Hikobial3：645-665.2002 Molecularphylo窪enyOfhypnobrJ/aleanmOssesasin化rredfroma lar淫e-scaledatasetofchlOroplastlｂｃＬ，withspecialre他rencetothe HypnaceaeandpOssiblyrelatedfnmilies1 HIRoMITsuBoTA，ＴｏＭｏＴｓｕＧｕＡＲＩＫＡｗＡ，ＨＩＲｏＹｕＫＩＡＫＩＹＡＭＡ，ＥＦＲＡＩＮＤＥＬｕＮＡ，DoLoREs GoNzALEz,MASANoBuHIGucHIANDHIRoNoRIDEGucHI TsuBoTA,Ｈ､,ARIKAwA,Ｔ,AKIYAMA,Ｈ,,ＤＥＬｕＮＡ,Ｅ,GoNzALEz,，.,HIGucHI,Ｍ ４ ＆DEGucHI，Ｈ、2002.Ｍolecularphylogenyofhypnobryaleanmossesasinferred fiPomalarge-scaledatasetofchloroplastr6cL,ｗｉｔｈspecialreferencetotheHypnaceae andpossiblyrelatedfamiliesl3:645-665． ▲ Phylogeneticrelationshipswithinthehypnobryaleanmosses(ｉｅ,theHypnales,Leuco- dontales,andHookeriales）havebeenthefbcusofmuchattentioninrecentyears Herewepresentphylogeneticinfierencesonthislargeclade，andespeciallyonthe Hypnaceaeandpossiblyrelatedftlmilies，basedonmaximumlikelihoodanalysisof l81r6cLsequences、Oursmdycorroboratesthat（１）theHypnales(ｓｓｔｒ.[＝sensu Vittl984])andLeucodontalesareeachnotmonophyleticentities、TheHypnalesand LeucodontalestogethercompriseawellsupportedsistercladetotheHookeriales;(2) theSematophyllaceae(s」at［＝sensuTsubotaetaL2000,2001a,ｂ])andPlagiothecia‐ ceae(s・str.［＝sensupresentDareeachresolvedasmonophyleticgroups,whileno particularcladeaccommodatesallmembersoftheHypnaceaeandCryphaeaceae;ａｎｄ (3)theHypnaceaeaswellasitstypegenusﾉﾘDlwz"川tselfwerepolyphyletioThese resultsdonotconcurwiththesystemsofVitt(1984)andBuckandVitt(1986),who suggestedthatthegroupswithasinglecostawouldhavedivergedfiFomthehypnalean ancestoratanearlyevolutionarystage，fbllowedbythegroupswithadoublecosta (seealsoTsubotaetall999;Bucketal2000）OurresultsfiPomlikelihoodanalyses ofalarger6cLdatasetconfinnthesuggestionbysmallerdatasetsfiommultipleloci firstbyDeLunaetal.(2000)andthenBucketaL(2000)thatthereisnoevidencefbr supportingtheHypnales(ｓｓｔｒ.[＝sensuVittl984])andLeucodontales・Adetailed reconsiderationofordinalclassihcationandfnmilialclassificationwithintheHyp- nidaeiｓｎｏｗnecessary． KeyWords:HypnalesLeucodontales,Hookeriales,hypnobryaleanmosses,molecular phylogeny,r6cLlargedataset,Hypnaceae,Sematophyllaceae,Plagiotheciaceae,Am- blystegiaceae 。 肋Ｄ'"ｊ７Ｍ６ｏ/ｑｄＭｒﾉﾉ℃"ＭＤｅｇ"ｃｈｊ,Depα""ze"ｔｑ/Ｂｊｏ/ogjcα/Ｓｂｊｅ"Ｃｅ,Ｇｒａｄ"α/e Sb/ioo/ｑ／血e"Ｃｅ,Ⅲ℃sﾉｶ〃α〔ﾉﾉｊｊｖｅｌＭ１ﾉ,hgamli〕ﾉα碗αノー3-/,〃jgZJSﾙﾉｰﾊjrosﾉi〃α- Ｗ,ﾉﾌﾞIﾉ･Csﾉi〃α739-852．ﾉﾋＷ/L 7bmojiFz`9ＭﾉWmwq,Depαﾉﾌﾞ'"e"'０/Ｂｊｏ/Ogjcα/Sbje"Ｃ＆Ｆ,Ｇ７α`"α/e此/ＩＯＣ/q/ｍｉｅ"Ｃｅ， U>ﾌﾙe'Ｍﾜﾉq/mtyo,〃O'7907-3-/,βz"7/bﾉo-k"’７bﾉb′o川-0033,上pα"・ 航rqMEMﾉhi〕′α',、,/Ｍｌｓａｌ''１ｑ／ﾉVb【/"'℃α'z`ﾉ仙加α'Ｍα〃jrjes,町ogo，YZlO′ojgaoAa 6-cho"ze,５位"dtJ-sAj,Hyogo669-/ｺﾞｲｄﾉﾋZpα〃 〃αｍＤｅＬ""α＆ＤＣ/oresOo'1Ｚ｡/ez,ＤｅＰａｒｔａｍｅ"ｊＭｅＳＭ伽/ｊｃａﾘﾉｾge/αＭＭｍｔｏ ｃ/ｅＥｃｏ/ｏｇｍｌＡＣ.,ＸＭＪｐａ，比７．９/000,ＭｄＸｊｃｏ、 MJSα"０６〃ｆﾉﾉgZlcAj，Depα"ｍｅ"／ｑ/ＤＣ/αノリＭＪ/jolZα/比ｊｅ"ｃｅＭ`Ｓｃ"〃，伽αAr"６０ ｲｰ/-ＩｍＭ"6α-sﾙﾉ,仏aMhH305-000LﾉﾋZpα",ＤＥＰα"ﾉ?Te"／ｑ/Ｂｉo/ogjcα/Ｓ℃/e"ＣＧＳ， G/ndZJα/ｅ比ﾉioo/ｑ/此ｊｅ"Ce,Ｕ>7〃e′Ｍﾜﾉｑ/、oﾉb′０，肋"ｇｏ７~3-/,Ｂ２`"ﾉtyo-ﾉ1Ｗ,Tbﾉbﾉ。 〃3-0033,上pα〃 'ContributionfiOmtheLaboratolyofPlantTaxonomyandEcology,HiroshimaUniversity・ＮＳｅｒ・No.S２Ｌ 646 HikobiaVoll3,No.４，２００２ Introduction talesandHookeriales）fbrcorroborationofthe phylogenetichypothesisofDeLunaetal.(2000） Thehypnobryaleanmosses，includingtheor- ａｎｄＢｕｃｋｅｔａＬ(2000);３)toclarifythefiamilial dersHypnales，LeucodontalesandHookeriales， relationships，especiallybetweentheHypnaceae areoneofthemostdiversifiedmossgroups、 andrelatedfamilies，suchastheSematophylla- Attentionhasbeenpaidtotheclassiflcationof ceaeandPlagiotheciaceae，withinthehypnobly- thehypnobryaleanmossessinceVitt（1984)pro‐ aleanmosses，giventhattheLeucodontalesand videdanewsystemfbrtheBryopsida、Within Hypnalesarenotmonophyletic;ａｎｄ４)totracein thepleurocarpshesuggestedthat“adoublecosta thephylogenythetransitionsofthecostafiDm ll groupasrepresentedbytheHypnaceae,Selnato- singletodouble，oneoftheimportantcharacters phyllaceaeandEntodontaceae，isdivergentfiDm fbrthefiamilialclassificationofthehypnobry- ０ agroupwithasinglecosta，ｗｈｉｃｈｃａｎｂｅｒｅ－ aleanmosses、 ｇａｒｄｅｄａｓｔｂｅｍｏｓｔｒecentevolutionarystageof Recent，molecularphylogeneticanalysesfbr theorderHypnales(seealsoTsubotaetall999； pleurocarpousmosseswerebasedonr6cLse- Bucketal2000）Subsequently,BuckandVitt quencesalongwithotherdatasets,ｓｕｃｈａｓａｍＢ， ￣ (1986)constluctedanotherclassificationfbrhyp- 伽Ｌ－Ｆａｎｄノアs４Ａｔｔｈｅｐｒｅｓｅｎｔｔｉｍｅ，tbere- noblyaleanmossesbasedonmorphologicaland sourcesofr6cLsequencesarethemostcompre- ecologicaldata、 hensivesamplingfbrthehypnaleanmosses・ Inrecentyears，phylogeneticrelationships Alignmentofthe'6cLsequencesisalsoeasybe- withinthehypnobryaleanmosseshavebeenthe causeoftheirdistinctionasaprotein-codingre‐ fbcusofmucbattentionatseveraltaxonomiclev- gioninthecodonpositions・Sqwepresenthere els(Arikawa＆Higuchil999,2002;BucketaL pbylogeneticin化rencesfbrthethreeordersHyp- 2000；ＤｅＬｕｎａ＆coworkersl999，2000；Maeda nales，LeucodontalesandHookeriales，basedon etaL200qTsubota&coworkersl999,2000, themaximum-likelihood（ＭＬ）analysesofl81 2001a,ｂ;VanderpoortenetaL2002)．Tberela- sequencesofthechloroplastribulosel,5-bisphos- tionshipswithinthehypnobryaleanmosseshave phatecarboxylase/oxygenaselargesubunit beenevaluatedbybothmorphologyandmolecu- (r6cL),togetherwithrecentinfbnnationonthe larstudies(Hedeniisl995;ＤｅＬｕｎａｅｔａＬ１９９９)． molecularphylogenyofhypnobryaleanmosses， Hedeniis（1995,1996)showedthemonophylyof anddiscusstheimplicationsofourfindingson theHookeriales-Sematophyllaceaeanddiscov- theHypnaceaeandtheirrelatedfamilies． eredtheLeucodontalesasagradetaxonbasedon morphologica1data、DeLunaetaL(1999)clari- Materialsandmethods fledthehomologyofpleurocarpyandthemono- phylyofallhypnobryaleanpleurocarpousmosses Phylogeneticanalysesbasedonther6cLgene (theLeucodontales,HypnalesandHookeriales)， sequenceswerebasicallyperfbnnedfbllowing basedonacombined'6cLsequencesandmor- Tsubotaandcoworkers（1999,2000,2001a,ｂ)． phologydataset・Furtherdetailswererecently Themethodconsistsoftwosteps：Dobtaining の revealedbyDeLunaetaL(2000)andBucketal． sequencedata(DNAextractionPCRamplifica- (2000)whoproposednewphylogeneticrelation- tionandDNAsequencing,orDNAdatabaseho- shipswithinthesepleurocarpsTheseworks mologysearch);and２)dataanalyses． suggestedthattheLeucodontalesandHypnalesas previouslyunderstoodarenotmonophyletic ８℃"7calα"c/Ｐ/α"ｒＭＪＺｅｒｊａん Familiesfroｍｔｈｅｓｅｔｗｏｏｒｄｅｒｓａｒｅｍｉｘｅｄｉｎａ Atotalofl81mossspecieswereexaminedfbr ｓｉｎｇlelargecladesistertothehookerialeanlin- theanalysesasshowninTablelandinAppendix eage、 A・Thelattercontainsvoucherinfbmationand Theobjectivesofthisstudyare:ｌ)toconstruct DNADatabaseAccessionNos・Voucherspeci- aphylogenetictreefbrhypnobryaleanmosses menssequencedinthepresentsmdyweredepos- basedonlarge-scaledataset;２)totestthemono- itedi、TNS,ＨＹｏ,orHIRoSomesequencesused phylyofthethreeorders（Hypnales,Leucodon- fbrtheanalyseswerealsoobtainedfi･omthe DNAdatabase． HIsuBoTA,Ｔ・ARIKAwA,Ｈ・AKlYAMA,ＥＤＥＬｕＮＡ,Ｄ､OoNzALEz,MHIGucHIANDH､DEGucHI ６４７ TableLListofspeciesandftlmiliesinvestigatedlbrl6cLgenewiththeaccessionnumberandvoucheror referenceofthesequences、TreatmentoffamiliesinthistablefbllowsVitt（1984)ｗｉｔｈsomemodificationsby BuckandVitt(1986)．FurtherinfbnnationfbrthesequencesobtainedinthepresentsmdyisshowninAppendix Ａ、 OrdelﾌﾟFamiIv/SDccies AccessionNo． Refcrcnccs/Origin HypnaleS(s・stⅨ） Hypnodendraceac A0lp"ocノセ"cjWmel2zjesjj(Hook.)Paris ＡＦ２３ＩＯ９３ MishIcrctal(onIyinDNA database） PIeuroziopsidaceae 雄"mzjqpsな'wﾉﾉze"たα(Weinm.)Kindb・cxBritt、 ＡＢＯ24683 ArikawaamdHiguchi(19”） Fontinalaceac Fo"""α/jFampW℃"ｃｑＨｅｄｗ ABO50949 msubotaetaL(2001a） ｡ Ｆｂ"'ｊ"αﾉ応α"伽ﾉｱ'eZjcqHedw、 AJ275183 Coxctal(2000） Ｆｂ"/、αﾉﾉＳ“んcar/ｊｃａＢｒｕｃｈ＆Schimp・ AF231074 DeLunaetaL(2000） Echinodiaceae Ebﾉｸmodmll2〃"61℃s"'?z(Mitt.)Ａ・JaCgcr AF233568 DcLunaetal(2000） Sbmrommm〃jcosjcJm”(SulL)Mitt、 ABO24677 Arikawa＆Higuchi(1999） Thuidiaceae j6jerj"eﾉﾉﾋJa6jerj"α(Hedw.)Ｍ・Fleisch ＡＦＯＯ５５１９ Goffinctctal(1998） ７１ﾉｶ､とﾉﾉ"ｍｃﾉｾﾉｊｃａｍｍｍ(Ｈｅｄｗ)Schimp、 ＡＦＩ５８１７７ DcLunactaL(2000） ７１ｌｂ､ﾋﾉmmplな'ocaOMMUⅡ､HaL)AJacgcr ＡＢＯ７１４Ｉ６ TSubotactal(200ｌｂ） nh"､１１"〃だcqg"""伽(Hedw.)Lindb． ＡＢＯ１９476 Akiyamactal.(2000;only inDNAdatabascXmsubota etal(2001a） Lcskcaceae Bom〃αｍｊ"e"ﾉﾉ(Broth.)Ｃａｒｄｏｔ ＡＢＯ２４９６３ TBubotactal(1999） Ｄｍ/tieﾉﾉﾋJSpecjosjFsjmqBrothexCardot ＡＢＯｌ９４６７ Akiyamaetal.(2000;only inDNAdatabasc);thiSstudy M〕′αbeαﾉﾘw"ＣｅﾉﾉﾋJ(Mitt.)Broth ＡＢＯｌ９４７５ Maedaetal(2000） OAcJm皿池ｅａｈａＡｏ"je"sjs(Mitt.)Broth ＡＢＯＩ９４７７ Macdactal(2000） Amblystegiaceac CtJ"je堰o"eﾉﾉﾋJc"sp此ｍね(Hedw.)Loeske ABO24678 Arikawa＆Higuchi(1999） C'、o"e"'℃"〃ｃｊ'､"(Hedw)Spruce ABO95270 thisstudy D'℃pα"ocmdi`sadi`"c"s(Hcdw.)Warnst． ABO24681 Arikawa＆Higuchi(1999） ｆbｲ91℃α''161ｌＷｅｇｊ"〃'e'､x(Hedw.)Jcnn AF23356５ DeLunactal.(２０００） Lembophyllaceae Doﾉﾉcﾉｶo､"rjqpsjM1ve応施'ｍな(Mitt.)Ｎｏｇ ABOI9465 Akiyamactal(2000;only inDNAdatabasc);thisstudy Ｌｅ"16qpﾉＭ/""'cﾉﾙ"/Sm"?(Hookf＆Wilson)Lindb， AF233570 DeLunactal.(2000） StcreophyIlaceac EmodO"'qpsハノＢＨＣＯＳ／２gα(Brid.)ＷＲＢｕｃｋ＆lreland ABO24635 Arikawa＆Higuchi(1999） S1e花qpﾉ!)ﾉﾉﾉ"加沢adIczjﾉCs"、(Hook)MitL ABO24637 Arikawa＆Higuchi(1999） Fabroniaceae A"αcα'Modb"spmch"o雄s(FroeLexBrid.)Ｂｒｉｄ AF231077 DeLunaetal.(2000） 〃ﾋﾞﾉﾉcodO"""maJpjﾉﾉﾋJ”(Hcdw)AJaeger AF23357］ DeLunaetal.(2000） Brachythcciaccac Bmc〃んecj"ｍｐ肋､Cs"、(Hedw.)Schimp、 AF233566 DeLunactaL(2000） Ｂｍｃﾉi〕ﾉﾉﾙecj"mrjw`ﾉα”Ｓｃｈｉｍｐ ABO24674 A｢ikawa＆Higuchi(1999） Ｂｍｃ/Ｍｈｅｃｍｍｓα/e6ms"、(HofYinexRWCber＆ AFl58I76 DeLunaetal(2000） DMohr)Schimp・ ノMb'Ｈｍｃﾉａｄｔｗｃｍｍｏｗｊｃｚｊｊ(Borszcz.)Stcere＆Ｓｃｈｏｆ ABO29389 Tsubotactal.（1999） Ｐﾉﾛﾘﾉ!〕や"jdm加吻α"o/“s(Hedw.)Dix， ABO29385 Tmbotactal(1999） PseWdOsc化mpodmmpⅢ'ｗｍ(Hcdw.)Ｍ・F1cisch． AF233567 DCLunaCtal(2000） Ｒ/i)′"c/zos/egm"'pαﾉﾉﾉ｡!/bﾉﾉz`"1(Mitt.)AJacgcr ABO24944 Tsubotactal(1999） ７bme"卯""〃〃/花"s(Hedw.)Loeske ABO24676 ArikawaandHiguchi(1999） Entodontaccae E7Troc！O〃cﾉｶαﾉﾉelZgelj(Paris)Cardot ABO50993 TBubotaetal.(2001a） E)?/odb"/”ｊｍ４ｓ(Gri征)AJacgcr ABO50994 T1subotactaL(2001a） 勘/odO""!)'”“(Hook)Hampe ABO24640 Arikawa＆Higuchi(1999） EmodO"'w6jc""c/bｲs(Mitt.)AJacger＆Sauerb・ ABO29386