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Distribution and population structure of the anther smut Microbotryum silenes-acaulis parasitizing an arctic-alpine plant Britta Bueker, Chris Eberlein, Pierre Gladieux, Angela Schaefer, Alodie Snirc, Dominic J. Bennett, Dominik Begerow, Michael E. Hood, Tatiana Giraud

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Britta Bueker, Chris Eberlein, Pierre Gladieux, Angela Schaefer, Alodie Snirc, et al.. Distribution and population structure of the anther smut Microbotryum silenes-acaulis parasitizing an arctic-alpine plant. Molecular Ecology, Wiley, 2016, 25 (3), pp.811–824. 10.1111/mec.13512. hal-01497390

HAL Id: hal-01497390 https://hal.archives-ouvertes.fr/hal-01497390 Submitted on 28 May 2020

HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. Version postprint Hood, M. E.(Auteurde correspondance), Giraud,T. (2016).Distribution andpopulation structure of the anther smut Microbotryum silenes-acaulis parasitizing an arctic-alpine plant. Molecular Bueker, B.,Eberlein,C., Gladieux,P., Schaefer,A., Snirc,A.,Bennett, D.J.,Begerow, D., G1V 0A6 G1V 0A6 Laval, PavillonCharles-Eugène-M

Bochum, Universitätsstraße 150, 44780 Bochum, Germany Germany 150,44780Bochum, Bochum, Universitätsstraße Dominic J. Bennett Dominic J. This article is protected Allrights bycopyright. reserved. 10.1111/mec.13512 of Record.versiondifferences and article Please between the cite this Version this as doi: through the copyediting, typesetting, pagination for publication accepted This articlehasbeen Running title:Plantparasitew nunatak, selfing prevalence, disease admixture, Keywords: equally tothemanuscript. * Theauthorscontributed +14135428538 Hood:[email protected], author: Michael Corresponding 6 5 4 3 2 1 Britta Bueker* Microbotryum silenes-acaulis smut the anther of population structure and Distribution Article Article type :Original Date:26-Nov-2015 Accepted Revised Date:02-Nov-2015 :08-Jul-2015 Date Received France; CIRAD, F-34398, Montpellier, France Montpellier,France F-34398, CIRAD, France; Department of Life Sciences Department of Evolution,CNRS,Univ. Ecologie Systématique Amhers Department ofBiology, Lehrstuhlund Biodiversität de fürEvolution Institut de BiologieIntégrative et des Syst INRA,InternBâtiment K,Campus UMRBGPI, Accepted France Saclay, 91400Orsay, Article Ecology, 25 (3), 811-824. DOI : 10.1111/mec.13512 1,2 , Chris Eberlein* 4,6 , DominikBegerow Comment citer cedocument: , Imperial College London, London,London, 2AZ,Kingdom College SW7 Imperial United , ith arctic-alpine distribution ith arctic-alpinedistribution t College, 220 South Pleasant Street, Amherst MA 01002, USA t College, PleasantStreet,AmherstMA01002,USA 220South 1,3 archand, 1030 Avenue de la M Avenue dela 1030 archand,

, PierreGladieux* parasitizing an arctic-alpine plant parasitizing anarctic-alpine 1 , Michael E.Hood , Michael Microbotryum lychnidis-dioicae, lychnidis-dioicae, Microbotryum è mes, D r Pflanzen, AG Geobotanik, Ruhr-Universität AGGeobotanik, r Pflanzen, and undergone full peer review buthasnotbeen review undergone fullpeer and and proofreading process, which may lead to ational deBaillarguet,ational é

partement de Biologie, PROTEO,Universit Biologie, partement de Paris-Sud, AgroParisTech, Universit AgroParisTech, Paris-Sud, 4,5 , Angela Schaefer 2 , TatianaGiraud é dicine, Quebec,QC, Canada F-34398 Montpellier, F-34398 1 , AlodieSnirc multiple infections, 4

4 ,

é Paris é

acial recolonization northwards fromsouthern northwards recolonization acial was present throughout the host’s geographic thehost’s present throughout was indicating aindicating major influe eys and populationgenetics eys northern Europe and s Europe northern footprints intheirpopulation structurethat footprints Silene acaulis Silene a during the last glaciaduring thelast a pathogen distribution. Genetic analyses with distribution.Geneticanalyses pathogen fungus parasitizing an arctic-alpine plant.A anarctic-alpine fungus parasitizing e distributionshavepersistedduring e thelast

M. silenes-acaulis were examined, and the anther anther andthe examined, were outhern Europe, likely Europe,likely outhern nce of environmental nce tion. The lower genetic tion. Thelower thus uniquely reveals thus individuals

revealed revealed Version postprint Hood, M. E.(Auteurde correspondance), Giraud,T. (2016).Distribution andpopulation structure of the anther smut Microbotryum silenes-acaulis parasitizing an arctic-alpine plant. Molecular Bueker, B.,Eberlein,C., Gladieux,P., Schaefer,A., Snirc,A.,Bennett, D.J.,Begerow, D., rangearctic-alpine of species was thusalso fragmented duringwhich the age, left ice footprints ice sheets during interglacial periods enable periods interglacial during sheets ice Westergaard Westergaard mountainscompletely “nunataks”, thatwere existence ofadditional, smaller refugia were ofte processes recolonization (Comes & 1998; Abbott& Kadereit Brochmann (Murray 1995;Abbott ice-sh edge ofthe atthe areas, more northern sp In contrast,cold-adapted diversity 2004). (Hewitt regions, whilesout genetic innorthern diversity differentiation (Bennett isolationofpopula Long-term southern regions. This article is protected Allrights bycopyright. reserved. Accepted Duringglacial the Pleistocene, periodsof cold-sensitive species ice-free persisted“refugia” in Article (Hewitt 2004). day been species has influenced strongly by the intemperature oscillations of the Quaternary ofpresent- structure genetic the that revealed have markers molecular using studies particular, and connectivity ofspecies´ extensions insealeveland changes (Hewitt 1996),as dispersal historical ability processesfrom and as climate continental such and changes drift distributionand geneticstruct The geographic INTRODUCTION et al. et Ecology, 25 (3), 811-824. DOI : 10.1111/mec.13512 2011), from which species dispersedwh fromwhich species 2011),

et al. et Comment citer cedocument: et al. 2000;Brochmann

habitats (Andersen &habitats Borns(Andersen 1997; 1991; Hewitt 1996, 2004). Increasing 1991;Hewitt1996,2004). n associated with foundereffect with n associated

in northern Europe and North America, America, EuropeandNorth in northern d range expansions and migration northwards andmigration expansions d range surrounded by glacial ice (e.g. Stehlik (e.g. ice glacial by surrounded eets, and also in mountains of southern areas andalsoinmountainsofsouthernareas eets, ure of present-day species result bothfrom result species of present-day ure et al. et hern regions remained reservoirs ofgenetic reservoirs remained regions hern tions in separate refu tions inseparate 2003). Expansions aspartofthese Expansions 2003). 2003). Molecular studies have supported the have supported 2003).Molecularstudies ecies were able to survive glacial maxima in maxima glacial able tosurvive ecies were of ice sheets have affected the availability the haveaffected oficesheets

en theclimateb Ehlers & Gibbard 2007). In 2007). & Gibbard Ehlers s, resulting inlowlevelsof temperatures and decline of anddecline temperatures gia fostered giapopulation fostered ecame warmer. ecame warmer. The e.g., et al. et in 2002;

in Version postprint Hood, M. E.(Auteurde correspondance), Giraud,T. (2016).Distribution andpopulation structure of the anther smut Microbotryum silenes-acaulis parasitizing an arctic-alpine plant. Molecular Bueker, B.,Eberlein,C., Gladieux,P., Schaefer,A., Snirc,A.,Bennett, D.J.,Begerow, D., This article is protected Allrights bycopyright. reserved. alpine environmentsinthenor silenes-acaulis, the lastglaciation. Th and after changes during Microbotryum silenes-acaulis thepopulationstruct studiedhere therefore We biodiversity. on warming temperatures from ofimpact issues andbroader andspread emergence change ondisease ofclimate effects the yetre & 2005); Eddington such (Cripps mycorrhizae onfu havefocused few studies instance, only leaving taxa, ast all focusonplantandanimal studies onthe geneticarctic-alpine diversity of distri depend inadditiononthe 2003; Stewart Brochmann factors, specific da ofpresent structure genetic the migration, of post-glacial impact the beside that, shown have species onarctic-alpine studies Recent Brysting & (Brochmann 2008). genetic variation gene betweendifferent fronted, withadmixtures broad- waslikely species cold-adapted ofmany recolonization postglacial the however, climates structures(Eidesen genetic population as strong M. silenes-acaulis lived cushionswithhundredsofshortstems(Jone AcceptedSilene latifolia Article Ecology, 25 (3), 811-824. DOI : 10.1111/mec.13512 , and comparable population genetics st , andcomparablepopulation isfoundontheplant e.g., , Microbotryum lychnidis-dioicae Microbotryum dispersal abilityand mating system (Taberlet Comment citer cedocument: et al. et for understanding itspopulationhistory forunderstanding toclimate relation in thern hemisphere (Körner 2003). 2003). (Körner thern hemisphere bution of the host (Nieberding bution ofthehost(Nieberding 2010). In organi 2010). pathogenic

Silene acaulis y species varies greatly depending onspecies- depending greatly y varies species ngal pathogens or fungi in symbiosis, insymbiosis, pathogens orfungi ngal riking lack of knowledge in other groups. For inothergroups.For ofknowledge lack riking is specialized basidiomycete fungal pathogen, fungalpathogen, basidiomycete is specialized ure of the arctic-alpine distributed pathogen distributed ofthearctic-alpine ure organisms has been accumulated, they virtually hasbeenaccumulated, organisms tic pools, and therefore pools,andtherefore tic et al. et s & Richards 1962). A well-studied relativeof Awell-studied s & Richards1962). , causes anther smut on the temperate host host temperate the on smut anther , causes (Lutz search is important if we are to understand aretounderstand isimportantifwe search 2013). Unlike species adapted to temperate adaptedtotemperate 2013).Unlikespecies udies indicate a clear patternof udies indicateaclear

et al. et sms, populationstructuremay et al. et 2008), which occurs inarctic- 2008),which Silene acaulis et al. et 2008). Although of awealth 2008). with littlelossofoverall with 1998;Abbott& formslarge,long- e.g ., M. Version postprint Hood, M. E.(Auteurde correspondance), Giraud,T. (2016).Distribution andpopulation structure of the anther smut Microbotryum silenes-acaulis parasitizing an arctic-alpine plant. Molecular Bueker, B.,Eberlein,C., Gladieux,P., Schaefer,A., Snirc,A.,Bennett, D.J.,Begerow, D., fungi can be detected among herbarium specimen herbarium among fungi bedetected can specimens of relation toglacial cycles? isthe 3)What spatial distributionofthe genetic at variation finer silenes-acaulis To study thedistributionof surveys Herbarium MATERIAL ANDMETHODS individuals? scales, including infections whether by multiple pathogen thegenotypes same in occur acaulis fo werethe questions specific Our markers. used a combination ofherbariumand genetic surveys population analysesusing microsatellite of phylogeography To investigate the Buono have important consequences ondisease dynamics (Lopez-Villavicencio Infections by multiple pathogengenotypes have in reported been (Vercken host its than refugia andsmaller numerous more in This article is protected Allrights bycopyright. reserved. Accepted Articlethat of (Vercken recolonizationItaly, fromsouthern European refugiaBalkans) (Spain, the after last glaciation Silene latifolia as compared to its host plant plant host its to ascompared et al. et et al. et 2014; Susi & Laine 2015). Laine 2014;Susi& 2015). S. acaulis Ecology, 25 (3), 811-824. DOI : 10.1111/mec.13512 acrossgeographical its does range, itallow and retracing histories migration in 2010). The 2010).The

for assessingspeciesdistribution for (Taylor & Keller 2007), although thatpa Keller 2007), (Taylor& were examined. Several previous previous st Several wereexamined. Comment citer cedocument: M. lychnidis-dioicae M. silenes-acaulis

S. acaulis M. silenes-acaulis llowing: isthedistributionof 1)What anditshost ? 2) isthepopulation structure of What ? recolonization history with recolonization congruent ismostly s of its hosts; because there islittleevidence s ofitshosts;becausethere

inrelation toitshost distribution,we

S. acaulis, udies have shown thatanther-smut udies haveshown et al. et thogen seemed tohavepersisted seemed thogen M. lychnidis-dioicae 2010; Gladieux 2010;Gladieux atotal of1437herbarium et al. et 2007,2011; M. silenes- et al. et and can andcan M. S. acaulis 2011).

Version postprint Hood, M. E.(Auteurde correspondance), Giraud,T. (2016).Distribution andpopulation structure of the anther smut Microbotryum silenes-acaulis parasitizing an arctic-alpine plant. Molecular Bueker, B.,Eberlein,C., Gladieux,P., Schaefer,A., Snirc,A.,Bennett, D.J.,Begerow, D., randomly selected healthy collection sites. On collection healthy randomly selected of50,100,or200km diseased sites(withinradii were translated intopairwise linear distances betweenof collection sites. The number sites near observedmaximum sitedensities, of82.For local defining latitude and longitude information sites usinggeneralized the linear modelwith Poissonlog approach linkfunction in th for analysed were term, and theirinteraction densities ofcollection sites near todiseased and healthyLatitude localities. and disease status, average between differences of significance assessstatistical to wereused sites collection norther quartiles fromsouthernto separatin by analysed of latitudewere effects analysed inthe context ofcollection site latitude anddensitycollection ofnearby sites. The S. acaulis specime The distributionsofhealthy anddiseased recorded. were specimens a Thelocation (BM). Museum ofNaturalHistory Minnesota Washington (MIN), State Washington University British (WTU), University (WS), of University (ISC), University Iowa State Iowa and of University (UC), Berkeley California, Museum National d'Histoire Naturelle State (P),Oregon University University (OSC), of Herbar Forest Service following U.S. herbaria: distribution aswell.Specimensof This article is protected Allrights bycopyright. reserved. Accepted Article (Rabeler 1993;Antonovics orsubsequentannot collectors that theoriginal map by Hultén and Fries (1986). Disease (1986). Disease andFries mapby Hultén Ecology, 25 (3), 811-824. DOI : 10.1111/mec.13512 Comment citer cedocument: et al. et 2003;Hood n latitudes; quartile la n latitudes;quartile S. acaulis

were examined intheexamined were generalcollections fromthe g the samplesintoequa g the et al. et e thousand randomly assemb randomly e thousand ator recognized the plants as being diseased theplantsas recognized ator ium (USFS), New York Botanical Garden (NY), (NY), Garden Botanical York New ium (USFS), e response variable of density variableof of hostcollection e response ) was compared to an equal number (N = 36) of number(N=36) toanequal ) wascompared ns were compared tothepreviously published compared ns were nd date of collection for healthy and diseased and diseased healthycollection for nd dateof 2010),thisallowsassess incidence among herbarium specimenswas herbarium among incidence titude boundariesof<44,49,62,andthe

l sized groupsl sized on based led sets of 36 healthy led setsof36healthy ment of the pathogen ment ofthepathogen SPSS Version postprint Hood, M. E.(Auteurde correspondance), Giraud,T. (2016).Distribution andpopulation structure of the anther smut Microbotryum silenes-acaulis parasitizing an arctic-alpine plant. Molecular Bueker, B.,Eberlein,C., Gladieux,P., Schaefer,A., Snirc,A.,Bennett, D.J.,Begerow, D., of the population structure an of thepopulationstructure power the increase to genotypes, local from developed specifically markers and with separately, inNorway density sampling Because ofhigher European dataset microsatelliteDevelopment markers of andgenotyping populations ofthe northern- 195individualsfrom39populations. represented which analyses, Only formore individuals withgenotype data previously available (Table S2) (Bucheli (Bucheli S2) (Table available previously (Giraud chelex using extracted andSvalba Norway including and northernEurope, individuals(frompopulati DNAs fromall255sampled Genotyping populations ofthe global dataset (Lopez-Villavicencio flower given sporesfroma populationand geographic 2001 to 2013. Diploid teliospores from host plants within an area of 2km 2001 to2013.Diploidteliosporesfromhostplantswithinanareaof natural,living populat individuals collectedfrom of The analyses Sample collection of sites. healthy selected number ofrandomly This article is protected Allrights bycopyright. reserved. Accepted Article STATISTICS 19 (SPSS, Inc, 19(SPSS, Chicago,Illinois); thisanalysis included the disease sites andequal an Ecology, 25 (3), 811-824. DOI : 10.1111/mec.13512 M. silenes-acaulis M. silenes-acaulis et al. et M. silenes-acaulis Comment citer cedocument: 2007). alyses in these regions. regions. inthese alyses et al. et population structure were performed on a sampleof on performed population structurewere

2004) and genotyped usinggenotyped 2004)and inliving populations for population genetic analyses et al. et than 70 % of the markers were retained for than 70%ofthe retained were markers for 1998;Giraud and Svalbard, we also an Svalbard,we and

ions across its range (Fig. 1, TableS1)from (Fig. acrossitsrange ions rd), hereafter calledrd), hereafterglobal the dataset, were were treated as a single diploidindividual as asingle treated were ons in North America, Iceland, southern ons inNorthAmerica,

et al. et 16 microsatellite markers 16 microsatellite 2002; Giraud 2002;Giraud alyzed thesepopulations alyzed 2 as were considered a et al. et 2008). Version postprint Hood, M. E.(Auteurde correspondance), Giraud,T. (2016).Distribution andpopulation structure of the anther smut Microbotryum silenes-acaulis parasitizing an arctic-alpine plant. Molecular Bueker, B.,Eberlein,C., Gladieux,P., Schaefer,A., Snirc,A.,Bennett, D.J.,Begerow, D., 350 bp (Table S3). Primer pairs were designed using using designed were pairs S3).Primer 350 bp(Table the sothat chosen Primer bindingsiteswere & 2011). Held theprobability (Leese maximizing ofpolymorphism motif,fordecreasingwere repeats ofthe perfect probabilityfor scoring allele wrong the and of least increasing fourrepeats, the for probability th 2) scorablemarkers, and for obtaining variable and nine length two between was markers 5-Carboxytetramethyl-rhodamine (TAM markers 5-Carboxytetramethyl-rhodamine markers at mostusingPCR, with multiplex different fragment lengthsadding andfluorescent the micros different After testing withmultipleamplifiedfragments 2011). Markers elongationtimeinth thefinal analyses, fragment in bands ofstutter occurrence the Toreduce Alaska. and Svalbard Norway, Alps, respectively: microsatellite markerswas tested bygenotyping fourdistinct regions, fourspecimens from by and prepared Only contigs subsequentretained larger3 kbwerefor the than detection microsatellites of using This article is protected Allrights bycopyright. reserved. Accepted ArticlePHOBOS program (GATC-Biotech Roche GSFLX+I the454sequencing using method popul (Jotunheimen, strain (CE008)fromNorway ofHoffm themethod by was extracted da forsix µg/ml) (100 (MYP) withkanamycin 137flowerswere For sporesfrom thisgoal, 3.3.12 (Mayer 2010). We retainedmicrosate We 2010). 3.3.12 (Mayer MIRA Ecology, 25 (3), 811-824. DOI : 10.1111/mec.13512 3.4.0.1 (Chevreux 3.4.0.1(Chevreux Sigma-Aldrich LifeScience Sigma-Aldrich Comment citer cedocument: atellite combinations using gel using wepooledtwo atellite combinations electrophoresis, et al. et bp, which is typically considered as the optimal length range length range optimal as the considered typically is bp, which

1999) was used toassemble 1999)was an & Winston (1987). The genome of a genome of (1987).The & Winston an germinated onMaltose germinated (Taufkirchen, Germany). Variation for the designed designed forthe Variation Germany). (Taufkirchen, amplified fragment would range between 100 and and 100 between range would amplified fragment ys at room temperature and DNA from cultures DNA fromcultures and ys roomtemperature at they of20bpand/orat aminimumlength had RA™), 5-Tetrachloro-Fluorescein (TET™) and (TET™) RA™), 5-Tetrachloro-Fluorescein at the and loci 3)they wouldbe polymorphic, ation 16) was sequenced (3X coverage) ona (3X coverage) ation 16)wassequenced e PCRs was set to 30 minutes (Leese & Held &e PCRswassetto30minutes Held (Leese were not retained for further genotyping. genotyping. further for were notretained llite loci only when: 1) their repeat motif theirrepeat llite locionly when:1) PRIMER

0.4.0 (Rozen &0.4.0 (Rozen Skaletsky 2000) -Yeast-Peptone medium -Yeast-Peptone single reads intocontigs.single reads , Konstanz, Germany). The The Germany). Konstanz, M. silenes-acaulis

rope was assessedby was rope 64 flowers from 16 plants, genotyped withthe16 genotyped from16plants, 64 flowers Sigma-Aldrich Sigma-Aldrich plant infourofthepopulationsfromsouthern plant ructure analysis of Norway and Svalbard ofNorway analysis ructure NIMALFARM NIMALFARM these markers, we checked for large allele large for we checked thesemarkers, S. acaulis et al.

Life Science to detect markers evolving under evolving markers todetect from 18populationswithnomore 2004) and removed locidisplaying2004) andremoved the geographic groupsNorth geographic the detect markers inlinkage markers detect allelic (A richness , we genotyped spores from , wegenotyped from spores 1.0 (Landry ) to the 5’-end forward ) tothe5’-endforward Applied Biosystems Applied et al. 2002). The 2002). The r ) (Petit et al. et ),

Accepted ArticleENALEX 6.5 (Peakall & Smouse 2005). Differences ingenetic (H diversity Smouse 2005).Differences & 6.5(Peakall

(Goudet 2014) wasusedtoestimateA (Goudet 2014) IS E Ecology, 25 (3), 811-824. DOI : 10.1111/mec.13512 coefficient wasestimatedwiththeco coefficient , H RSTUDIO RSTUDIO O TRUCTURE TRUCTURE , and G´ , and E TRUCTURE TRUCTURE Comment citer cedocument: , A 3.0.2 (RStudioTeam2015). ST r ) using rank Spearman´s coefficient. the Allstatistical tests were K 2.3 (Pritchard . The significance of the heterozygosity deficit compared to the the to compared deficit heterozygosity the of significance . The were investigated with the Bayesian assignment method theBayesian assignment with investigated were > 9.Barplots withcluster-membership coefficients forall ST fections and population genetic di fections andpopulation ) (Nei 1987; Meirmans & Hedrick 2011). The R & 2011). Hedrick (Nei 1987;Meirmans ) H

ARVESTER after a burn-in periodof100,000.S a burn-in after were generated with the program withtheprogram generated were et al. et Wilcoxon signed rank tests. The relationship signed ranktests.The Wilcoxon E utations, and the Greedy algorithm for algorithm utations, andtheGreedy ), observed heterozygosity (H heterozygosity ), observed (Earl & von Holdt 2012).C & von (Earl 2000) using Ten thediploiddatasets. 2000) of K (number of clusters), with K ranging from from withKranging clusters), of of K(number r , andG independent runswith rrection of Nei and rrection ofNei ENALEX

6.5 (Peakall & Smouse2005) (Peakall 6.5 versity was assessed by testing testing assessedby was versity Chesser (1983) using (1983)using Chesser DISTRUCT the same clustering sameclustering the TRUCTURE LUMPP O E ), and Nei´s ), andNei´s , A ENEPOP r ) between ) between

1.1.2 package package 1.1 K outputs 4.2 ≤

9 andthe (Rousset Version postprint Hood, M. E.(Auteurde correspondance), Giraud,T. (2016).Distribution andpopulation structure of the anther smut Microbotryum silenes-acaulis parasitizing an arctic-alpine plant. Molecular Bueker, B.,Eberlein,C., Gladieux,P., Schaefer,A., Snirc,A.,Bennett, D.J.,Begerow, D., was no significant difference inthedistributionof difference was nosignificant conformed tothespecies Herbarium specimens of Distribution and demographic data RESULTS in thiscluster was than higher 0.7. twototen,indi value ofKfrom (Oksanen genetica to visualize withoutany patterns Inaddition tothesemodel-base 1930s, consistentwithprevious studi plant ofthe period collection Peak disease. anther-smut the by affected examin specimenswere A totalof1,437herbarium agapinthenorther the Rocky Mountains,and distribution,includingalpine/circum-polar mores populations were estimated using G populations wereestimated and visualized with neighbour-joining trees using the R package “ape” withneighbour-joining usingtheRpackageand visualized trees et al. et This article is protected Allrights bycopyright. reserved. Accepted Article “RgoogleMaps” package geographic distributionsof (Rosenberg 2003).The 2007), respectively. F 2007),respectively. et al. et Ecology, 25 (3), 811-824. DOI : 10.1111/mec.13512 2015), “rgl” (Adler (Adler 2015),“rgl” Comment citer cedocument: ( S. acaulis map provided by Hultén a Hultén map providedby Loecher 2015). Genetic diversity Loecher 2015).Genetic ST values among genetic clusterswe among values d methods, we performed a principal component analysis (PCA) analysis component aprincipal d methods,weperformed viduals were assigned to a cluster if their membership coefficient coefficient membership their if acluster to wereassigned viduals were examined from throughout the species range and largely andlargely range thespecies fromthroughout examined were et al et

ENALEX es on other plant species (Antonovics es onotherplantspecies (Antonovics . 2015) and “scatterplot3d” (Ligges (Ligges “scatterplot3d” . 2015)and ssumption on panmixia (R packages “vegan” (Rpackages“vegan” ssumption onpanmixia 6.5 (Peakall & Smouse 2005) and INSTRUCT& (Gao Smouse2005)and 6.5(Peakall n Eurasia regions of the Sakha Republic. theSakha of regions n Eurasia outhern extensions in mountain ranges, such as inmountain such ranges, outhern extensions collection times (years) between diseased and diseased between (years) times collection genetic clusters were visualized using R visualized were geneticclusters ed, including 41 (2.9%) that were found tobe that were ed, including 41(2.9%) nd Fries (1986) (Fig. 1), 1), (1986) (Fig. nd Fries

and selfing rates ingenetic rates and selfing re calculated at K = 3 to K=8 K=3to at calculated re ( S. acaulis Paradis et al. et et al. et i.e., 2003).There et al. et an 2015). For each 2015).For was inthe 2015 ) . Version postprint Hood, M. E.(Auteurde correspondance), Giraud,T. (2016).Distribution andpopulation structure of the anther smut Microbotryum silenes-acaulis parasitizing an arctic-alpine plant. Molecular Bueker, B.,Eberlein,C., Gladieux,P., Schaefer,A., Snirc,A.,Bennett, D.J.,Begerow, D., more often lowdensities inregions of with collection sitesin significant the among patterns infections.However, of free being major region foundtobewidespread was Anther-smut disease were including identified, 36withanther smutinfection. or ambiguousspecimens withillegible locality particular, mentionofdiseased nowritten recorded was onherbariumExcluding sheets. havebiasedco status oftheplantsandwould This article is protected Allrights bycopyright. reserved. Acceptedsites withinaradiusof100km wasindeedpr of inareas being found of latitude,withdisease nearby collection sites was significantly determined hostcollectionsiteWhile a strong density had quartiles, respectively). two southern latitudinal quartiles, samples/site and 1.4andmore 1.3 northern forthe two pers numbersofsamples greater greaterThis pattern of disease incidence latitudes inthe northern was the despite occurrence of diseasedplants proportion ofsitesthatcontained thesetof observedfor or 200km,aslowthat healthy setsof randomly sampled Article(Antonovics As incidence. disease in change healthy (Kolmogorov-Sm samples et al. et Ecology, 25 (3), 811-824. DOI : 10.1111/mec.13512 2003), there was no indication thatcoll therewasnoindication 2003), Comment citer cedocument: previously observed forother observed previously ite in more southern localities(1 ite inmoresouthern sites had average densitiesofnear sites hadaverage irnov twosampletest

S. acaulis edicted from the generalized linear model by modelby linear generalized fromthe edicted llection towards healthyIn ordiseased plants. latitudinal component (Fig. 3), thenumberof latitudinalcomponent(Fig. 3), information, 790distinctcollectionlocalities information, lower host site density. The number of nearby number of nearby hostsitedensity. The lower diseased sites (p-value < 0.001). Second,the <0.001). sites(p-value diseased increased in more northern regions (Fig. 3). (Fig. regions inmorenorthern 3). increased two ways. First, the disease tendedtobefound two ways. First, thedisease the distributionofanther the within the host’s geographic range, withno range, within thehost’sgeographic by disease status inde status by disease collectionsites(Fig.2); noneofthe1,000 , p-value = 0.545), indicating norecent , p-value=0.545),

ectors had been aware of the disease thedisease of aware ectors hadbeen Silene .7 samples/site for each of the for eachof .7 samples/site by sites, inradiiat50,100 sites, by herbarium specimens specimens herbarium smut diseaseshowed smut pendently of the effect theeffect pendently of Version postprint Hood, M. E.(Auteurde correspondance), Giraud,T. (2016).Distribution andpopulation structure of the anther smut Microbotryum silenes-acaulis parasitizing an arctic-alpine plant. Molecular Bueker, B.,Eberlein,C., Gladieux,P., Schaefer,A., Snirc,A.,Bennett, D.J.,Begerow, D., 2.65) and intermediate in North America (H inNorthAmerica 2.65) andintermediate Expected heterozygosity (H heterozygosity Expected significantly lowervalues than the expected heterozygosity (H value =0.244,A displayedalleles S2).For all 24 (Table markers, heterozygosity (H observed Typically, three tonine alleles per marker, locus werethe observed,E18 except which for dataset, including detectedWe alleles 99different overall across16 microsatellite the markersglobal used onthe Genetic variation inpopulations and within individual host plants 0.017), while the interaction te 0.017), whiletheinteraction northern Europe (H northern Europe southern Europe and North Americawe and North southern Europe (H differences significant indicated tests signed rank This article is protected Allrights bycopyright. reserved. latitude (Wald populations (H heterozygotewas also indicated and deficiency high by positive G There was no significant correlation between th between correlation nosignificant There was per plant, detected were genotypes from multiple stems hadgenotyped been(mean = ofstems analyzed4).Amean number of3.25 detected inthe16plan Multiple infectionswere

AcceptedE Article : W = 4, p-value < 0.001, A =4,p-value<0.001, : W Ecology, 25 (3), 811-824. DOI : 10.1111/mec.13512 E X = 0.45,A 2 r : W = 118, p-value = 0.008). p-value=0.008). =118, : W = 113.32, p-value < 0.001) =113.32,p-value<0.001) M. silenes-acaulis E : W = 4, p-value = 0.009;A =4,p-value : W Comment citer cedocument: r = 4.89), lowest in northern European populations (H populations European = 4.89),lowestinnorthern E ) and allelic(A richness rm was non-significant (Wald (Wald rm wasnon-significant r : W = 136, p-value = 0.031) and between North America and North America and between =0.031) =136,p-value : W i.e.,

samples from 195different samplesfrom nearly every stem analyzed bore a specific genotype. genotype. aspecific bore analyzed stem every nearly re significantly different only for A re significantly for different only E and disease status (Wald disease status(Wald and = 0.35, A = 0.35, e average number of genotypes perindividual genotypes of number e average ts from populations in the Alps for which spores from populationsintheAlpsfor whichspores ts r : W = 129, p-value < 0.001), whilethevaluesof =129,p-value<0.001), W : between southern Europe and northern Europe andnorthernEurope Europe between southern r ) were highest in southern European insouthernEuropean highest ) were r = 3.76) (Table 1). Paired Wilcoxon Wilcoxon 1).Paired = 3.76)(Table

X E 2 ) (p < 0.001); thistendency of (p <0.001); ) = 0.43, p-value = 0.514). =0.514). =0.43,p-value plants across 42populations. across plants IS values (Table 1, Table S2). S2). 1,Table (Table values X 2 =5.66,p-value r (H O ) showed E E : W =81,p- : W = 0.18,A r = Version postprint Hood, M. E.(Auteurde correspondance), Giraud,T. (2016).Distribution andpopulation structure of the anther smut Microbotryum silenes-acaulis parasitizing an arctic-alpine plant. Molecular Bueker, B.,Eberlein,C., Gladieux,P., Schaefer,A., Snirc,A.,Bennett, D.J.,Begerow, D., We first used the Bayesian methodsimplementedintheS firstusedtheBayesian We Population structure value =0.368). one being restricted to this area (light green), (light tothisarea restricted one being Individuals individuals fromthewesternAlps. from some with also violet), and orange (red, clusters three to assigned mainly were Alps central from and brown),s found(yellowtwo clusterswere Colorado, while thecluster orange occurred mostly In butalso inthein Alaska Alps. Norway, ex The blueclusterwas 4and5). (Figs. orange) populationswere AtK=8theNorthAmerican assignedAlps were toeith singlegreenyelloweach clusterand (blue, respectively), clusters, while individuals fromthe NorthAm Europe. Populationsfrom between wasobserved separation At K=3,aclear w we coulddetect thisindicat Together a genuinepopulationstructure. complete appeared cluster each new that forK>8 = 8the rate increase probability of in became markedlyIn smaller. the barplots addition, showed rangin (K) ofclusters forthenumber Ln(Pr(X|K)) dataset.Th population structureintheglobal Iceland and Pyrenees (dark green). Pyrenees and Pyrenees green). (dark and Pyrenees Iceland This article is protected Allrights bycopyright. reserved. Accepted Article and geneticdiversity (H Ecology, 25 (3), 811-824. DOI : 10.1111/mec.13512 ith our dataset. Comment citer cedocument: E andA er one of these three clusters (Figs. 4 and 5). 4and5). (Figs. clusters three er oneofthese r ) of a population(H ) of

erica, Norway and the Pyrenees were mostly toa assigned were and thePyrenees Norway erica, ere was a constant increase of the likelihood of aconstantincrease ere was while the other cluster could be alsofoundin could while the othercluster cattered across all Norwegian regions. Samples allNorwegian across cattered clusively found in North America, and mostly in andmostly America, clusively foundinNorth mainly assigned to two clusters (blueand totwo assigned mainly Iceland populations were highly admixed, with highly admixed, Iceland populationswere g twoto20clusters from ly admixed and therefore didnotcorrespond to andtherefore ly admixed North America, northern Europe and southern Europeandsouthern northern NorthAmerica, E ed that K = 8 was the finest genetic structure genetic structure thefinest ed thatK=8was southern Alps were assigned totwoclusters, assigned southern Alpswere : r = 0.80, p-value = 0.333; A = 0.333; : r=0.80,p-value

TRUCTURE TRUCTURE software toinvestigate the ; nevertheless, after K ; nevertheless,after r : r=0.63,p- Version postprint Hood, M. E.(Auteurde correspondance), Giraud,T. (2016).Distribution andpopulation structure of the anther smut Microbotryum silenes-acaulis parasitizing an arctic-alpine plant. Molecular Bueker, B.,Eberlein,C., Gladieux,P., Schaefer,A., Snirc,A.,Bennett, D.J.,Begerow, D., Observed heterozygosity (H Observed heterozygosity 110 individualscollectedinthe18populati markers. showed similargenetic levels of variability of and Svalbard,th Norway For the populationsfrom populations Norway/Svalbard of structure Genetic genetical populations, appeared American North the in mostly occurred which cluster, Theblue Alps. central in cluster the twoclusters identified inNorway/Svalbard, also which appearedgeneticallyclose tothe red in the southern Alps,while the highest selfingrate was inferred (0.67),in inthe cluster red ra selfing thelowest wasclear, cluster assignment and pairwise F and pairwise PCA analysespatterns largely displayed consistent with the Bayesianclustering(Fig. S2).PCA central (A Alps.Allelic richness and the tendency of homozygosity excess was also indicated by high G high indicatedby also was excess ofhomozygosity and thetendency S S2). of 0.65(Table1,Table highest in the dark green cluster in the southern Alps (A cluster inthesouthernAlps greenhighest inthedark This article is protected Allrights bycopyright. reserved. I substantial membership population structure as the S population structureasthe 9, but the increase became smaller above K = smallerabove became 9, buttheincrease NSTRUCT Accepted TRUCTURE Article

The average number of alleles per population population ofallelesper number average The , which jointly estimates selfing rates an rates estimates selfing , whichjointly results displayed an increase of thelike of results displayedan increase Ecology, 25 (3), 811-824. DOI : 10.1111/mec.13512 ST values (Table 3;Figs. 5andS2)furt values(Table Comment citer cedocument: proportions in northern and southern proportions innorthernand O TRUCTURE ) was lower than expected heterozygosity (H heterozygosity expected than lower ) was ly the most distant. ly themostdistant. r ) was lowest in the Norway yellow lowest inthe cluster) was (A Norway

software. For K = 5, i.e., the highest K-value for which K-value for K=5,i.e.,thehighest For software. ons from Norway and Svalbard populations. ons fromNorway andSvalbard M. silenes-acaulis 4. The Evanno-method thus indicatedthatthe The Evanno-method thus 4. tes were found in the dark green cluster (0.37), green cluster foundinthedark were tes d population structure, inferred asimilar d populationstructure,inferred e set of the seven new e setoftheseven lihood Ln(Pr(X|K) for increasing K up to K = Kupto Ln(Pr(X|K)for increasing lihood her revealed a close relationship between between relationship a close her revealed indeed ranged from 1.6to2.14acrossthe from indeed ranged r = 3.51) (Table 2). 2). = 3.51)(Table

European populations. populations. European as the previously developed asthepreviously IS values, with an average anaverage with values, microsatellite markers markers microsatellite E ) for all seven loci, ) forallseven r = 1.69) and = 1.69) Version postprint Hood, M. E.(Auteurde correspondance), Giraud,T. (2016).Distribution andpopulation structure of the anther smut Microbotryum silenes-acaulis parasitizing an arctic-alpine plant. Molecular Bueker, B.,Eberlein,C., Gladieux,P., Schaefer,A., Snirc,A.,Bennett, D.J.,Begerow, D., infections on the plants, which has been also infections ontheplants, which hasbeen confir surveys further inherbarium specimens (Antonovics hosts their with preserved pathogens fungal for including ranges, species worldwide estimating Th Hultén andFries(1986). acaulis pathogen Herbarium surveys allowed mapping the distributionoftheplant mapping surveysHerbarium allowed the Distribution of an with plant pathogenspecies effects of climate and envir ofclimateand effects survey herbarium combined The presentstudy DISCUSSION (Table 4). S4). The selfing rates inferred by I S4). The selfing rates inferred by This article is protected Allrights bycopyright. reserved. Accepted ArticleS4). Thiscluster appeared tobe th yellow re clusterwasmainly regions, although the (Figs. S3 regions geographical with associated genetic offour theexistence analysis supported clusters was thestronges subdivision infourgenetic was largely in agreement with the herbarium and records-based species map provided by by provided map species andrecords-based herbarium the with agreement in waslargely M. silenes-acaulis et al. et Ecology, 25 (3), 811-824. DOI : 10.1111/mec.13512 Silene acaulis 2003;Hood Comment citer cedocument: onmental factors on the distribu onmental factorsonthe is supports the utility of herbarium collections as resources for for as resources is supportstheutility collections of herbarium at a broad geographic scale. The geographical range inferred for inferred range geographical The scale. geographic abroad at arctic-alpine distribution. arctic-alpine andanther-smutdisease et al. et e most genetically distant based on F e mostgenetically distantbasedon NSTRUCT

2010). The absence of notes about anther-smut disease on aboutanther-smut ofnotes 2010).Theabsence varied from 0.72 to 0.81 for the four genetic clusters four from0.72to0.81forthe varied shown by other studies (Hood & Antonovics2003), by studies(Hood shown other med that collectors do not appear to detect detect to appear donot collectors that med s and population genetics for understanding the understanding populationgenetics for s and and S4). Each cluster was indeed present in all all in present was indeed Eachcluster S4). and clusters, that were however notstrongly however thatwere clusters, stricted to Svalbard populations (Figs.stricted toSvalbardpopulations S3and t structure (Fig. S1).Resultsofthestructure t structure(Fig.

tion andpopulationsubdivisiona S. acaulis ST values (Table values S4;Fig. andofitsanthersmut S. Version postprint Hood, M. E.(Auteurde correspondance), Giraud,T. (2016).Distribution andpopulation structure of the anther smut Microbotryum silenes-acaulis parasitizing an arctic-alpine plant. Molecular Bueker, B.,Eberlein,C., Gladieux,P., Schaefer,A., Snirc,A.,Bennett, D.J.,Begerow, D., also found higher disease prevalence inregions prevalence disease also foundhigher (Anderson & May 1979). However, field studies of fieldstudiesof However, & May 1979). (Anderson higherpopulations hostdensity isusually thought topromotesustainability pathogen of of hostsitedensity onanth effect 2003). Anegative In addition, there was significantly greater incidence of anther-smut disease in more northern northern more in disease ofanther-smut incidence greater significantly was there addition, In collected. are materials whenherbarium sampled withinpopulations),whichwouldre are diseased associated diseaseequilibrium withhigher prevalence ( levels of localized endemicity (Hood endemicity levels oflocalized higher exhibiting species host in common more was disease the where prevalence, on disease surveys ha herbarium previous sampling efforts, possibility lowerde regions have northern thatfar on hostdemographic structure factors and/or latitudes and locations where the were host less de dynamics of anther smut transmission, Brunsdynamics anthersmut of This article is protected Allrights bycopyright. reserved. Accepted Articlewhich in years;Forbis & Doak2004) hundred (several on the high ofdisease levels (Hood surveys smuts estimatedfrombroaderherbarium other to compared rate average the twice is which of2.9%, incidence of range geographic the foundthroughout was Anther-smut pathogen rates and range disease inferring mi been haslikely bias collection that suggesting S. acaulis Ecology, 25 (3), 811-824. DOI : 10.1111/mec.13512 typically rarely recovers (Marr (Marr recovers typicallyrarely Comment citer cedocument: S. acaulis of disease in nature (Antonovics innature of disease

et al. et mayfrom the verynature long-lived come ofthe plants 2010) or inmoreisolatedlocalities(Antonovics 2010)or the pathogen distribution.While thereisa the pathogen et al. et of more rarefied host populations (Carlsson- hostpopulations more rarefied of combination with a systemic infectionfrom withasystemic combination ve suggested the importance ofhostdemography importance suggestedve the et al. sult in the disease being more likely to be morelikely inthediseasebeing tobe sult nimal. Thus, herbaria surveys are useful for useful surveys are nimal. Thus,herbaria nsities of collection sites because of biasesin nsities ofcollectionsitesbecause nse, indicating an influence of environmental ofenvironmental an influence nse, indicating (2015) recently showed that hostlongevity is showedthat (2015)recently er smut incidence may appear surprising, as appear surprising, er smutincidencemay Microbotryum 2005).Using that incorporates amodel i.e., et al. et

proportion of host individualsthat ofhost proportion 2010). Part of the explanation for theexplanation 2010).Partof et al. et on the plant on theplant Silene S. acaulis 2003). hosts ofanther Viscaria alpine withan et al. et

and effects oflatitudemay effects play hosts et al. et hosts (Biere & Honders 1996; Cafuir & 1996;Cafuir hosts(Biere Honders M. silenes-dioicae M. ity of the pathogen to warmer conditions. conditions. towarmer pathogen ity ofthe in comparison to other non-arctic-alpine species species non-arctic-alpine toother comparison in was low overall was other compared to 2014). This was explained by population lower 2014).Thiswasexplained S. latifolia eas wouldhavelessof eas ronments. In comparison, population size and In and populationsize comparison, ronments. selection for resistant host genotypes. Ingenotypes. forresistanthost selection study by Abbate and Antonovics (2014)actually andAntonovics Abbate by study species (Kaltz (Kaltz species have occurred due to founder effects during during effects duetofounder have occurred S. vulgaris on between high frequencies of disease and ofdisease frequencies on betweenhigh and

( population size, due to a high degree degree a high dueto size, population S. dioica Vercken et et al. Vercken were more likely more tobefoundin were et al. et a direct rolea indetermining direct an impactoninfection havegeneralist much more 1999); it would therefore 1999);itwouldtherefore 2010). This could be 2010).Thiscouldbe et al. et Microbotryum 2007)while S.

M. lychnidis-dioicae M. lychnidis-dioicae M. lychnidis-dioicae 2011). Opportunities to encounter a mating partner for for partner 2011).Opportunitiestoencounteramating S. acaulis past geological data,and past (Marr & Delph 2005). Alternatively, arctic alpine arctic Alternatively, & Delph2005). (Marr population under-study coul population under-study genotypes. The long lifespan and the high number number high andthe lifespan Thelong genotypes. et al. et persistence of host populations, andlowdisease ofhostpopulations, persistence may also be expected to affect pathogen genetic genetic pathogen affect to beexpected also may than inother occupy different stems. Marr & Delph (2005) & Delph (2005) stems.Marr occupy different expected tofavor theoccurrence ofmultiple me samples from the Alps moststemswithiname samplesfromtheAlps thin individualhostplants through multiple et al. et 2015) may affect mating system and system mating 2015)mayaffect 2008), estimated selfing not were rates 2008),estimated (Hood 2003; Lopez-Villavicencio Lopez-Villavicencio (Hood2003; and

M.silenes-dioicae Microbotryum Microbotryum M. silenes-acaulis S. acaulis d have persisted for d havepersistedfor cushions can bear a a canbear cushions species due to the the dueto species , for example example , for . Although et al. et

Accepted Article 2003;Alsos Ecology, 25 (3), 811-824. DOI : 10.1111/mec.13512 et al. e.g., lleles withinsmalle lleles 2007; Westergaard Comment citer cedocument: Lira-Noriega Lira-Noriega et al. et dispersal of seeds of dispersal ofseeds Microbotryum M. lychnidis-dioicae 2013; Gladieux 2013;Gladieux e.g.

, (Abbott& Brochmann2003;Eidesen oceans or mountains) are known tolimitdispersaland areknown ormountains) oceans as North American individuals assigned to European to assigned individuals American North as et al et M. silenes-acaulis M. silenes-acaulis r populations(Karron et al. et concerning long-distance dispersal similarlyraise d by importing plants as ornamentals ( d by importing plantsasornamentals . 2015). . 2015). ersal (Brochmann &ersal (Brochmann Brysting It 2008). remains genetic differentiation differentiation genetic 2011). Genetic footprin Genetic 2011). S. acaulis S. acaulis et al. et other arctic-alpine distributed species (Abbott (Abbott species distributed arctic-alpine other pathogens that are neither transmitted by the neither pathogensthatare

– 2015).

that wasNorth America introduced in – occurs more regular e.g. by seed transfer transfer seed by e.g. regular occurs more

between populations fromNorth betweenpopulations et al. et et al. et between southern and between 2012;Levin ts oflong-distance 2014). However, the 2014).However, et al. et 2013).Our e.g., et al. et Farr Farr 2012). 2012). et al. et et et

2006). High genetic diversity and clus genetic diversity 2006).High levels ofgenetic levels è (Schönswetter s et al. et Comment citer cedocument: bott & Brochmann 2003; Eidesen bott & Brochmann 2003; 2008) distinct glacial refugia in Europe (Hewitt 1996,2004),including (Hewitt inEurope distinctglacial refugia rring in southern Europe in the NJ trees.Europe inthe Similar insouthernrring NJ patterns have in Europe would result fromdemographic inEurope bottlenecks would pheral refugia or even“nunataks” or refugia pheral S. acaulis .

et al. et variation in variation

found subdivision between continents, betweencontinents, foundsubdivision 2003). 2003). thepopulationstructureofitshost (Abbott 1995;Mikhaylova (Abbott large scales in natural plant pathogens ( scales innaturalplantpathogens large within small areas as well as admixture admixture as areasaswell small within on differentiation.Multiple at populations haveremain at of distinct clusters intheof more northern l distinct genepools in separated refugia also genepools refugia l distinct inseparated M. silenes-acaulis a. This is supported by the northern clusters the northernclusters a. Thisissupportedby onditions in central Europe were not were onditions incentralEurope

ter richness in southern Europe are insouthernEurope ter richness Arabis alpina et al. et 2013). Only few studieshave few 2013).Only insouthernEurope.Post-

would then have resulted in would thenhaveresulted (Koch et al. et ed separated for long for ed separated previous studieshave S. acaulis e.g., 2010)andother et al. et in i.e., Melampsora 2006)and

non- Version postprint Hood, M. E.(Auteurde correspondance), Giraud,T. (2016).Distribution andpopulation structure of the anther smut Microbotryum silenes-acaulis parasitizing an arctic-alpine plant. Molecular Bueker, B.,Eberlein,C., Gladieux,P., Schaefer,A., Snirc,A.,Bennett, D.J.,Begerow, D., Foundation (SCHA 1723/1-1) and a grant from the DAAD program “Promos” DAAD program grant fromthe anda Foundation (SCHA 1723/1-1) genotyping. Thisres part ofthemicrosatellite Ecophysiologie des Céréales (UMR1095), Gentyane Céréales Ecophysiologie des glaciation. hostthroughout integrat very long-livedherbaceous long-term associati and supports thevery close This article is protected Allrights bycopyright. reserved. Accepted thankthe 1115765. We DEB- Foundation award grant ANR-13-BSV7-0008 the TG acknowledges ACKNOWLEDGMENTS consistent withthose inthe reported host multiple differentiated and recent populations migration events. These patterns overall are acaulis, Articleevidence additionally We found connectedness. to post-glacial recol northern regionsdue agricultural ordomesticated systems. Thismaygenetic bedue tolower forresistance variation in indis variation a majorwithin-species reveals locations host rarefied and more northern more in pathogen ofthe incidence greater significantly thehosts by inregions occupied ubiquitouspresence to haveanearly withar fungalpathogen a of global biogeography This study combinedsurveys naturalhistor of CONCLUSION suggesting persistence indistinct glacial refugia,recolonization from followed by Ecology, 25 (3), 811-824. DOI : 10.1111/mec.13512 Comment citer cedocument:

onization and a paucity of host population andapaucity ofhostpopulation onization S. acaulis earch was also supported by the German Science GermanScience the wasalsosupportedby earch y andpopulationgenetics toinferthe collections ease risk that has been rarely observed innon- risk thathasbeenrarely ease on of the obligate and specialized pathogen ona andspecialized of theobligate on for strong population structurewithin strong population for INRA Research Unit Génétique, Diversité INRA UnitGénétique, Research ctic-alpine distribution. The pathogen appears appears distribution.Thepathogen ctic-alpine . MEH acknowledges the NationalScience the . MEHacknowledges (Abbott1995;Mikhaylova ed processes of recolonization afterthelast ofrecolonization ed processes platform(Clermont-F

S. acaulis errand, France) for errand, France)

(German (German et al. et .

The 2010).This M. silenes-

et et Version postprint Hood, M. E.(Auteurde correspondance), Giraud,T. (2016).Distribution andpopulation structure of the anther smut Microbotryum silenes-acaulis parasitizing an arctic-alpine plant. Molecular Bueker, B.,Eberlein,C., Gladieux,P., Schaefer,A., Snirc,A.,Bennett, D.J.,Begerow, D., Bennett KD, Tzedakis PC, Willis KJ (1991) Quaternary refugia of north European trees. trees. northEuropean of refugia (1991) Quaternary KJ Willis PC, Tzedakis Bennett KD, Systematics KellnerR,YurkovA, AM, Begerow D,Schäfer and evoluti This article is protected Allrights bycopyright. reserved. smutfungus theanther in Hostadaptation S(1996) Honders A, Biere theirhost. evolution ofsmutfungi on Onthe Lutz M,StollM (2004) M, Göker Begerow D, (2014) J Abbate JL, Antonovics REFERENCES collecting and samples. providing R surveysand Oliver herbarium with assistance thankErin We Service). Academic Exchange Abbott RJ, BrochmannAbbott RJ, C(2003)Histor pl of (2000)Molecularanalysis Abbott RJ Accepted RA (1996)EvaluatingBeaumont loci MA,Nichols A, C,Andrieux F,B, Dutech Barrès Halkett JY, HoodME,ThrallPH,Abrams Antonovics J, Anderson RM,May RM(1979)Populationbiol anintroductiontoquaternary world: (1994)Theiceage Borns HW Andersen GB, Availablefro version0.93.1098. package IG,Alsos I, D,Skrede We EidesenPB, Ehrich Article others(2014).rgl: 3D M,and Adler D,Murdoch Chapman FoAbbott RJ, HM, CrawfordRMM, 587. poplar rustfungus Europe and recent founder effects overseas. overseas. effects founder Europe andrecent Ustilaginomycotina 280 years.last 2.5million Scandinavian Oslo: Univ.Press. with emphasis on Nort historyresearch and system: insights from changes across host populations andclimate. hostpopulations across system:changes insights from of Biogeography Frontiers inBasidiomycoteMycology 1343–1346. 1136. Science Frequent long-distance C(2007) P, Brochmann Arctic and s more populationsof derivations of Hultén. structure. structure. 1531. States. United eastern the in (Caryophyllaceae) distribution ofanther-smutfungus ( : 361–367. Ecology, 25 (3), 811-824. DOI : 10.1111/mec.13512 , Molecular Ecology Molecular 316 Proceedings of the Royal Society B: BiologicalSciences theRoyalSocietyB: of Proceedings , 1606–1609. , 1606–1609. Comment citer cedocument: , outherly latitudes. 18 Melampsora larici-populina Melampsora . In: McLaughlin DJ, Spatafora JW (eds) The Mycota. 7. Part A. 7. Part (eds)TheMycota. JW Spatafora DJ, In:McLaughlin . , 103. on, 2ndedn. Springer.

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Version postprint Hood, M. E.(Auteurde correspondance), Giraud,T. (2016).Distribution andpopulation structure of the anther smut Microbotryum silenes-acaulis parasitizing an arctic-alpine plant. Molecular Bueker, B.,Eberlein,C., Gladieux,P., Schaefer,A., Snirc,A.,Bennett, D.J.,Begerow, D., Gao H, Williamson S,Bustamante CD(2007)A Gao H,Williamson Brochmann C, BrystingBrochmann Forbis TA, DoakDF Seedling (2004) establishment P,HoodME,GiraudT(2013) Fontaine MC,Gladieux Fungi onplantsandplantproduc (Ed.) (1989) Farr DF Bakkestuen V D, Eidesen PB, Ehrich This article is protected Allrights bycopyright. reserved. Eidesen PB, Alsos IG, Popp M IG, Alsos Eidesen PB, an (2007) Theextent GibbardPL Ehlers J, Earl DA,vonHoldtBM(2012)STRUCTURE I,Landvik TM,Nordal C,Gabrielsen Brochmann Bruns E, Hood ME, Antonovics J (2015) Rate of (2015)Rate J ME,Antonovics Hood Bruns E, Bucheli E, Gautschi B, Shykoff JA (1998) Isolati (1998) Shykoff JA B, Gautschi Bucheli E, Buono L, López-Villavicencio M, Shykoff JA, Snirc A, Giraud T (2014) Influence of multiple multiple Influence of T(2014) SnircA,Giraud JA, M,Shykoff L,López-Villavicencio Buono Cafuir L, Antonovics J, L,Hood ME(2007)Tissu Cafuir AntonovicsJ, Comes HP, Kadereit JW (1998) The effect of Quat of effect (1998)The Comes HP,KadereitJW Accepted Distributionof LH (2005) Eddington Cripps CL, Suhai,S(1999) B, Tand Wetter, Chevreux, Article of hostlongevity ondisease PH(2006) Theimpact U,Thrall Carlsson-Granér BE, PH(2014) Thrall U,Giles Carlsson-Granér Ecology Research, dynamics host-pathogen transmission: German Conference on Bioinformatics (GCB), on German Conference information and additionalsequence highways, traffic barriers and capitals of diversity. andcapitalsofdiversity. barriers highways, traffic 176 andinbr ofpopulationstructure inference American JournalofBotany pathogen on traits ontwohostspeciesandtheirF1-hybrid. Diversity ( history of a circumpolar key history species. a circumpolar of Quaternary International Genetics Resources visualizing STRUCTUREoutputandim patterns inarctic–alpinehabitats. rasa? Thehistory ofNorth rasa? the anther smut fungus fungus smut anther the and short-livedhosts:Brief communication. population and its castrating parasite. parasite. population anditscastrating onvi infection andrelatedness Sciences anther to resistance families on the Beartooth Plateau, Rocky Mountai Plateau,Rocky Beartooth families onthe and evolution. spatially systems. structured Microbotryum violaceum , 1635–1651. Ecology, 25 (3), 811-824. DOI : 10.1111/mec.13512 , , 168 1 , 181–195. , 181–195. Silene latifolia , 415–419. , 415–419. Trends inPlantScience Comment citer cedocument: AK (2008)TheArctic–

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Molecular Ecology 9 45-56. New Phytologist Taxon , e98526. ns, U.S.A., in reference to large-scale tolarge-scale reference ns, U.S.A.,in and AlpineResearch , 69 . , , , International JournalofPlant 16 107 , 551–560. 52 , 3902–3925. , 3902–3925. , 417. tion and alteration of floral tion andalterationoffloral , 307–320. , the Arctic: plant dispersal Arctic: plantdispersal the ation ofindividual 198 ogy: Proceedingsofthe , , 138 , 946–956. 200 , 7 Evolutionary , 499–511. , 665–666. , 665–666. Plant Ecology &Plant Ecology , , 898–910. 37 , 177–188. Conservation Conservation level Genetics , Version postprint Hood, M. E.(Auteurde correspondance), Giraud,T. (2016).Distribution andpopulation structure of the anther smut Microbotryum silenes-acaulis parasitizing an arctic-alpine plant. Molecular Bueker, B.,Eberlein,C., Gladieux,P., Schaefer,A., Snirc,A.,Bennett, D.J.,Begerow, D., This article is protected Allrights bycopyright. reserved. withintheme (2004) Mating Hood ME,AntonovicsJ (2003) Plantspeciesdescripti Hood ME,AntonovicsJ Hood ME(2003)Dynamics of fungus. smut anther the in species cryptic of Giraud T(2004)Patterns of host-specific evidence Phylogenetic T(2007) E,Giraud Hood ME,Fournier LeGac M, Hood ME,Mena-Alí M, D,Solignac Vautrin E, Fournier Giraud T, Jakobsson M,Jakobsson RosenbergNA(2007)CLUMPP: aclus (1986) Fries M Hultén E, (2005)Selfing O,Shykoff JA Giraud T,Jonot Kaltz O,GandonS,Kaltz Michalakis Y,Shykoff JA (1962) V,RichardsPW Jones Giraud T, Yockteng R,Lopez-Villavicencio M, Giraud T,Yockteng Goudet J (2014)hierfstat:Estima Goudet J Hood ME P,FeurteyA, Gladieux AguiletaGladieux P,Byrnes G EJ, Marthey S,ChiapelloH, R, Giraud T,Yockteng Hewitt GM (2004) Genetic cons Genetic Hewitt GM(2004) Hewitt GM(1996)Somege AcceptedIntratetrad (2000) mating,Hood ME,AntonovicsJ Article 241. Anther heterozygosity. Sciences, Biological B: Society Royal violaceum microsatellite loci hostraces inthree thephytopathogenic of fungus violaceum 3: commentary to the maps: totalindex3: commentary tothe maps: Microbotryum Bioinformatics dealing withlabelswitchin artificial inoculations. fungus, Eukaryotic Cell of theanthersmutfungus Molecular Ecology andanimals. plants in Resources complex fungal phytopathogenic ofthe species Isolationresources: polymorphic microsat of60 Gendrault-J G, Refregier DM, HoodME, deleterious alleles in Philosophical Transactions oftheRo http://www2.unil.ch/popgen/softwares/hierfstat.htm. speciation. speciation.

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Polymorphism in Southern Europe Ecology, 25 (3), 811-824. DOI : 10.1111/mec.13512 0.32** 0.19 4.89 0.45 et al. et 53 Comment citer cedocument:

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r and Version postprint Hood, M. E.(Auteurde correspondance), Giraud,T. (2016).Distribution andpopulation structure of the anther smut Microbotryum silenes-acaulis parasitizing an arctic-alpine plant. Molecular Bueker, B.,Eberlein,C., Gladieux,P., Schaefer,A., Snirc,A.,Bennett, D.J.,Begerow, D., individual and the colors indicate the proportion of indicate theproportionof individual andthecolors fromK=3to assumptions the set (N=195)under Microbotryum silenes-acaulis. andgenetic selfingfor the ratesat clusters Figure 4. collecti herbarium and density of loca northern latitudes.Herbarium 3 Figure healthy sites shown). sele aroundrandomly line)and diseased sites(red numberofothe the sites. Sitesaresortedby Figure 2. sampled. representregions Blue points shown ingreen. Thedistributionma inherbaria. specimen observed Figure 1. FIGURE LEGENDS heterozygosity. This article is protected Allrights bycopyright. reserved. + Table 4. individuals analyzed, Number of different alleles per locus; A per locus; alleles ofdifferent Number analyzed, individuals

Averageacross microsatellite seven markers developed inthe study; present N:Numberof Red cluster N Blue cluster Accepted ArticleGreen cluster Yellow cluster Numberofindividuals,di . Demographic and disease incidence variation in Silene acaulis across southern to to southern across acaulis Silene in variation incidence disease and Demographic . Genetic clustering Geneticclustering of Micr Density Silene of acaulis localities neardiseased and healthy herbarium collection points) (green mapofhealthy Distribution

Ecology, 25 (3), 811-824. DOI : 10.1111/mec.13512

Comment citer cedocument: 28 23 22 25 on sites was based upon100kmradii. on siteswasbased

lities (N = 790) were dividedinto lities (N=790) versity (allelic richness A

obotryum silenes-acaulis genotypes for theworldwidedata for genotypes obotryum silenes-acaulis K=4 in the Norway/Svalbard populations of populations the Norway/Svalbard K=4in r collections siteswithina100kmradiusaround r collections from where populations for present study were were present study wherepopulationsfor from 2.02 1.77 2.04 2.12 cted healthy sites (grey lines; first40random sites(grey cted healthy A p of S. acaulis after Hulten Fries (1986) is Fries Hulten acaulisafter p ofS. its genotype assigned to the different clusters. clusters. the different its genotype to assigned r and diseased (red points) Silene acaulis acaulis Silene (red points) anddiseased = 8 clusters. Each verti = 8clusters.Each

r and expected heterozygosity heterozygosity and H expected r : Allelic richness; H latitude quartiles for analysis analysis latitudequartilesfor 0.22 0.16 0.18 0.19 H E

cal bar represents an cal barrepresents E Selfing rate : Expected Expected : 0.81 0.74 0.72 0.77

E ) Version postprint Hood, M. E.(Auteurde correspondance), Giraud,T. (2016).Distribution andpopulation structure of the anther smut Microbotryum silenes-acaulis parasitizing an arctic-alpine plant. Molecular Bueker, B.,Eberlein,C., Gladieux,P., Schaefer,A., Snirc,A.,Bennett, D.J.,Begerow, D., This article is protected Allrights bycopyright. reserved. Accepted Article based onFST distances. clusters distances among onthethreelocalitiesinAlps. is azoom = 3 K at analyses STRUCTURE the from estimated Figure 5. Maps showing the mean assign mean the showing Maps Ecology, 25 (3), 811-824. DOI : 10.1111/mec.13512 Comment citer cedocument:

ment proportions per locality perlocality ment proportions Neighbour-joining trees illustrate the genetic illustratethe trees Neighbour-joining (a) and K = 8 (b). The square at the bottom at thebottom square (b). The and K=8 (a)

Healthy Sites within Radius (100km) 10 20 30 40 50 60 CollectionSites Sorted by Nearby Densityof Sites 0 11 51 92 32 72 13 35 33 31 29 27 25 23 21 19 17 15 13 11 9 7 5 3 1

Density of Collection Sites 10 15 20 25 0 5

urie1Qatl urie3Quartile 4 Quartile 3 Quartile2 Quartile 1 Latitude Quartilesnorthern) (southern to

0.00 0.02 0.04 0.06 0.08 0.10 Proportion of Sites Diseased Sites of Proportion Version postprint Hood, M. E.(Auteurde correspondance), Giraud,T. (2016).Distribution andpopulation structure of the anther smut Microbotryum silenes-acaulis parasitizing an arctic-alpine plant. Molecular Bueker, B.,Eberlein,C., Gladieux,P., Schaefer,A., Snirc,A.,Bennett, D.J.,Begerow, D.,

This article is protected Allrights bycopyright. reserved. Accepted Article Ecology, 25 (3), 811-824. DOI : 10.1111/mec.13512 Comment citer cedocument: