A comprehensive molecular phylogeny of the Mortierellales ... · 3HUVRRQLD±9ROXPH Fig. 1 Typical morphological structures of different isolates of the Mortierellales ZKLFK DUH VXLWDEOH

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A comprehensive molecular phylogeny of the Mortierellales (Mortierellomycotina) based onnuclear ribosomal DNAWagner, L.; Stielow, B.; Hoffmann, K.; Petkovits, T.; Papp, T.; Vágvölgyi, C.; de Hoog, G.S.;Verkley, G.; Voigt, K.Published in:Persoonia - Molecular Phylogeny and Evolution of Fungi

DOI:10.3767/003158513X666268

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Citation for published version (APA):Wagner, L., Stielow, B., Hoffmann, K., Petkovits, T., Papp, T., Vágvölgyi, C., ... Voigt, K. (2013). Acomprehensive molecular phylogeny of the Mortierellales (Mortierellomycotina) based on nuclear ribosomalDNA. Persoonia - Molecular Phylogeny and Evolution of Fungi, 30, 77-93.https://doi.org/10.3767/003158513X666268

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Persoonia 30, 2013: 77–93www.ingentaconnect.com/content/nhn/pimj http://dx.doi.org/10.3767/003158513X666268RESEARCH ARTICLE

INTRODUCTION

The order Mortierellales – from historical aspects on morphology and systematics to modern approaches in fungal identificationThe Mortierellales are a long known, species rich order of the basalfungi.Withnearly100describedspecies,theMortierellalesisoneofthelargestbasalfungalorders.However,only13genera are described in one family, the Mortierellaceae(Kirketal.2008,andSpeciesFungorumJanuary2013).Outofthesegenera six are currently accepted with one potential additional genusrecentlydescribed(Kirketal.2008,Jiangetal.2011,Table1).ThefirstspeciesofthetypegenuswasdescribedbyCoemans(1863)asMortierella polycephala, originally isolated fromamushroom.ThenameMortierella was given in tribute toM.DuMortier,thepresidentoftheSociétédeBotaniquedeBelgique (Coemans1863).Nevertheless, the common life-style of those fungi is as soil inhabiting saprobic organisms ondecayingorganicmatter.Onlyonespeciesisoccasionallydescribedfromanimalfungalinfections(deHoogetal.2009).Many mortierellean species possess the ability to produce poly- unsaturated fatty acids or to convert organic compounds, making them highly interesting organisms for biotransforma-tionsandotherbiotechnologicalapplications (Holland2001,Higashiyamaetal.2002).As many basal fungal species, the Mortierellales possess a reduced macro- and micromorphology with only few morpho-

logical characters available for differentiation.Examples ofmicromorphologicalfeaturesareshowninFig.1and2.Overallappearance of the colonies is the typical zonate, rosette-like growth(Fig.1a)andtheoftenoccurringgarlic-likeodour.Colo-nies are in general white to light-grey, young mycelium is coeno-cyticandseptateinagedcultures.Asexualsporesareproducedinsporangiaorsporangiolaandarepassivelyreleased(e.g.,Fig. 1h, s).The sporangiophores could bewidened at thebase(e.g.,Fig.1o)andvariouslybranched(e.g.Fig.1h, l). Acolumella isneverprotruding intothesporangium.Sexualreproductive structures (zygospores, Fig. 2r) are often sur-roundedbyahyphalsheat.Variouslyshapedchlamydosporesandstylosporesarealsopossible(Fig.1w,2l) (Zychaetal.1969,Gams1977).Morphologicalidentificationbasedsolelyon asexual features, leading to the aforementioned traditional classification.Mortierella was furthermore divided into nine sec-tions based on morphology: Actinomortierella, Alpina, Haplosporangium, Hygrophila, Mortierella, Schmuckeri, Simplex, Spinosa and Stylospora(Gams1977).Judgingfromtheproposedtotalnumberoffungiwith1.5millionspecies and the current number of described and registered specieswith75000 (Hawksworth2001) it seems likely thatalso for the order Mortierellales an unknown percentage of undescribed species may exists, a fact which might influence phylogeneticanalyses.Yet,arecentstudychallengedpreviousestimations of the potential number of undescribed fungal spe-cies and proposed that, at least for Mortierella, nearly all species aremostlikelydescribedalready(Nagyetal.2011).Basedonthisknowledge,phylogeneticanalysesincludingsequencesofan extensive amount of type and reference strains could reveal thenaturalevolutionaryrelationships.Nevertheless, the phylogenetic position of the Mortierellales iscontroversialdiscussed.Theyareeitherplacedwithinthesubphylum Mucoromycotina(Hibbettetal.2007)orelevatedto an own subphylum, the Mortierellomycotina (Hoffmannetal.2011).Furthermore,relationshipswithinthisorderarealso

A comprehensive molecular phylogeny of the Mortierellales (Mortierellomycotina) based on nuclear ribosomal DNAL.Wagner1,2,B.Stielow3,K.Hoffmann1,2,T.Petkovits4,T.Papp4,C.Vágvölgyi4, G.S.deHoog3,G.Verkley3,K.Voigt1,2*

1 UniversityofJena,SchoolofBiologyandPharmacy,InstituteofMicrobio-logy,DepartmentofMicrobiologyandMolecularBiology,JenaMicrobialResourceCollection,Neugasse25,07743Jena,Germany;

correspondingauthore-mail:[email protected] Leibniz-InstituteforNaturalProductResearchandInfectionBiology(HKI),DepartmentofMolecularandAppliedMicrobiology,Beutenbergstrasse11a,07745Jena,Germany.

3 CentraalbureauvoorSchimmelcultures,CBS-KNAWFungalBiodiversityCentre,P.O.Box85167,3508ADUtrecht,TheNetherlands.

4 UniversityofSzeged,FacultyofScienceandInformatics,DepartmentofMicrobiology,6726Szeged,Középfasor52,Hungary.

Key words

internal transcribed spacerlargesubunitribosomalDNAtaxonomic revisionZygomycetesZygomycota

Abstract The basal fungal order Mortierellalesconstitutesoneofthelargestordersinthebasallineages.Thisgroupconsistsofonefamilyandsixgenera.Mostspeciesaresaprobicsoilinhabitingfungiwiththeabilityofdiversebiotransformations or the accumulation of unsaturated fatty acids, making them attractive for biotechnological ap-plications.Onlyfewstudiesexistaimingattherevelationoftheevolutionaryrelationshipsofthisinterestingfungalgroup.ThisstudyincludesthelargestdatasetofLSUandITSsequencesformorethan400specimenscontaining63typeorreferencestrains.BasedonaLSUphylogram,fungalgroupsweredefinedandevaluatedusingITSse-quencesandmorphologicalfeatures.Traditionalmorphology-basedclassificationschemeswererejected,becausethe morphology of the Mortierellalesseemstodependoncultureconditions,afact,whichmakestheidentificationofsynapomorphiccharacterstedious.Thisstudybelongstothemostcomprehensivemolecularphylogeneticanalysesfor the Mortierellalesuptodateandrevealsunresolvedspeciesandspeciescomplexes.

Article infoReceived:17June2012;Accepted:2January2013;Published:13March2013.

78 Persoonia–Volume30,2013

Fig. 1 Typical morphological structures of different isolates of the Mortierellales,whicharesuitableforspeciesdelimitation.a.M. zychae CBS316.52,macro-scopicshapeofagrowingculturewiththetypicalzonategrowth;b.M. hypsicladiaCBS116202,acrotonousbranchingofasporangiophore;c.M. epicladia CBS 355.76,sporangiophoreandsporangiospores;d.M. zonataCBS228.35,basitonousbranchedsporangiophorewithsporangioles;e.Gamsiella multidivaricata CBS227.78,typicalbranchedsporangiophores;f.M. elongata FSU9721,basitonousbranchedsporangiophore;g.M. alpina FSU2698,sporangiophore;h.M. polycephala FSU867,sporangiosporeswithsporangia(arrow)andsporangiospores;i.Mortierellacf.wolfii CBS614.70,sporangiophorewithelongated

79L.Wagneretal.:MolecularphylogenyoftheMortierellales

Fig.1(cont.)sporangiospores;j.M. parvispora FSU10759,sporangiophores;k.M. hypsicladia CBS116202,typicalsporangiophorewithrhizoid;l.Mortierellacf.wolfii CBS 614.70,acrotonousbranchingofasporangiophore;m.Mortierellasp.FSU10557,sporangiophoreandsporangiospores;n.M. paraensis CBS547.89,tipsofasporangiophorewithapseudocolumellaandsporangiospores;o.M. alpina FSU2698,sporangiophorewithunmaturedsporangia;p.M. nanthalensis CBS 610.70,typicalrhizoidofasporangiophore;q.M. wolfii CBS651.93,sporangiosporeswithunusualremainofthesporangiacover(arrow);r.M. strangulata CBS455.67,rhizoidofthesporangiophore;s.Gamsiella multidivaricataCBS227.78,sporangiophoreswithsporangioles;t.Lobosporangium transversale CBS357.67,typicalsporangia,arrangedinclusters,containingnumeroussphericalsporangiospores;u.M. gamsii FSU10538,acrotonousbranchingofasporangiophoreandsporangiospores;v.Dissophora decumbens CBS592.88,septatesporangiophoresalongahyphaandsporangiospore(arrow);w.M. polycephala FSU867,stylospores;x.Gamsiella multidivaricataCBS227.78,sporangiolacontainingspores;y.M. kuhlmanii CBS157.71,branchingpatternofthebasitonouspartofthesporangiophoreandelongatedsporangiospores,pseudocolumella.—Scalebars:b,c,s–u,x=30µm;d,e,i=20µm;f,j,k,p=100µm;g,n,o,w=10µm;h,l,m,q,r,v,y=50µm.

Year Genus Synonyms Typespecies Numberof MycoBank describedspecies no.

1863 Mortierella Coem. Actinomortierella Chalab. 1968 Carnoya Dewèvre 1893 Haplosporangium Thaxt. 1914 Azygozygum Chesters 1933 Naumoviella Novot. 1950 M. polycephala 91 MB#20345

1914 Dissophora Thaxt. none D. decumbens 3 MB#20187

1936 Modicella Kanouse none M. malleola 2 MB#20336

1967 Aquamortierella Embree & Indoh none A. elegans 1 MB#20047

2004 Gamsiella (R.K.Benj.) Benny & M.Blackw. none G. multidivaricata 1 MB#28820

2004 Lobosporangium M.Blackw.&Benny Echinosporangium Malloch 1967 L. transversale 1 MB#28819

2011 Echinochlamydosporium X.Z.Jiang,X.Y.Liu&XingZ.Liu none E. variabile 1 MB#511829

MB=Mycobank:http://www.mycobank.org;IndexFungorum:http://www.indexfungorum.org.

Table 1 Chronological overview of descriptions and name changes for accepted genera in the order Mortierellales Caval.-Sm. 1998[MB#90555].Theorderconsists of several genera and one family, the MortierellaceaeA.Fisch. 1892[MB#81029].DatabasedonMycoBankandIndexFungorum(accessed7Janu-ary2013).

poorly understood and were extensively analysed only in few studiesuntilnow(Nagyetal.2011,Petkovitsetal.2011).Ourstudy contributes to the effort to elucidate natural phylogenetic relationships based on one of the largest datasets assembled sofar.Thisstudyconcernstheextensionofpreviousdatasetsandfacilitatesanapproachtomolecular identificationof theMortierellales.WesurveyedthediversityoftheMortierellales in-cludingare-evaluationofthemorphologybasedclassifications.This study based on the broad sampling of specimens which aremaintainedatthefungalculturecollectionsCBS(Centraal-bureauvoorSchimmelcultures,Utrecht,TheNetherlands)andJMRC(JenaMicrobialResourceCollection,Jena,Germany).

MATERIALS AND METHODS

Taxon sampling, culture conditions and light microscopic investigationsFor this study, a total of 421 isolates were obtained from the Centraalbureau voorSchimmelcultures (CBS,Utrecht,TheNetherlands) and the JenaMicrobial ResourceCollection(JMRC,Jena,Germany)(Table2).Strainswerecultivatedonmalt-extractmedium(3%maltextract,0.5%yeastextract)forDNAisolationandonoatmealagar(OA,3%),soilextractagar(Gams1969)orsyntheticnutrientdeficientagar(SNA,Nirenberg1981)formorphologicalexaminations.Cultivationwas done at 20–37 °C for 7–20 days depending on the re-quirementsofthefungustosporulate.ThelightmicroscopicalexaminationsshowninFig.1and2wereperformedwithanAxiophot(Zeiss,Germany).Thebestmethodtoobservemicro-scopicfeaturesistogrowculturesdirectlyoncoverslips.

Preparation of genomic DNA, PCR amplification and DNA sequencingGenomicDNAwaspreparedfrommyceliagroundedtoafinepowderinliquidnitrogenfollowedbypurification(Cenis1992)orlivingculturesalternatively,usingtheJetquickgeneralDNAclean up kit (Genomed) or a high-throughput 96-well plate

extraction(Ivanovaetal.2006)followingthegivenprotocols.ThePCRfortheamplificationoftheITS1-5.8S-ITS2nuclearribosomalDNAregionusesITS5/ITS1andITS4understand-ardorsemi-nestedconditions(Whiteetal.1990,Stielowetal.2009).PCRforamplifyingthepartial28SrDNA(LSU)wasdoneusingthestandardprimersLR0RandLR5ortheNL-primer(http://www.biology.duke.edu/fungi /mycolab/primers.htm).Theprimersdifferonlyintheirannealingtemperature(55°Cor 60°C). Increasing cycle extension time (90s/cycle)wasdoneinsomecasestoimproveamplification.PCRproductsweredirectlypurifiedusingFastAP thermosensitivealkalinephosphataseand shrimpalkalinephosphatase (Fermentas,ThermoScientific)orusingtheGeneCleanprotocol(Vogelstein&Gillespie1979).Thecycle-sequencingreactionwassetupusingABIbigdyeterminatorv.3.1,followingthemanufacturesinstructionsorbyusingaquarterof thesuggestedvolumes(modifiedmanufacturesprotocol),followedbybidirectionalse-quencingwithalaboratorycapillaryelectrophoresissystem(LifeTechnologies3730XLDNAanalyser).Sequenceswereevalu-atedwithChromasLite(TechnelysiumPty.Ltd.).SequencingprimerswerethesameasusedforPCR.ManuallycorrectionandassemblingofforwardandreversesequenceswasdoneusingtheBiolomicsdatabase(www.bio-aware.com)(Vuetal.2012)orSeqman(v.7.2.1).SequencesweredepositedatNCBIGenBank(Table2).

Alignments and phylogenetic analysesAtotalof364sequencesofITSand213sequencesofLSUweregeneratedinthisstudy.FortheextensionofthedatasetadditionalsequenceswereretrievedfromGenBank(Table2).Atotalof15sequenceswereexcludedand562weresubjectedtofurtheranalyses(298ITSand263LSUsequences).Align-mentswereperformedwithMAFFTv.6.833(Katoh2008)asimplementedinEPoS(Griebeletal.2008).MaximumLikelihoodanalyseswere carriedout usingRAxML (Stamatakis 2006)providedbytheCIPRESScienceGatewayv.3.2(http://www.phylo.org).RAxMLwasrununderthedefaultsettingswiththe


Fig. 2 Typical morphological structures of different isolates of the Mortierellales,whicharesuitableforspeciesdelimitation.a.M. verticillata CBS315.52,sporangiophorewithasporangiola;b.M. elongata FSU9721,elongatedsporangiosporescontainingcentraloildroplets;c.M. wolfii CBS651.93,crackedsporangiareleasingsporangiospores,onacrotonousbranchedtipofthesporangiophore;d.M. indohii CBS720.71,stylospores;e.M. schmuckeri CBS295.59,sporangiophoresalongsideahyphawithsporangiola;f.M. claussenii CBS294.59,sporangiophoresalongahyphawithsporangiola;g.M. clonocystis CBS 357.76,typicalswollenhyphae;h.M. zychae FSU719,typicalswollenhyphaearrangedinclusters;i.M. parvispora FSU10759,tipofasporangiophore,


followingadjustments:GTRGAMMAforbootstrappingandfinaltreeinferencewith1000bootstrapiterations.Theresultingphy-logenetictreeswhichbasedontheLSUsequenceswereusedtoidentifyclustersofstrains.FortheseclustersMAFFTalign-mentsoftheITSregionwerecomputedandRAxMLanalysesperformed.SubsequentalignmentsarecrucialsinceITSisingeneralhighlydiverseonhigherlevelclassification.Ifagroupofsequencescontainsahighnumberofarepetitivespeciesnotallsequenceswere includedintheITStree.AlignmentsandtreesaredepositedinTreeBASE2underhttp://purl.org/phylo/treebase/phylows/study/TB2:S13827.

RESULTS AND DISCUSSION

Phylogenetic analyses and relationships within the Mortierellales based on single-locus analysesAccordingtopreviousstudies(Whiteetal.2006,Petkovitsetal. 2011), themajorgenusof theMortierellales, Mortierella, appears as paraphyletic genus since the genera, Dissophora, Gamsiella and Lobosporangium arenestedwithin.SincethereisnosequencedataorlivingmaterialavailableforAquamortierella and Modicella (Whiteetal.2006)thesegenerawerenotincluded.Duetolackingspeciesmaterialthenewlyproposedand described genus Echinochlamydosporium (Jiang et al.2011)wasalsoexcludedfromthecurrentanalysis.Althoughthe pre-molecular classification schemes definedmorpho-logicallywell-supportedclades(Linnemann1941,Zychaetal.1969,Gams1977)thesecladescouldnotberetainedinanymolecularbasedanalyses(Whiteetal.2006,Petkovitsetal.2011,thisstudy).Thepresentstudyextendedapreviousstudybyadditionofsequenceinformationfor407specimens.Oneisolate, Mortierella mutabilis,wasexcludedduetomiss-fittingmorphologicalcharacteristics.ThemorphologyofM. mutabilis isincontradictionwithitsoriginaldescription(Linnemann1941)and resembles Gamsiella multidivaricata in all morphological featuresaswellitsmoleculardata.Sinceonlyoneisolateisavailable, we postpone its phylogenetically analysis till addi-tionalmaterialisavailable.Nineteenspecieswereadditionallyincludedwithatotalof115sequences.Outofthesesequences57sequencesweregeneratedforITS,58forLSUand1ITSsequencewasretrievedfromGenBank.Outof421specimens in total,213sequences forLSUand364sequencesforITSweregenerated.Thedatasetwassup-plementedwithadditionalsequencesformGenBank(69LSUand11ITSsequences)(Table2).AfirstphylogenetictreebasedonLSUsequencesfrom266taxawas generated to define placement and relationshipsofallsequencesgeneratedinthisstudy(datanotshown).Asubset of all relevant groups and isolates was taken for the finaltreeoftheLSUdataset(Fig.3,justforbetteroverview).Thefinalalignmentcontains781charactersand101taxa.Forsubsequentdeep-levelanalysessevenartificialsubsetsoutofeightcladesofthistreeweredefinedreferringtothepreviouslypublishedgroupdelimitations(Petkovitsetal.2011).ForeachgrouptheITS1-5.8SrDNA-ITS2sequenceswerealignedandanalysed with Maximum Likelihood although the backbone of theunderlying LSU tree is not resolved (Fig. 3).Groups

aremainlylocatedononebranch(‘monophyletic’)exceptforthe under-represented chienii /selenospora-group which was combined and aligned togetherwith themost basal group.Taking these groups as single taxa sets allows alignments providing phylogenetic signals with higher resolution on deep levelclassification.Thealignmentsofthesubsetsconsistsofthefollowingnumbersoftaxaandcharacters:subset1:58/816(means58taxaand816characters,Fig.4);subset2:36/636(Fig.5);subset3:38/701(Fig.6);subset4:17/710(Fig.7);subset5:18/761(Fig.8);subset6:60/703(Fig.9);subset7:73/688(Fig.10).Ourresultsdonotallowfortherevelationofthenaturalrelation-ships between different species or between groups of species since the cladesare poorly supported in theLSU tree.Butdefinitionofboundariesbetweenthespecies/speciesgroupsis possible and the presented species groups are in full accord-ance with the twelve large clades distinguished in a previous study(Petkovitsetal.2011).Becausethecurrentdatasetismore comprehensive, we will keep, but also extend some of thegroups.

Group 1 ‒ selenospora and parvispora (Fig. 4, somemorphological features are displayed inFig.1j, 2i) containsthetwomostbasalgroupsoftheLSUtree(Fig3).Mortierella selenospora clusters well with M. chienii (BootstrapsupportBS=100%). Mortierella chienii was not included in the previous study(Petkovitsetal.2011).IncaseswherethemorphologicalidentificationdoesnotmatchthepositionofthestrainintheITStreethestrainsweredesignatedasMortierellasp.withtheepithetinquotationmarks.Strainswhichareverydistinct,notpartofacladeandconsequentlymightrepresentundescribedspeciesarehighlighted inblue.TheselenosporacladealsocontainsthequestionableM. wolfii CBS614.70whichshowsdifferentcharacteristics(e.g.nothermotolerance)totheoriginalM. wolfii strains although the sporangiospores are ellipsoidal to kidney-shaped like those of M. wolfii.Adetailedanalysisof the morphology and several molecular markers is needed toclarify thestatusof thisparticularstrain.Theothergrouptermed ‘parvispora’ contains also the speciesM. alliacea, M. basiparvispora, M. fimbricystis, M. jenkinii, M. macrocystis, M. macrocystopsis, M. sossauensis in addition to the previously includedspecies (M. cystojenkinii, M. dichotoma, M. elongatula, M. parvispora, M. pulchella, M. turficola;Petko-vitsetal.2011).Mortierella alliacea, M. chienii, M. cystojenkinii, M. elongatula, M. macrocystis, M. macrocystopsis, M. pul chella and M. sossauensis form well-supported clades and the mor-phologicallydefinedspeciesboundariesarewellreflectedintheITStree(Fig.4).Theparvispora-jenkinii-complexconsistspredominantlyofstrainsmorphologicallyidentifiedasM. jenkinii or M. parvispora.Thesetwospeciesdiffermainlybytheshapeof their sporangiospores: ellipsoidal for M. jenkinii and globose for M. parvispora.ThisdistinctionisnotsupportedbytheITStree,mixingbothtypesofspores.ThestrainM. basiparvispora CBS517.72isalsoclusteringinthiscomplex,butisdifferingmorphologically from the ex-type strain of this species, which wasnotincludedinthisstudy(Gams1976).Adetailedrevisionof this species in relation to Mortierellawillbeneeded.

Fig.2(cont.)sporangialeavingacollar(arrow),globosesporangiospores;j.M. lignicola CBS207.37,sporangiophores,sporangiola(arrow1),stylospores(arrow2);k.M. exigua CBS655.68,chlamydosporeswith typicaloutgrowinghyphae; l.M. gemmifera CBS134.45,chlamydospores;m.M. hypsicladia CBS116202,stylosporeswithprojections;n.M. polygonia CBS685.71,stylospores;o.M. nanthalensis CBS610.70,acrotonousbranchingpartofasporangiophore;p.M. alpina FSU2698,oildropletscontaininghypha;q.M. camargensis CBS221.58,sporangiophoresalongahyphawithsporangiola;r.M. epigama CBS489.70,zygospores;s.M. echinosphaera CBS575.75,chlamydospores;t.M. microszygospora CBS880.97,microzygospore;u.M. camargensis CBS221.58,oildropletscontainingsphericsporangiola;v.Dissophora decumbens CBS592.88,sporangiophoreswithsporangia;w.M. paraensis CBS547.89,twosporan-giophoreswithtypicalbasitonousbranchings(arrowsmarkthebasalpart).—Scalebars:a,b,i,n,p,r,u=10µm;c,j,q=20µm;d,e,g,h,m,v=30µm;f,k,l,s,t=15µm;o=250µm;w=100µm.

(textcontinuesonp.88)


Originalnam

eStrainnum

bers

Microscopic

Typestatus

Locality

Substrate

Accession

Accession

identification

no.ITS

no.LSU

Dis

soph

ora

decu

mbe

ns

CBS301.87,FSU9780

D

. dec

umbe

ns

Kingston,RhodeIsland

ground-uplitterofQ

uercus-Acerw

ood-

JX976001

HQ667354.1

la

nd, i

ncub

ated

at 0

°C fo

r tw

o m

onth

s

CBS592.88,FSU801

D. d

ecum

bens

RhodeIsland

ground-upQuercusandAcerleaves,

HQ630276.1H

Q667355.1

in

cuba

ted

at 0

°C fo

r 21

mon

ths

Dis

soph

ora

orna

ta

CBS347.77,FSU9782

–

HolotypeofM

ortie

rella

orn

ata

CordilleraCentral,CaucaenHuila,

soil,inmountainforestunderW

ein-

HQ630278.1H

Q667357.1

ParqueNacionaldelPuracé,Colom

bia

mannia,Clusiaetc.,alt.3100m

CBS348.77,FSU9783

–

HolotypeofM

ortie

rella

orn

ata

CordilleraCentral,CaucaenHuila,

soil,inmountainforestunderW

ein-

JX976036

HQ667356.1

ParqueNacionaldelPuracé,Colom

bia

mannia,Clusiaetc.,alt.3100m

Gam

siel

la m

ultid

ivar

icat

a CBS227.78,FSU9784

G

. mul

tidiv

aric

ata

IsotypeofM

ortie

rella

mul

tidiv

aric

ata

Moskva,SokolnikiPark,Russia

decayingstump

JX975871

HQ667355.1

Lobo

spor

angi

um tr

ansv

ersa

le

CBS357.67,FSU9785

–

Type

of E

chin

ospo

rang

ium

tran

sver

sale

Nevada,VirginiaCity

soil

–HQ667404.1

Mor

tiere

lla a

crot

ona

CBS386.71,FSU9788

–

Type

of M

ortie

rella

acr

oton

aJaipur,R

ambaghPalaceHotel,R

ajasthansoil

JX975921

HQ667405.1

Mor

tiere

lla a

lliac

ea

CBS106.78

–

France

gymnospermlitter

JX976019

KC018349

CBS894.68

–

Tirol,Obergurgl,A

ustria

alpinerawhum

ussoil

JX975990

JX976148

Mor

tiere

lla a

lpin

a CBS110518

–

SouthAfrica

soil,drysandyhighveldgrassland

JX975906

–

CBS210.32,FSU9789

M

. alp

ina

Aut

hent

ic s

train

of M

ortie

rella

reni

spor

aVictoria

sandyloam

soil

JX975853

HQ667421.1

CBS219.35

–

–

JX976018

KC018359

CBS250.53

–

–

JX975955

KC018184

CBS384.71C

–

Jaipur,R

ambaghPalaceHotel,R

ajasthansoil

JX976098

JX976154

CBS387.71

–

GranCanaria,S

pain

soil,underP

inus

can

arie

nsis

JX976038

KC018378

CBS396.91

–

Washington

airbladderofjuvenilefish

JX975994

KC018375

CBS529.72

–

NorthCarolina

pasturesoil

JX976124

KC018320

CBS585.81

M

. kuh

lman

ii

Netherlands

agriculturalsoil

JX976132

JX976152

CBS608.70

–

Netherlands

agriculturalsoil

JX976046

KC018438

CBS696.70

M

. cys

toje

nkin

ii

Wageningen,Mansholtlaan,N

etherlands

agriculturalsoil

JX975947

KC018328

FS

U2698

M

. alp

ina

Argentinia

JX976004

KC018272

FS

U6524

M

. alp

ina

Geisenheim,G

ermany

JX976045

KC018273

Mor

tiere

lla a

mbi

gua

CBS373.96

–

Fukiageham

a,Kagoshima,Japan

soilofsaltm

arsh

JX976062

JX976147

CBS450.88

–

–

JX976067

KC018411

CBS457.66

–

Armenia

soil

JX976041

KC018398

CBS474.96

–

Ootom

i,Iriom

otejimaIsland,O

kinawa,

calcareoussoilinditch

JX976056

KC018416

Japan

CBS521.80

–

Delhi,India

dung

JX976120

KC018423

Mor

tiere

lla a

moe

boid

ea

CBS889.72,FSU9790

M

. alp

ina

Type

of M

ortie

rella

am

oebo

idea

TeutoburgerW

ald,BellerH

olz,Germany

JX976073

HQ667422.1

Mor

tiere

lla a

ngus

ta

CBS293.61,FSU9791

M

. ang

usta

N

eoty

pe o

f Mor

tiere

lla p

olyc

epha

lavar.

Chesh.,Delam

ereForest,E

ngland

podzolsoil,pH

upto2.8

JX976061

HQ667358.1

an

gust

aM

ortie

rella

ant

arct

ica

CBS194.89

–

NorthernFoothills,N

orthernVictoria

soil

JX976087

KC018345

Land

, Ant

arct

ica

CBS195.89

–

NorthernVictoriaLand,Edm

onsonPoint,

soil

JX975843

–

A

ntar

ctic

a

CBS196.89

–

NorthernVictoriaLand,CapeKing,

soil

JX976059

–

A

ntar

ctic

a

CBS609.70,FSU9792

–

Type

of M

ortie

rella

ant

arct

ica

nearHallettStation,Antarctica

soil,rockcrevicenearglacier

JX975907

HQ667503.1

Mor

tiere

lla a

rmill

ariic

ola

CBS105.78

–

Putten,Schovenhorst,Netherlands

JX976100

KC018432

CBS914.73,FSU9793

–

Type

of M

ortie

rella

arm

illar

iicol

aBaarn,G

roeneveld,Netherlands

attackedby

Dip

odas

cus

arm

illar

iae

JX975924

HQ667446.1

Mor

tiere

lla b

aini

eri

CBS220.35

–

formerW

est-G

ermany

JX975901

KC018324

CBS272.71

M

. kuh

lman

ii

Sou

th C

arol

ina

soil

unde

r Pin

us ta

eda

JX975964

JX976155

CBS273.71

M

. kuh

lman

ii

Sou

th C

arol

ina

soil

unde

r Pin

us ta

eda

JX975920

KC018355

CBS442.68

–

Georgia

soilfrompineforest

JX975864

KC018331

CBS508.81

–

GetzbachnearEupen,B

elgium

JX975844

KC018393

CBS552.80

–

Eifel,HundsbachtalnearG

erolstein,

JX975850

JX976174

Germany

Mor

tiere

lla b

asip

arvi

spor

a CBS517.72

–

Valdivia,C

ordilleraPelada,Chile

soil,underF

itzro

ya c

upre

ssoi

des

JX976048

JX976167

Tabl

e 2S

trainsusedinthisstudy.

83L.Wagneretal.:MolecularphylogenyoftheMortierellalesM

ortie

rella

bel

jako

vae

CBS102878

–

TorontoHighPark,Ontario

infrabuccalpelletofC

ampo

notu

sJX976090

KC018350

pe

nnsy

lvan

icus(carpenterant)onPinus

CBS109594

–

Toronto,HighPark,Ontario


ampo

notu

sJX975848

KC018449

pe

nnsy

lvan

icus

, in

mat

ure

Pin

us tr

ee

CBS109595

–

Zw

eifallerW

aldnearAachen,Germany

infrabuccalpelletofF

orm

ica

rufa

JX976129

KC018358

CBS109596

–

St.Andrews,AnnesleyHouse,


ampo

notu

sJX975971

JX976170

New

Bru

nsw

ick

penn

sylv

anic

us, i

n P

inus

tree

CBS109597

–

Scarborough,O

ntario


ampo

notu

sJX975918

KC018433

pe

nnsy

lvan

icus

, in

mat

ure

Pin

us tr

ee

CBS109655

–

BayerischerW

ald,PfahlbeiViechtach,


ampo

notu

sJX975869

JX976171

Germany

herc

ulea

nus,

in P

icea

abi

es

CBS109658

–

Zw

eifallerW

aldnearAachen,Germany


orm

ica

rufa

JX976051

KC018376

CBS109659

–

Utrecht,LageVuursche,Netherlands


orm

ica

rufa

JX975998

KC018340

CBS123.72,FSU9794

M

. bel

jako

vae

Type

of M

ortie

rella

bel

jako

vae

Rovenskregion,S

arna,U

kraine

soil,coniferousforest

JX976126

HQ667428.1

CBS267.71

–

N

orth

Car

olin

a se

edlin

g, P

inus

tead

aJX976072

KC018346

CBS268.71

–

N

orth

Car

olin

a se

edlin

g, P

inus

tead

aJX976043

KC018323

CBS274.71

–

S

outh

Car

olin

a ro

ot, P

inus

taed

aJX976011

KC018388

CBS275.71

–

S

outh

Car

olin

a ro

ot, P

inus

taed

aJX975913

KC018401

CBS276.71

–

S

outh

Car

olin

a ro

ot, P

inus

taed

aJX975937

KC018442

CBS806.68

–

NorthCarolina

barkofroot,Pinus

JX975987

KC018397

Mor

tiere

lla b

iram

osa

CBS370.95,FSU9795

M

. bira

mos

a Ty

pe o

f Mor

tiere

lla w

uyis

hane

nsis

Wuyi,Fujian,China

forestsoil

JX976094

HQ667389.1

CBS506.81

–

Odenw

ald,ObererB

untsandstein,

decayingfineroot,30yrold,on

JX975963

KC018407

Germany

acidicloam

ysoil

CBS550.80

–

Odenw

ald,Germany

rootlet

JX976064

KC018419

Mor

tiere

lla b

ispo

ralis

CBS145.69

–

Italy

JX975857

KC018377

FS

U9675

M

. bis

pora

lis

–

JX975953

JX976176

Mor

tiere

lla c

amar

gens

is

CBS110638

–

Soest,S

mickel,Netherlands

thatchofroof

JX976024

–

CBS221.58,FSU9796

M

. cam

arge

nsis

Ty

pe o

f Mor

tiere

lla c

amar

gens

is

Cam

argue,BoisdesRièges,France

sandysoil

JX975949

HQ667408.1

Mor

tiere

lla c

apita

ta

CBS110640

–

Berlin,K

önigin-Luise-Straße,nearB

BA,

soilwithArmadillidium

JX975923

JX976163

Germany

CBS293.96

–

Naganohara,Gunma,Japan

gardensoil

JX976123

KC018334

CBS859.70

–

NorthCarolina

pillbuggut

JX976008

KC018395

Mor

tiere

lla c

hien

ii CBS287.96

–

Amakubo,Tsukuba,Ibaraki,Japan

soilunderQ

uerc

us m

irsin

ifoliaforest

JX976013

KC018427

CBS289.96

–

Nanam

agari,Yokohama,Kanagaw

a,

soilunderC

asta

nops

is s

iebo

ldiiforest

JX975898

JX976161

Japan

CBS290.96

–

soil

unde

r Mis

cant

hus

sine

nsis

JX976075

KC018373

CBS292.96

M

. sel

enos

pora

Shitoko,YakushimaIsland,K

agoshima,

soilunderF

icus

mic

roca

rpaforest

JX975951

JX976153

Japan

CBS554.73

M

. sel

enos

pora

Kuang-MiauCo.,16km

EofTainan,

soilfrombam

boogrove

JX975912

KC018381

Taiw

anM

ortie

rella

chl

amyd

ospo

ra

CBS120.34,FSU9799

–

Syn

type

of A

zygo

zygu

m c

hlam

ydos

poru

m

infe

cted

by

Rhi

zoct

onia

sol

ani

JX975942

HQ667430.1

CBS529.75

–

Netherlands

soil

JX975927

–M

ortie

rella

cla

usse

nii

CBS790.85

–

–

JX976012

JX976159

Mor

tiere

lla c

lono

cyst

is

CBS357.76,FSU9801

M

. clo

nocy

stis

Ty

pe o

f Mor

tiere

lla c

lono

cyst

is

GranCanaria,S

pain

soil,under

Apo

lloni

as c

anar

iens

is

JX975899

HQ667395.1

Mor

tiere

lla c

ogita

ns

CBS879.97,FSU9802

–

Type

of M

ortie

rella

cog

itans

Nagano,Sanada,Sugadaira

decayingtreebark

JX976017

HQ667360.1

M.R.C.,Japan

Mor

tiere

lla c

ysto

jenk

inii

CBS456.71,FSU9803

M

. cys

toje

nkin

ii Ty

pe o

f Mor

tiere

lla c

ysto

jenk

inii

Wageningen,Netherlands

agriculturalsoil

JX976030

HQ667504.1

CBS660.82

–

Bakkeveen,N

etherlands

Pinusforest

JX975868

KC018325

Mor

tiere

lla d

ecip

iens

CBS873.68

–

Kiel-K

itzeberg,Germany

wheatfieldsoil

–JX976173

Mor

tiere

lla d

icho

tom

a CBS221.35,FSU9804

M

. dic

hoto

ma

Syn

type

of M

ortie

rella

dic

hoto

ma

formerW

est-G

ermany

dungofm

ouse

JX975842

HQ667393.1

Mor

tiere

lla e

chin

osph

aera

CBS574.75

–

nearW

ageningen,Netherlands

soil

JX976060

KC018370

CBS575.75,FSU9805

M

. ech

inos

phae

raHolotypeofM

ortie

rella

ech

inos

phae

ra

Aalsm

eer,Netherlands

JX976015

HQ667431.1

Mor

tiere

lla e

chin

ula

CBS282.71

–

Iceland

soil

JX975948

–M

ortie

rella

elo

ngat

a CBS110517

–

AltiMountains,S

outhAfrica

soil,grassland,sum

merrainfallregion

JX976042

KC018348

CBS122.71

–

Georgia,M

onroe,USA

soil,undergolfturf-grass

JX976000

KC018396

CBS126.71,FSU823

M. e

long

ata


agriculturalsoil

JX976101

KC018279

CBS208.71

–

Netherlands

greenhousesoil

JX975995

JX976135


CBS276.89

–

Quebec

(blackfly)

JX976111

KC018452

CBS279.62

–

Kiel-K

itzeberg,Germany

wheatfieldsoil

JX976089

KC018417

CBS344.66

–

Alaska

tundrasoil

JX976081

KC018322

FS

U532

M. e

long

ata

–

JX975976

KC018281

FS

U822,CBS125.71

M

. elo

ngat

a


agriculturalsoil

JX975978

KC018282

FS

U97

21

M. e

long

ata

Münchenroda,G

ermany

JX975894

KC018284

Mor

tiere

lla e

long

atul

a CBS488.70,FSU9808

–

Type

of M

ortie

rella

elo

ngat

ula

formerW

est-G

ermany

municipalwaste

JX975967

HQ667425.1

CBS661.70

–

Braunschw

eig,Germany

municipalwaste

JX976069

KC018431

Mor

tiere

lla e

picl

adia

CBS246.75

–

S

urin

ame

soil,

und

er E

laei

s gu

inee

nsis

JX975890

KC018361

CBS355.76,FSU9809

M

. epi

clad

a Ty

pe o

f Mor

tiere

lla e

picl

adia

GranCanaria,S

pain

soil,underA

pollo

nias

can

arie

nsis

JX976130

HQ667396.1

CBS356.76

–

GranCanaria,S

pain

soil,underA

pollo

nias

can

arie

nsis

JX975972

–

CBS555.89

–

Pará,200kmSEfrom

Belém

,Capitâo

rainforestsoil

JX975991

JX976150

Poç

o, B

rasi

lM

ortie

rella

epi

gam

a CBS161.76

M

. epi

gam

a

Exeter,HatherlyLaboratories,England

compostheap

JX976109

JX976158

CBS489.70,FSU9810

M

. epi

gam

a Ty

pe o

f Mor

tiere

lla e

piga

ma

formerW

est-G

ermany

municipalwaste

JX976057

HQ667367.1

CBS881.97

–

Kagoshima,Kam

ei,Tokunoshima-Island,

olddungofcow

JX976053

KC018445

Japan

Mor

tiere

lla e

xigu

a CBS358.76

–

GranCanaria,S

pain

soil,underA

pollo

nias

can

arie

nsis

JX976113

KC018439

CBS510.63

–

Kiel-K

itzeberg

agriculturalsoil

JX975863

JX976134

CBS655.68,FSU9811

M

. exi

gua

Type

of M

ortie

rella

ste

rilis

Allahabad,India

farmsoil

JX976047

HQ667406.1

CBS865.68

–

Kiel-K

itzeberg,Germany

wheatfieldsoil

JX976070

–M

ortie

rella

fats

hede

rae

CBS388.71

–

GranCanaria

soil,underP

inus

can

arie

nsis

JX976003

JX976136

Mor

tiere

lla fi

mbr

icys

tis

CBS943.70

–

Type

of M

ortie

rella

fim

bric

ystis

SouthPatagonia,P

uertoEdw

ardsnear

centreofm

osscushion,inverywetbogGU559986.1JX976172

Bea

gle

Can

al, A

rgen

tinia

Mor

tiere

lla fo

rmic

icol

a CBS109589

–

Brampton,O

ntario


ampo

notu

sJX975933

JX976140

pe

nnsy

lvan

icus,inhouse(windowsill)

Mor

tiere

lla g

amsi

i CBS110630

–

Boekrijk,B

elgium

soilwithPorcellio

JX976106

KC018410

CBS253.36,FSU9813

M

. gam

sii

Syn

type

of M

ortie

rella

spi

nosa

formerW

est-G

ermany

forestsoil

JX975968

HQ667415.1

CBS314.52,FSU9814

M

.cf.

gam

sii

Syn

type

of M

ortie

rella

spi

nosa

formerW

est-G

ermany

forestsoil

JX975892

HQ667384.1

CBS551.73,FSU824

M. g

amsi

i

NorthCarolina

pasturesoil

JX976079

JX976177

CBS552.73,FSU825

M. g

amsi

i

AlleghanyCounty,NorthCarolina

pasturesoil

JX975984

KC018285

CBS749.68,FSU9812

M

. gam

sii

Type

of M

ortie

rella

gam

sii

Baarn,M

aarschalksbos,Netherlands

soil

HQ667416.1

FS

U2057

M

. gam

sii

–

JX976118

KC018287

Mor

tiere

lla g

emm

ifera

CBS124.72

–

MeerdinkbosnearW

interswijk,

soil,hum

uslayer

JX975909

KC018390

Net

herla

nds

CBS134.45,FSU9815

M

. gem

mife

ra

Type

of M

ortie

rella

gem

mife

ra

nearNottingham

,England

soilfrompineforest

JX975931

HQ667371.1

CBS383.85

–

Spandersw

oudnearBussum,N

etherlands

soil,inpineforest

JX976121

JX976157

CBS661.82

–

B

akke

veen

, Net

herla

nds

End

ogon

e la

ctifl

ua,P

inusforest

JX975989

KC018360

Mor

tiere

lla g

loba

lpin

a CBS226.78

–

Katwijk,N

etherlands

sanddunesoil

JX976006

JX976160

CBS718.88

–

Japan

JX975925

–M

ortie

rella

glo

bulif

era

CBS108.68

–

Schweden

JX975847

KC018332

CBS746.68

–

Netherlands

agriculturalsoil

JX976026

KC018371

CBS857.70,FSU826

–

England

decayingneedle

JX975910

HQ667369

CBS858.70,FSU9817

M

. glo

bulif

era

Neo

type

of M

ortie

rella

glo

bulif

era

England

decayingroot

JX975915

HQ667368.1

CBS867.68

–

Tirol,Obergurgl,A

ustria

alpinerawhum

ussoil

JX976107

JX976165

Mor

tiere

lla h

isto

plas

mat

oide

s CBS321.78,FSU9819

–

Type

of M

ortie

rella

his

topl

asm

atoi

des

Louisiana

dung

HQ630309.1H

Q667386.1

Mor

tiere

lla h

ortic

ola

CBS305.52,FSU9820

M

. hor

ticol

a S

ynty

pe o

f Mor

tiere

lla h

ortic

ola

formerW

est-G

ermany

JX975874

HQ667399.1

CBS869.68

–

Kiel-K

itzeberg

wheatfieldsoil

JX976058

JX976138

CBS254.76

–


agriculturalsoil

JX976021

JX976166

Mor

tiere

lla h

umili

s CBS180.72

–

Piedm

ont,NorthCarolina

forestsoil

JX976125

KC018436

CBS181.72

–

Piedm

ont,NorthCarolina

soil

JX975887

KC018405

CBS222.35,FSU9821

–

Syn

type

of M

ortie

rella

hum

ilis

Mexico

soilfromPinusforest

HQ630325.1H

Q667401.1

CBS363.95

–

Shennongjia,H

ubei,C

hina

forestsoil

JX976097

KC018443

Originalnam

eStrainnum

bers

Microscopic

Typestatus

Locality

Substrate

Accession

Accession

identification

no.ITS

no.LSU

Tabl

e 2(cont.)


CBS443.68,FSU828

M. h

umili

s

SouthCarolina

barkofstump

JX976002

HQ667402

CBS745.68,FSU829

M. h

umili

s

Baarn,E

emnesserweg90,Netherlands

soil

JX975867

HQ667403

Mor

tiere

lla h

yalin

a CBS100563

–

SchoharieCo.,N

ewYork

JX976023

KC018356

CBS115655,FSU9822

M

. hya

lina

IsotypeofH

ydro

phor

a hy

alin

aNorthofLondon,Rothamsted,E

ngland

roots

HQ630355.1H

Q667432.1

CBS117.74

–

Boekesteynnear’s-Graveland,

JX976083

KC018392

Net

herla

nds

CBS117152

–

Graz,Austria

soilandcheesmixtureusedasfood

JX975977

KC018394

form

itesbyE.E

bermann

CBS166.25

–

Netherlands

seed

JX975928

–

CBS167.25

–

–

JX975895

KC018406

FS

U10532

M. h

yalin

a

Austria

JX975992

KC018289

FS

U509

M. h

yalin

a

–

JX975981

KC018291

Mor

tiere

lla h

ypsi

clad

ia

CBS116202,FSU9825

M

. hyp

sicl

adia

Ty

pe o

f Mor

tiere

lla h

ypsi

clad

ia

KyushuIsl.,KariuCave,Japan

batdungincave

JX975866

HQ667379.1

CBS116203

Aut

hent

ic s

train

of M

ortie

rella

hyp

sicl

adiaKyushuIsl.,KariuCave,Japan

batdungincave

JX975872

KC018369

Mor

tiere

lla in

dohi

i CBS220.72

Naaldwijk,N

etherlands

greenhousesoil

JX975993

KC018408

CBS331.74,FSU830

M. i

ndoh

ii

Lienden,Netherlands

root

JX975860

KC018292

CBS460.75,FSU831

M. i

ndoh

ii

Athens,Georgia

dungofanimal

JX975878

HQ667438

CBS478.95

Chengdu,S

ichuan,C

hina

soil

JX975903

KC018347

CBS528.75

SouthAfrica

bagasseinchickenfarm

JX976044

KC018451

CBS665.70


agriculturalsoil

JX975956

KC018357

CBS720.71,FSU9826

M

. ind

ohii

IsotypeofM

ortie

rella

indo

hii

Athens,Georgia

dungofanimal

JX975856

HQ667377.1

Mor

tiere

lla je

nkin

ii CBS188.73

Nottingham

,England

turflayerofgolfgreen,received

JX975999

KC018389

fu

ngic

idal

trea

tmen

t for

long

per

iod

CBS666.75C

S

wed

en

soil

unde

r Pic

ea a

bies

–JX975873

CBS667.70


agriculturalsoil

JX976088

KC018422

CBS850.70


agriculturalsoil

JX975849

KC018352

CBS965.73C

Sweden

forestsoil

JX976117

JX976139

Mor

tiere

lla k

uhlm

anii

CBS157.71,FSU9827

M

. kuh

lman

ii Ty

pe o

f Mor

tiere

lla k

uhlm

anii

SouthCarolina,Miley

stum

pJX975846

HQ667372.1

CBS269.71

stum

p, P

inus

taed

aJX975935

KC018384

CBS270.71

Patrick,SouthCarolina

stum

pJX975851

JX976142

CBS271.71

SouthCarolina

seedling

JX975883

KC018338

Mor

tiere

lla li

gnic

ola

CBS100594

–

JX975889

–

CBS116.65


blacksoil

JX975965

KC018402

CBS207.37,FSU9828

M

. lig

nico

la

Type

of H

aplo

spor

angi

um li

gnic

ola

SierraNevadadeSantaMarta,C

olom

bia

rottenwood

JX976095

HQ667435.1

CBS313.52,FSU9829

M

. lig

nico

la

Type

of M

ortie

rella

sep

edon

ioid

es

formerW

est-G

ermany

soilunderP

inus

syl

vest

ris

JX976127

HQ667434.1

Mor

tiere

lla lo

ngig

emm

ata

CBS653.93

Höglwald,Germany

soil

JX976055

JX976162

Mor

tiere

lla m

acro

cyst

is

CBS110716

DeVeluwe

oakforestsoil

JX976084

–

CBS314.85

formerW

est-G

ermany

rootletofgym

nosperm

JX975974

JX976169

CBS431.81

Cundinamarca,páram

oCruzVe

rde,

soil

JX975897

KC018437

Col

ombi

a

CBS482.73

formerW

est-G

ermany

soil

JX975862

–

CBS937.69

Baarn,P

ekingtuin,Netherlands

soil

JX975881

KC018341

Mor

tiere

lla m

acro

cyst

opsi

s CBS302.87

SouthKingstown,RhodeIsland

soilunderP

inus

resi

nosaand

JX975908

KC018362

P

inus

stro

bus

CBS387.91

M

. cys

toje

nkin

ii

Norway

soil

JX976105

JX976144

CBS520.88

RhodeIsland

soil

JX976078

–

CBS528.87

SouthKingstown,RhodeIsland

forestsoil,underP

inus

resi

nosaand

JX975946

JX976164

P

inus

stro

bus

Mor

tiere

lla m

icro

zygo

spor

a CBS880.97,FSU9831

M

. mic

rozy

gosp

ora

Type

of M

ortie

rella

mic

rozy

gosp

ora

Shiga,M

aibara,Japan

soilinhedge

JX976027

HQ667394.1

Mor

tiere

lla m

inut

issi

ma

CBS226.35

formerW

est-G

ermany

JX976092

JX976168

CBS277.71,FSU832

M. m

inut

issi

ma

Georgia

forestsoil

JX975938

KC018293

FS

U2735

M

. zon

ata

–

JX976103

KC018318

Mor

tiere

lla m

inut

issi

mavar.

dubi

a CBS307.52,FSU9832

Syn

type

of M

ortie

rella

min

utis

sim

avar.

formerW

est-G

ermany

soil

JX976122

HQ667400.1

du

bia

Mor

tiere

lla n

anta

hale

nsis

CBS610.70,FSU9834

M

. nan

taha

lens

is

Type

of M

ortie

rella

nan

taha

lens

is

JoyceKilm

erMem

orialForestinthe

soil

JX976022

HQ667388.1

Nan

taha

la N

atio

nal F

ores

t, N

orth

Car

olin

aM

ortie

rella

olig

ospo

ra

CBS101758

Pennsylvania

supplementtomushroomculture

JX976032

KC018327

CBS191.79

ElephantW

hiteNileIsland,S

udan

soil

JX975966

JX976151


CBS381.71

Jaipur,R

ambaghPalaceHotel,R

ajasthansoil

JX976033

KC018368

Mor

tiere

lla p

arae

nsis

CBS343.89

Pará,CapitãoPoço,Brazil

forestsoil,virginforest

JX975944

KC018329

CBS547.89,FSU9835

M

. par

aens

is

Type

of M

ortie

rella

par

aens

is

Pará,200kmSEfrom

Belém

,Capitão

rainforestsoil

HQ630353

HQ667429.1

Poç

o, B

rasi

lM

ortie

rella

par

azyc

hae

CBS868.71,FSU9836

M

. par

azyc

hae

Type

of M

ortie

rella

par

azyc

hae

Tree

k ne

ar A

mer

sfoo

rt, N

ethe

rland

s de

cayi

ng w

ood,

with

Bot

ryob

asid

ium

JX975985

HQ667362.1

su

bcor

onat

umM

ortie

rella

par

visp

ora

CBS304.52,FSU9837

M

. par

visp

ora

Syn

type

of M

ortie

rella

gra

cilis

formerW

est-G

ermany

soil

JX975859

–

CBS311.52,FSU9838

Syn

type

of M

ortie

rella

par

visp

ora

formerW

est-G

ermany

soil

JX976076

HQ667373.1

CBS315.61,FSU834

M. p

arvi

spor

a

Cheshire,D

elam

ereForest,E

ngland

soil,iron-hum

uspodzol

JX976104

HQ667374.1

CBS316.61,FSU835

M. p

arvi

spor

a

Cheshire,D

elam

ereForest,E

ngland

soil,iron-hum

uspodzol

JX976029

HQ667375.1

CBS445.68


beet-fieldsoil

JX976049

KC018414

FS

U2736

M

. jen

kini

i

–

JX976093

KC018295

Mor

tiere

lla p

olyc

epha

la

CBS227.35

–

JX976096

KC018321

CBS293.34

M

. hya

lina

Netherlands

JX976050

JX976137

CBS327.72,FSU866

M. p

olyc

epha

la

Lincs.,G

ibraltarP

oint,E

ngland

salt-marshsoilunderS

parti

na to

wns

endi

iJX976085

JX976175

CBS328.72,FSU867

M. p

olyc

epha

la

UK

soil

JX976102

KC018296

CBS456.66,FSU759

M. p

olyc

epha

la

nearKiev,Ukraine

dungofw

oodmouse

JX976034

KC018297

FS

U696

M. p

olyc

epha

la

–

JX976035

KC018298

Mor

tiere

lla p

olyg

onia

CBS248.81

S

exbi

erum

, Net

herla

nds

clay

soi

l und

er S

olan

um tu

bero

sum

JX975891

JX976145

CBS685.71,FSU9839

Type

of M

ortie

rella

pol

ygon

ia


agriculturalsoil

JX975900

HQ667378.1

Mor

tiere

lla p

seud

ozyg

ospo

ra

CBS779.86

Kingston,NorthW

oods,U

niv.ofR

hode

soilunderQ

uercus-Acerw

oodland,

JX975960

KC018353

IslandCam

pus,RhodeIsland

aboutsealevel,upper5cmdepth

CBS780.86

PeaceDale,HazardTract,RhodeIsland

soil,underP

inus

stro

bus

and

Pin

us

JX975880

JX976143

re

sino

sawoodland,from

upper5cm

depth,soiltem

p.2.5°C

Mor

tiere

lla p

ulch

ella

CBS205.86

Netherlands

root

JX976031

KC018366

CBS312.52,FSU9840

Aut

hent

ic s

train

of M

ortie

rella

pul

chel

la

formerW

est-G

ermany

root

JX976054

HQ667427.1

CBS675.88

Berlin,G

runewald,Jagen91,Germany

soil,litterlayer

JX976082

KC018440

Mor

tiere

lla re

ticul

ata

CB

S11

0044

LanarknearB

ranxholme,Victoria

dungofP

eram

eles

gun

nii

JX975980

–

CBS223.29

–

JX975973

–

CBS241.33

–

JX976116

JX976133

CBS415.81

Toronto,Ontario

dungofm

ouse,collectedinahouse

JX975877

–M

ortie

rella

rish

ikes

ha

CBS652.68,FSU9842

Type

of M

ortie

rella

rish

ikes

ha

Rishikesh,India

forestsoil

JX976110

HQ667385.1

Mor

tiere

lla ro

staf

insk

ii CBS522.70,FSU9844

Neo

type

of M

ortie

rella

rost

afin

skii

nearBainbridge,G

eorgia

soilunderP

inus

elli

ottiivar.e

lliot

tii

JX975885

HQ667436.1

Mor

tiere

lla s

arny

ensi

s CBS122.72,FSU9845

M

. sar

nyen

sis

Type

of M

ortie

rella

sar

nyen

sis

Rovenskregion,nearS

arny,U

kraine

coniferousforest

JX975957

HQ667390.1

Mor

tiere

lla s

chm

ucke

ri CBS156.78

MadhyaPradeshandUttarP

radesh

soil,from

ravines

JX975854

KC018372

regions,India

CBS295.59,FSU9846

M

. sch

muc

keri

Syn

type

of M

ortie

rella

sch

muc

keri

Queretaro,M

exico

soil,underO

punt

iasp.,pH6.7

JX976112

HQ667414.1

CBS777.86

ShoshoneNationalForest,HorseCreek

soil,upper10cm

,underP

seud

otsu

ga

JX976099

KC018413

Cam

pground,W

yoming

men

zies

ii,alt.2500m

Mor

tiere

lla s

cler

otie

lla

CBS529.68,FSU9847

M

. scl

erot

iella

Ty

pe o

f Mor

tiere

lla s

cler

otie

lla

Ukraine

dungofm

ouse

JX975988

HQ667387.1

Mor

tiere

lla s

elen

ospo

ra

CBS452.88

Cibodas,Indonesia

soil

JX976037

KC018429

CBS811.68,FSU9848

M

. sel

enos

pora

Ty

pe o

f Mor

tiere

lla s

elen

ospo

ra

Horst,N

etherlands

mushroomcom

post,togetherw

ith

JX975875

HQ667419.1

E

ntom

opht

hora

cor

onat

a an

d

Aph

anoc

ladi

um a

lbum

Mor

tiere

lla s

impl

ex

CBS110.68


oat-fieldsoil

JX975982

–

CBS243.82

Baarn,C

.Dopperlaan18,Netherlands

compostheap

JX975870

JX976156

Mor

tiere

lla s

ossa

uens

is

CBS153.76C

S

chw

eden

fo

rest

soi

l und

er P

icea

abi

es

JX976063

JX976146

CBS176.74

M

. clo

nocy

stis

Athens,Georgia

Greenhousesoile

JX975926

KC018428

CBS281.71

SouthCarolina

root

JX975911

KC018447

CBS890.72

Ireland

peatsoil

JX975865

KC018385

CBS898.68

Lincs.,G

ibraltarP

oint,E

ngland

salt-marshsoil

JX975970

KC018374

Mor

tiere

llasp.

FS

U10519

M. a

lpin

a

Austria

JX975959

KC018258

Originalnam

eStrainnum

bers

Microscopic

Typestatus

Locality

Substrate

Accession

Accession

identification

no.ITS

no.LSU

Tabl

e 2(cont.)


FSU10520

M. a

lpin

a

Austria

JX975969

KC018259

FS

U10522

M. a

lpin

a

Austria

JX975930

KC018261

FS

U10523

M. a

lpin

a

Austria

JX976114

KC018262

FS

U10551

M. a

lpin

a

Austria

JX975852

KC018269

FS

U10555

M. a

lpin

a

Austria

JX975996

KC018315

FS

U10558

M. a

lpin

a

Austria

JX975884

KC018271

FS

U10683

M. a

lpin

a

Austria

JX976039

–

FSU10696

M. a

lpin

a

Austria

JX976108

–

FSU10706

M. a

lpin

a

Austria

JX976068

–

FSU10715

M. a

lpin

a

Austria

JX976080

–

FSU10716

M. a

lpin

a

Austria

JX975879

–

FSU8712

M

. alp

ina

Wehlen,Mosel,G

ermany

JX975845

KC018274

FS

U8722

M

. alp

ina

Wehlen,Mosel,G

ermany

JX975961

KC018275

FS

U8736

M

. alp

ina

Wehlen,Mosel,G

ermany

JX976119

KC018276

FS

U8737

M

. alp

ina

Wehlen,Mosel,G

ermany

JX975902

KC018277

FS

U8738

M

. alp

ina

Wehlen,Mosel,G

ermany

JX976010

KC018278

CBS118520

Græ

se,Zealand,D

enmark

agriculturalsoil

JX975936

JX976149

FS

U10767

Austria

JX975929

–

FSU

1079

2

Austria

JX976014

–

FSU

1079

7

Austria

JX975950

–

FSU10541

M. e

long

ata

Austria

JX975876

KC018310

FS

U10

771

M. e

long

ata

Austria

JX976131

–

FSU8711

M

. elo

ngat

a

Wehlen,Mosel,G

ermany

JX976071

KC018283

FS

U10538

M. g

amsi

i

Austria

JX975858

KC018286

FS

U10535

M. h

umili

s

Austria

JX976052

–

FSU1954

M

. hya

lina

–

JX975861

KC018290

FS

U10804

M. m

inut

issi

ma

Austria

JX976020

–

FSU10552

M. p

arvi

spor

a

Austria

JX976009

KC018294

FS

U10

712

Austria

JX975941

–

FSU

1073

0

Austria

JX975916

–

FSU10753

Austria

JX976016

–

FSU10758

Austria

JX976005

–

FSU10759

M. p

arvi

spor

a

Austria

JX975934

–

FSU10789

Austria

JX976065

–

FSU10530

Austria

JX975893

KC018306

FS

U10540

Austria

JX975986

KC018309

FS

U10557

Austria

JX975932

–

FSU2188

–

JX975945

KC018316

FS

U10534

M. v

ertic

illat

a

Austria

JX975914

KC018317

Mor

tiere

lla s

trang

ulat

a CBS455.67,FSU9849

M

. stra

ngul

ata

Neo

type

of M

ortie

rella

stra

ngul

ata

Baarn,G

roeneveld,Nmetherlands

dung,offox?

JX975997

HQ667437.1

Mor

tiere

lla s

tylo

spor

a CBS211.32,FSU9850

M

. sty

losp

ora

Type

of M

ortie

rella

sty

losp

ora

Victoria

sandyloam

JX976086

HQ667359.1

Mor

tiere

lla tu

rfico

la

CBS430.76

HeseperVeennearCoevorden,

decayingS

phag

num

recu

rvum

JX975919

KC018444

Net

herla

nds

CBS431.76


decayingS

phag

num

recu

rvum

JX976025

KC018333

Net

herla

nds

CBS432.76,FSU9851

M

. tur

ficol

a N

eoty

pe o

f Mor

tiere

lla tu

rfico

la


decayingS

phag

num

recu

rvum

JX975952

HQ667426.1

Net

herla

nds

CBS433.76


decayingS

phag

num

recu

rvum

JX975939

KC018424

Net

herla

nds

CBS547.76

CaucaenHuila,C

ordilleraCentral,

soilfrommountainforestunder

JX975896

KC018339

ParqueNacionaldelPuracé,3100malt.,

Wei

nman

niaetc.

Col

ombi

a

CBS581.80

Netherlands

Triocom

post

JX976040

KC018409

Mor

tiere

lla v

ertic

illat

a CBS130.66

Lancashire,Freshfield,E

ngland

sandyforestsoil

JX976007

KC018326

CBS131.66

Lancashire,Freshfield,E

ngland

sandyforestsoil

JX975886

KC018446

CBS220.58,FSU9853

M

. ver

ticill

ata

Type

of H

aplo

spor

angi

um fa

scic

ulat

um

Font

aine

blea

u, F

ranc

e so

il un

der B

etul

asp.

JX975905

JN940873.1

CBS225.35,FSU9854

M

. ver

ticill

ata

Syn

type

of M

ortie

rella

mar

burg

ensi

sformerW

est-G

ermany

JX975940

JQ040251.1

CBS279.71

SouthCarolina

root

JX975917

KC018426


CBS280.71

SouthCarolina

root

JX976066

KC018404

CBS315.52,FSU9856

M

. ver

ticill

ata

Syn

type

of M

ortie

rella

mar

burg

ensi

sformerW

est-G

ermany

forestsoil

JX975943

–

CBS346.66,FSU9852

Alaska

tundrasoil

JX975855

HQ667397.1

CBS374.95,FSU9855

M

. ver

ticill

ata

Type

of H

aplo

spor

angi

um a

ttenu

atis

Wuyi,Fujian,China

forestsoil

JX976077

HQ667398.1

si

mum

Mor

tiere

lla w

olfii

CBS614.70,FSU9860

M

.cf.

wol

fi

Matam

ata,New

Zealand

decayedhay

JX975975

HQ667420.1

CBS209.69,FSU9858

M

. wol

fii

Keele,E

ngland

coalspoiltipsoil

HQ630303.1H

Q667380.1

CBS611.70,FSU9857

Morrinsville,New

Zealand

lung,dyingfrom

mycoticpneum

onia

HQ630306.1H

Q667383.1

CBS612.70,FSU9859

New

Zealand

decayedhay

HQ630304.1H

Q667381.1

CBS651.93,FSU9862

M

. wol

fii

Limburg,H

orst,N

etherlands

compostform

ushrooms

JX975904

HQ667382.1

Mor

tiere

lla z

onat

a CBS228.35,FSU9863

M

. zon

ata

Type

of M

ortie

rella

zon

ata

formerW

est-G

ermany

JX975983

HQ667433.1

CBS615.70

Braunschw

eig-Völkenrode,Germany

soil

JX975958

KC018434

CBS617.76

Cordillera,C

entralParqueNacional

páramosoil,openvegetationwith

JX976028

JX976141

delPuracé,3900malt.

extensivepasture

CBS863.68

Ringw

ood,New

Forest,UK

forestsoil

JX975888

KC018335

Mor

tiere

lla z

ycha

e CBS102879

TorontoHighPark,Ontario

pelletofC

ampo

notu

s pe

nnsy

lvan

icus

JX976074

(carpenterant)

CBS109599

ElYunque,RioBlancoTrail,PuertoRico

infrabuccalpelletofant

JX975882

CBS143.91

formerW

est-G

ermany

JX976091

CBS316.52,FSU9864

M

. zyc

hae

Type

of M

ortie

rella

zyc

hae

Allgäu,G

ermany

decayingwood

JX975979

HQ667407.1

CBS531.81

formerW

est-G

ermany

mushroomcasingsoil

JX975962

KC018421

FS

U71

9 M

. zyc

hae

–

JX976128

KC018319

Um

belo

psis

isab

ellin

a NRRL1757,CBS100559

Wisconsin

soil

JN943789.1JN

940879.1

Originalnam

eStrainnum

bers

Microscopic

Typestatus

Locality

Substrate

Accession

Accession

identification

no.ITS

no.LSU

Tabl

e 2(cont.)

Group 2 – verticillata-humilis (Fig.5,somemorphologicalfeaturesaredisplayedinFig.1c,2a,g,r)isagroupthatalsocontains the genera M. clonocystis, M. epicladia, M. epigama, M. horticola and M. minutissima.Thetopologyissimilartotheonepreviouslypublished(Petkovitsetal.2011)butincludessomemorphologicallymisidentified specimens.Mortierella zonata CBS863.68andM. sossauensis CBS898.68arewellseparat-edfromanyothermembersoftheirspecies.Themainclusterof M. sossauensis is closely related to the parvispora-jenkiniicomplex(Fig.4)whilethetypestrainofM. zonata is related to M. hyalina and M. bainieri (Fig.10).Afteraprofoundmorpho-logical revision M. zonata CBS863.68andM. sossauensis CBS 898.68shouldberenamedandincludedintheM.minutissima-M.horticolacomplex,whichmakesthisphylogeneticgroupofM. minutissimaM. horticola indistinguishablebyITSsequencesalthough both species could be distinguished by the number of theirsporesinthesporangiola.WhileM. minutissima develops few-spored sporangiola, M. horticola produces single-spored sporangiola.This suggests that the number of spores persporangiumisnotstrictlyfixedinthisgroupandisthereforenotoftaxonomicrelevance.ThesinglespecimenCBS246.75resembles M. epicladia but it clusters distantly from the ex-type materialCBS355.76whichisclosetoM. clonocystis (Fig.5).SincenootherknownspeciesgrouptogetherwithCBS246.75,thismightbeasofarundescribedspecies.CBS226.78wasoriginally deposited as M. globalpina andCBS 226.35 as M. minutissima but molecular data of both species currently re-sembles M. clonocystis, indicating an original misapplication or acontamination. Morphology of both species was checked twice andbothspecieswerefinallyassignedtoM. clonocystis. The morphospecies M. clonocystis, M. epicladia and M. epigama arewell recognizedby the ITStreewhileM. verticillata and M. humilis formanotherspeciescomplex.Anotherapparentcluster, the M. verticillataM. humilis cluster, contains strains includingtypestrainsofbothspecies.BasedonITSsequences,adifferentiationisnotpossible.Sequencesaresimilarbetween98–100%.Bothspeciesaremorphologicallysimilarwithoutanysignificantdifferences.Consequentlybothspeciesshouldbesynonymized.

Group 3 – lignicola (Fig.6,somemorphologicalfeaturesaredisplayedinFig.1n,y,2j,l,s,w).Thisgroupcontainsthespecies Mortierella beljakovae, M. chlamydospora, M. echinosphaera, M. formicicola, M. gemmifera, M. kuhlmanii, M. ligni cola and M. paraensis.Severalofthemorphologicallydefinedspecies, namely M. beljakovae, M. chlamydospora, M. echinosphaera, M. formicicola, M. lignicola and M. paraensis, are nicelydetectedbythemoleculardata.Mortierella chlamydospora and M. echinosphaera appear to be closely related as theyaresistergroups(BS=100%).ThespeciesM. gemmifera and M. kuhlmanii aremorphologically very similar (complexissupportedbyBS=85%)anddifferjustgraduallybysporeshapeandchlamydospores.Theex-typestrainsofbothspe-ciesdifferjustby12differentbasepairsintheITSsequences(=98%).TheoriginalmorphologicalidentificationofstrainCBS268.71couldnotbeverifiedbecauseitdidnotsporulateunderdifferent conditions, but its molecular data places it between the gemmifera-complex, M. chlamydospora and M. echinosphaera.ThestrainsCBS109659andCBS555.89werenotexaminedmorphologically and assigned as Mortierella sp.sincetheirorigi-naldescriptionsdoesnotcorrespondwiththemoleculardata.

Group 4 – mutabilis, globulifera and angusta (Fig. 7,somemorphologicalfeaturesaredisplayedinFig.1e,s,v,x,2v).This group contains two of the three included non-Mortierella genera: Gamsiella and Dissophora.ThegenusGamsiella does not cluster with any other mortierellean species, although it was reported to be sister with M. mutabilis(Petkovitsetal.2011).


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Fig. 3MaximumLikelihoodanalysisbasedon781alignednucleotidesoftheD1/D2domainofthelargesubunit(LSU,28S)rDNAfrom101taxa(100ingrouptaxaoftheMortierellales and 1 outgroup taxon Umbelopsis as member of the Mucorales,Meyer&Gams2003).ThephylogrambasedonaMAFFT-Alignment(L-ins-I).Nodesupportsabove75%isgiven.Thetreedefines7groups:groups1–7,whicharemore profoundly analysed in individual analyses based on theITS1-5.8S-ITS2showninFig.4–10.ThestrainsnamedMortierellasp.‘epithet’arestrainswithanoriginallydifferentassignmentbasedonmorphology.Bluemarkedstrainsarepotentialnewspecies.


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A revision of the morphology revealed different features for M. mutabilis asoriginallydescribed.Mortierella mutabilis should develop explicitly branched sporangiophores with globose sporangia containing globose to subglobose sporangiospores, forexample.ButtheobservedmorphologyresemblesthatofGamsiella.Furthermore,LSUandITSsequencesaresimilarwith100and99.8%,respectively.Basedonthesedata,weare rejecting thepreviousgroupnamedmutabilis (Petkovitsetal.2011).ForthefinalplacementofM. mutabilis, additional strainmaterialisnecessary.

The angusta group is extended by M. simplex and consists of the subclades M. angustaM. simplex (BS=88%)andthesubclade Dissophora with D. decumbens and D. ornata (BS=100%).Mortierella simplex could not by differentiated from M. angusta bysignificant features,suggestinganupcomingsynonymizationofbothspecies.Theglobuliferagroupcontainsexclusively M. globulifera (BS=94%).ThestrainCBS254.76formerlyidentifiedasM. horticola might represent a new spe-ciesbecauseofitsdistinctITSsequence.TheITSsequencesof true M. horticolastrainsbelongtogroup2(Fig.5)wheretheex-syntypeofthisspeciesislocated.


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Strainnumber Originalname Revisedname

CBS585.81 M. alpina M. kuhlmaniiCBS696.70 M. alpina M. cystojenkiniiCBS272.71 M. bainieri M. kuhlmaniiCBS273.71 M. bainieri M. kuhlmaniiCBS292.96 M. chienii M. selenosporaCBS554.73 M. chienii M. selenosporaCBS387.91 M. macrocystopsis M. cystojenkiniiFSU2736 M. parvispora M. jenkinii CBS293.34 M. polycephala M. hyalinaCBS176.74 M. sossauensis M. clonocystis

Table 3 Summary of isolates which were revised and assigned to different specieswithinthisstudy.

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Group 5 – strangulata and wolfii (Fig.8,somemorpho-logicalfeaturesaredisplayedinFig.1q,r,2c,t)containsonlyfewspecies,whichcouldallbeidentifiedbymoleculardata.Thewolfii group (BS=100%) is extended in this study by M. ambigua (cladesupportBS=99%).Mortierella ambigua is sisterclade(BS=81%)toM. capitata (BS=98%)andbothclades are sister group to M. wolfii (BS=96%).Thestrangulatagroup is retained, containing M. strangulata and M. rostafinskii (BS=100%).Mortierella microzygospora, M. parazychae and M. pseudozygospora werenotassignedtoanydefinedgroup.

Group 6 – alpina and polycephala (Fig.9,somemorpho-logicalfeaturesaredisplayedinFig.1b,g,h,k,o,w,2d,m,n,p).Thepolycephalagroupharboursthetypespeciesofthewhole genus Mortierella: M. polycephala.Therefore,thiscladeresembles the core group of the genus Mortierella.RelatedtoM. polycephalaandwellsupported inLSU(BS=99%)andITS(BS=100%)arethespeciesM. bisporalis, M. hypsicladia, M. indohii, M. oligospora, M. polygonia and M. reticulata.Ex-cept for the ex-type strain of M. polygoniaCBS685.71whichclusters within the M. polycephala, all species form well sup-portedclades(Fig.9).Butjudgingfromthedifferentobservedmorphology of M. polygonia, which is that of M. polycephala insteadof thatoriginallydescribed (Gams1976), this strainshouldbetreatedassuch.Althoughthestrainissterile,itshowsthe typical stylospores of M. polycephala.AsecondisolateofM. polygonia (CBS248.81)couldnotbeconfirmedas ‘true’ M. polygonia since it does not sporulate, displaying only untypi-calstylosporesandclusterswithinthealpina-complex(Fig.9).Thereforethestatusofthisspeciesseemsdoubtful.Mortierella alpinaisoneofthemajorspeciesisolatedandidentifiedfromour environmental samples collected inAustria.Mortierella alpina forms a heterogeneous cluster with the two species M. antarctica and M. amoeboidea.ForM. amoeboidea again is the observed morphology not identical with the described oneandresemblesthespeciesindicatedbymoleculardata.ThisjustifiesM. amoeboidea W.Gams 1976 to be treated as synonym of M. alpina Peyronel 1913. OneisolateofM. globalpina (CBS718.88)isplacedwithinthealpinacomplexandoneisolate(CBS226.78)islocatedintheM. clonocystis clade (Fig.5).Verificationbyinclusionofthetypestrainisnotpossiblesincethisparticularstrainseemstobedeadnow.

Group 7 – gamsii (Fig.10,somemorphological featuresaredisplayedinFig.1a,d,f,p,u,2b,e,f,h,k,o,q,u)isthelargestgroupinthisandourpreviousstudycontaining73taxa.Thepreviousdataset(Petkovitsetal.2011)withthespeciesMortierella acrotona, M. armillariicola, M. biramosa, M. camargensis, M. cogitans, M. elongata, M. exigua, M. gamsii, M. his toplasmatoides, M. hyalina, M. nantahalensis, M. rishikesha, M. sarnyensis, M. schmuckeri, M. sclerotiella, M. zonata and M. zychae was extended by M. bainieri, M. claussenii, M. fat shederae and M. longigemmata.Mortierella armillariicola, M. bainieri, M. fatshederae, M. hyalina and M. zychae form monophyletic clades supported by the coherence of several strains(Fig.10).Mortierella exigua, M. gamsii and M. zonata arepolyphyletic.Strainsidentifiedasthesespeciesappearindifferentplacesof the tree.Noneof thestrainsofM. exigua clusterstogetherwiththeex-typestrain.ForM. gamsii at least threedividedclustersarepresent.Onesequenceofanex-typestrainisplacedintheelongata-complex.Mortierella schmuckeri forms one monophyletic clade together with M. claussenii and M. camargensis (BS=97%).Duetoalackofsufficientamountsof strains neither the phylogenetic position nor the species coherence of M. acrotona, M. cogitans, M. histoplasmatoides, M. longigemmata, M. nantahalensis, M. sclerotiella and M. zonata couldbeconfirmed.

CONCLUSIONS

InordertostudyandevaluatethemonophylyofMortierella, and to address the phylogenetic relationships of other genera in the Mortierellales, we analysed one of the largest datasets of LSUandITSsequencesforthisorder.ThegeneraDissophora, Gamsiella and Lobosporangium are placed within the genus Mortierella.ThissuggestseitherapolyphylyofMortierella with the necessity to establish additional genera or the necessity to reducetheexistinggeneratoone.Althoughourstudycontainsa comprehensive dataset it is still not possible to elucidate all species and species groups of the Mortierellales.Itwasalreadyproposed that additional molecular markers are necessary for aprofoundphylogeneticstudy(Petkovitsetal.2011).Butourstudy supports existing and reveals new contradictions to the traditionalmorphologybasedclassifications(Linnemann1941,Zychaetal.1969,Gams1977).Severalspecies,originallyiden- tifiedasone,appearondifferentplaces in thephylogeneticanalyses.Thismightoriginateeitherfromsimplemisapplica-tions or from the observed phenomenon of dependency of thephenotypeon culture conditions (Petkovits et al. 2011).Furthermore,namesofnewgeneraandspeciespublishedjustrecently may be superfluous at a nomenclatural level because their respective phylogenetic markers were not compared with the full molecular dataset of the Mortierellales,e.g.Echinochlamydosporium variabile(Jiangetal.2011),whichmayturnoutto be a micromorphologically degenerate Mortierella stylospora.Herewepresentthemostcomprehensivemoleculardatasetofthe Mortierellales which is available up to date and facilitates revisionofexistingandvalidationofupcomingnames.Finally,all these actions will lead to several species name changes andsynonymizations.Nevertheless,severalspeciesorevengroups of species seem to be distinguishable by morphol-ogy and phylogeny.Themonophyletic clade ofMortierella s.str. contains the type species of the genus, M. polycephala Coem. 1863.Whetheradditional speciesare related to thisgroup and therefore belonging to the genus Mortierella needs tobeevaluatedinfurtherstudies.Currentdata(Petkovitsetal.2011)arecontradictorywithregardtorelationshipsofspe-ciesandspeciesgroups.Duetothelackofsuitablemorpho-logical criteria the following species and species groups were misapplied and require taxonomic revision,where indicatednomenclaturalsynonymization.Theseare:M. angusta, M. basi parvispora, M. carmagensis, M. fimbricystis, M. gamsii, M. gem mifera, M. globalpina, M. horticola, M. humilis, M. jenkinii, M. kuhlmanii, M. minutissima, M. parvispora, M. rishikesha, M. schmuckeri, M. simplex, M. sossauensis, M. turficola, M. verti cillata and M. zonata.Underrepresented in this study, but due to the lack of compre-hensive additional material, are the species: M. acrotona, M. an gusta, M. dichotoma, M. epicladia, M. exigua, M. fimbricystis, M. formicicola, M. longigemmata, M. microzygospora, M. nanta halensis, M. parazychae, M. rishikesha, M. rostafinskii, M. scle rotiella and M. strangulata.

Acknowledgements This research was supported by an international co-operationgrantoftheGermanandHungarianResearchFoundations(DFGVo772/9-1andOTKANN106394)andtheHungariangrantTÉT_10-1-2011-0747.TamasPetkovitswassupportedbytheEuropeanUnionandbytheEuropeanSocialFund(projectnumber:TÁMOP-4.2.2/B-10/1-2010-0012).WeliketoexpressourgratitudetoMartinKirchmair(UniversityofInnsbruck,Austria)forcollectingandprovidingenvironmentalstrainsofMortierella from thealpineregion.AlsowewouldliketothankDomenicaSchnabelrauch(MaxPlanckInstituteforChemicalEcologyJena,Germany)andthemembersofthe molecular barcoding team at the CBS Utrecht for technical support in DNAsequencing.


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