to"manipulate"the"expression"of"transcriptional"regulators."Previous"work"in"our"laboratory"has"shown"that"placing"the"transcription"factor"

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of"a"given"gene"cluster"under"the"control"of"an"inducible"promoter"can"be"a"successful"strategy"to"induce"or"increase"expression"of"a"cryptic"gene"cluster."However,"some"gene"clusters"were"not"activated"when"this"approach"was"taken,"suggesting"the"need"for"an"improved"strategy"to"induce"the"expression"of"gene"clusters"of"interest."The"focus"of"this"project"is"to"generate"hybrid"transcription"factors"to"activate"a"silent"gene"clusters."This"was"accomplished"by"maintaining"the"native"DNA/binding"domain,"but"replacing"the"native"activation"domain"of"a"transcription"factor"with"one"that"is"more"efficient"in"the"recruitment"of"the"transcriptional"machinery"necessary"for"transcribing"a"given"gene."Preliminary"data"from"this"project,"in"which"hybrid"transcription"factors"were"generated"for"three"putative"gene"clusters"in"Aspergillus'nidulans,"suggests"that"this"approach"is"successful"in"turning"on"silent"gene"clusters."$*73.$A$Systems$Biology$Approach$to$Signal$Transduction$Gene$Regulation$Response$to$an$Antifungal$Drug$in$Aspergillus*nidulans"""S$Reese1,"C"Chelius2,"M"Marten2,"R"Srivastava3,"W"Riekhof1,"S"Harris4.""1)'School'of'Biology,'University'of'NebraskaQLincoln,'Lincoln,'NE6'2)'Chemical,'Biochemical'and'Environmental'Engineering,'University'of'MarylandQBaltimore'County,'Baltimore,'MD6'3)'Chemical,'Materials,'and'Bimolecular'Engineering,'University'of'Connecticut,'Mansfield,'CT6'4)'Department'of'Biological'Sciences,'University'of'Manitoba,'Winnipeg,'Canada"""When"challenged"by"environmental"stress,"filamentous"fungi"activate"specific"responses"that"are"transmitted"through"signaling"pathways"from"the"cell"surface"to"the"nucleus."For"example,"Aspergillus'nidulans'responds"to"cell"wall"damage"by"activating"the"Cell"Wall"Integrity"Signaling"(CWIS)"pathway"featuring"the"MAP"kinase"MpkA"to"repair"the"cell"wall"and"restore"its"integrity."Anti/fungals"such"as"micafungin"are"typically"used"to"activate"the"CWIS"pathway"because"they"inhibit"the"production"of"beta/1,3/glucan,"which"is"essential"for"cell"wall"formation."As"part"of"a"larger"effort"to"generate"a"mathematical"model"for"a"canonical"signaling"pathway"in"A."nidulans,"we"are"employing"phosphoproteomics"and"transcriptomics"to"dynamically"characterize"CWIS"activation"and"output"in"response"to"treatment"of"both"wildtype"and"mpkAmutants"with"micafungin."Here,"using"RNA/Seq,"we"observed"a"shift"in"gene"expression"in"response"to"cell"wall"damage"and"repair.'Two"main"phases"of"gene"expression"were"observedS"a"cell"stress"response"that"ends"about"50"minutes"post/treatment,"followed"by"an"adaptation"response"that"extended"to"the"final"timepoint"and"presumably"prepares"hyphae"for"resumption"of"growth."Notably,"the"later"phase"showed"a"coupling"of"light/induced"genes"with"secondary"metabolite"production."This"phase"also"featured"a"dramatic"increase"in"brlA'expression,"and"suggests"a"previously"unknown"role"for"BrlA"in"the"response"to"cell"wall"stress."Our"investigation"looks"to"fully"characterize"the"CWIS"pathway"in"filamentous"fungi"and"to"fully"understand"the"role"of"MpkA"in"this"response."""*74.$Quantitative$gene$expression$of$Aspergillus*bidirectional$histone$promoters$and$application$as$gene$expression$platform""J.K.$Rendsvig,"J.B."Hoof.""Department'of'Biotechnology'and'Biomedicine,'Technical'University'of'Denmark,'DK"""Fungal"promoters"are"an"indispensable"tool"for"heterologous"expression"and"product"elucidation"for"silent"biosynthetic"gene"clusters"–"however"constitutive"expression"of"such"pathways"requires"multiple"promoters"without"need"for"induction."In"this"study,"we"set"out"to"characterize"and"exploit"the"intergenic"region"between"the"histone"genes"H3"and"H4"of"five"Aspergilli"in"order"to"function"as"bidirectional"promoters"facilitating"the"construction"of"gene/expression"platform"to"build"one/locus"biosynthetic"pathways."The"0.8kb"bidirectional"promoter,"Ph3h4,"of"five"Aspergillus"species"(A.'nidulans,"A.'niger,"A.'flavus,"A.'clavatus"and"A.'terreus),"representing"five"sections"of"genus"Aspergillus,"were"each"applied"for"simultaneous"expression"of"two"fluorescent"proteins"(mRFP"and"mCitrine)"in"an"A.'nidulans"integration"site,"and"analyzed"by"fluorescence"microscopy"confirming"functionality"of"all""heterologous"promoters"in"A.'nidulans."The"relative"gene/expression"levels"and"strengths"of"promoters"were"analyzed"by"qRT/PCR"for"each"reporter"gene"with"comparison"to"two"conventional"fungal"promoters,"PgpdA"and"Ptef1"of"A.'nidulans."Next,"we"used"the"Ph3h4"of"A.'niger"and"A.'clavatus"to"heterologously"and"successfully"express"the"biosynthetic"gene"cluster"of"the"valuable"compound"family"malformin"from"A.'brasiliensis"in"A.'

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nidulans."The"final"malformin"producing"strain"showed"no"morphological"change"compared"to"the"parental"strain,"indicating"the"application"of"heterologous"Ph3h4s"do"not"affect"growth"negatively."Moreover,"by"verifying"the"continued"presence"of"the"complete"construct,"we"confirmed"that"the"presence"of"two"heterologous"Ph3h4"in"the"same"locus"did"not"result"in"genetic"instability."The"authors"aspire"that"these"bidirectional"promoters"will"be"a"valuable"aid"in"the"construction"of"various"pathways"for"constitutive"expression"commencing"in"the"non/stationary"growth"phases,"thus"facilitating"heterologous"expression"and"pathway"elucidation.""75.$Cell$wall$integrity$pathway$controls$the$asexual$development$of$Aspergillus*fumigatus""M.C.$Rocha1,"J.H.T.M"Fabri1,"R."Silva/Rocha2,"A.F"Cunha1,"D."Canovas3,"I."Malavazi1.""1)'Department'of'Genetics'and'Evolution,'Federal'University'of'São'Carlos,'Sao'Carlos,'São'Paulo,'BR6'2)'Department'of'Pharmacology,'School'of'Medicine'of'Ribeirão'Preto,'University'of'São'Paulo,'Ribeirão'Preto,'São'Paulo,'BR6'3)'Department'of'Genetics,'University'of'Seville,'Seville,'ES""'Aspergillus'fumigatus"is"an"opportunistic"human"pathogen"that"causes"systemic"infections,"including"invasive"pulmonary"aspergillosis"in"immunocompromised"individuals."Survival"of"this"fungus"is"highly"dependent"on"the"cell"wall"organization"and"function"of"its"structural"components."Cell"wall"integrity"pathway"(CWIP)"is"the"primary"signaling"cascade"that"controls"de'novo"synthesis"of"the"cell"wall."In"this"work,"we"investigated"the"function"of"the"CWIP"components"PkcA,"MpkA"and"RlmA,"in"the"asexual"development"of"A.'fumigatus."The"mutants"of"CWIP,"pkcAG579R,"ΔmpkA,'and'ΔrlmA"exhibit"reduced"asexual"sporulation,"and"it"was"possible"to"observe"that"pkcA"and"rlmA"are"induced"during"synchronized"asexual"development"in"A.'fumigatus."In"addition,"it"was"found"increased"phosphorylation"of"MAP/kinase"MpkA"in"this"same"condition"in"the"wild/type"strain,"but"these"genes"had"their"expression"reduced"in"mutants"of"CWIP,"as"well"as"the"reduction"of"MpkA"phosphorylation"in"these"mutants,"which"leads"to"the"conclusion"that"CWIP"is"induced"both"at"protein"and"transcriptional"levels"during"the"conidiation."It"was"also"possible"to"observe"that"CWIP,"through"the"transcription"factor"RlmA,"regulates"transcription"of"key"transcription"factors"involved"in"conidiogenesis"in"Aspergillus"ssp.,"i.e"brlA"and"abaA,"besides"regulating"also"transcription"of"other"important"genes"involved"in"the"asexual"development,"including"flbB,"flbC"and"rasB."Overall,"RlmA"works"as"a"transcription"factor"associated"with"the"CWIP"acting"downstream"of"PkcA/MpkA,"signaling"and"regulating"the"conidiation"in"A.'fumigatus.""*76.$Increased$Hypoxia$fitness$of$clinical$Aspergillus*fumigatus$isolates$from$chronic$Aspergillosis$patients""B."Ross1,"J."Stajich2,"R."Cramer1.""1)'Department'of'Microbiology'and'Immunology,'Geisel'School'of'Medicine'at'Dartmouth,'Hanover,'NH6'2)'Department'of'Microbiology'and'Plant'Pathology,'University'of'California'Riverside,'Riverside,'CA"""Adaptation"mechanisms"of"human"fungal"pathogens"to"long/term"colonization"in'vivo"are"poorly"understood."Moreover,"how"in'vivo'adaptation"to"the"infection"microenvironment"affects"disease"outcomes"is"unknown."The"ubiquitous"environmental"mold"Aspergillus'fumigatus"causes"a"spectrum"of"disease"in"humans."In"particular,"patients"with"the"genetic"disorder"Cystic"Fibrosis"(CF)"have"been"associated"with"long/term"fungal"colonization."We"are"studying"a"series"of"clinical"isolates"obtained"from"a"single"CF"patient"over"~4.5"years."Genotypic"and"phenotypic"analyses"using"this"series"of">50"A.'fumigatus'isolates"has"yielded"several"observations,"including"diverse"colony"and"hyphal"morphologies"and"stress"resistance"profiles."An"interesting"subset"of"9"isolates,"spanning"~1.5"years,"display"a"unique"“compact”"colony"morphology,"inducible"in"the"presence"of"glucose"and"preferred"nitrogen"sources"in"ambient"oxygen."It"is"characterized"by"reduced"radial"growth,"a"dense"colony"of"hyper/branching"hyphae,"and"robust"secretion"of"an"extracellular"matrix."Compact"isolates"also"display"increased"tolerance"to"voriconazole"and"Congo"Red,"but"poor"oxidative"and"osmotic"stress"tolerance."Most"strikingly,"the"compact"isolates"show"an"upwards"of"55%"increased"radial"growth"in"1%"oxygen."This"finding"suggests"that"the"compact"isolates"have"adapted"to"low"oxygen"conditions"within"the"CF"lung."We"hypothesize"that"prolonged"colonization"of"the"CF"lung"results"in"adaptation"and"mutation"in"genes"important"for"increased"growth"in"low"oxygen."To"determine"whether"this"phenomenon"is"specific"to"this"series"of"isolates,"analyses"of"isolates"collected"from"other"CF"patients"are"ongoing."Current"work"is"focused"on"identifying"specific"mutations"that"contribute"to"the"increased"growth"using"whole"genome"sequencing"as"well"as"classical"and"forward"genetic"approaches."This"work"will"provide"important"knowledge"to"aid"in"diagnosis"

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and"treatment"and"will"contribute"to"answering"long/sought/after"clinical"questions"related"to"isolation"of"fungal"pathogens"from"colonized"and"infected"tissue.""*77.$What’s$in$a$spore?$Comparative$proteomics$of$the$mycelia$to$the$spore""K.$RothschildLMancinelli,"L."Bundgaard,"T."Vesth,"U."auf"dem"Keller,"M.R."Andersen.""Bioengineering,'Technical'University'of'Denmark,'Lyngby,'DK"""One"of"the"hardiest"organismal"forms"in"life"is"the"spore,"and"yet"there"is"surprisingly"little"known"about"it"outside"Bacillus'subtilis."A"spore"is"inherently"thought"of"as"inert,"yet"we"know"nothing"about"how"inert"it"really"is"so"to"study"this"we"have"done"a"comparative"whole/proteome"study"of"the"Aspergillus'niger"spore"grown"in"liquid"vs"agar"as"compared"to"the"mycelia"grown"in"liquid"vs"agar"as"well"as"the"secreted"proteome"in"both"conditions."This"was"done"to"better"understand"the"physiological"stresses"these"different"environments"put"on"A.'niger,"and"how"this"knowledge"can"be"applied"for"furthering"academic"and"industrial"interests."Initially,"A.'niger"was"grown"on"Minimal"Media"as"a"controlled"media,"then"the"spores,"mycelia,"and"media"were"isolated"from"each"other,"broken"apart"in"a"hydrolic"press,"and"analyzed"by"mass"spectrometry."The"data"suggests"that"there"is"a"proteomic"shift"as"well"as"a"morphological"shift"from"spore"to"mycelia."This"has"also"allowed"us"to"expand"upon"our"knowledge"of"the"proteomes"of"the"spore"and"mycelia,"which"has"applications"far"beyond"our"direct"protein"research.""*78.$Unexpected$production$of$rhizobium$signaling$molecules$across$the$Fungal$Kingdom""T.A."Rush1,2,3,"A."Bascaules"4,"P."Jargeat4,"F."Maillet5,"K.R."Cope1,2,"C."Johnson6,"B."Kleven1,2,"A."QuyManh/Maës4,"Q.J."Choudhury7,"J."Labbé7,"C."Swift8,"M.A."O’Malley8,"J.W."Bok1,6,"S."Cottaz9,"S."Fort9,"J."Nett6,"N.P."Keller1,6,"V."Puech/Pagès"4,"G."Bécard"4,"J.M."Ané"1,2.""1)'Department'of'Bacteriology,'University'of'Wisconsin–Madison,'Madison,'WI,'USA6'2)'Department'of'Agronomy,'University'of'Wisconsin–Madison,'Madison,'WI,'USA6'3)'Department'of'Plant'Pathology,'University'of'Wisconsin–Madison,'Madison,'WI,'USA6'4)'Laboratoire'de'Recherche'en'Sciences'Végétales,'Toulouse,'France6'5)'Laboratoire'des'Interactions'Plantes–Microorganismes,'Castanet–Tolosan,'France6'6)'Department'of'Medical'Microbiology'and'Immunology,'University'of'Wisconsin–Madison,'Madison,'WI,'USA6'7)'Biosciences'Division,'Oak'Ridge'National'Laboratory,'Oak'Ridge,'TN,'USA6'8)'Department'of'Chemical'Engineering,'University'of'California,'Santa'Barbara,'CA,'USA6'9)'Centre'de'Recherche'sur'les'Macromolécules'Végétales,'CERMAV–CNRS'UPR,'Grenoble,'France"""Lipo/chitooligosaccharides"are"short"chitooligosaccharides"(polymers"of"three"to"five"β/1,4/linked"N/acetyl/D/glucosamine"residues)"substituted"by"a"fatty"acyl"chain"at"the"non/reducing"end."The"lipid"chains"can"vary"in"length"and"saturation,"and"the"chitooligosaccharidic"backbone"can"carry"various"chemical"substitutions"(sulfate,"arabinose,"and"fucose)."Such"molecules"were"first"identified"as"the"primary"signaling"molecules"produced"by"nitrogen/fixing"bacteria"called"rhizobia"and"perceived"by"their"legume"hosts."More"recently,"lipo/chitooligosaccharides"and"short"chitooligosaccharides"were"also"identified"in"plant/associated"symbiotic"fungi"such"as"the"arbuscular"mycorrhizal"fungus,"Rhizophagus'irregularis'(syn:"Glomus'intraradices)"and"the"ectomycorrhizal"fungus,"Laccaria'bicolor."Given"the"discovery"of"lipo/chitooligosaccharides"being"produced"by"such"different"fungi,"we"tested"if"other"fungi"can"produce"lipo/chitooligosaccharides."Using"both"biological"assays"(legume"root"hair"branching"and"ENOD11"expression)"and"mass"spectrometry,"we"analyzed"fungal"exudates"of"55"fungi"across"five"phyla"in"the"fungal"kingdom"and"three"species"of"oomycetes."We"were"able"to"detect"lipo/chitooligosaccharides"in"fungal"exudates"in"47"fungi"across"the"fungal"kingdom"with"various"lifestyles"and"growth"habits"but"not"in"non/dimorphic"yeasts"(Saccharomyces'cerevisiae"and"Candida'glabrata)"and"oomycetes."In"contrast,"chitooligosaccharides"were"detected"in"all"fungi"and"oomycetes"tested.""$ $

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*79.$The$monothiol$glutaredoxin$GrxD$interacts$with$the$iron$regulator$HapX$and$CmtA,$a$protein$with$yet$unknown$function,$in$Aspergillus*fumigatus""M.T.$MLL."Scheven1,"2,"M."Misslinger3,"P."Hortschansky1,"T."Krüger1,"J."Weber1,2,"H."Haas3,"A."A."Brakhage1,2.""1)'Leibniz'Institute'for'Natural'Product'Research'and'Infection'Biology'Q'Hans'Knöll'Institute'(HKI),'Department'Molecular'and'Applied'Microbiology,'Jena,'DE6'2)'Friedrich'Schiller'University'Jena,'Institute'for'Microbiology,'Jena,'DE6'3)'Innsbruck'Medical'University,'Biocenter,'Division'of'Molecular'Biology,'Innsbruck,'A""'Aspergillus'fumigatus"is"a"ubiquitous"saprophytic"mold,"which"causes"life/threatening"diseases"in"immunocompromised"patients."Infection"by"A.'fumigatus"depends"on"iron"accessibility"in"the"human"host."Iron"is"a"vital"nutrient,"but"can"be"harmful"in"excess"by"triggering"the"formation"of"reactive"oxygen"species."As"a"result,"A.'fumigatus'has"evolved"fine/tuned"mechanisms"to"regulate"its"iron"homeostasis."Adaptation"to"iron"limitation"and"excess"is"mediated"by"the"bZIP"transcription"factor"HapX,"which"functions"via"physical"interaction"with"the"heterotrimeric"CCAAT/binding"complex."During"iron"starvation,"iron"consumption"is"repressed"and"iron"uptake"is"activated."During"iron"overload,"the"cell"is"detoxified"from"iron"by"activation"of"vacuolar"iron"storage"[1,"2]."Currently,"the"molecular"mechanisms"of"iron"sensing"by"HapX"are"unknown."HapX"senses"the"cellular"iron"level"most"likely"by"interaction"with"other"proteins,"like"monothiol"glutaredoxin"(GrxD)."For"identification"of"possible"GrxD"as"well"as"HapX"interaction"partners,"a"co/immunoprecipitation"approach"was"applied."VENUS/tagged"GrxD"and"MYC/tagged"HapX"proteins"were"enriched"from"crude"cell"extracts"by"GFP/Trap"and"MYC/Trap,"respectively."Proteins"were"subsequently"identified"and"quantified"by"nano"LC/MS/MS"analysis."HapX"co/purified"highly"abundant"during"GrxDVENUS"enrichment"under"iron"limitation"and"vice'versa."In"addition,"a"protein"with"yet"unknown"function,"CmtA"(orthologs"are"copper"metallothioneins)"was"co/enriched"during"GrxDVENUS"trap"with"high"abundance"under"iron"excess."In"line"with"the"in'vivo"results,"recombinant"A.'fumigatus'GrxD"and"HapX"proteins"as"well"as"GrxD"and"CmtA"proteins"were"co/purified"in'vitro"from"Escherichia'coli."These"data"provide"first"evidence"that"the"iron/dependent"regulatory"function"of"HapX"in"A.'fumigatus'requires"the"interaction"with"GrxD."The"functional"relevance"of"the"CmtA/GrxD"interaction"is"subject"of"current"studies."References"[1]"Gsaller,"F.S"Hortschansky,"P."et'al.'EMBO'J'2014,$33,"2261/2276."[2]"Hortschansky,"P."et'al.'Biochim'Biophys'Acta'Gene'Regul'Mech'2017,$1860,"560/570.""*80."Examination$of$ploidy$in$Aspergillus*fumigatus""B.A.$Shuman,"E."Scopel,"S.E."Kang,"D."Bensasson,"M."Momany.""Fungal'Biology'Group'and'Plant'Biology'Department,'University'of'Georgia,'Athens,'GA""'Aspergillus'fumigatus'is"commonly"found"as"a"haploid"saprobe"in"a"variety"of"environments,"and"is"also"a"human"fungal"pathogen"that"can"be"fatal"for"immunocompromised"individuals."A.'fumigatus'strains"that"are"resistant"to"antifungal"drugs"are"being"reported"at"increasing"frequencies.""Ploidy"changes"have"been"shown"to"be"a"mechanism"of"antifungal"resistance"in"the"human"fungal"pathogens"Candida'albicans'and"Cryptococcus'neoformans."While"large/scale"intrachromosomal"deletions"have"been"reported"in"A.'fumigatus,"not"much"is"known"about"ploidy"in"wild"isolates."We"are"studying"the"ploidy"of"A.'fumigatus'by"plotting"B/allele"frequency"and"read/depth"using"whole"genome"sequences"from"a"variety"of"clinical"and"environmental"isolates.""$ $

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*81."Extensive$loss$of$cell$cycle$and$DNA$repair$genes$in$an$ancient$lineage$of$bipolar$budding$yeasts""J.L."Steenwyk1,"D.A."Opulente3,"J."Kominek3,"X./X."Shen1,"X."Zhou2,"A.L."Labella1,"N.P."Bradley1,"B.F."Eichman1,"N."Cadez4,"D."Libkind5,"J."DeVirgilio6,"A.B."Hulfachor3,"C.P."Kurtzman6,"C.T."Hittinger3,"A."Rokas1.""1)'Biological'Sciences,'Vanderbilt'University,'Nashville,'TN'37235,'USA6'2)'Department'of'Plant'Pathology,'College'of'Agriculture,'South'China'Agricultural'University,'Guangzhou'510642,'Guangdong'Province,'China6'3)'Laboratory'of'Genetics,'Genome'Center'of'Wisconsin,'Department'of'Energy'Great'Lakes'Bioenergy'Research'Center,'Wisconsin'Energy'Institute,'J.'F.'Crow'Institute'for'the'Study'of'Evolution,'University'of'Wisconsin–Madison,'Wisconsin'53706,'USA6'4)'University'of'Ljubljana,'Biotechnical'Faculty,'Department'of'Food'Science'and'Technology,'Jamnikarjeva'101,'1000'Ljubljana,'Slovenia6'5)'Laboratorio'de'Microbiología'Aplicada'y'Biotecnología,'Instituto'de'Investigaciones'en'Biodiversidad'y'MedioQambiente,'Consejo'Nacional'de'Investigaciones'Científicas'y'Técnicas'(CONICET)QUniversidad'Nacional'del'Comahue,'8400'Bariloche,'Argentina6'6)'Mycotoxin'Prevention'and'Applied'Microbiology'Research'Unit,'National'Center'for'Agricultural'Utilization'Research,'Agricultural'Research'Service,'US'Department'of'Agriculture,'Peoria,'Illinois'61604,'USA"""Cell"cycle"checkpoints"and"DNA"repair"processes"protect"organisms"from"potentially"ruinous"mutational"damage."We"noticed"that"a"group"of"species"in"the"genus"Hanseniaspora"exhibited"very"high"evolutionary"rates,"low"GC"content,"small"genome"sizes,"and"lower"gene"numbers"compared"to"other"budding"yeasts'(subphylum"Saccharomycotina)."To"better"understand"Hanseniaspora'evolution,"we"sequenced"11"new"genomes"and"analyzed"them"jointly"with"14"publicly"available"ones,"a"sample"that"represents"18"/"20"known"species"in"the"genus."Our"phylogenomic"analyses"identify"two"Hanseniaspora"lineages,"the"fast/evolving"lineage"(FEL),"which"originated"~87"million"years"ago"(mya)"and"the"slow/evolving"lineage"(SEL),"which"originated"~54"mya."Remarkably,"both"lineages"have"lost"genes"associated"with"the"cell"cycle"and"genome"integrity"but"these"losses"are"greater"in"the"FEL."For"example,"all"species"have"lost"the"cell"cycle"regulator"WHI5,"and"the"FEL"lost"components"of"the"spindle"checkpoint"pathway"(e.g.,"MAD1,"MAD2)"and"DNA"damage"checkpoint"pathway"(e.g.,"MEC3,"RAD9)."Similarly,"both"lineages"lack"genes"involved"in"DNA"repair"pathways,"including"the"DNA"glycosylase"gene"MAG1,"which"is"part"of"the"base"excision"repair"pathway,"and"the"DNA"photolyase"gene"PHR1,"which"is"involved"in"pyrimidine"dimer"repair."Strikingly,"the"FEL"has"lost"31"additional"genes,"including"those"encoding"DNA"polymerases"(i.e.,"POL4'and"POL32)"and"telomere/associated"genes"(e.g.,"RIF1,"RFA3)."Molecular"evolutionary"analyses"reveal"that"FEL"species"harbor"greater"mutational"loads,"homopolymer"instabilities,"and"fractions"of"mutations"associated"with"the"common"endogenously"damaged"base,"8/oxoguanine."We"conclude"that"Hanseniaspora"is"an"ancient"lineage"that"has"diversified,"despite"the"apparent"lack"of"many"otherwise"universally"conserved"cell"cycle"and"genome"integrity"genes"and"pathways,"and"may"represent"a"novel"system"for"studying"the"limits"of"a"reduced"capacity"for"these"pathways"or"their"novel"rewiring.""*82.$A$robust$phylogenomic$timetree$for$biotechnologically$and$medically$important$fungi$in$the$genera$Aspergillus*and$Penicillium$Jacob$L.$Steenwyk1,"Xing/Xing"Shen1,"Abigail"L."Lind2,3,"Gustavo"H."Goldman4,"and"Antonis"Rokas1,2$1"Department"of"Biological"Sciences,"Vanderbilt"University,"Nashville,"TN,"37235,"USA$2"Department"of"Biomedical"Informatics,"Vanderbilt"University"School"of"Medicine,"Nashville,"Tennessee,"United"States"of"America$3"Gladstone"Institute"for"Data"Science"and"Biotechnology,"San"Francisco,"California,"USA$4"Departamento"de"Ciências"Farmacêuticas,"Faculdade"de"Ciências"Farmacêuticas"de"Ribeirão"Prêto,"Bloco"Q,"Universidade"de"São"Paulo,"São"Paulo,"Brazil$"The"filamentous"fungal"family"Aspergillaceae"contains">"1,000"known"species,"mostly"in"the"genera"Aspergillus'and"Penicillium."Several"species"are"used"in"the"food,"biotechnology,"and"drug"industries"(e.g.,"Aspergillus'oryzae,'Penicillium'camemberti),"while"others"are"dangerous"human"and"plant"pathogens"(e.g.,"Aspergillus'fumigatus,'Penicillium'digitatum)."To"infer"a"robust"phylogeny"and"pinpoint"poorly"resolved"branches"and"their"likely"underlying"contributors,"we"used"81"genomes"spanning"the"diversity"of"Aspergillus'and"Penicillium"to"construct"a"1,668/gene"data"matrix."Phylogenies"of"the"nucleotide"and"amino"acid"versions"of"this"full"data"matrix"as"well"as"of"five"additional"834/gene"data"matrices"constructed"by"subsampling"the"top"50%"of"genes"according"to"different"criteria"associated"with"

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strong"phylogenetic"signal"were"generated"using"three"different"maximum"likelihood"schemes"(i.e.,"gene/partitioned,"unpartitioned,"and"coalescence)."Examination"of"the"topological"agreement"among"these"36"phylogenies"and"measures"of"internode"certainty"identified"12"/"78"(15.4%)"bipartitions"that"were"incongruent"and"pinpoint"the"likely"underlying"contributing"factors"(incomplete"lineage"sorting,"hybridization"or"introgression,"and"reconstruction"artifacts"associated"with"poor"taxon"sampling)."Relaxed"molecular"clock"analyses"suggest"that"Aspergillaceae"likely"originated"in"the"lower"Cretaceous"and"the"Aspergillus'and"Penicillium"genera"in"the"upper"Cretaceous."Our"results"shed"light"on"the"ongoing"debate"on"Aspergillus'systematics"and"taxonomy"and"provide"a"robust"evolutionary"and"temporal"framework"for"comparative"genomic"analyses"in"Aspergillaceae."More"broadly,"our"approach"provides"a"general"template"for"phylogenomic"identification"of"resolved"and"contentious"branches"in"densely"genome/sequenced"lineages"across"the"tree"of"life."$*83.$Regulation$of$Leucine$Biosynthesis$and$a$Key$Nitrogen$Assimilation$Gene$by$the$Transcription$Factor$LeuR$in$Aspergillus*nidulans""Joel$T.$Steyer,"Damien"J."Downes,"Cameron"C."Hunter,"Richard"B."Todd.""Kansas'State'University,'Manhattan'KS'""Branched"chain"amino"acids"(BCAAs)"leucine,"isoleucine,"and"valine"are"essential"amino"acids,"unable"to"be"synthesized"by"animals"and"must"be"obtained"through"their"diet."Many"fungal"species,"including"Aspergillus,'Candida,'and"Saccharomyces,"synthesize"BCAAs."Proper"regulation"of"BCAA"metabolism"is"important"for"protein"synthesis,"growth,"and"virulence."In"Aspergillus'nidulans,'the"Zn(II)2Cys6"transcription"factor"LeuB"regulates"leucine"biosynthesis"in"response"to"leucine"levels."LeuB"also"regulates"expression"of"gdhA,"which"encodes"the"key"nitrogen"assimilation"enzyme"NADP/glutamate"dehydrogenase,"NADP/GDH."We"have"identified"a"paralog"of"LeuB,"LeuR,"and"examined"its"role"in"leucine"biosynthesis"and"gdhA'regulation."Phylogenetic"analysis"shows"that"while"LeuB"is"conserved"in"Ascomycetes,"LeuR"is"conserved"only"within"Eurotiomycetes.The"leuB∆"mutant"is"a"leaky"leucine"auxotroph."We"deleted"leuR'and"found"the"leuR∆"mutant"to"be"a"prototroph."However,"the"leuB∆"leuR∆"double"mutant"is"a"tight"leucine"auxotroph,"indicating"a"role"for"LeuR"in"regulating"leucine"biosynthesis."Using"a"gdhAQlacZ'translational"fusion"reporter"gene"and"exogenous"leucine,"we"show"that"LeuR"regulates"gdhA'expression."Previously,"we"used"a"series"of"deletions"in"the"gdhA'promoter"to"show"that"LeuB"acts"through"two"sites"of"action."We"now"are"using"these"promoter"deletions"to"compare"the"wildtype"strain"to"leuB∆,"leuR∆,"and"leuB∆"leuR∆"mutants"to"identify"where"LeuR"acts"in"the"gdhA'promoter."Our"experiments"show"that"the"transcription"factors"LeuB"and"LeuR"link"regulation"of"nitrogen"assimilation"and"leucine"biosynthesis."""*84.$FungalLbacterial$coLculture$induces$biosynthesis$of$cryptic$natural$products$in$the$genus$Aspergillus*and$leads$to$identification$of$BasR$as$key$regulatory$node$for$translating$bacterial$signals""Maria$C."Stroe1,2,"Tina"Netzker1,"Kirstin"Scherlach1,"Vito"Valiante3,"Nils"Jaeger5,"Juliane"Fischer1,"Volker"Schroeckh1,"Christian"Hertweck2,4,"Thorsten"Heinzel5,"Axel"Brakhage1,2.""1)'Department'of'Molecular'and'Applied'Microbiology,'Leibniz'Institute'for'Natural'Product'Research'and'Infection'Biology'(HKI),'Jena,'Germany6'2)'Institute'of'Microbiology,'Friedrich'Schiller'University'Jena6'3)'Research'Group'Biobricks'of'Microbial'Natural'Product'Synthetases,'Leibniz'Institute'for'Natural'Product'Research'and'Infection'Biology'(HKI),'Jena,'Germany6'4)'Department'of'Biomolecular'Chemistry,'Leibniz'Institute'for'Natural'Product'Research'and'Infection'Biology'(HKI),'Jena,'Germany6'5)'Department'of'Biochemistry,'FriedrichQSchillerQUniversity'Jena,'Jena,'Germany"""Fungi"are"well/known"producers"of"secondary"metabolites"(SMs),"which"are"low/molecular"mass"compounds"often"with"antimicrobial,"anticancer"or"cholesterol/lowering"activities."However,"their"ecological"function"remains"obscure."We"propose"that"SMs"serve'as"communication"signals"during"coexistence"of"microorganisms"(1)."Recent"advances"in"sequencing"and"genome"mining"have"revealed"that"the"biosynthetic"potential"of"fungi"is"far"greater"than"the"number"of"currently"identified"compounds"(2)."However,"many"SM"biosynthetic"gene"clusters"are"silent,"i.e.,"they"are"not"expressed"under"standard"laboratory"conditions"because"their"environmental"or"developmental"triggers"are"not"met"in"an"artificial"culture"setting."One"successful"approach"to"awaken"microbial"gene"clusters"is"to"simulate"natural"environmental"conditions"through"co/cultivation"of"microorganisms"(3)."Here,"we"discuss"two"examples"of"

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inter/kingdom"microbial"interactions"between"Aspergillus"and"Streptomyces"in"which"the"bacterial"partner"triggers"the"production"of"fungal"metabolites"otherwise"not"observed"in"axenic"cultures."Furthermore,"we"report"on"the"ecological"function"of"one"newly/identified"fungal"SM"in"co/culture,"as"well"as"the"identification"of"a"fungal"transcription"factor"acting"as"a"key"regulatory"node"for"‘translating’"bacterial"signals"leading"to"regulation"of"SM"gene"clusters"(4).""(1)"Brakhage"(2013)"Nature,"(2)"Macheleidt"et"al.,"(2016)"Annu"Rev"Genet,"(3)"Netzker"et"al.,"(2018)"Curr"Opin"Microbiol,"(4)"Fischer"et"al.,"(2018)"eLIFE""*85.$Comparative$genomics$of$endoparasitic$fungi,$Esteya*vermicola""Jongoh"Shin1,"Jaemin$Sung1,"Byungkwan"Cho1,2."1)'Department'of'Biological'Sciences'and'KAIST'Institute'for'the'BioCentury,'Korea'Advanced'Institute'of'Science'and'Technology,'Daejeon'305Q701,'Republic'of'Korea6'2)'Intelligent'Synthetic'Biology'Center,'Daejeon'305Q701,'Republic'of'Korea""Esteya'vermicola"is"the"first"reported"endoparasitic"nematophagous"fungus"that"attacks"the"pinewood"nematode,"the"Bursaphylenchus'xylophilus."Although"the"infection"activities"of"E.'vermicola"had"been"verified"as"a"biocontrol"agent"of"pine"wilt"disease,"their"infection"mechanism"has"not"been"unraveled."To"understand"their"unique"pathogenic"properties,"we"compared"6"nematophagous"fungi"including"E.'vermicola"to"identify"pathogenic"genes"that"are"unique"and"conserved"among"them."A"targeted"comparison"with"nematophagous"fungi"highlighted"nematophagous"fungi"were"equipped"with"diverse"pathogenic"genes"which"hypothetically"play"role"in"penetration"and"digestion"processes."Particularly,"genes"encoding"glycoside/degrading"enzymes"and"membrane"transporters"were"highly"expanded"in"E.'vermiocla."It"also"permitted"the"initial"characterization"of"several"secondary"metabolite"gene"clusters"in"E.'vermicola,"which"show"the"great"potential"of"virulence."These"results"provide"a"useful"genetic"information"for"the"understanding"of"infection"mechanism"of"nematode/destroying"fungi"and"open"the"way"for"molecular"dissection"of"E.'vermicola"physiology.""*86."IntraLspecific$aflatoxin$inhibition$profile$by$atoxigenic$Aspergillus*flavus$reveals$touchLindependent$inhibition""R.R.$Sweany,"C.D."DeRobertis,"K.E."Damann"Jr..""Plant'Pathology'and'Crop'Physiology,'Louisiana'State'University,'Baton'Rouge,'LA"""Atoxigenic"biocontrol"strains"competitively"prevent"aflatoxin"contamination"by"direct"replacement"enhanced"by"thigmoregulated"“touch”"inhibition."This"study"was"conducted"to"determine"the"extent"of"aflatoxin"inhibition"against"diverse"toxigenic"strains"and"if"growth"rates"contribute"to"successful"inhibition."To"assess"intra/specific"inhibition,"aflatoxin"was"extracted"and"quantified"from"20"toxigenic"A.'flavus"isolates"representing"both"S"(sclerotia"<"400"µm)"and"L"(sclerotia">"400"μm)"strains"and"both"mating"types"(M1Q1"and"M1Q2)"grown"in"co/culture"with"7"atoxigenic"biocontrol"strains"(L,"M1Q2)"for"4"days"at"25°C."Atoxigenic"growth"rates"were"estimated"with"quadratic"linear"growth"curves"for"dry"weights"of"mycelia"harvested"after"36,"48"and"72"hours"of"growth.""Atoxigenic"strains"differentially"inhibited"toxigenic"strains:"M1Q1"(S"or"L)"or"small"strains"were"less"inhibited"than"M1Q2,"L"strains.""When"atoxigenic"17"(from"LA"soils)"was"co/cultured"with"L,"M1Q2"strains"aflatoxin"below"the"safe"consumption"limit"(20ppb)"was"produced.""Atoxigenic"strains"with"faster"biomass"growth"rates"were"the"best"inhibitors,"suggesting"more"touch"leads"to"greater"inhibition."In"contrast,"two"strains"inhibited"aflatoxin"substantially"in"spite"of"relatively"slow"growth,"therefore"inhibition"through"a"touch/independent"mechanism"was"investigated.""Atoxigenic"strains"were"grown"in"24/well"plates"separated"from"a"common"toxigenic"strain"(53)"by"0.4"μm"filter"inserts."Additionally,"53"was"grown"in"medium"amended"(1:1)"with"2"and"4"day"culture"filtrates"of"the"atoxigenic"strains."Strain"53"produced"low"aflatoxin"when"separated"from"fastest/growing"strain,"17,"and"slow/growing,"highly/inhibitory"strains"K49"and"Af36."The"addition"of"4"day"AF36,"K49"and"17"filtrates"to"medium"reduced"53’s"aflatoxin"production."Filtrate"from"17"resulted"in">90%"inhibition.""Filtrate"of"51"did"not"inhibit"production,"eliminating"the"possibility"of"a"nutrient"depletion"effect."Aflatoxin"reduction"by"filtrate"and"across"inserts"provides"evidence"of""secreted"soluble"inhibitor(s)"of"aflatoxin"production."Expansion"of"inhibition"specificity"profile,"not"only"revealed"limited"inhibition"of"Mat1Q1"and"S"strains,"but"revealed"extracellular"chemicals"inhibit"aflatoxin"production"therefore"touch"and"direct"replacement"are"not"the"sole"causes"of"aflatoxin"elimination"by"biocontrol"strains.""$

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87.$Live$cell$imaging$analysis$of$fungalLbacterial$interaction""N.$Takeshita,"M."Kuchira,"G."Abeysinghe,"D."Hagiwara,"N."Nomura,"N."Takaya,"N."Obana.""Microbiology'Research'Center'for'Sustainability'(MiCS),'University'of'Tsukuba,'Tsukuba,'JP"""Physical"spaces"to"inhabit"and"nutrients"are"prerequisites"for"surviving"organisms"while"no"interspecies"mutual"strategy"is"documented"that"satisfies"them."Here"we"found"that"co/cultured"bacterium"Bacillus'subtiliscells"travel"along"mycelia"of"the"fungus"Aspergillus'nidulansat"a"rate"of"~30"μm/s"and"disperse"with"fungal"colony"expansion."These"values"largely"exceed"those"observed"in"single/cultured"bacterium."This"travel/dispersal"require"intact"bacterial"flagella,"and"result"in"bacterial"colonization,"indicating"that"the"fungal"hyphae"are"beneficial"“highways”"for"the"bacterium"to"explore"spatial"niche"to"inhabit."Besides,"transcriptome"analysis"indicates"the"interspecies"interaction"of"vitamin"biosynthesis."These"evoke"a"novel"mechanism"for"mutualistic"strategy"that"facilitates"the"communicating"species"to"obtain"nutrient"and"environmental"niche."""88."Heterologous$production$and$the$productivity$enhancement$of$polyunsaturated$free$fatty$acids$by$metabolic$engineering$of$Aspergillus*oryzae""K."Tamano1,2,"A."Itoh3,"Y."Yasunaka3,"A."Miura1,"E."Kan4,"Y."Koyama4,"T."Tamura1,2.""1)'National'Institute'of'Advanced'Industrial'Science'and'Technology'(AIST)'BPRI,'Sapporo,'Hokkaido,'JP6'2)'National'Institute'of'Advanced'Industrial'Science'and'Technology'(AIST)'CBBDQOIL,'Sapporo,'Hokkaido,'JP6'3)'Hokkaido'High'Technology'College,'Eniwa,'Hokkaido,'JP6'4)'Noda'Institute'for'Scientific'Research,'Noda,'Chiba,'JP"""""""Aspergillus'oryzae"is"well/known"as"an"industrial"filamentous"fungus"by"the"high"enzyme"production"and"safety,"and"thus"has"been"utilized"for"manufacturing"Japanese"alcohol,"soy"sauce,"etc."We"focused"on"such"superior"characteristics"of"this"fungus"and"have"studied"for"applying"it"to"production"of"valuable"compounds."Several"free"fatty"acids"(FFAs)"and"the"derivatives"produced"by"microorganisms"are"known"to"be"useful"as"pharmaceuticals,"dietary"supplements,"and"their"feedstocks."Thus,"we"attempted"to"make"A.'oryzae"produce"such"useful"FFAs"in"a"heterologous"expression"manner"as"well"as"enhance"the"productivity"by"the"metabolic"engineering."To"date,"FAA"productivity"was"successfully"enhanced"at"9.2/fold"more"than"the"wild/type"strain"by"disrupting"one"acyl/CoA"synthetase"gene"faaA"out"of"the"six"genes."Also,"the"cDNA"fragments"of"three"enzyme"genes"responsible"for"fatty"acid"conversion"of"linoleic"acid"(C18:2,"LA)"to"arachidonic"acid"(C20:4ω6,"ARA)"were"prepared"from"the"other"filamentous"fungus"Mortierella'alpina"and"expressed"in"the"faaA"disruptant."However,"the"constructed"strain"named"ARA1"produced"only"free"fatty"acids"up"to"dihomo/ɤ/linolenic"acid"(C20:3ω6,"DGLA)"that"was"a"precursor"of"free"ARA,"and"did"not"produce"free"ARA."It"remains"to"be"elucidated"why"free"ARA"cannot"be"produced."On"the"other"hand,"free"DGLA"is"also"useful"as"a"feedstock"of"some"pharmaceuticals,"and"then"we"attempted"to"enhance"productivity"of"free"DGLA"as"increasing"the"content"ratio."""""In"the"ARA1"strain,"saturated"FFAs"that"are"palmitic"acid"(C16:0,"PA)"and"stearic"acid"(C18:0,"SA)"exist"at"about"70%"content"ratio"in"the"total"FFA."It"is"predicted"that"ten"desaturase"genes"involved"in"the"conversion"of"SA"to"LA"originally"exist"in"the"A.'oryzae'genome."They"are"unknown"about"which"encode"Δ9/desaturase"or"Δ12/desaturase,"and"also"about"which"are"the"predominant"desaturase"genes."Thus,"these"ten"desaturase"genes"were"individually"overexpressed"in"the"ARA1"strain,"and"their"overexpression"effect"on"content"ratio"of"polyunsaturated"FFAs"such"as"free"DGLA"was"observed."As"a"result,"two"desaturase"genes"out"of"ten"only"showed"increased"effect"on"desaturation"by"overexpression."One"accelerated"conversion"of"SA"to"oleic"acid"(C18:1,"OA),"and"the"other"did"conversion"of"OA"to"LA."Consequently,"the"free"DGLA"content"ratio"was"maximally"attained"at"9.5%"in"the"total"FFA,"and"the"maximal"free"DGLA"yield"is"190"mg/L"for"5/day"culture."We"are"currently"attempting"combination"of"the"overexpression"of"these"two"selected"desaturase"genes"in"the"ARA1"strain,"and"aiming"at"further"increase"of"free"DGLA"production.""$ $

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89.$Identification$and$distinct$regulation$of$oligopeptide$transporter$genes$in$Aspergillus*oryzae$"M.$Tanaka1,"K."Ito1,"Y."Kawarasaki1,"K."Gomi2"1)"School"of"Food"and"Nutritional"Science,"University"of"Shizuoka,"Shizuoka,"JPS"2)"Graduate"School"of"Agricultural"Science,"Tohoku"University,"Sendai,"JP"'Aspergillus'oryzae"possesses"over"130"proteinase"genes"in"the"genome,"and"expression"of"these"genes"is"induced"by"soy"peptides."In"eukaryotic"cells,"the"proton/dependent"oligopeptide"transporter"(POT)"family"incorporates"mainly"di/"and"tri/peptide,"and"Ptr2p"is"the"sole"founding"POT"member"in"budding"yeast."Interestingly,"15"putative"transporters"having"homology"to"the"Ptr2p"were"found"from"A.'oryzae"genome."However,"these"putative"transporters"have"not"been"characterized."In"this"study,"we"identified"three"transporters"having"the"ability"to"uptake"of"di/"and"tri/peptides,"and"we"further"examined"the"regulatory"mechanism"of"these"transporter"genes"expression."""RT/PCR"analysis"revealed"that"four"putative"transporter"genes"were"highly"expressed"under"the"culture"condition"using"soy"peptides"as"nitrogen"source."Among"them,"three"transporters"were"able"to"complement"the"ability"of"di/"and"tri/peptides"uptake"in"yeast"PTR2∆"mutant."Therefore,"we"designated"these"transporters"as"PotA,"PotB,"and"PotC,"respectively."Quantitative"RT/PCR"analysis"showed"that"potB"and"potC"constitutively"expressed,"whereas"potA"expression"was"only"induced"by"soy"peptides."In"budding"yeast,"expression"of"PTR2"was"repressed"by"a"transcription"factor"Cup9,"and"Ubr1/dependent"proteolytic"degradation"of"Cup9"induces"PTR2"expression"when"tryptophan"or"di/peptides"are"added."In"A.'oryzae,"expression"of"all"POT"genes"was"not"up/regulated"by"disruption"of"cup9"ortholog,"although"potA"and"potC"expression"in"the"medium"containing"soy"peptides"was"repressed"by"disruption"of"ubr1"ortholog."Disruption"of"prtR"gene,"encoding"a"transcription"factor"for"secretory"proteinase"genes,"repressed"expression"level"of"potA"and"potB."In"addition,"overexpression"of"prtR"induced"potA"and"potB"expression"in"the"medium"lacking"soy"peptides."In"contrast,"expression"of"potC"was"neither"affected"by"disruption"nor"overexpression"of"prtR."These"results"suggest"that"expression"of"each"oligopeptide"transporter"gene"is"controlled"by"a"different"regulatory"mechanism."$90."The$secondary$metabolism$regulator$LaeA$in$Penicillium*sp.$influences$the$cheese$rind$community$assembly""J."Tannous1,"C."Cosetta2,"T."Rush3,"J."Jiler2,"N."Keller1,4,"B."Wolfe2.""1)'Department'of'Medical'Microbiology'and'Immunology,'University'of'Wisconsin'–'Madison,'Madison,'WI,'USA6'2)'Department'of'Biology,'Tufts'University,'Medford,'MA,'USA'6'3)'Department'of'Plant'Pathology,'University'of'Wisconsin'Q'Madison,'Madison,'WI,'USA6'4)'Department'of'Bacteriology,'University'of'Wisconsin'QMadison,'Madison,'WI,'USA"""Species"within"the"genus"Penicillium"are"widely"known"producers"of"secondary"metabolites,"including"polyketides"(e.g."patulin),"non/ribosomal"peptides"(e.g."roquefortine)"and"terpenes"(e.g."gibberellin)."Among"these"metabolites"there"many"are"valued"as"pharmaceuticals,"whereas"others"termed"as"mycotoxins"are"detrimental"due"to"their"acute"and"chronic"toxicities"and"yield"losses"they"engender."Over"the"past"years,"a"significant"body"of"work"has"emphasized"the"role"of"LaeA"as"a"global"regulator"of"fungal"secondary"metabolites."Yet,"the"impact"of"LaeA"on"microbial"communities"is"poorly"understood."In"this"study,"cheese"rinds"were"shown"to"be"an"ideal"system"to"test"the"ecological"impacts"of"LaeA"in"polymicrobial"interactions."The"deletion"of"this"gene"in"a"cheese"Penicillium"species,"closely"related"to"P.'cyclopium,"influences"ecologically"relevant"phenotypic"and"metabolomic"properties,"including"spore"production,"germination"rate"and"secondary"metabolite"biosynthesis."Moreover,"the"lack"of'laeA"in"this"species"has"an"impact"on"the"composition"of"cheese"rind"bacterial"communities"associated"with"altered"production"of"metabolites"with"antimicrobial"properties."This"study"highlights"the"role"of"fungal"secondary"metabolism"in"and"identifies"a"strategy"to"find"novel"compounds"associated"with"useful"biologic"activities."$$ $

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91.$Peptidyl$compound$synthetic$factors$widely$conserved$and$highly$diverse$in$the$Fungi$kingdom""Maiko"Umemura1,2,"Linh"Dao1,2,"Yuki"Nakashima1,2.""1)'Bioproduction'Research'Institute,'AIST,'Tsukuba,'Ibaraki,'Japan6'2)'Computational'Bio'Big'Data'Open'Innovation'Laboratory'(CBBDQOIL),'AIST,'Tsukuba,'Ibaraki,'Japan""""Repertories"of"secretory"peptides"have"been"found"in"plants"and"insects"as"signaling"molecules,"but"knowledge"of"those"in"fungi"are"limited."Recently,"we"identified"the"gene"cluster"responsible"for"biosynthesis"of"ustiloxin,"which"is"a"cyclic"peptide"composed"of"five"amino"acids,"from"Aspergillus'flavus"as"the"first"case"of"ribosomally"synthesized"and"post/translationally"modified"peptide"(RiPP)"synthetic"pathway"in"Ascomycetes."RiPPs"are"biosynthesized"from"precursor"peptides,"which"contains"peptide"sequences"used"for"compound"backbone"structures"(core"peptides)."In"the"ustiloxin"case,"its"precursor"peptide"had"another"two"distinctive"featuresS"a"signal"peptide"to"endoplasmic"reticulum,"and"highly"repeated"core"peptide"sequences"processed"by"Kex2"protease"in"Golgi"apparatus."Based"on"these"characteristics,"we"revealed"that"ustiloxin/type"RiPPs"precursor"peptide"(ust/RiPPs)"genes"were"widely"found"among"the"genus"Aspergillus,"meanwhile"the"core"peptide"sequences"were"diversified"into"more"than"40"distinct"types"(Nagano"et"al.,"FGB,"86:58,"2016)."From"the"detected"gene"information,"we"also"identified"a"novel"cyclic"peptide,"asperipin/2a,"from"Aspergillus'flavus."Even"though"both"cyclic"peptides"are"secreted"by"cells,"their"role"in"fungal"ecology"is"still"uncharacterized.""Here,"to"obtain"information"for"ust/RiPPs"functions,"we"mined"entire"genomic"data"of"857"strains"belonging"to"the"Fungi"kingdom"for"ust/RiPPs"genes."As"a"result,"2,423"ust/RiPPs"genes"were"found"from"84%"strains"in"entire"phylums"of"Fungi,"including"Ascomycota"and"Basidiomycota."Their"core"peptide"sequences,"which"are"in"length"from"6"to"45"a.a."and"repeated"more"than"three"times,"were"classified"into"more"than"1,500"types"using"the"thresholds"of"65%"identity"and"90%"bidirectional"coverage."Although"the"largest"group"of"type"contained"473"members,"the"size"of"group"rapidly"declined,"and"the"22nd"largest"group"of"type"contained"just"50"members."One/third"of"types"including"the"most"major"one"had"the"motif"of"yeast"mating"factor,"whilst"another"one/third"had"gene(s)"with"the"motif"of"cyclization"factor"specific"to"ust/RiPPs"pathway"within"three"genes.""These"results"illustrate"that"ust/RiPPs"genes"are"widely"conserved"in"the"Fungi"kingdom,"but"core"peptide"sequences"are"highly"diverse."It"can"be"suggested"that"the"large"group"of"ust/RiPPs"factors"plays"important"roles"as"fungal"secretory"peptides"like"pheromones"or"hormones"to"communicate"among"cells"in"the"same"and/or"other"organisms."We"will"try"to"obtain"a"wide"variety"of"ust/RiPPs"compounds"by"heterologous"expression"to"unclear"their"actual"role"in"fungi,"as"well"as"their"effects"on"mammalian"cells"as"acceptor"of"these"factors."$92.$Surface$analysis$tools$identify$how$Aspergillus*niger*and$its$enzymes$modify$lignocellulose""J.M.$van"Munster1,"T.V."Vuong2,"S.E."Marcus3,"J.P."Knox3,"E.R."Master2.""1)'Manchester'Institute'of'Biotechnology,'University'of'Manchester,'Manchester,'UK6'2)'Department'of'Chemical'Engineering'and'Applied'Chemistry,'University'of'Toronto,'Toronto,'Canada6'3)'Centre'for'Plant'Sciences,'Faculty'of'Biological'Sciences,'University'of'Leeds,'Leeds,'UK"""Fungal"carbohydrate"active"enzymes"are"exploited"on"an"industrial"scale"as"biocatalyst"that"convert"plant"lignocellulose"to"simple"sugars,"which"can"subsequently"be"converted"to"platform"chemicals"and"biofuels."However,"we"have"poor"knowledge"of"the"effect"of"fungi"and"their"enzymes"on"the"actual"insoluble"complex"substrate,"while"such"understanding"underpins"exploitation"of"enzymes"to"make"designer"polysaccharide"materials,"effective"degradative"enzyme"cocktails"and"to"engineer"fungal"production"strains.""We"investigated"how"exposure"and"accessibility"of"polysaccharides"and"lignin"on"lignocellulose"surface"changes"during"cultivation"with"industrially"relevant"fungus"Aspergillus'niger,"with"the"aim"to"understand"how"this"fungus"and"its"enzymes"interacts"with"lignocellulose."""Analysis"of"time/staged"changes"of"lignocellulose"using"mass/spectrometry"based"imaging"identified"surface"exposure"of"lignin"was"increased"over"time."We"identified"differential"degradation"of"hemicellulose"and"pectin"polysaccharides"using"immunohistochemistry"and"fractionation"followed"by"ELISAs"employing"antibodies"specific"for"carbohydrate"epitopes."Degradation"of"specific"polysaccharides"was"not"always"

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linked"to"presence"of"known"corresponding"degradative"enzymes,"suggesting"lack"of"substrate"access"or"absence"of"essential"accessory"enzymes.""Our"results"highlight"that"full"understanding"of"fungal"and"enzymatic"lignocellulose"degradation"requires"a"combination"of"enzyme"biochemical"data"with"identification"of"modifications"in"real,"complex"lignocellulose"materials."The"understanding"underpins"engineering"of"more"effective"biocatalysts"and"their"exploitation"in"either"break"down"of"lignocellulose"or"modification"to"glyco/materials."$93."Discovery$and$elucidation$of$the$biosynthesis$of$aspernidgulenes,$novel$polyenes$from$Aspergillus*nidulans,$using$serial$promoter$replacement"""Tzu/Shyang"Lin1,"Bethany"Chen1,"Yi/Ming"Chiang1,2,"Clay$C.$C.$Wang1,3.""1)'Pharmacology'&'Pharmaceutical'Science,'University'of'Southern'California,'Los'Angeles,'CA6'2)'Department'of'Pharmacy,'Chia'Nan'University'of'Pharmacy'and'Science,'Tainan,'Taiwan6'3)'Department'of'Chemistry,'University'of'Southern'California,'Los'Angles,'CA"""Through"serial"promoter"exchanges,"several"novel"polyenes,"named"aspernidgulenes,"were"isolated"from"Aspergillus'nidulans."The"biosynthesis"pathway"involving"only"four"enzymes,"highlighting"the"efficiency"of"fungal"tailoring"enzymes"in"chemical"transformation."Comparison"with"citreoviridin"and"aurovertin"biosynthesis"reveals"that"α/pyrone"methylation"is"the"branching"point"where"the"biosynthesis"of"methylated"α/pyrone"polyenes"and"dimethylatedγ/lactone/"or"oxabicyclo[2.2.1]heptane"ring/containing"polyenes"diverge."Thus,"the"presence"or"absence"of"methyltransferase"is"a"potential"predictor"of"product"structure"in"homologous"clusters.""""""$*94."Elucidating$biosynthetic$pathway$of$a$compound$isolated$from$Aureobasidiumpullulans""Jingyi"Wang1,"Adriana"Blachowicz1,2,"Ngan"Pham1,"Su"Jeung"Lim1,"Tamas"Torok3,"Kasthuri"Venkateswaran2,"Clay"C."C."Wang1,4.""1)'Department'of'Pharmacology'and'Pharmaceutical'Sciences,'School'of'Pharmacy,'University'of'Southern'California,'Los'Angeles,'CA6'2)'Biotechnology'and'Planetary'Protection'Group,'Jet'Propulsion'Laboratory,'California'Institute'of'Technology,'Pasadena,'CA6'3)'Ecology'Department,'Lawrence'Berkeley'National'Laboratory,'Berkeley,'CA6'4)'Department'of'Chemistry,'Dornsife'College'of'Letters,'Arts'and'Sciences,'University'of'Southern'California,'Los'Angeles,'CA"""Fungal"secondary"metabolites"(SMs)"are"a"rich"source"of"medically"useful"compounds"due"to"their"varying"biological"activities."Up"to"date,"biosynthesis"of"many"SM"is"still"not"fully"understood."However,"the"rapid"increase"in"the"number"of"sequenced"fungal"genomes"and"development"of"versatile"gene"targeting"methods"enabled"genetic"manipulation"of"non/model"filamentous"fungi"leading"to"the"discovery"of"novel"SMs."SMs"are"not"directly"involved"in"fungal"survival,"however"may"confer"environmental"advantage"that"enables"adaptation"to"extreme"habitats."For"example,"it"was"proposed"that"melanins"help"to"shield"the"fungi"from"harmful"irradiation"and"help"to"convert"irradiation"into"an"energy"source."Chernobyl/isolated"fungi"are"extremophiles"that"have"adapted"to"the"high"irradiation"environment"and"shown"the"unknown"phenomenon"of"growing"towards"the"radiation"source,"referred"as"radiotropism."Aureobasidiumpullulans'IMV00882"is"one"of"the"nuclear"power"plant"accident/associated"fungi"that"were"selected"for"SM"characterization."SM"analysis"of"IMV"00882"via"LCMS"showed"high"production"yield"of"a"compound"whose"biosynthetic"pathway"remains"cryptic."To"elucidate"the"unknown"biosynthetic"pathway,"we"are"going"to"use"CRISPR/Cas9"system"provided"by"Professor"Uffe"Mortensen.CRISPR/Cas9"the"bacterial"and"archaeal"immune"mechanism"consists"of"two"components,"the"Cas9"nuclease"and"a"single"chimeric"guide"RNA"(sgRNA),"which"allows"the"introduction"of"specific"DNA"double/strand"breaks,"and"in"turn,"efficient"gene"targeting."The"first"step"in"creating"CRISPR/Cas9/based"genetic"system"for"the"IMV"00882"strain"was"optimizing"the"protoplasting"protocol"followed"by"the"selection"of"the"marker"gene."To"easily"identify"the"transformants"with"mutations"in"the"target"cluster,"we"will"use"phleomycin"as"the"marker."Upon"disrupting"genes"involved"in"the"production"of"the"compound"of"interest"we"will"use"HPLC"and"NMR"to"analyze"the"structure"of"intermediates"and"propose"a"biosynthetic"pathway."

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Unraveling"this"uncharacterized"biosynthetic"pathway"may"lead"to"optimizing"the"production"process"of"this"chemically"interesting"compound."$*95.$The$function$of$long$nonLcoding$RNA$lncR6731$in$a$plant$pathogenic$fungus$Fusarium*graminearum""J."Wang1,2,"W"Zeng1,2,"Y"Fu2,"D"Jiang1,2,"J"Cheng1,2.""1)'State'Key'Laboratory'of'Agricultural'Microbiology,'Huazhong'Agricultural'University,'Wuhan'430070,'Hubei'Province,'China6'2)'The'Provincial'Key'Lab'of'Plant'Pathology'of'Hubei'Province,'College'of'Plant'Science'and'Technology,'Huazhong'Agricultural'University,'Wuhan'430070,'Hubei'Province,'China""""""""""Fusarium'graminerum'(teleomorph"Gibberella"zeae)"is"one"of"the"major"pathogens"that"cause"Fusarium"head"blight"(FHB)"in"wheat,barley,maize"and"other"grains.Non/coding"RNA"plays"an"important"role"in"the"growth"and"development"of"plants"and"animals."In"previous"research"we"predicted"and"analyzed"long"non/coding"RNA(lncRNAs)"in"F.'graminearum"by"strand/specific"sequencing."We"found"that"a"majority"of"lncRNAs"located"upstream"or"downstream"of"the"protein/coding"genes."In"the"present"study,"the"biological"function"of"a"lncR6731"in"F.'graminearum'was"explored.""''''''''lncR6731"is"located"in"the"upstream"of"a"hypothetical"protein"encoding"gene"FGSG_05042."FGSG_05042"encodes"534"amino"acids"with"two"MFS"structural"domain"belonging"to"the"sugar"transporter"sub/family"of"the"MFS"and"is"predicted"to"contain"14"trans/membrane/spanning"segments."A"Split/Marker"system"was"used"for"lncR6731'and"FGSG_05042"disruption."then"the"phenotype"of"different"strains"were"compared."The"results"showed"that"no"obvious"changes"was"observed"in"hyphal"growth,"conidiation,"and"conidia"morphology"between"the"wild"type"strain"PH/1"and"knockout"strains"ΔlncR6731'and"ΔFGSG_05042."During"sexual"development,"normal/shaped"perithecia"were"obtained"from"these"mutants"at"7"days"after"fertilization,"but"few"cirrhi"were"produced"in"ΔlncR6731"and"ΔFGSG_05042."The"discharged"ascospores"were"also"significantly"decreased"in"ΔlncR6731'and"ΔFGSG_05042."The"virulence"of"different"strains"were"detected"with"flowering"wheat"heads,"corn"silks"and"coleoptiles"inoculation."Results"showed"that"the"virulence"of"ΔFGSG_05042"and"ΔlncR6731"mutants"was"significantly"reduced"when"inoculated"with"corn"silks"and"coleoptiles."However,"only"a"slight"decrease"in"virulence"was"observed"when"inoculated"with"flowering"wheat"heads."These"results"suggested"that"lncR6731"and"FGSG_05042"play"important"roles"in"virulence"and"the"sexual"development"stage"in"F.'graminearum."Furthermore,"comparison"with"the"wild"type"PH/1,"the"DON"production"in"the"inoculated"floret"inoculated"with"the"ΔFGSG_05042'and'ΔlncR6731"mutants"were"remarkable"increased"and"the"expression"level"of"TRI"family"gene"were"also"upregulated"significantly."These"result"indicates"that"FGSG_05042"negatively"controls"DON"production"in"F.'graminearum."""""""""""Taken"together,"our"findings"indicate"that"FGSG_05042'and'lncR6731'plays"a"critical"role"in"the"regulation"of"sexual"development,"pathogenesis,"and"DON"biosynthesis"in"F.'graminearum.'But"the"regulation"mechanism"and"the"relationship"between"FGSG_05042'and'lncR6731'needs"further"study.""96."FineLtuning$gene$expression:$Pantothenic$acid$inducible$promoters$in$Trichoderma*reesei""Franziska"Wanka1,"Robert"H."Bischof1,"Benjamin"Metz1,"Christian"Gamauf2,"Georg"Schirrmacher2,"Christian"P."Kubicek1,3","Bernhard"Seiboth1,3.""1)'Austrian'Centre'of'Industrial'Biotechnology'(ACIB)'GmbH'c/o'Institute'of'Chemical,'Environmental'and'Biological'Engineering,'TU'Wien,'Vienna,'Austria6'2)'Group'Biotechnology,'Clariant'Produkte'(Deutschland)'GmbH,'Planegg,'Germany6'3)'Synthetic'Biology'and'Molecular'Biotechnology,'Research'Division'Biochemical'Technology,'Institute'of'Chemical,'Environmental'and'Bioscience'Engineering,'TU'Wien,'Vienna,'Austria""'T.'reesei"is"the"most"widely"employed"producer"of"cellulases"and"hemicellulases."Inducible"recombinant"protein"expression"in"this"organism"is"currently"hampered"by"the"relatively"small"repertoire"of"suitable"promoters,"which"are"derived"from"(hemi)cellulase"encoding"genes."Therefore"there"is"an"urgent"demand"for"novel"tunable"promoters"acting"in"a"(hemi)cellulase"independent"fashion."Several"different"substances"as"pantothenic"acid,"thiamine,"L/serine,"niacin"ect."were"tested"in"wheat"straw"cultivation"for"their"influence"on"T.'reesei’s"biomass"or"(hemi)cellulase"production."Addition"of"up"to"100"µM"pantothenic"acid"to"T.'reesei"did"not"alter"growth"or"the"expression"profile"of"(hemi)cellulase."Using"whole"transcriptome"analysis,"five"T.'reesei"genes"were"found"that"were"strongly"induced"by"the"addition"of"pantothenic"acid."The"promoter"regions"of"two"of"those"pantothenic"acid"induced"genes"were"fused"to"the"green"fluorescent"

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protein"gene"(gfp)"as"reporter."Transcription"analysis"showed"that"expression"of"gfp"was"pantothenic"acid"inducible"and"surpassed"the"expression"of"the"reference"strain"with"the"strong"tef1"promoter"in"one"case."Fluorescence"emission"measurements"of"GFP"using"a"plate"reader"indicated"that"the"inducibility"of"the"pantothenic"acid"promoters"is"not"specific"to"wheat"straw"as"carbon"source,"also"works"in"glycerol"medium."The"protein"expression"values"confirmed"the"transcription"analyses,"but"under"these"conditions"more"GFP"is"present"in"the"Ptef1_gfp"strain"as"GFP"is"constitutively"formed."This"inducible"system"is"inexpensive"and"adjustable,"and"can"therefore"be"used"as"valuable"tool"for"strain"engineering"of"industrial"T.'reesei"strains."$97.$The$unfolded$protein$response$is$linked$with$Ca2+$homeostasis$to$mediate$stress$adaptation$and$cell$wall$integrity$in$Aspergillus*fumigatus""M."Weichert1,"J.P."Guirao/Abad1,"K."Krishnan1,"V."Aimanianda2,"C."Grisham1,"R."Abbu1,"A."Sheehan1,"D.S."Askew1.""1)'Department'of'Pathology'and'Laboratory'Medicine,'University'of'Cincinnati,'Cincinnati,'OH,'USA6'2)'Unité'des'Aspergillus,'Institut'Pasteur,'Paris,'France"""The"unfolded"protein"response"(UPR)"is"an"adaptive"stress"response"in"eukaryotic"cells"that"orchestrates"changes"in"gene"expression"to"restore"proteostasis"under"conditions"of"increased"stress"in"the"endoplasmic"reticulum"(ER)."Although"we"have"previously"shown"that"the"human/pathogen"fungus"Aspergillus'fumigatus"heavily"relies"on"this"pathway"for"both"virulence"and"antifungal"drug"tolerance,"the"mechanistic"details"involved"are"still"poorly"understood."Here"we"demonstrate"that"ER"stress"increases"the"expression"of"pmrA,"encoding"a"Ca2+"ATPase"previously"localized"to"the"Golgi"apparatus,"as"well"as"srcA,"which"codes"for"a"homolog"of"mammalian"SERCA"(sarco/endoplasmic"reticulum"Ca2+"ATPase)"pumps"that"is"uncharacterized"in"A.'fumigatus."Since"these"proteins"transfer"cytosolic"Ca2+"into"the"secretory"pathway"to"support"protein"folding"and"sorting,"we"sought"to"determine"the"contribution"of"PmrA"and"SrcA"to"ER"stress"responses."Surprisingly,"gene"deletion"mutants"lacking"either"pmrA"or"srcA"were"resistant"to"the"ER"stress"agents"dithiothreitol,"tunicamycin"and"brefeldin"A."However,"thermal"stress"delayed"germination"rates"of"ΔpmrA"and"ΔsrcA,"and"both"mutants"showed"increased"sensitivity"to"carvacrol,"a"natural"antifungal"compound"that"disrupts"ER"morphology"and"impairs"Ca2+"homeostasis."Unlike"the"single"gene"knockout"strains,"deleting"both"pmrA"and"srcA"severely"impaired"growth"in"the"absence"of"exogenous"stress"and"resulted"in"compact"colonies"with"a"popcorn/like"morphology,"hyperbranching"filaments,"and"poor"conidiation."Evaluation"of"the"double"mutant"by"transmission"electron"microscopy"revealed"a"thickened"cell"wall,"and"biochemical"analysis"showed"an"increase"of"the"alkali/soluble"fraction"of"the"wall"and"a"marked"depletion"of"galactomannan."Since"the"increased"expression"of"srcA"during"ER"stress"in"A.'fumigatus"has"been"shown"to"be"dependent"on"the"UPR,"we"hypothesized"that"failure"to"induce"this"pathway"would"impair"growth"under"conditions"that"disrupt"Ca2+"homeostasis."Consistent"with"this"notion,"we"found"that"the"deletion"of"the"transcription"factor"required"for"UPR"activation,"HacA,"not"only"caused"hypersensitivity"to"Ca2+"stress"agents,"including"BAPTA,"carvacrol,"amiodarone,"and"cyclopiazonic"acid,"but"even"synergized"with"the"calcineurin"inhibitors"cyclosporine"A"and"FK506."Interestingly,"while"the"osmotic"stabilization"with"sorbitol"protected"ΔhacA"from"compounds"that"affect"Ca2+"homeostasis"and"signaling,"the"addition"of"exogenous"Ca2+"significantly"rescued"this"UPR"mutant"from"both"ER"and"cell"wall"stress."Taken"together,"these"studies"establish"a"novel"link"between"HacA"and"Ca2+"homeostasis"in"A.'fumigatus,"and"indicate"partially"overlapping"roles"of"the"pumps"SrcA"and"PmrA"in"regulating"the"cytosolic"and"ER/Golgi"luminal"Ca2+"levels"during"the"stress"adaptation"of"this"pathogenic"fungus."$$ $

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*98.$Discovery$and$activation$of$a$tetracyclineLlike$producing$gene$cluster$in$Aspergillus*sydowii""P.$B.$Wolff1,"K."Subko1,"S."Theobald1,"J."B."Hoof1,"J."C."Frisvad1,"C."H."Gotfredsen2,"M."R."Andersen1,"U."H."Mortensen1,"T."O."Larsen1.""1)'DTU'Bioengineering,'Technical'University'of'Denmark,'Lyngby,'DK6'2)'DTU'Chemistry,'Technical'University'of'Denmark,'Lyngby,'DK"""Polycyclic"aromatic"polyketides"represent"an"essential"group"of"Natural"Products"(NPs)"exhibiting"a"broad"variety"of"antimicrobial"and"cytotoxic"activities."Among"these"cyclic"compounds,"tetracycline"stands"out"for"its"bioactivity"in"various"applications,"i.e.,"namely"anti/inflammatory,"anticarcinogenic,"but"accurately"known"as"a"pharmaceutical"agent"with"a"broad"spectrum"of"activity"against"bacteria."With"the"ever/rising"resistance"against"all"antibiotics"on"a"global"scale,"it"has"never"been"more"important"to"locate"new"reservoirs"to"gene"mine"for"novel"antibiotics."In"this"study,"we"research"the"genus"of"Aspergillus"and"determine"potential"tetracycline"gene"cluster"(GC)"candidates"using"known"fungal"tetracycline"PKSs"for"comparison."Since"most"secondary"metabolite"(SM)"GCs"are"not"expressed"under"standard"laboratory"conditions,"we"were"able"to"activate"a"silent"putative"tetracycline/like"GC"in"Aspergillus'sydowii"by"overexpression"of"a"pathway/specific"transcription"factor"(TF)."We"show"here"the"production"of"multiple"SMs,"several"with"tetracyclic"structure."We"use"both"inducible"and"constitutive"promoters"to"control"the"expression"level"of"the"regulatory"protein,"and"hence"the"production"of"SMs"related"to"the"GC."By"the"construction"of"deletion"strains"and"supportive"real/time"quantification"PCR,"we"suggest"a"biosynthetic"pathway"for"several"of"the"discovered"SMs"and"show"that"the"Aspergillus"genus"has"the"potential"to"contribute"in"the"search"for"new"antibiotics"$*99.$The$Zn2Cys6Ltype$transcription$factor$LeuBLinvolved$interconnection$between$BCAA$biosynthesis$and$the$Ca2+$signaling$pathways$in$Aspergillus*fumigatus""J.$Ye,"N."Long,"L."Gao,"L."Lu.""Nanjing'Normal'University,'Nanjing,'Jiangsu,'CN"""Aspergillus'fumigatus"is"an"opportunistic"pathogen"and"allergen"of"mammals."Previously,"we"identified"that"the"Zn2Cys6/type"transcription"factor"LeuB"in"A.'fumigatus"is"a"multiple"functions"regulon"which"involves"in"regulation"of"not"only"branched/chain"amino"acids"(BCAA)"biosynthesis"and"nitrogen"metabolism"but"also"iron"acquisition"including"siderophore"metabolism."The"metabolism"of"leuBQinvolved"network"is"most"likely"based"on"the"requirement"of"nitrogen"and"iron"for"biosynthesis"of"BCAA."However,"how"these"pathways"interconnect"remain"inquisitive."In"this"study,"by"searching"putative"transcription"targets"obtained"by"chromatin"immunoprecipitation"DNA"sequencing,"we"found"that"47"new"LeuB"putatively"regulated"genes"which"have"not"reported"in"LeuB"homolog"Leu3p"in"yeasts."Among"them,"PmaA"(AFUB_041460),'a'plasma'membrane'H+QATPase'and'Ena1'(AFUB_094610),'a'cationQtransporting'ATPase'have'the'DNA'binding'site'of'LeuB'in'the'promoters'and'have'been'identified'to'be'bound'by'LeuB'by'electrophoretic'mobility'shift'assay.'Interestingly,'colony'phenotypic'assay'displayed'that'compared'to'that'its'parental'wild'type'with'normal'colony'growth,'lack'of'LeuB'showed'hypersensitive'to'high'concentrations'of'extracellular'calcium'(>10'mM)'showing'remarkable'defects'in'hyphal'growth'and'conidiation'which'is'similar'defected'phenotypes'to'that'of'calcineurinQCrzA'related'mutants.'Moreover,'overexpression'of'PmaA'or'additions'of'leucine'or'iron'were'able'to'significantly'rescue'defects'of'LeuB'deletion'mutant'induced'by'high'concentrations'of'calcium.'These'data'suggest'lack'of'LeuB'may'affect'normal'functions'of'CrzAQcalcineurin'during'regulating'of'Ca2+'homeostasis'via'disturbing'transcription'of'PmaA,'and'also'inferring'that'the'normal'calcium'signaling'is'required'for'iron'acquisition'and'BCAA'biosynthesis."$$ $

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100.$In*silico$screened$compound$ZL705$specifically$inhibits$protein$kinase$C$of$filamentous$fungi.""A."Yoshimi1,"A."Sugahara2,"F."Shoji2,"T."Fujioka3,"K."Kawai3,"H."Umeyama4,"K."Komatsu5,"K."Miyazawa2,"K."Abe1,2.""1)'New'Industry'Creation'Hatchery'Center,'Tohoku'University,'Sendai,'JP6'2)'Grad.'Sch.'Agric.'Sci.,'Tohoku'University,'Sendai,'JP6'3)'Kumiai'Chemical'Industry'Co.'Ltd.,'Tokyo,'JP6'4)'Sch.'of'Pharma.'Sci.,'Kitasato'University,'Tokyo,'JP6'5)'InQSilico'Sciences,'Inc.,'Tokyo,'JP"""The"cell"wall"integrity"signaling"(CWIS)"pathway"is"involved"in"fungal"cell/wall"biogenesis."This"pathway"is"composed"of"sensor"proteins,"protein"kinase"C"(PKC),"and"the"mitogen/activated"protein"(MAP)"kinase"pathway,"and"controls"the"transcription"of"many"cell/wall"related"genes."Among"their"signaling"factors,"PKC"plays"a"pivotal"role"in"this"pathwayS"deficiency"in"PkcA"in"the"model"filamentous"fungus"Aspergillus'nidulans'and"in"MgPkc1"in"the"rice"blast"fungus"Magnaporthe'grisea'is"thought"to"be"lethal."This"suggests"that"PKC"in"filamentous"fungi"is"a"potential"target"for"antifungal"agents."In"this"study,"to"search"for"MgPkc1"inhibitors,"we"carried"out"in'silico'screening"by"three/dimensional"structural"modeling"and"performed"growth"inhibition"tests"for"M.'grisea'on"agar"plates."From"approximately"800,000"compounds"in"databases,"we"identified"a"candidate"inhibitor"of"filamentous"fungal"PKC,"Z/705."To"evaluate"the"effects"of"fungal"PKC"inhibitors"in"Saccharomyces'cerevisiae,"we"constructed"a"screening"system"which"can"evaluate"its"inhibitory"activity"against"chimeric"PKC"in"which"the"kinase"domain"of"native"S.'cerevisiae'PKC"was"replaced"with"those"of"PKCs"of"filamentous"fungi."Transcriptional"analysis"of"MLP1,"which"encodes"a"downstream"factor"of"PKC"in"S.'cerevisiae,"and"phosphorylation"analysis"of"the"MAP"kinase"Mpk1p,"which"is"activated"downstream"of"PKC,"revealed"that"Z/705"specifically"inhibited"PKCs"of"filamentous"fungi."Moreover,"the"analysis"of"the"AGS1'transcript"level,"which"is"regulated"by"Mps1p"MAPK"via"PKC,"in"M.'grisea'revealed"that"Z/705"had"a"PKC"inhibitory"effect"comparable"to"that"of"a"well/known"PKC"inhibitor,"staurosporine."Interestingly,"the"β/1,3/glucan"synthase"inhibitor"micafungin"induced"hyphal"melanization"in"M.'grisea,"and"the"melanization,"which"is"required"for"pathogenicity"of"M.'grisea,'was"inhibited"by"PKC"inhibition"by"both"Z/705"and"staurosporine."The"transcript"levels"of"4HNR,"3HNR,"and"SCD1,"which"are"essential"for"melanization"in"M.'grisea,"were"suppressed"by"both"PKC"inhibitors."In"conclusion,"PKC"is"a"promising"target"for"antifungal"drugs,"and"Z/705"is"a"candidate"fungicide"for"agricultural"use."$*101.$Genome$Mining$of$the$Biosynthetic$Gene$Cluster$of$Citrinalin$A$in$Penicillium*citrinum*using$CRISPRLCas9""B."Yuan1,"J"van"Dijk1,"Y"Ye2,"C"Wang1,3,"D"Sherman2,4,5,6.""1)'Department'of'Pharmacology'and'Pharmaceutical'Sciences,'School'of'Pharmacy,'University'of'Southern'California,'Los'Angeles,'CA6'2)'Life'Sciences'Institute,'University'of'Michigan,'Ann'Arbor,'MI6'3)'Department'of'Chemistry,'University'of'Southern'California,'Dornsife'College'of'Letters,'Arts,'and'Sciences,'Los'Angeles,'CA6'4)'Department'of'Medicinal'Chemistry,'University'of'Michigan,'Ann'Arbor,'MI6'5)'Department'of'Chemistry,'University'of'Michigan,'Ann'Arbor,'MI6'6)'Department'of'Microbiology'and'Immunology,'University'of'Michigan,'Ann'Arbor,'MI"""Secondary"metabolite"(SM)"analysis"of"Penicillium'citrinum'showed"that"it"is"a"potent"producer"of"citrinalin,"an"interesting"nitro"compound."Based"on"a"structurally"similar"compound"with"a"known"biosynthetic"pathway,"the"putative"gene"cluster"of"citrinalin"was"proposed."To"confirm"the"genes"involved"in"citrinalin"biosynthesis,"CRISPR/Cas9"technology"was"used"to"create"a"genetic"system"for"P.'citrinum."Plasmids"carrying"Cas9"cassette,"kindly"donated"by"the"Mortensen"lab,"were"used"to"target"the"genes"of"interest."The"first"step"was"to"modify"the"protospacer"of"gRNA"constructs"and"reassemble"the"plasmid,"that"was"further"cloned"in"Escherichia'coli."After"isolation,"the"plasmids"were"linearized"and"used"for"transformation"of"P.'citrinum."In"order"to"increase"the"success"rate"of"the"P.'citrinum'transformation,"protoplasting"conditions"were"further"optimized."Correct"transformants"were"then"selected"using"phleomycin"resistance"marker"gene"and"screened"for"SM"production."Since"genes"within"the"citrinalin"cluster"were"disrupted"individually,"the"function"of"each"gene"was"analyzed"by"culturing"the"fungal"mutants"under"citrinalin/producing"conditions,"and"further"analyzed"via"LCMS"and"NMR."By"this"approach,"the"critical"enzymes,"intermediates,"and"the"biosynthetic"pathway"of"citrinalin"will"be"discovered."$

! 65"

102.$Towards$minimal$Aspergillus$genomes:$editing$Aspergillus*niger$mycotoxin$clusters$using$an$optimized$CRISPR/Cas$system""X."Zheng1,"2,"P."Zheng1,"2,"T."Cairns1,"2,"V."Meyer3,"Y."Ma1.""1)'Tianjin'Institute'of'Industrial'Biotechnology,'Chinese'Academy'of'Sciences,'Tianjin,'CN6'2)'Key'Laboratory'of'Systems'Microbial'Biotechnology,'Chinese'Academy'of'Sciences,'Tianjin,'CN6'3)'Technische'Universität'Berlin,'Institute'of'Biotechnology,'Department'Applied'and'Molecular'Microbiology,'Berlin,'DE""'Aspergillus'niger"is"used"for"bulk/scale"production"of"proteins"and"acids,"and"is"a"promising"platform"for"synthesis"of"new/to/nature"pharmaceuticals."However,"a"significant"challenge"to"biotechnological"applications"of"A.'niger'is"the"capacity"to"produce"a"diverse"arsenal"of"mycotoxins."Consequently,"a"major"goal"is"the"production"of"minimalized"A.'niger"genomes"that"do"not"encode"mycotoxin"biosynthetic"gene"clusters"(BGCs)."Here,"we"developed"a"highly"efficient"gene"editing"system,"and"applied"this"technology"to"remove"the"entire"BGC"for"the"mycotoxin"fumonisin"B1."Uniquely,"our"CRISPR/Cas9"system"utilised"the"5S"rRNA"gene"as"promoter"to"drive"guide"RNA"expression,"which"is"both"highly"conserved"and"efficiently"expressed"in"fungi."This"novel"system"enabled"100%"rates"of"precision"gene"modifications"using"short"(40/bp)"homologous"donor"DNA"at"an"individual"gene"level."Moreover,"in"a"proof"of"principle"experiment"for"chromosomes"minimalization,"we"deleted"the"48"kb"gene"cluster"required"for"biosynthesis"of"the"mycotoxin"fumonisin"B1"at"very"high"efficiency"to"generate"a"strain"that"was"incapable"of"producing"this"molecule."We"believe"that"this"novel"gRNA"expression"based"on"5S"rRNA"gene"can"broadly"be"utilized"for"engineering"highly"efficient"fungal"CRISPR/Cas9"toolkits."Additionally,"the"system"reported"here"will"enable"development"of"mycotoxin"free"production"strains,"and"designer"chromosomes"in"model,"pathogenic,"and"industrially"important"fungi.""$103.$A$manually$curated$resource$for$the$completely$finished$Aspergillus*nidulans$A4$genome$Erin"McDonnell1,"Kimchi"Strasser1,"Ian"Reid1,"Alexandre"Beaudoin1,"Nicholas"Geoffrion1,"Debra"Fulton1,"Sylvester"Palys1,"Gregory"Butler1,"Isabelle"Benoit"Gelber1,"Thi"Truc"Minh"Nguyen1,"Justin"Powlowski1,"Kenneth"Dewar2,"Jessica"Wasserscheid2,"Elizabeth"Oakley3,"Damien"Downes4,"Chris"Wong5,"Tomohiro"Akashi6,"Arthur"Ram7,"Xin"Xiang8,"Gustavo"Goldman9,"Ronald"de"Vries10,"Richard"Todd4,"Berl"Oakley3,"Adrian$Tsang1."1)'Centre'for'Structural'and'Functional'Genomics,'Concordia'University,'Montreal,'Canada6'2)'McGill'University'Genome'Quebec'Innovation'Centre,'Montreal,'Canada6'3)'Department'of'Molecular'Biosciences,'University'of'Kansas,'Lawrence,'United'States6'4)'Department'of'Plant'Pathology,'Kansas'State'University,'Manhattan,'United'States6'5)'Faculty'of'Health'Sciences,'Macau'University,'Macau6'6)'Center'for'Neurological'Diseases'and'Cancer,'Nagoya'University,'Nagoya,'Japan6'7)'Institute'of'Biology,'Leiden'University,'Leiden,'the'Netherlands6'8)'Department'of'Biochemistry'and'Molecular'Biology,'The'Uniformed'Services'University,'Bethesda,'United'States6'9)'Faculdade'de'Medicina'de'Ribeirão'Preto,'Universidade'de'São'Paulo,'Brazil6'10)'Westerdijk'Institute,'Utrecht,'the'Netherlands.''"104."Pharmacologic$inhibition$of$the$UPR$sensor$IreA$has$antifungal$effects$in$Aspergillus*fumigatus.$"Jose$P.$Guirao$Abad,$Aaron"Albee,"Martin"Weichert,"David"S."Askew$"Department'of'Pathology'and'Laboratory'Medicine,'University'of'Cincinnati,'Cincinnati,'OH.'''Aspergillus'fumigatus'(AF)"is"one"of"the"most"important"pathogenic"fungi"in"humans,"responsible"for"life/threatening"infections"in"immunocompromised"patients."The"high"incidence"of"this"infection"is"due,"in"part,"to"the"limited"number"of"treatment"options,"making"the"search"for"new"antifungal"targets"a"high"priority."The"unfolded"protein"response"(UPR)"is"an"intracellular"signaling"network"that"maintains"protein"homeostasis"of"the"endoplasmic"reticulum"(ER)."The"UPR"in"AF"is"triggered"by"IreA,"an"ER/transmembrane"protein"comprised"of"a"luminal"stress"sensing"domain"and"a"cytosolic"effector"region"containing"both"kinase"and"RNAse"domains."Genetic"ablation"of"IreA"induces"an"avirulent"phenotype,"as"well"as"increased"sensitivity"to"several"physiological"stresses"and"clinical"antifungals."Interest"in"targeting"this"pathway"in"humans"has"resulted"in"the"identification"of"selective"inhibitors"of"the"RNAse"and/or"kinase."However,"these"compounds"are"untested"in"AF."In"this"study,"we"determined"the"biological"effects"of"treating"AF"with"4μ8c,"a"small"molecule"that"was"identified"as"an"inhibitor"of"the"RNAse"domain"of"the"human"ER"stress"sensor"Ire1."This"compound"inhibited"the"growth"of"AF"with"a"MIC"of"40"mg/L"and"

! 66"

triggered"cytoplasmic"granulation"and"shrinkage"at"10"mg/L,"indicative"of"hyphal"damage."Phenotypic"screening"revealed"that"the"IreA"inhibitor"increased"sensitivity"to"calcofluor"white"and"NaCl,"suggesting"cell"wall"and/or"membrane"disruption."4μ8c"inhibited"the"growth"of"AF"on"a"complex"carbon"source,"similar"to"the"phenotype"of"UPR/deficient"strains."Gene"expression"analysis"by"qPCR"demonstrated"that"4μ8c"prevented"UPR"activation"of"target"genes,"confirming"target"specificity"of"the"compound."In"addition,"4μ8c"blocked"the"induction"of"catalase"mRNA"during"ER"stress,"which"correlated"with"increased"sensitivity"to"H202."Finally,"compounds"that"disrupt"calcium"homeostasis"enhanced"the"antifungal"activity"of"4μ8c."Together,"these"results"demonstrate"that"4μ8c"inhibits"the"AF"UPR,"raising"the"possibility"of"harnessing"these"effects"for"future"antifungal"strategies."$ $

! 67"

List$of$Registrants$

Name$ Institute$ Email$Keietsu"Abe" " Grad."Sch."Agricult."Sci.,"

Tohoku"Univ."kabe@niche.tohoku.ac.jp"

Sheba"Agarwal/Jans" Elsevier"BV" s.agarwal/jans@elsevier.com"

Maria"Victoria"Aguilar"Pontes"

Westerdijk"Fungal"Biodiversity"Institute"

v.aguilar@westerdijkinstitute.nl"

Qusai"Al"Abdallah" UTHSC" qalabdal@uthsc.edu"

Mikael"Andersen" Technical"University"of"Denmark"

mr@bio.dtu.dk"

Yasuko"Araki" Kikkoman"Corporation" yaaraki@mail.kikkoman.co.jp"Scott"Baker" Pacific"Northwest"Natl"Lab" scott.baker@charter.net"

Amelia"Barber" Leibniz"Institute"of"Natural"Product"Research"and"Infection"Biology"–"Hans"Knöll"Institute"

amelia.barber@hki/jena.de"

Annie"Bellemare" Concordia"University" annie.bellemare@concordia.ca"

Isabelle"Benoit" Concordia"University" isabelle.benoit@concordia.ca"Adriana"Blachowicz" University"of"Southern"

California"blachowi@usc.edu"

Gerhard"Braus" Georg/August"Univ" gbraus@gwdg.de"

Michael"Bromley" The"University"of"Manchester" mike.bromley@manchester.ac.uk"Kenneth"Bruno" Zymergen" bruno@zymergen.com"

Gregorio"Budkewitsch" Concordia"university" gregorio.budkewitsch@concordia.ca"Gregory"Bulmer" Manchester"Institute"of"

Biotechnology"gregory.bulmer@outlook.com"

Caroline"Burks" Texas"A&M"University" cburks817@gmail.com"

Mark"Caddick" Univ"Liverpool" caddick@liv.ac.uk"David"Canovas" University"of"Sevilla" davidc@us.es"

Wanping"Chen" Georg"August"University"Göttingen"

chenwanping1@foxmail.com"

Tsokyi"Choera" Hexagon"Bio" choera@wisc.edu"

Yit"Heng"Chooi" University"of"Western"Australia" yitheng.chooi@uwa.edu.au"Tania"Chroumpi" Westerdijk"Fungal"Biodiversity"

Institute"t.chroumpi@westerdijkinstitute.nl"

Blake"Commer" Texas"A&M"University" blakecommer@tamu.edu"

Robert"Cramer" Geisel"School"of"Medicine"at"Dartmouth"

robert.a.cramer.jr@dartmouth.edu"

Tiezhong"Cui" Modern"Meadow,"Inc." elect.haven@gmail.com"Kurt"Dahlstrom" California"Institute"of"

Technology"kdahlstr@caltech.edu"

Paul"Daly" Westerdijk"Fungal"Biodiversity"Institute"

p.daly@westerdijkinstitute.nl"

Elise"de"Reus" Technical"University"Denmark" elisedr@bio.dtu.dk"Ronald"de"Vries" Westerdijk"Fungal"Biodiversity"

Institute"r.devries@westerdijkinstitute.nl"

Adiphol"Dilokpimol" Westerdijk"Fungal"Biodiversity"Institute"

a.dilokpimol@westerdijkinstitute.nl"

Milton"Drott" University"of"Wisconsin"Madison"

mdrott@wisc.edu"

! 68"

Wenlong"Du" Nanjing"Normal"University" 1225920538@qq.com"

Nigel"Dunn"Coleman" FunGeneX.USA" ndunncoleman.fungenex@gmail.com"Valeria"Ellena" University"of"Natural"Resources"

and"Life"Sciences"valeria.ellena@boku.ac.at"

João"Henrique"Fabri" Federal"University"of"São"Carlos"

joao.fabri@hotmail.com"

Moran"Farhi" ADM" moran.farhi@adm.com"Erzsébet"Fekete" University"of"Debrecen" kicsizsoka@yahoo.com"

Jarrod"Fortwendel" UTHSC/Memphis" jfortwen@uthsc.edu"Jens"Frisvad" Technical"University"of"

Denmark"jcf@bio.dtu.dk"

Malgorzata"Futyma" Technical"University"of"Denmark"

malfu@dtu.dk"

Jennifer"Gerke" Georg/August"University"Goettingen"

jgerke@gwdg.de"

Cameron"Gilchrist" The"University"of"Western"Australia"

cameron.gilchrist@research.uwa.edu.au"

Katsuya"Gomi" Tohoku"Univ" katsuya.gomi.a6@tohoku.ac.jp"

Michelle"Grau" University"of"Southern"California"School"of"Pharmacy"

mgrau@usc.edu"

Claudio"Greco" University"of"Wisconsin/Madison"

greco4@wisc.edu"

Markus"Gressler" Friedrich"Schiller"University"Jena"

markus.gressler@leibniz/hki.de"

Igor"Grigoriev" US"DOE"Joint"Genome"Institute" ivgrigoriev@lbl.gov"

Jose"P."Guirao"Abad" University"of"Cincinnati" guiraojo@ucmail.uc.edu"Amy"Gundersen" Novozymes,"Inc." aygm@novozymes.com"

Daisuke"Hagiwara" University"of"Tsukuba" hagiwara.daisuke.gb@u.tsukuba.ac.jp"Kap/Hoon"Han" Woosuk"University" khhan@woosuk.ac.kr"

Omar"Harb" University"of"Pennsylvania" oharb@sas.upenn.edu"Rebekka"Harting" Georg"August"University" rhartin@gwdg.de"

Thorsten"Heinekamp" Leibniz"Institute"for"Natural"Product"Research"and"Infection"Biology"

thorsten.heinekamp@leibniz/hki.de"

Kristiina"Hilden" University"of"Helsinki" kristiina.s.hilden@helsinki.fi"Terry"Hill" Rhodes"College" hill@rhodes.edu"

Kui"Hong" Wuhan"University" kuihong31@whu.edu.cn"Cameron"Hunter" Kansas"State"University" Cameron.Hunter15@gmail.com"

Mikel"Iradi/Serrano" University"of"the"Basque"Country"

mikel.iradi@ehu.eus"

Sachin"Jain" Zymergen" sjain@zymergen.com"

Zofia"Dorota"Jarczynska" DTU" zofja@bio.dtu.dk"Chihiro"Kadooka" Kagoshima"university" chihiro19940224@yahoo.co.jp"

Earl"Kang" University"of"Georgia" sekang2@uga.edu"Levente"Karaffa" University"of"Debrecen" karaffa.levente@science.unideb.hu"

Takuya"Katayama" The"University"of"Tokyo" akata@mail.ecc.u/tokyo.ac.jp"Nancy"Keller" Univ"Wisconsin,"Madison" npkeller@wisc.edu"

Joshua"Kerkaert" Dartmouth"College" joshua.d.kerkaert.gr@dartmouth.edu"

! 69"

Olaf"Kniemeyer" Leibniz"Institute"for"Natural"Product"Research"and"Infection"Biology"(HKI)"

olaf.kniemeyer@hki/jena.de"

Benjamin"Knox" Zymergen" bpknox@zymergen.com"Joanna"Kowalczyk" University"of"Helsinki" joanna.kowalczyk@helsinki.fi"

Caitlin"Kowalski" Geisel"School"of"Medicine"at"Dartmouth"

caitlin.h.kowalski.GR@dartmouth.edu"

Roland"Kun" Westerdijk"Institute" r.kun@westerdijkinstitute.nl"Emi"Kunitake" Mie"University" kunitake@bio.mie/u.ac.jp"

Mi"Kyung"LEE" Korea"Research"Institute"of"Bioscience"and"Biotechnology"

miklee1010@kribb.re.kr"

Patrick"Lehman" University"of"Southern"California"

plehman@usc.edu"

Éva"Leiter" University"of"Debrecen" leiter.eva@science.unideb.hu"

Su"Jeung"Lim" University"of"Southern"California"

sujeungl@usc.edu"

Jun"Lin" Leibniz"Institute"for"Natural"Product"Research"and"Infection"Biology"/"Hans"Knöll"Institute"

jun.lin@hki/jena.de"

Alfredo"Lopez"De"Leon" Novozymes,"Inc" allo@novozymes.com"

Dona"Love" NIAID,"National"Institutes"of"Health"

donalove@mail.nih.gov"

Ling"Lu" Nanjing"Normal"University" linglu@njnu.edu.cn"Ronnie"Lubbers" Westerdijk"fungal"biodiversity"

institute"r.lubbers@westerdijkinstitute.nl"

Christina"Lyra" University"of"Helsinki" christina.lyra@helsinki.fi"Silke"Machata" Leibniz"Institute"for"Natural"

Product"Research"and"Infection"Biology/"Hans"Knöll"Institute"

silke.machata@hki/jena.de"

Raj"Majumdar" USDA/ARS/SRRC" raj.majumdar@ars.usda.gov"

Miia"Mäkelä" University"of"Helsinki" miia.r.makela@helsinki.fi"Md."Abdulla"Al"Mamun" The"University"of"Tokyo" mamungeb26@gmail.com"

Sandrine"Marqueteau" Concordia"University" sandrine.marqueteau@concordia.ca"Adela"Martin/Vicente" University"of"Tennessee"Health"

Science"Center"amartinv@uthsc.edu"

Jun/ichi"Maruyama" The"University"of"Tokyo" amarujun@mail.ecc.u/tokyo.ac.jp"Sarah"McFarland" Novozymes"Inc" samf@novozymes.com"

Matthew"Mead" Vanderbilt"University" matthew.mead@vanderbilt.edu"Alexander"Mela" University"of"Georgia" apm61506@uga.edu"

Vera"Meyer" TU"Berlin" vera.meyer@tu/berlin.de"Ken"Miyazawa" Tohoku"University" ken.m.tubbt@gmail.com"

Michelle"Momany" Univ"Georgia" mmomany@uga.edu"Uffe"Mortensen" Technical"University"of"

Denmark"um@bio.dtu.dk"

Scott"Moye/Rowley" Univ"Iowa" scott/moye/rowley@uiowa.edu"

Gloria"Muzzi/Erichsen" Novozymes"Inc" gmer@novozymes.com"Akihiro"Ninomiya" University"of"Tsukuba" ninomiya.akihiro.gb@u.tsukuba.ac.jp"

Berl"Oakley" Univ"Kansas" boakley@ku.edu"Ana"Oliveira"Souza" UTHSC" aolivei5@uthsc.edu"

! 70"

Jean/Paul"Ouedraogo" Centre"for"Structural"and"Functional"Genomics"

jean/paul.ouedraogo@concordia.ca"

Norman"Paege" TU"Berlin" norman.paege@tu/berlin.de"Miguel"Penalva" CSIC"Centro"de"

Investigaciones"Biológicas"penalva@cib.csic.es"

Brandon"Pfannenstiel" Zymergen" bruno@zymergen.com"

Ngan"Le"Hong"Pham" University"of"Southern"California"

nganph21@gmail.com"

István"Pócsi" University"of"Debrecen" pocsi.istvan@science.unideb.hu"

Peter"Punt" Dutch"DNA" peter.punt@ddna/biotech.com"Christian"Rabot" University"of"Southern"

California"crabot@usc.edu"

Alice"Rassinger" University"of"Natural"Resources"and"Life"Sciences"

alice.rassinger@boku.ac.at"

Samantha"Reese" University"of"Nebraska/Lincoln" reesesammy50@gmail.com"

Jakob"Rendsvig" Technical"University"of"Denmark"

Jakren@dtu.dk"

Judith"Rhodes" Univ"Cincinnati" judith.rhodes@uc.edu"

Marina"Rocha" Federal"University"of"São"Carlos"

marinacamposrocha@gmail.com"

David"Roos" Univ"Pennsylvania" droos@sas.upenn.edu"Brandon"Ross" Dartmouth"College" tlights@optonline.net"

Kyle"Rothschild/Mancinelli"

Technical"University"of"Denmark"

kyroma@dtu.dk"

Tomas"Rush" University"of"Wisconsin/Madison"

trush@wisc.edu"

Mareike"Scheven" Leibniz"Institute"for"Natural"Product"Research"and"Infection"Biology"/"Hans"Knöll"Institute"

mareike.scheven@leibniz/hki.de"

Monika"Schmoll" Austrian"Institute"of"Technology"AIT"

monika.schmoll@ait.ac.at"

Julia"Schumacher" BAM"/"Federal"Institute"for"Materials"Research"&"Testing"

julia.schumacher@wwu.de"

Achchuthan"Shanmugasundram"

University"of"Liverpool" achchu@liverpool.ac.uk"

Brian"Shaw" Texas"A&M"Univ" bdshaw@tamu.edu"Brent"Shuman" University"of"Georgia" brent.shuman@uga.edu"

Joe"Spraker" Hexagon"Bio" joespraker@gmail.com"Jason"Stajich" Univ"California,"Riverside" jason.stajich@ucr.edu"

Jacob"Steenwyk" Vanderbilt"University" jacob.steenwyk@vanderbilt.edu"Joel"Steyer" Kansas"State"University" jsteyer@ksu.edu"

Maria"Stroe" Leibniz"Institute"for"Natural"Product"Research"/"Hans"Knöll"Institute"

maria.stroe@leibniz/hki.de"

Rebecca"Sweany" Louisiana"State"University" rswean1@gmail.com"

Edyta"Szewczyk" Bolt"Threads" eszewczyk@boltthreads.com"Norio"Takeshita" University"of"Tsukuba" takeshita.norio.gf@u.tsukuba.ac.jp"

Koichi"Tamano" AIST" tamano/k@aist.go.jp"Mizuki"Tanaka" University"of"Shizuoka" mizuki.tanaka@u/shizuoka/ken.ac.jp"

! 71"

Joanna"Tannous" University"of"Wisconsin/Madison"

jtannous@wisc.edu"

Dominik"Terfehr" evoxx"technologies"GmbH" d.terfehr@evoxx.com"Dawn"Thompson" Ginkgo"Bioworks" dawnt@ginkgobioworks.com"

Sheryl"Thompson" Novozymes" shth@novozymes.com"Chaoguang"Tian" Tianjin"Institute"of"Industrial"

Biotechnology"tian_cg@tib.cas.cn"

Richard"Todd" Kansas"State"Univ" rbtodd@ksu.edu"

Adrian"Tsang" Concordia"Univ" adrian.tsang@concordia.ca"Maiko"Umemura" National"Institute"of"Industrial"

Science"and"Technology"(AIST)"umemura/m@aist.go.jp"

Shunichi"Urayama" University"of"Tsukuba" syun.ura@gmail.com"Hans"van"den"Brink" Chr"Hansen"A/S" DKHVB@CHR/HANSEN.COM"

Johannes"van"Dijk" Zymergen" jvandijk@zymergen.com"Jolanda"van"Munster" University"of"Manchester" jolanda.vanmunster@manchester.ac.uk"

Noel"van"Peij" DSM"Biotechnology"Center" noel.peij/van@dsm.com"Joseph"Vasselli" Texas"A&M" joe723@tamu.edu"

Andreas"Vestergaard" Technical"University"of"Denmark"

amve@dtu.dk"

Clay"Wang" Univ"Southern"California" clayw@usc.edu"Jingyi"Wang" University"of"Southern"

California"wang710@usc.edu"

Tristan"Wang" Cornell"University" tww57@cornell.edu"

Franziska"Wanka" TU"Wien" franziska.wanka@gmail.com"Martin"Weichert" Medical"Sciences"Building,"

University"of"Cincinnati"weichemn@ucmail.uc.edu"

Philipp"Wiemann" Hexagon"Bio" philipp@hexagonbio.com"Peter"Wolff" Technical"university"of"

Denmark"pewol@dtu.dk"

Ming/Yueh"Wu" Ginkgo"Bioworks" kiky530@gmail.com"

Jing"Ye" Nanjing"Normal"University" 2257691723@qq.com"Akira"Yoshimi" NICHe,"Tohoku"Univ." a/yoshimi@niche.tohoku.ac.jp"

Bo"Yuan" University"of"Southern"California"Pharmacy"school"

yuan984@usc.edu"

Xiaomei"Zheng" Tianjin"Institute"of"Industrial"Biotechnology,"Chinese"Academy"of"Sciences"

zheng_xm@tib.cas.cn"

Yun"Zheng" Concordia"University" yun.zheng@concordia.ca"

Kolea"Zimmerman" Ginkgo"Bioworks" kolea@ginkgobioworks.com"$