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KellermaniawasestablishedbyEllis&Everhart(1885)toac-commodate an unusual coelomycete, K. yuccigena, with large, cylindrical, and septate conidia that were considered stipitate, and it occurred on dead leaves of Yucca(Asparagaceae, sub-family AgavoideaesensuAPGIII(2009)=Agavaceae in earlier classifications).ThestipesoftheconidiadescribedbyEllis&Everhart(1885)areinterpretedasapicalappendages(Sutton1968).Höhnel(1918a)adoptedthegenericnamePiptarthron that was provisionally suggested by Montagne for Septoria macrospora, a coelomycetous species without appendages that occurred on senescing leaves of Agave(Asparagaceae, subfamily Agavoideae=Agavaceaeinearlierclassifications).He provided a generic diagnosis and comparison to Keller-mania.Subramanian&Ramakrishnan(1954)establishedthecoelomycetous genus Alpakesa, which was characterised in part by conidia with multiple apical appendages, when their study revealed that Neottiospora yuccifolia, a species found on dead leaves of Yucca, was not congeneric with the type of Neottiospora.Several other studies have added new species to Alpakesa, Kellermania, and Piptarthron and/or provided revised generic
circumscriptions.Sutton(1968)restrictedKellermania to spe-cies with simple, blastic conidiophores; septate, hyaline, and appendage-bearing conidia; and sclerotioid, pycnidial conidio-mata, while discussing segregate genera and excluding many namesclassifiedinKellermania.Morgan-Jonesetal.(1972a)re-examined the type species of Alpakesa,modifiedthegenericcircumscription,andaddedtwospecies.Morgan-Jonesetal.(1972b)revisedtheconceptsofthetwoKellermania species thatwereacceptedbySutton(1968),addedtwonewspecies,and included taxa with or without a single apical conidial ap-pendage.Sutton (1977) synonymised theYucca-inhabiting genus Septoplaca with Piptarthron, but he was unable to de-termine which species was represented by the type, S. limbata.Sutton(1980)latertreatedS. limbata as Piptarthron limbatum and subsequently as P. yuccae(Sutton1983).Sutton(1980)maintained Alpakesa, Kellermania, and Piptarthron for genera havingmultiple,single,ornoconidialappendages,respectively.Sutton(1980)alsonotedthepossibilityofabroadlyexpandedKellermania for all species in the complex.NagRaj (1993)treated Alpakesa as a later synonym of Kellermania and listed theunexaminedorexcludedtaxa.Ramaley(1991,1992,1993,1995,1998)providedthenextmajor advances in the study of this group of fungi by discovering and describing the sexual states of Kellermania and Piptarthron, adding new species, and reviewing several coelomycetous species.ThegenusPlanistroma, the sexual state of Piptarthron, was characterised by subepidermal, ostiolate ascomata in multi-locular stromata with bitunicate asci and lackling paraphyses (Ramaley1991).Ramaley (1992)addedanotherspeciesofPlanistroma, P. obtusilunatum, with unilocular conidiomata more typical of Kellermania.Shenotedthe intermediatena-ture of the asexual state in that the lack of apical appendages
Phylogeny and taxonomic revision of the Planistromellaceae including its coelomycetous anamorphs: contributions towards a monograph of the genus KellermaniaA.M.Minnis1a,A.H.Kennedy2a,D.B.Grenier2,M.E.Palm3,A.Y.Rossman4
1 Center forForestMycologyResearch,NorthernResearchStation,USForestService,OneGiffordPinochetDr.,Madison,WI53726,USA;
Abstract ThecorespeciesofthefamilyPlanistromellaceae are included in the teleomorphic genera Planistroma and Planistromella and the connected anamorphic, coelomycetous genera Alpakesa, Kellermania, and Piptarthron.Thesegenerahavebeendefinedprimarilyonthebasisofascosporeseptationornumberofconidialappendages.Due to a lack of DNA sequence data, phylogenetic placement of these genera within the Dothideomycetes, evaluation ofmonophyly,andquestionsaboutgenericboundariescouldnotbeadequatelyaddressedinthepast.Isolatesofnearlyalloftheknownspeciesinthesegenerawerestudiedgeneticallyandmorphologically.DNAsequencedataweregeneratedforthenSSU,ITS,nLSU,andRPB1markersandanalysedphylogenetically.Theseresultsplacedthe Planistromellaceae, herein recognised as a distinct family, in an unresolved position relative to other genera within the order Botryosphaeriales.SpeciesrepresentingthecoregeneraofthePlanistromellaceae formed a clade andevaluationofitstopologyrevealedthatpreviousmorphology-baseddefinitionsofgeneraresultedinanartificialclassificationsystem.Alpakesa, Kellermania, Piptarthron, Planistroma, and Planistromella are herein recognised as belonging to the single genus Kellermania.Thefollowingnewcombinationsareproposed:Kellermania crassispora, K. dasylirionis, K. macrospora, K. plurilocularis, and K. unilocularis.Fivenewspeciesaredescribed,namelyK. con- fusa, K. dasylirionicola, K. micranthae, K. ramaleyae, and K. rostratae.Descriptionsofspeciesinvitroandakeytospeciesknownfromcultureareprovided.
Article info Received:18May2012;Accepted:6August2012;Published:25October2012.
12 Persoonia – Volume 29, 2012
GenBankaccesionno.2
Species Isolatecultureno.1 Host Locality SSU LSU ITS RPB1
suggested placement in Piptarthron while the presence of unilocular conidiomata supported placement in Kellermania.Ramaley(1993)circumscribedPlanistromella, the sexual state of Kellermania, and noted that it was similar to Planistroma but distinct in having septate ascospores.Thegenus originallyincluded two new species that were correlated with one new and one existing species of Kellermania(Ramaley1993).Shedid not embrace synonymising the asexual and sexual states intoabroadlydefinedPlanistroma with Kellermania anamorphs. Ramaley(1995)addedfivenewspeciesbasedonbothasexualand sexual states and provided a key to the species of Keller-mania and Piptarthron.Mostnotablyamongthenewspecieswas Planistromella torsifolium, the first sexual state havingAlpakesa-type conidia, which provided further support for the
synonymy of Alpakesa and Kellermania(Ramaley1995).Lastly, Ramaley(1998)namedtwomoreteleomorphicspeciescon-nected with known species of Kellermania and illustrated another undescribed species with both Piptarthron and Plani-stromastates.Planistroma kellermaniae, which has aseptate ascospores, was connected to the anamorphic Kellermania nolinae, a species with Alpakesa-type conidia, providing further confusioninregardtogenericboundariesandRamaley(1998)suggestedthatare-evaluationwasnecessary.Barr(1996)observedthatseveralgeneraintheDothideales werenotclassifiedeasilyintoanyfamily,andsheestablishedthe family Planistromellaceae to accommodate “taxa having ascostromata,interthecialtissues,andschizogenouslyformed,periphysateostioles”.Planistromella and Planistroma were in-
13A.M.Minnisetal.:Planistromellaceae - genus Kellermania
cluded and three species were transferred to the former genus along with circumstantial links to anamorphs for two species (Barr1996).ThegeneraLoratospora, Eruptio, Microcyclus, and MycosphaerellopsiswerealsoclassifiedinPlanistromellaceae.DuetotheavailabilityofanumberofRamaley’sculturesthatrepresent nearly all of the known species of Alpakesa, Keller-mania, and Piptarthron and additional cultures obtained from plantdiseaseinterceptionsatU.S.portsofentry,asystematicstudywasmadetoaddressthefollowingquestions:1)Whatare the phylogenetic relationships between members of the Planistromellaceae and the Dothideomycetes?; 2) are thePlanistromellaceae and its generamonophyletic?; 3) doesmorphology of conidial appendages or ascospore septation correlatewith phylogeny?; and 4) are slightmorphologicaldifferences among otherwise similar isolates, which are often obtained from different hosts, indications of distinct phylogenetic species?Toanswer thesequestions,nuclearprotein-codingDNA and nuclear ribosomal DNA sequence data were gener-atedforseverallociandanalysedphylogenetically.Additionally,detailed studies were made of these species in culture, including micromorphological characters.Herbariumspecimenswereexaminedwheneverpossibleand/ornecessary.
MATERIALS AND METHODS
Morphology and herbarium materialDriedherbariummaterialwasrehydratedandviewedin3%KOH(Largentetal.1977),andmicroscopicobservationsofculturesweremadeofmaterialmountedin3%KOHorbuff-eredShear’smountingfluid(Graham1959).LengthtowidthratiosaregivenasQ.Meanvaluesforlength,widthandQareindicated by Lm,Wm,andQm, respectively,basedonn=30.HerbariumacronymsfollowThiers(2012).SeeFarr&Rossman(2012)foradditionalinformationaboutcollectionshousedattheU.S.NationalFungusCollections(BPI).NewspecimensweredepositedatBPI.TheRamaleycollectionsfromwhichmanyof the cultures utilized in this studywere obtained, ifextant,aremissing.
CulturesIsolates were grown in plastic Petri plates on Difco potato-dextroseagar (PDA),whichwaspreparedaccording to themanufacturers’ instructions.Growth conditionswere 24°Cwitha12hlight/darkregimen.Culturesweremeasuredandphotographed after 2wk.Notes on colour and general ap-pearanceweremadeafter 2–3wk.Terminology for colourincludes general terms as well as standard terminology with thesamplereferencecode inparenthesesfromKornerup&Wanscher(1967).Culturesthathadnotsporulatedafter3wkwere continuously incubated under the same conditions for up to severalmonths andperiodically re-examined.Referencecultures were deposited at the Centraalbureau voor Schimmel-cultures(CBS-KNAWFungalBiodiversityCentre,Utrecht,TheNetherlands;Table1).
DNA extraction, PCR amplification, and sequencingDNAwas extracted from freshmycelium usingQiagen’sDNeasyPlantMiniKit (Germantown,MD).RibosomalDNAfromthenuclearsmallsubunit(SSU),theinternaltranscribedspacerregion(ITS;ITS1,5.8S,ITS2),andthenuclearlargesubunit(LSU)werePCRamplifiedusingtheprimerpairsNS1andNS4(Whiteetal.1990),ITS5andITS4(Whiteetal.1990),LROR (Moncalvo et al. 2000) and LR7or LR5 (Vilgalys&Hester1990),respectively.Additionally,aportionofthelargestsubunitoftheRNApolymeraseII(RPB1)wasamplifiedusingtheprimerpairRPB1-AcandRPB1-Cr(Mathenyetal.2002).EachregionwasamplifiedusingGoTaq(Promega,Madison,
WI)andassociatedstandardreagentsfollowingthemanufac-turer’srecommendationsincluding2.0mMMgCl2and1.5µMofeachprimer.ThermalcyclingconditionsforRPB1andLSUwereaccordingtoMalkusetal.(2006)andReebetal.(2004),respectively.ThermalcyclingconditionsfortheSSUandITSwere:95°Cfor60s;35cyclesat95°Cfor15s,50°C(55°CforITS)for20s,and72°Cfor60s;and72°Cfor180s.Cyclesequencing and fluorescent labelling was conducted with the samecorrespondingPCRprimersthatwereusedforeachlocusandtheBigDyeTerminatorv.3.1CycleSequencingkit(Ap-pliedBiosystems,FosterCity,CA).ThepurifiedproductswerethensequencedonanABI3730automatedDNAsequencer.GeneiousProv.5(Drummondetal.2010)wasusedtoeditelectropherograms and to build consensus sequences that were submittedtoGenBank(http://www.ncbi.nlm.nih.gov;Table1).
Data matrix and phylogenetic analysisFor the purpose of determining the phylogenetic position of Planistromellaceae among the Dothideomycetes, sequences of theSSUandLSU from the type speciesof Kellermania, Piptarthron, Planistroma, and Planistromella were manually incorporated into thealignment ofSchochet al. (2009) us-ing theprogramGeneiousProv.5 (Drummondetal.2010;i.e.,Dothideomycetes alignment). Identical sequenceswereremoved from this alignment.AMaximumLikelihood (ML)analysiswasconductedinRAxMLv.7.3.0(Stamatakis2006)usingthe‘RAxML-HPC2onXSEDE’toolviatheCIPRESSci-enceGateway(Milleretal.2010),selectingtheGTRGAMMAnucleotide substitution model and Opegrapha dolomitica and Schismatomma decolorans asoutgroups(Schochetal.2009).Aseparaterapidbootstrap(bs)analysisof1000iterationswasconductedwithidenticalsettings.Multiple-sequence alignments were conducted for each of the datasets from the PlanistromellaceaeinGeneiousProv.5(Drum- mondetal.2010)usingMUSCLEv.3.6(Edgar2004),adjustedmanually,andthenconcatenated(i.e.,Kellermaniaalignment).Congruenceamongthesefourdatapartitions(SSU,ITS,LSU,RPB1)wasevaluatedbycomparingtheirtopologiesinsearchofwell-supported clades (posterior probability >0.85)withconflicting compositions, and separately with the Incongruence LengthDifferencetest(ILD;Farrisetal.1994).TheindividualtopologieswereconstructedwithBayesian inference (BI) inMrBayesv.3.1.2(Huelsenbeck&Ronquist2001,Ronquist&Huelsenbeck2003)wherebest-fittingmodelsforeachdatasetweredeterminedinMrModeltestv.2.2(Nylander2004)bytheAkaikeInformationCriterion(AIC;Posada&Buckley2004).Allotherparameterswereleftasdefault.Theposteriorprobability(pp)distributionoftreeswasestimatedfromthosecollecteduntil the standard deviation of split frequencies reached less than0.01 (1million generations)minus the burn-in (10%),whichwasdeterminedintheprogramTracerv.1.5(Rambaut&Drummond2007).TheILDtestwasimplementedinPAUPv.4.0b10(Swofford2003)asthePartitionHomogeneityTestwith1000homogeneity replicates.Each replicatesearchedtreespacewith100randomadditionsequences(RAS)sav-ing10treesperRASwhileignoringuninformativecharacters.ThemonophylyofcorePlanistromellaceae was tested by con-ductingaphylogeneticanalysisofamatrixcomprisedofSSU,ITS,andLSUfromawiderangeofBotryosphaeriaceae se-lectedfromtheanalysesofCrousetal.(2006),Phillips&Alves(2009),andSchochetal.(2009);plustypespeciesofgeneraof Kellermania, Piptarthron, Planistroma, and Planistromella as wellasatleastonerepresentativeofeachmajorcladeofcorePlanistromellaceae; and Helicomyces roseus(CBS283.51)asoutgroup(i.e.,Botryosphaeriaceaealignment).ThismatrixwasalignedwithMUSCLEv.3.6(Edgar2004)andanalysedwithBI,asdescribedabove.
ThemajorcladesofthecorePlanistromellaceaewereidentifiedfromtheresultsofamixed-modelBIanalysisoftheKellermania alignment plus Helicomyces roseus(CBS283.51)inMrBayesv. 3.1.2 using themethodsoutlined above.This phylogenywasalsousedtoi)testthemonophylyofgenericconceptsinPlanistromellaceae;ii)determinerelationshipsamongitsspe-ciesincludingnoveltaxa;andiii)evaluatetheevolutionofkeymorphologicalcharactersandhostrelationships.AllalignmentsandresultingtreesweredepositedintoTreeBASE(S13234),andnomenclaturalnoveltiesinMycoBank(Crousetal.2004).
RESULTS
Data matrix and phylogenetic analysesSequencing was successful for all core Planistromellaceae taxawith living cultures except for theSSUofKellermania
Phylogenetic relationships of the PlanistromellaceaeMaximum Likelihood analysis of Dothideomycetes revealed highsupport(bs=91%)foracladecomprisedstrictlyofBotryo-sphaeriaceaethatwascongruentwithSchochetal.(2009)with
15A.M.Minnisetal.:Planistromellaceae - genus Kellermania
theexceptionofawell-supportedclade(bs=99%)containingthe type species of the genera Kellermania, Piptarthron, Plani-stroma, and Planistromella of Planistromellaceae(phylogramnot shown).Bayesian inference of theBotryosphaeriaceae alignment suggested that Planistromellaceae is one of fivemonophyletic members of Botryosphaeriaceae s.l. (seeDis-cussion):i)Bagnisella examinans;ii)Saccharata proteae;iii)Melanops(pp=1.0);iv)Planistromellaceae(pp=1.0);andv)the remaining members of the family including the core mem-bers of Botryosphaeriaceae(pp=1.0;Fig.1).
Phylogenetic diversity and relationships within KellermaniaInspection of the individual phylogenies resulting fromBIanalysesofSSU,ITS,LSU,andRPB1revealedasinglewell-supported incongruence.This incongruencewas betweentheSSUandRPB1treeswhereKellermania macrosporaAR3468was sisterwith a clade containingK.crassispora AR3463,K. dasylirionicolaAR3465,K. micranthaeAR3474,andK. nolinaeAR3475intheSSUanalysis(pp=0.94);whereasK. macrosporawaspartofapolytomy(pp=1.0)intheRPB1analysis containing two other clades, one comprised of K.uni-septataAR3476,K.yuccifoliorumAR3472,andK.yuccigena AR3470,andanothercontainingK.anomalaAR3471andK.nolinifoliorumAR3473.Resultsof the ILD test (Farrisetal.1994)suggestedthattheSSU,ITS,LSU,andRPB1datafrom Planistromellaceaewerecongruent(P=0.230)andthussuitableforcombinedanalysis.OurBI analysis of theKellermania alignment resulted in a consensusphylogram (Fig. 2) comprisedof fivemajor line-ages: i)Kellermania macrosporaAR3468,K.confusaAR3469,K.ramaleyaeMEP1260,andK. rostrataeJB5.16.11-01(pp=0.89);ii)K. yuccigenaAR3470,K.uniseptataAR3476,and K.yuccifoliorumAR3472(pp=0.95);iii)K.anomalaAR
3471,K.nolinifoliorumAR3473(pp=1.0);iv)K.micranthaeAR3474,K.dasylirionisAR3464,K.crassisporaAR3463,K.da-sylirionicolaAR3465,andK.pluilocularis AR3467(pp=0.74);andv)K.nolinaeAR3475(Fig.2).Lineagev,whichisbasedupon a single isolate, is herein labelled as distinct because its position among the other lineages is unresolved and varied in differentanalyses.Therelationshipsamongthesefivelineagesarenotwellresolvedthoughsomemajorgroupingsareweaklysupported and a sister relationship of lineages ii– iii receives somesupport(pp=0.70).Membersoflineagesi– iiiwereallisolated from either Agave or Yucca(Asparagaceae, subfamily Agavoideae),withtheexceptionofK.nolinifoliorumAR3473on Nolina(Asparagaceae, subfamily Nolinoideae).Membersof lineages iv–v were isolated from members of Nolina or Da-sylirionis(subfamilyNolinoideae)withtheexceptionofK.pluri- locularis AR3467onYucca (Fig.2).Lineages i– iiipossessseptate ascospores and lineages iv–v possess aseptate as-cospores, but see K. unilocularisbelowandinFig.3.Lineagei possesses conidia without conidial appendages, lineage ii pos-sesses conidia with single or multiple appendages, lineage iii possesses conidia with single appendages, lineage iv pos-sesses conidia without appendages, and lineage v possesses conidiawithmultipleappendages.
Colony characteristics and micromorphology of KellermaniaColony colour typically ranged from greenish to greyish tones, but K. crassisporauniquely remainedpink.Differenceswereobserved in growth rates, but in general slight differences in growth rates and overall colony appearances were observed fromsubculturetosubculture.Thus,tendenciestowardslowervsfastergrowthratesaremoreusefulthanexactmeasurements.Conidiomata were produced in culture by all species except K. crassispora.Theyarecharacterisedas:superficialorimmersed,
scattered to densely gregarious, at times confluent becoming fused, discrete or associated with stromatal growths including irregularly column-like and rarely mat-like structures, frequently more or less round in shape, with or without necks, black, at times coveredwithwhitish,greenish,orgreengreyhyphae.Conidiog-enous cells lining the inner conidiomatal walls were holoblastic, determinate, discrete or integrated on short single-celled conidi-ophores,doliiformtocylindrical,hyaline,andsmooth.Conidia werevariableinsizeandshape,aseptate,singletomultiseptate,andbear0–multipleapicalappendages.Alessusefulcharacterincludedthepresenceofafrillatthebaseofconidia.Conidia,which developed within weeks or sometimes only after months, wereexudedfromconidiomatainwhitish,mucilaginousmasses.Conidia often germinate quickly and, in some several month old cultures, nearly all conidia in the mucilaginous masses had eithergerminatedordesiccated.Inthedescriptionsofcultures,colony appearance, growth rate, and conidia are the most valu-ablecharactersfordistinguishingspecies.
TAxONOMy
PlanistromellaceaeM.E.Barr,Mycotaxon60:433.1996. Type genus.PlanistromellaA.W.Ramaley.
Kellermania Ellis&Everh.,J.Mycol.1:153.1885.
Type species.Kellermania yuccigenaEllis&Everh. =Piptarthron Mont.exHöhn.,Hedwigia:60:203.1918. Type species.Piptarthron macrosporum(Durieu&Mont.)Höhn. =Alpakesa Subram.&K.Ramakr.,J.IndianBot.Soc.33:204.1954. Type species.Alpakesa yuccifolia(J.G.Hall)Subram.&K.Ramakr.as‘yuccaefolia’. ?=SeptoplacaPetr.,Sydowia17:271.1964‘1963’. Type species.Septoplaca limbataPetr. =Planistroma A.W.Ramaley,Mycotaxon42:69.1991. Type species.Planistroma yuccigenumA.W.Ramaleyas‘yuccigena’. =Planistromella A.W.Ramaley,Mycotaxon47:260.1993. Type species.Planistromella yuccifoliorumA.W.Ramaley.
Culturecharacteristics—ColoniesonPDA43–44mmaf-ter2wkat24°Cwitha12hlight/darkregimen;surfaceneardullgreen(30E4),coveredwithlow,cobwebbytovelutinous,greenish hyphae; margin uneven, whitish; reverse black with margin uncoloured.Conidia 40–69× 5–8µm,Q=5–14.3(Lm=52.3µm,Wm=7.3µm,Qm=7.4),cylindricaltofusiform,apices tapering to a relatively acute point, tapering towards and typically truncate at bases that may bear an indistinct frill,1–2(–3)-septate;wallssmooth,thinandhyaline,andnotsignificantlyconstrictedatsepta;contentshyaline;appendagesabsentorpresent,attimesscarcelyvisible,upto11µmlong,apical,single,appearingasashort,filiform,hyalinemucro. Habitat&Distribution—DeadleavesofYuccaspp.(Morgan-Jonesetal.1972b,NagRaj1993,Farr&Rossman2012).Thetype was found on Y. draconis(Cooke1878),probablycorrectlynamed Y. brevifolia.ThisfungalspeciesiswidelydistributedinthewesternhalfoftheUSA(Morgan-Jonesetal.1972b,Farr&Rossman2012).
Notes—Ramaley(1993)notedthattheappendagemaybehard to observe or absent and that the middle cell in conidia with twoseptaisregularlyshorter.Weobservedthattheappendage(whenpresentinculture)wasshorterthaninpreviousreportsfrommaterialinnature(Morgan-Jonesetal.1972b,NagRaj1993).Shoemaker&Babcock(1987)redescribedP. yuccigena on Yucca glauca.Barr (1996)provisionally linkedPlanistro-mella yuccigena to K. anomala based on the circumstantial occurrenceofbothonthesameleaves.Thislinkisindirectlysupported by the morphological similarity of the sexual state to that of K. nolinifoliorum,asisterspecies.
Culturecharacteristics—ColoniesonPDA35–40mmafter 2wk at 24°Cwith a 12h light/dark regimen; surface nearpalegrey(30B1),coveredwithlow,whitecobwebbytoveluti-nous hyphae; margin uneven, whitish; reverse grey with black conidiomatavisible.Conidia38.5–64×6.5–9.5µm,Q=4.8–8(Lm=49.9µm,Wm=8.3 µm,Qm=6.0), obclavate, at timescurved, apices tapering towards and slightly acute, typically truncateatbases,(1–)2(–3)-septate;wallssmooth,thin,hya-line, and not constricted at septa; contents hyaline; append-agesabsent. Habitat&Distribution—DeadleavesofYucca thornberi.ItisknownonlyfromthetypelocalityintheUSA:AZ.
pendages.Ramaley(inlitt.)notedthattwoisolatesidentifiedas Piptarthron macrosporum, this one from Yucca and another from Agave,hadconsistentdifferencesinconidialmorphology.Morphological and DNA sequence data support the separation of this species from Kellermania macrospora on Agave.
Culturecharacteristics— ColoniesonPDA30–36mmafter 2wkat24°Cwitha12hlight/darkregimen;surfacenearpink-ishwhite(10A2)tocottoncandy-pink,coveredwithlow,veluti-nous hyphae; margin uneven, whitish to pale pink; reverse near palered(7A3).Noconidiomataobserved.Noconidiaobserved.
Habitat&Distribution—DeadleavesofNolina micrantha, Nolina sp. (Ramaley1995, thisstudy). It isknown from theUSA:NM,TX(Ramaley1995,thisstudy).
Specimen examined.USA,Texas,CulbersonCo.,GuadalupeMtns.NationalPark,5.6milesfromhighwayalongWilliamsRanchRd.,ondeadleaves of Nolina micrantha,23Oct.1995,coll.A.W. Ramaley,AR3463(CBS131714)isolatedbyA.W.RamaleyfromAWR9536,driedcultureonPDA(BPI882815).
Notes—TheculturethatwestudieddidnotsporulatethoughRamaley(1995)observedconidiainculture.Ramaley(1995)statedthatconidiaonthehostwere56.8–78.4×12.8–14.4µm,cylindrical with rounded apices, aseptate, and without append-ages.Thelinkbetweentheasexualandsexualstatesisbasedon cultural similarities thatwerenotedpreviously (Ramaley1995).
Fig. 5a.ConidiaofKellermania anomalafromculture(AR3471);b.conidiaofK. confusafromculture(AR3469);c.conidiaofK. dasylirionicola from culture (AR3465);d.conidiaofK. dasylirionisfromculture(AR3464);e.conidiaofK. macrosporafromculture(AR3468);f.conidiaofK. micranthae from culture (AR3474);g.conidiaofK. nolinaefromculture(AR3475);h.conidiaofK. nolinifoliorumfromculture(AR3473);i.conidiaofK. plurilocularisfromculture(AR3467).—Scalebars=30µmforall.
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19A.M.Minnisetal.:Planistromellaceae - genus Kellermania
Culturecharacteristics—ColoniesonPDA22–25mmafter2wkat24°Cwitha12hlight/darkregimen;surfacewhitishto near pinkishwhite (10A2), coveredwith low, tomentoseto velutinous hyphae; margin uneven, whitish; reverse near orangewhite(5A2).Conidia32–45×3–5µm,Q=6.7–12.5(Lm=39.4µm,Wm=4.5µm,Qm=9.0),moreorlesscylindri-cal with irregularly curved shape, flexuous, apices tapering to a somewhat acute point, typically truncate at bases, aseptate; walls smooth, thin, and hyaline; contents hyaline; appendages absent. Habitat&Distribution—DeadleavesofDasylirion leiophyl-lum.ItisknownonlyfromthetypelocalityintheUSA:TX.
Notes—Althoughsimilar toKellermania dasylirionis, this second species on Dasylirion is distinguished by its aseptate conidia with irregular, curved shapes and slower growth on PDA.Ramaley(inlitt.)notedaseptate,curlyconidiainmaterialfrom the host in nature and speculated that the collection may representanundescribedspecies.DNAsequencedataandassociatedanalysesconfirmthisasanewspecies.
Culturecharacteristics—ColoniesonPDA45–52mmaf-ter2wkat24°Cwitha12hlight/darkregimen;surfaceneargreenish grey (30C2), coveredwith low, velutinoushyphaethat often covers conidiomata; margin uneven, whitish; reverse near brownish grey (7E2) to greyish orange (5B3).Conidia 45–64×5–9.5µm,Q=6.3–12.3(Lm=57.1µm,Wm=7.2µm,Qm=8.1),cylindricaltonarrowlyfusiformwithsomedegreeofcurvature, apices tapering to a relatively acute point, tapering towards and typically truncate at bases, 0–1-septate, septa approx.medianwhenpresent;wallssmooth,thin,hyaline,andnot constricted at septa; contents hyaline, at times granular; appendagesabsent. Habitat&Distribution—DeadleavesofDasylirion leiophyl-lum, D. wheeleri, Dasylirionsp.(Ramaley1995,thisstudy).ThisspeciesisknownfromMexicoandtheUSA:AZ,TX(Ramaley1995,thisstudy).
Specimen examined.USA,Texas,BrewsterCo., BigBendNationalPark, Sotol Vista, on dead leaves of Dasylirion leiophyllum,25Oct.1994,coll.A.W. Ramaley,AR3464(CBS131715)isolatedbyA.W.RamaleyfromAWR9441,driedcultureonPDA(BPI882816).
Basionym.Septoria macrosporaDurieu&Mont.,Expl.Sci.Algerie1:589.1849. ≡Hendersonia montagneiCooke,NuovoGiorn.Bot.Ital.10:19.1878.Note:Thisnom.nov.wasestablishedsincetheepithet ‘macrospora’wasoccupied by H. macrospora Berk.&Broome1850. ≡Hendersonia piptarthraSacc.,Michelia2:111.1880.Note:Thisnom.nov.wasestablishedsincetheepithet‘macrospora’isoccupiedbyH. mac-rospora Berk.&Broome1850.Itisanom.illeg.viasuperfluous,ICBNArt.52
(McNeilletal.2006),sinceH. montagnei was already published as a re-placementname. ≡Stagonospora macrospora(Durieu&Mont.)Sacc.,Syll.Fung.3:450.1883. ≡Piptarthron macrosporum(Durieu&Mont.)Höhn.,Hedwigia60:203.1918.
Culture characteristics—Colonies onPDA52mmafter2wkat24°Cwitha12hlight/darkregimen;surfacenearlightgrey(30C1),withscatteredlow,hyphaebetweenconidiomata;margin uneven, whitish; reverse uncoloured, black, to near grey-ishbrown(6D3).Conidia54.5–93×6.5–11µm,Q=4.9–13(Lm=72.2µm,Wm=9.0µm,Qm=8.3),cylindricaltoobclavate,at times slightly curved, apices tapering towards and obtuse to slightly acute, typically truncate at bases that frequently bear a marginalfrill,3–5(–7)-septate;wallssmooth,thin,hyaline,andnotconstrictedatsepta;contentshyaline;appendagesabsent. Habitat&Distribution—DeadleavesofAgave americana, Agave sp. (Sutton1980,Farr&Rossman2012).Basedonscatteredreports,thisspeciesisknownfromAfrica(Algeria),Europe,andNorthAmerica(USA)(Höhnel1918a,Sutton1980,Farr&Rossman2012).Thisspecieshasalsobeenreportedfrom species of Yucca fromvarious locations (Sutton1980,Farr&Rossman2012),butwehavefoundnospecimensonYucca.
Notes—ThisspecieswasdescribedoriginallyfromAgave.Collections on Yucca, including the type of Kellermania multi-septata,havebeentreatedasconspecific(e.g.Sutton1980),but it appears that K. macrospora is limited to Agave.Plani-stromella parryi, described originally from Agave shawii(Cooke1885),Plowrightia agaves, described from Agavesp.(Maublanc1903),andPlowrightia williamsoniana, described from Agave americana (Kellerman1906), are potential synonyms (Barr1996)aswellaspossiblythesexualstateofK. macrospora, buttheseteleomorphsarepoorlyknown.
Culturecharacteristics—ColoniesonPDA38–48mmafter2wkat24°Cwitha12hlight/darkregimen;surfaceneargrey-ishgreentogreenishgrey(30B6–30B2)togreyishgreentodullgreen(30E5–30E4),attimesportionscoveredwithlow,white,cottony hyphae; margin more or less even, whitish; reverse neargreyishgreen togreenishgrey (30C3–30C2).Conidia 43–61× 6.5–9.5µm,Q=4.5–7.8 (Lm=49.2µm,Wm=8.6µm,Qm=5.8),cylindricaltoobclavate,attimescurved,apicestapering towards and obtuse, rounded or truncate at bases, aseptate; walls smooth, thin, hyaline; contents hyaline, at times granular;appendagesabsent. Habitat&Distribution—DeadleavesofNolina micrantha.ThisspeciesisknownonlyfromthetypelocalityintheUSA:TX.
Specimen examined.USA,Texas,CulbersonCo.,GuadalupeMtns.NationalPark,5.6milesfromhighwayalongWilliamsRanchRd.,ondeadleaves of Nolina micrantha,23Oct.1995,coll.A.W. Ramaley,AR3474(CBS131724)isolatedbyA.W.RamaleyfromAWR9536,driedcultureonPDA(holotype,BPI882825).
Notes—Thisspeciesisdistinguishedfromtheothersknownfrom Nolina by the combination of its aseptate conidia that lack appendagesanditsgreenishcolourationinculture.Ramaley(inlitt.)statedthatthiswasanundescribedspeciesandgaveittheprovisional name, Piptarthron sotoli.Anundescribedspecies
21A.M.Minnisetal.:Planistromellaceae - genus Kellermania
(Ramaley1998)withaseptate,Planistroma-type ascospores, which was found on the type specimen of P. kellermaniae on Nolina erumpens inTexas,hasananamorphthatissimilartoK. micranthaeandthetwospeciesmaybeconspecific.
Culturecharacteristics—ColoniesonPDA9–20mmafter2wk at 24°Cwith a 12hlight/dark regimen; surface neargreenishgrey(25C2),formingtiersofraisedmounds,smoothtograiny, at times portions covered with scattered white, velutinous, aerial hyphae; margin uneven, whitish; reverse near greyish white(25B1).Conidia35–48×8–11µm,Q=3.7–6(Lm=40.5µm,Wm=8.8µm,Qm=4.7), fusiform toobclavate, at timesslightly curved, tapering or not towards generally obtuse apices, tapering towards and typically truncate at bases that frequently bearafrill,2–3-septate;wallssmooth,thinandhyaline,andnotconstricted at septa; contents hyaline; appendages present, 8–24µmlong,apical,3–5,filiform,unbranched,hyaline. Habitat&Distribution—DeadleavesofNolina erumpens, N. microcarpa(Pollack1947,Ramaley1998,thisstudy).ItisknownfromtheUSA:AZ,TX(Pollack1947,Ramaley1998,thisstudy).
Culturecharacteristics—ColoniesonPDA23–26mmafter2wkat24°Cwitha12hlight/darkregimen;surfaceneardarkgrey(28F1),at timesportionscoveredwithscatteredwhite,cobwebby, aerial hyphae; margin uneven, whitish, somewhat slimyinappearance;reversenearlightgreytodarkgrey(28D1–28E1)with offwhitemargin.Conidia 37–57.5× 5–9.5µm,Q=5.6–9.3(Lm=49.1µm,Wm=7.1µm,Qm=7.0),fusiform,straight, slightly curved to somewhat sigmoid, apices tapering to a relatively acute point, tapering towards and typically truncate atbasesthatmaybearmucilaginousmaterial,approx.medianly1-septate;wallssmooth,thinandhyaline,andnotsignificantlyconstricted at septa; contents hyaline; appendages present or scarcelyvisible,perhapsabsent,upto5µmlong,apical,single,appearingasablunt,mucilaginousmucro. Habitat&Distribution—DeadleavesofNolina microcarpa, N. micrantha, Nolinasp.(Ramaley1995,1998,thisstudy).ThisfungusisknownfromtheUSA:AZ,NM,TX(Ramaley1995,1998,thisstudy).
Notes—Thesizerangeof theconidiaof theholotypeofPiptarthron plurilocularewasreportedas(48–)59–76(–98)× (4–)5.5–7(–8)µm(Ramaley1991).Theconidiallengthofthepresentisolate(AR3467)inculturewasmuchshorter,buttheoverall appearance of the conidia was basically the same as thoseoftheholotype.Thelinkbetweentheasexualandsexualstatesisbasedonculturalsimilarities(Ramaley1991).
Etymology.ThisspeciesisnamedinhonourofAnnetteW.Ramaleyforher outstanding contributions towards the study of Planistromellaceae and coelomycetousanamorphs.
Onhost(holotype):Foliicolous,stromataapprox.0.5–0.75mmdiam, scattered to gregarious, round in top view, subepidermal, immersed becoming erumpent after peeling back of disc-shapedepidermal tissue, black,multilocular.Conidiophores absent.Conidiogenous cells lining the basal and lateral, locu-larwalls,moreorlessdoliiformtoslightlycylindrical.Conidia 48–70.5×11–16µm,Q=4.3–5.3(Lm=58.8µm,Wm=12.6µm,Qm=4.7),cylindricaltoslightlyclavateorrarelycurved,apicesobtuse, typically truncateatbases,2–4(–5)-septate;wallssmooth,thin,hyaline,andnotsignificantlyconstrictedatsepta;contentshyaline,attimesgranular;appendagesabsent. Culturecharacteristics—ColoniesonPDA7–21mmafter2wkat24°Cwitha12hlight/darkregimen;surfaceneardullgreen(26D3,29E4,30E4,)todarkgreen(29F4),coveredwithlow, cottony to velutinous hyphae; margin uneven, whitish; re-verse multi-coloured: uncoloured, black, near greyish turquoise (24D5),anddullgreen(27E3).Conidia35–51×9.5–16µm,Q=2.6–4.6(Lm=42.7µm,Wm=12.3µm,Qm=3.5),cylindri-cal to slightly clavate or rarely curved, apices obtuse, typically truncateatbases,2–4-septate;wallssmooth,thin,hyaline,andnot constricted at septa; contents hyaline, at times granular; appendagesabsent. Habitat&Distribution—DeadleavesofYucca sp.Itisknownonly from the material from Mexico intercepted at the Laredo PlantInspectionStation.
Specimen examined. Mexico,InterceptedatLaredo,Texas,USAondeadleaves of Yucca sp.,2Dec.1985,coll.S. Vesper,MEP1260(CBS131722)isolatedbyM.E.PalmfromBPI525045(holotype,as ‘P. macrosporum’),driedcultureonPDA(BPI882823).
23A.M.Minnisetal.:Planistromellaceae - genus Kellermania
Notes—Thisspeciesisdistinguishedfromtheothersthatoccur on Yuccabyitsconidiainculturewith2–4septa,Q=2.6–4.6,andappendageslacking.Somedifferenceswereobservedindimensionsofconidiaproducedonthehostvsinculture.Aloculoascomycete producing tardily septate ascospores that wasfoundonthehostmaybethesexualstateofthisspecies.
Onhost(holotype):Foliicolous,stromataapprox.0.5–1mmdiam, scattered to gregarious, round to ellipsoid in top view, subepidermal, immersed becoming erumpent after peeling back of disc-shaped epidermal tissue, black,multilocular.Conidiophores absent.Conidiogenous cells lining the basal and lateral, locular walls, more or less doliiform to slightly cy-
lindrical.Conidia30.5–48×5–6.5µm,Q=6.3–9.3(Lm=41.3µm,Wm=5.3µm,Qm=7.8),cylindricaltoobclavate,attimescurved, apices tapering towards and obtuse to slightly acute, typically truncate at bases, aseptate; walls smooth, thin, and hy-aline;contentshyaline,attimesgranular;appendagesabsent. Culturecharacteristics—ColoniesonPDA52mmafter2wkat24°Cwitha12hlight/darkregimen;surfacewhitishtoneargreenishgrey(30C2),attimesportionscoveredwithscatteredlow, wispy, white hyphae; margin uneven, whitish; reverse uncol-ouredwithconidiomatavisible.Conidia33.5–62.5×5–9.5µm,Q=4.2–9(Lm=44.1µm,Wm=7.1µm,Qm=6.3),cylindricaltoobclavate, at times curved, apices tapering towards and obtuse toslightlyacute,typicallytruncateatbases,0–1(–3)-septate;walls smooth, thin, hyaline, and not constricted at septa; con-tentshyaline,attimesgranular;appendagesabsent. Habitat&Distribution—DeadleavesofYucca rostrata.ThisspeciesisknownonlyfromaninterceptionofMexicanmaterial.
Specimen examined.Mexico,InterceptedatLosIndios,Texas,USAondead leaves of Yucca rostrata,12May2011,coll.A. Garza,JB5.16.11-01(CBS131721)isolatedbyJ.F.BischofffromBPI884092(holotype),driedcultureonPDA(BPI882822).
Notes—Thisspeciesisdistinguishedfromtheothersthatoc-cur on Yuccabyitstypicallycylindricaltoobclavate,0–1(–3)-sep-tateconidiainculturethatlackappendages.Conidiaproducedonthehostwereaseptate.Inculture,septawereobservedonasmallpercentageofconidia.Somedifferencesinsizeofconidiawereobservedbetweenmaterialfromhostandinculture.
Onhost(isotypeofPiptarthron uniloculare):Conidia48–72× 8–9.5µm,Q=5–7.6(Lm=56.4µm,Wm=9.0µm,Qm=6.3),cylindrical, at times curved, apices straight, obtuse, less com-monly tapering towards, typically rounded to inconspicuously truncateatbases,2–4(mostly3)-septate;wallssmooth,thin,hyaline, andnot significantly constricted at septa; contentshyaline;appendagesabsent. Culture characteristics—Only sterile, irregular stromatalgrowthsobservedinthedriedculture. Habitat&Distribution—DeadleavesofseveralYuccaspp.(Ramaley1992).The typesof both theasexual andsexualstates were found on Yucca baccata (Ramaley 1992).ThisspecieshasbeenreportedfromtheUSA:CO,NV(Ramaley1992,thisstudy).
Specimens examined.USA,Colorado, LaPlataCo.,Durango, alongbiketrailbetweenChapmanHillskislopeandLion’sDen,ondeadleavesofYucca baccata,8June2000,coll.A.W. Ramaley,AR3466isolatedbyA.W.RamaleyfromAWR2001a,driedculture(BPI883224);RidgeS.ofSmelterMountain, Cactus ridge, on dead leaves of Yucca baccata,10Feb.1992,coll.A.W. Ramaley,AWR9018(isotypeofPiptarthron uniloculare,BPI1110167).
Notes—Theconidiaof the isotypewereobserved tobeslightlyshorterthanreportedbyRamaley(1992),andnoconidiawereobservedwith5septaalthoughRamaley(1992)reportedthatsomehad5septa.Thecultureofthisspeciesisdeadandonlyadriedcultureofthisisolateisextant,butRamaley(1992)reportedthattypicalconidiawereformedinculture.Thelinkbetween the asexual and sexual states is based on the produc-tionoftypicalconidiainculturesobtainedfromasci(Ramaley1992).An additional analysis was performed as the Kellermania analy-sisabovebutwiththeadditionofanexistingITSsequencefromK. unilocularisAR3466.Theresultssuggestedasisterrela-tionship between K. unilocularisandlineagei(pp=0.82).Thisexpanded clade with lineage i was member of a larger clade (pp=0.89)alsocontaining lineage ii (Fig.3).Theunilocularstromata of K. unilocularis is shared by members of lineage ii havingconidiawithappendages.Itslackofconidialappendagesissharedbymembersoflineageihavingplurilocularstromata.Itsaseptateascosporesdistinguishitfrombothlineages.Thisistheonlyspeciesinorcloselyrelatedtolineagesi–iii(Fig.2)withaseptateascospores.Itwasexcludedfromotherphylogeneticanalysesduetomissingdata.
Culture characteristics—Colonies on PDA 10–13mmafter 2wk at 24°Cwith a 12h light/dark regimen; surfacenearorangewhite(6A2),slightlyslimyinappearance,attimesportions covered with low, cottony hyphae; margin uneven, whitish; reverse off white to nearly concolourous with upper surface.Conidia27–45×8–17.5µm,Q=2–4.2 (Lm=35.8µm,Wm=12.4µm,Qm=3.0),cylindrical,fusiform,obclavate,to ellipsoid, apices tapering or not, obtuse, tapering or not towards and typically truncate at bases bearing indistinct frills, approx.medianly1-septate;wallssmooth, thinandhyaline,andnotsignificantlyconstrictedatsepta;contentshyaline,attimesgranular;appendagespresent,13–41.5µmlong,apical,3–5(–8),filiform,unbranched,flexuous,hyaline. Habitat &Distribution—Dead leaves ofYucca rupicola (Morgan-Jonesetal.1972a,Ramaley1995,thisstudy).ThisspeciesisknownonlyfromtheUSA:TX(Morgan-Jonesetal.1972a,Ramaley1995,thisstudy).
Culturecharacteristics—Thisspecies isnotknown fromculture.Itischaracterisedbyitsaseptateconidiawithmultipleappendages(Hall1915,Morgan-Jonesetal.1972a,NagRaj1993). Habitat&Distribution—DeadleavesofYucca filamentosa, Y. gloriosa, and Yuccasp.(Morgan-Jonesetal.1972a,Sutton1980).ThisspeciesiswidespreadintheUSAbasedonscat-teredreports(Morgan-Jonesetal.1972a,Sutton1980,Farr&Rossman2012).ThetypewascollectedonYuccasp.intheUSA:WA(Hall1915).
Specimen examined —None.
Notes—NoDNA sequence data exist for this species,which is the type of the genus Alpakesa(Subramanian&Ram-akrishnan1954).Consideringthatthedatapresentedhereforother species of Kellermania indicate that conidial septation and appendage number are not important for distinguishing genera, this species belongs in the genus Kellermania.Theoccurrenceof this species on Yucca and its multiple conidial appendages suggestaprobablephylogeneticplacementinlineageii.
Culturecharacteristics—ColoniesonPDA27–40mmafter2wkat24°Cwitha12hlight/darkregimen;surfaceneardarkgreen(30F5–30F3),coveredwith low,cottony tovelutinoushyphae;marginuneven,whitish;reversegreytoblack.Conidia 40–65.5×8–11µm,Q=3.9–7.4(Lm=52.6µm,Wm=9.8µm,Qm=5.4),cylindricaltoobclavate,apicestaperingtowardsandobtuse, typically truncate to less commonly somewhat rounded atbases,2–3-septate;wallssmooth,thinandhyaline,andnotsignificantlyconstrictedatsepta;contentshyaline;appendages
25A.M.Minnisetal.:Planistromellaceae - genus Kellermania
present,attimesscarcelyvisible,possiblyabsent,upto1.5µmlong,apical,single,appearingasashort,dome-likemound. Habitat&Distribution—Dead leavesofYucca baccata, Y. brevifolia(holotypesofbothnames),Y. schidigera, Y. thorn-beri(Ramaley1993,thisstudy).ThisspecieshasbeenreportedfromtheUSA:AZ,CA,UT(Ramaley1993,thisstudy).
Notes—Ramaley(1993)reportedvariationinconidialsizesbetween specimens found on different hosts and between fresh anddriedconidia.Giventhediversityfoundinthisstudy,itisdifficulttostatewithcertaintythatallspecimensfromallhostsareconspecific.Theconidiafromthecultureexaminedhereweresmallerthanthosereportedfromthehostinnature(Ra-maley1993).Theconnectionoftheasexualandsexualstatesiscircumstantial(Ramaley1993).
Culturecharacteristics—ColoniesonPDA19–25mmafter2wkat24°Cwitha12hlight/darkregimen;surfacewhitishtonearpalegrey(30B1),coveredwithcottonyhyphae;marginuneven, whitish; reverse greyish with black conidiomata vis-ible.Conidia37–56×8–13µm,Q=3.3–6.2(Lm=48.2µm,Wm=10.1µm,Qm=4.8),cylindricaltosomewhatobclavate,apices tapering towards or not and typically obtuse but some-times slightly acute, tapering or not, typically truncate at bases, approx.medianly1(–2)-septate;wallssmooth,thinandhyaline,and not or slightly constricted at septa; contents hyaline; ap-pendagespresent,5–35µm(Lm=19.6µm)long,apical,single,filiform,unbranchedtorarelybranchedwithasinglebifurcation,flexuous,hyaline. Habitat&Distribution—DeadleavesofseveralspeciesofYucca(Morgan-Jonesetal.1972b,NagRaj1993,Ramaley1993,Farr&Rossman2012).Thetypeoftheasexualstatewasfound on Y. glauca(asY. angustifolia)(Ellis&Everhart1885)and the type of the sexual state was found on Y. elata(Ramaley1993).ThisspeciesiswidelydistributedinthewesternhalfoftheUSA(Morgan-Jonesetal.1972b,Ramaley1993,Farr&Rossman2012).
Notes—ThisspeciesdescribedfromYucca filamentosa in NewJersey(Ellis&Everhart1888)couldbethesexualstateof a Kellermania, but confusion around a species complex as well as a Stigmina anamorph have hindered the development of a proper species concept (Barr 1996).Other homotypicsynonymslistedinMycoBankarenotlistedhere.
Notes—ThisspeciesfoundonAgave shawii(Cooke1885)hasplurilocularstromatawithseptateascospores(Barr1996).It may represent a distinct species of Kellermania or the sexual state of K. macrospora.OtherhomotypicsynonymslistedinMycoBankarenotlistedhere.
Notes—Thisspeciesbasedonmaterial fromAgave sp.inMexico(Maublanc1903)maybethesexualstateofaKel-lermania, possibly K. macrospora. It hasbeen treatedasasynonym of Planistromella parryi(Barr1996).
Notes—Intheprotologue,thisspecieswasreportedtohaveconidia70–85×3–5µm,cylindrical,aseptate,appendageslacking (Morgan-Jonesetal.1972b).The type fromMexicooccurs on Yucca filifera, but other collections from widespread North American localities occur on other Yuccaspp.(Morgan-Jonesetal.1972b).Basedonthefigureoftheconidiaintheprotologue(Morgan-Jonesetal.1972b), the length towidthratio is12–15.ThoughtreatedasasynonymofPiptarthron limbatumbySutton(1980),wholatertreatedbothoftheseassynonyms of Piptarthron yuccae(Sutton1983),thisseemstobe a distinct species on Yucca.
Notes—Intheprotologue,thisspecieswasreportedtohaveconidia55–75×11–14µm,2-septatewiththemiddlecellhalfthesizeoftheendcellswith1–2appendages,15–18×3–4µm(Dearness1924).AccordingtoDearness(1924),thetypefromtheUSA:CAoccursonHesperoyucca whipplei(asYucca whipplei;subfam.Agavoideae).Thereareafewotherreportsof this species on Yuccaspp.fromthewesternUSA(Farr&Rossman2012).ThoughtreatedasasynonymofKellermania anomalabyMorgan-Jonesetal.(1972b),Sutton(1980),andNagRaj(1993),thisisalmostcertainlyadistinctspeciesgiventhe importance of host associations since it is the only species described originally from the host genus Hesperoyucca.
Notes—In the protologue, this plurilocular specieswasreportedtohaveconidia50–68×6–7.5µm,cylindricaltoob-clavate,aseptate,3–4-septate,appendageslacking(Morgan-Jonesetal. 1972b).The type from theUSA:TXoccursonYucca macrocarpa and an additional collection was reported from theUSA:AZonYucca brevifolia (Morgan-Jonesetal.1972b).Basedonthefigureoftheconidiaintheprotologue(Morgan-Jonesetal.1972b),thelengthtowidthratiois8.2–12.ThoughtreatedasasynonymofPiptarthron macrosporum by Sutton(1980),thisseemstobeadistinctspeciesonYucca.
Notes—In the protologue, this specieswas reported tobe an immature ascomycete and a question mark was placed nexttothegenustoindicateuncertaintyaboutitsclassification(Ellis&Everhart1895).According to theirobservations, theimmatureasci, if interpretedasconidia,aregivenas50–60×7–8µm,cylindrical,taperedtowardsapices,aseptate.Thetype from Mexico occurs on Yucca glauca (asY. angustifolia)(Ellis&Everhart1895).BasedoncorrespondencewithM.E.Barr,Sutton(1983)determinedthatthisspeciesrepresenteda Piptarthron and that P. limbatum and K. attenuata were latersynonyms.Ifthetypematerialisacoelomycetebearingaseptate conidia that lack appendages, this would likely be a distinctspeciescorrectlyclassifiedinKellermania.However,M.E.Barr’sannotationofthetypespecimenfrom1970(NY)states that the specimen is an immature K. anomala.
Notes—Kellerman(1906)describedthisspeciesfromGua-temalan material found on Agave americana.Thecombinationof host, plurilocular stromata, and septate ascospores suggests a possible link with Kellermania macrospora.Ithasbeentreatedas a synonym of Planistromella parryi and Plowrightia agaves (Barr1996).
Notes—Intheprotologue,thisspecieswasdesignatedasthe type of the genus Septoplaca, and it was reported to have indistinctly pseudoseptate conidiameasuring 35–60(–78)× 3–3.5 (Petrak 1964).The type from theUSA:AZoccurson Yucca macrocarpa (Petrak1964).Sutton (1977) treatedSeptoplaca as a synonym of Piptarthron, but he was unable to placethetypespecies.Assumingtheconidiamayactuallybeaseptate and after considering conidial measurements, Sutton (1977)suggestedthatS. limbata was an earlier synonym for Kellermania attenuata.Later,Sutton(1980)formallytransferredS. limbata to Piptarthron, treated K. attenuata as a synonym, and considered the species to be aseptate. Sutton (1983)treated these names as later synonyms of P. yuccae.AlthoughPetrak(1964)mayhaveconfusedpseudoseptateconidiawiththe euseptate conidia in Piptarthron, it seems unlikely that he wouldconfuseaseptatewithseptateconidia.
*Not known from culture **Basedonstudyofisotypeonhost(BPI1110167)
DISCUSSION
Phylogenetic analyses support the placement and monophyly of the core Planistromellaceae in the order Botryosphaeriales of class Dothideomycetes where the Planistromellaceae rep-resent a well-supported clade comprised of the coelomycetous genera Alpakesa, Kellermania, and Piptarthron with associated teleomorphic genera Planistroma and Planistromella.WithinBotryosphaeriales, it is positioned in a polytomy with four other lineages of Botryosphaeriaceaes.l.,threeofwhichoccupybasalpositionsinpreviousstudies(Philips&Alves2009,Schochetal.2009)relativetothefourththatincludesthecorediversityof Botryosphaeriaceae(Fig.1).ThiscoreBotryosphaeriaceae lineage(pp=1.0)iscomprisedoftwomainclades,onewithBotryosphaeria(pp=1.0)andanother(pp=0.75)withPseudo-fusicoccum stromaticum and Guignardia(pp=1.0).Theposi-tion of Guignardiaissignificantduetoitsunclearpositionbasedonpreviousstudies(Crousetal.2006,Philips&Alves2009).ThoughthefamilyBotryosphaeriaceae has been recognised withabroadcircumscription, i.e.sensulato(Philips&Alves2009),andPlanistromellaceae could well be treated as a later synonym, we prefer to maintain the family Planistromellaceae as distinct from Botryosphaeriaceae s.str. and suggest thatfurthersamplingisneededtoclarifyfamilialclassificationwithinthe Botryosphaeriales.ThePlanistromellaceaeissignificantlyand phylogenetically distinct from the Botryosphaeriaceae.All genera of core Planistromellaceae are here considered to constitute one genus, namely Kellermania.Fourothergeneraclassified inPlanistromellaceae (Barr 1996) are redisposedas follows: Eruptio has been shown to be a member of Myco-sphaerellaceae (Verkleyetal.2004);Loratospora has been shown to be a member of the Phaeosphaeriaceae (Schochetal.2009);andnoneofthespeciesofMycosphaerellopsis including the type species, M. myricariae(Höhnel1918b),havebeen placed phylogenetically; Microcyclus,typifiedbyMicro-cyclus angolensis(Sydow&Sydow1904),isonlyrepresentedby DNA sequence data of Microcyclus ulei, and based on BLASTsearchesofGenBankusingavailableITSsequences,it does not belong in the order Botryosphaeriales.TheclosesthitsresultingfromtheBLASTsearchesaremembersoftheMycosphaerellaceae,butitsfamilialclassificationisuncertain.
27A.M.Minnisetal.:Planistromellaceae - genus Kellermania
Among the Planistromellaceae(Kellermania)arefivelineagesthat,withtheexceptionoflineageiv,areeachwellsupported.Lineagevistreatedseparatelyasitspositionisunresolved.These lineageswere recovered inall analysesof individualgeneswiththeexceptionoftheSSU.ThelackofresolutionintheSSUtreewasduetoalowlevelofsequencedivergence,and thusa lackof informativecharacters (Table2).Overall,the relationshipsamong the five lineageswerenot stronglysupported.In general, the phylogenetic groupings within the Planistromel-laceae (Kellermania) do not correspondwith the presenceor absence and numbers of conidial appendages nor with conidialseptation(e.g.multipleappendagesinlineagesiiandv;appendagesabsentinlineagesiandiv).Previousgenericcircumscriptions of these coelomycetes based on conidial ap-pendagesalonearenotwellsupported,afindingwhichwasalso recently observed among other genera of appendaged coelomycetes (Barber et al. 2011,Crous et al. 2012).Thetype species of Kellermania, K. yuccigena, and Piptarthron, P. macrosporum(≡K. macrospora),belongtoseparatelineagesiiandi,respectively,thatnotablyformaclade(pp=0.89)withK. unilocularisintheITSanalysis(Fig.3).Lineageii,whichin-cludes K. yuccigena, also includes three well-known species of Planistromella for which DNA sequence data exist including the type species, Planistromella yuccifoliorum(=K. yuccifoliorum).Thoughlineagesi–iiiinFig.2containonlyspecieswithseptateascospores, Planistroma obtusilunatum, linked to P. uniloculare (≡K. unilocularis), asmentioned above appears to belongamong these lineages (Fig. 3). It has aseptate ascospores(Ramaley1992).Otherspecieslackingascosporeseptation,namely Planistroma yuccigenum, the type of the genus and linked to P. pluriloculare(≡K. plurilocularis)andPlanistroma nolinae, linked to P. crassisporum(≡K. crassispora),belongtolineage iv and Planistroma kellermaniae, linked to Kellermania nolinae,representslineagev.Thus,Planistroma and Planis-tromella do not form well-supported clades if circumscribed byascosporeseptationalone.Consideringthebiologicalandphylogeneticsimilaritiesofthespeciesinthefivemajorline-ages and the lack of resolution among the lineages, there is little pragmatic value in recognising any subclades or lineages asdistinctgenera.Thus,theentirecladeishereinrecognisedas a broadly definedgenusKellermania, the generic name having priority. In accordancewith the changes enacted intheMelbourneCodeinregardtoArt.59(Norvell2011),wedonotrecogniseseparateanamorphicandteleomorphicnames.Morphological variation of conidia among isolates from different hosts or even the same host, especially in regard to append-ages, septation, and shape of conidia, is frequently an indicator ofthepresenceofdistincttaxa.However,slightdifferencesinsize, septation, andappendagesof conidia amongmaterialfrom fresh collections, herbarium collections, and culture were frequentlyobservedforagivenspecies(Ramaley1993,1995,thisstudy).Inspiteofthesedifferences,theoverallappearanceof conidia on the host and in culture is similar for a given species and species differences tend to be obvious when considering within species andamong speciesmorphological variation.DNA sequence data are required to distinguish species and to confirmspeciesrecognitioninambiguouscases.From an ecological and evolutionary point of view, it is interest-ing to note that several lineages including lineages i, sisters ii– iii,andiv(Fig.2)arelargelycorrelatedwithplanthostsfromdifferent host clades in the Asparagaceae, subfamilies Agavoid-eae and NolinoideaesensuAPGIII(2009).Thissuggeststhathost phylogeny is correlated with the phylogeny of these fungi tosomedegree.However,samplingwithinthesefungiremainslimitedanditistooearlytoformulatefinalconclusionsgiventhelackofresolutionamongthelineages.Althoughanumber
of Kellermania species have been recognised historically as oc-curring on numerous hosts, we expect that there are few, if any, plurivorousspeciesgiventheevidenceofhostspecificityandnumbersofcryptictaxafoundinthisstudy.However,multipleisolates were not available for species of Kellermania reported from numerous hosts, thus, this hypothesis needs further study as many members of the Botryosphaeriales are known to inhabit multiplehosts(vanNiekerketal.2004,Glienkeetal.2011).
AcknowledgementsThe authors extend heartfelt gratitude toAnnetteW.Ramaleyfordetailedobservationsoftheseunusualfungiaswellashercontribution of the cultures used in this study and her notes without which this studywouldnothavebeenpossible.TheauthorsthankJohnH.Wiersemafor providing the nomenclaturally correct names for some species of Yucca thatwere identified by others. JosephF.Bischoff kindly provided fungalmaterialsandothersupportfromtheinitiationofthisstudytoitslaterstages.LisaA.CastleburyiswarmlyacknowledgedforprovidinganITSsequenceof Kellermania unilocularis.PedroW.Crousgenerouslyassistedwiththepublicationofthismanuscriptandadvisedonmattersoffamilialtaxonomy.
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