Table Of ContentElyt・,a,「olive,27(2):643-649,November13,1999
OriginandEvolutionoftheApotomopterusGroundBeetles
(Coleoptera,Carabidae)asDeducedfromMitochondrial
ND5GeneSequences
Cheong-GenKIM,Zhi-HuiSU,YukiIMURAl)andSyozoOsAwA
JTBiohistoryResearchHall,1-1Murasaki-Cho,Takatsuki,Osaka,569-1125Japan,and
1)DepartmentofGynecology,TokyuGeneralHospital,Kita-senzoku1-45-6,
Ota-ku,Tokyo,145-0062Japan
Abstract Phylogenetic trees have been constructed from the mitochondrial
NADHdehydrogenasesubunit5(ND5)genesequencesof13species(27specimens)of
Ca,abus(Apoto,,1opte1-11s)withC.(Llnlnoca,・abus)clat11,atusandC.(L),naackiasout-
group.TheND5 treesindicatethat thereare threemainlineages. 0neofthespecies,C.
(A)sauteri,ismorphologicallydividedintoseveralsubspeciesbyslightdiffeirences.Seven
specimensoffoursatlte,-1subspeciesfromvariouslocalitiesinmainlandChinaandTaiwan
havebeenclassifiedbytheND5genesequencesintothreecladeswiththeirancientsepara-
tionca 22MYA.Ca,abus(A)1/upeensls,whichshowsconsiderablemorphologicaldiffer-
encefromC.(A)saute1・i,branchedofffromoneofthesa1lteristems.
Introduction
ThesubgenusApotomopterusofthegenusCa,・abus(s lat)comprisesnearly100
speciesdistributedinChinaandtheadjacentregionsincludingTaiwan. Intheprevious
paper(IMURAeta1., l998),aphylogenetictreeoftheNADHdehydrogenasesubunit5
(ND5)genesequencesfrom9taxaofthesubgenusApotomopteruswasreported. In
thepresentstudy,revisedphylogenetictreeshavebeenconstructedbyadding10taxa,
usingtwo umnocarabussequencesasoutgroup.Basedonthetrees,evolutionaryhis-
toryanddiversificationpatternwithinthesubgenusApotomopterusarediscussed.
Materials and Methods
Thespeciesanalysed for theND5genesequences including thepreviously re-
portedones(IMURAeta1.,1998)arelistedinTable t.Fortheanalyticalmethodsand
constructionofthephylogenetictrees,seeSUetal. (1996,1998)andKIM,ZHoUetal.
(1999).Usingtheneighbor-joining(NJ)method(SAITou&NEI,1987)andunweighted
pair-groupmethodwitharithmeticmean(UPGMA)(KUMAReta1.,1993),thephyloge-
netictreeswereconstructedusingthesequencescontainingale69bp3'-regionofthe
ND5geneof19taxaof thesubgenusApotomopterus. Bootstrapanalysiswasper-
formedforthetreesbasedon500resamplings(FELsENsTEIN, l985).Theevolutionary
644 Cheong-GenKIMofaf
.一一一し ,s)‘s一一S加 ' 加erycs,:.“ 2.一一ee-S・ s・〇fly,t- 1
-一一s)一一 'M 加M e,・s3
jCC一「一一as))),M.,==一a“ 'M.'Esp yy60s“,--,M・一一 ・45s“ 7 Lineage1(●ｻ
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CC s))a“ m, p・-e‘al7a-s16 L.meage2
C ) ,o ngm ao9en918
C .)to - ,2o
C ) =.-:=・ sp2l -
C s) 在ao 22
CC .)a「 。 ・ m or一一.oha「,-・24一一S23 L.mea 3 ■
C ):!r 25
C )c s“ m 26
C ) P 27
C .mno ,、・一al n‘s
C )ma 一一商.s
0.06
Distance
MYA
distances(D)werecomputedbyKIMURAs' two-parametermethod(KIMURA, l980).For
settingthetimescale,a0.01Dunitcorrespondingto3.6millionyears(MYR)was
used(SUeta1.,1999).
ResultsandDiscussion
OriginandPhylogeny.
ApotomopterusisthesolesubgenusinthedivisionSpinulatiofthegrandgenus
Carabus,anditsmembersconstituteawell-definedmonophyleticclusterontheND5
phylogenetictrees(IMURAeta1., l998).Thepresentphylogenetictreesindicatethat
therearethreemainlineagesoftheApotomopterusspeciesthathavebeenexamined
(Fig. l).ThefirstlineageincludesC.(A)kouich1i fromNorthVietnam,C.(A)sauteri'
andC.(A)hupeensisfromtheChineseContinentandTaiwan.Carabus(A) kouichii
andC.(A)sauteriseparatedlongtimeago[ca32millionyearsago(MYA)].Follow-
ingthis,threeC.(4.)sauterisublineagesdivergedataboutthesametimeca22MYA.
OneofthemcontainingC. (4.)s dayaoshanlcusandC.(,4.)s.ssp also includes five
specimensofC.(A)hupeensls,indicatingthatC.(4.)hupeensisfromvariouslocali-
tiesbranched off fromoneofthesauteristems. Thesecond lineagecontainsC. (4.)
tonkinenstsfromNorth VietnamandC. (A) toulgoeti,C. (‘4.) delavay1,andC. (A)
tuxeni from Central and South China. Carabus (‘4.) toulgoet1 and C. (A) ton-
kinensls/C.(A)tux:enl/C.(A)delavayiseparatedca25MYA, followedbydiversifi-
EvolutionofApotomopte''fisGroundBeetles 645
Lineage1ｫ●)
]
u・nea9e2 ▲)
l
unea9e3 ●)
D:0.01
cationofthelatterthreespeciesca. l4MYA.ThethirdlineageconsistsofC.(4.)cler-
montianusfromNorthVietnam,andC.(A)cyanlpennlsssp.,C.(A) iris,C.(A)sp.
(alliedtoC breunlngianus; IMURAeta1.,1999),C.(A)art・owl,C. (A) laoshantcus
1iaorumandC.(A)1. laoshamcusfromCentralandSouthChina.Carabus(A)cler-
montianusandC. (A)sp.separated fromacommonancestorca26MYA,andthere-
mainingspecies diverged ca. 18 MYA into two sublineages, i.e., C. (A) cyan1-
ds)/ . . pemzs/C.( ) faosﾍamcMsffaorum C (4. aos ﾝamcu an C )arrowl/C.( .)
iris.Furtherdiversificationintothreeandtwodescendantspeciestookplacewithinthe
respectivesublineageslessthan5MYA.
TheoriginofApotomopterusisoldandcanbetracedbacktothetimeoftheex-
plosiveradiationof theCarabina50-40MYA(seeSuet al., 1999).Thus, Apoto-
mopterus may beconsideredasone of thegroupsoriginatedupon the radiation.
Shortlyaftertheradiation,threelineagesshowninFig.1anddiscussedabovediverged
almostsimultaneouslypresumablysomewhereinthesoutheasternregionoftheChi-
Fig.1(onpp.61'1 645). PhylogenetictreesofthemitochondrialND5gene(1,069bpupstreamfrom3'
terminalstopcodon)fromtheApoto'nopterusspecies.TheUPGMAtree(A)andtheNJtree(B).The
bootstrapconfidencelevel(%)(basedon500resampIings)isshownateachbranchingpoint.Distance
(D)denotesKIMURA'stwo-parameterevolutionarydistance(KIMURA,1980).Thesequencesofthe
specimens8-10,andthoseof19and20wereidentical,respectively.
646 Cheong-GenKIMelal
Fjg 2 MapshowjngthecollectingsitesoftheApoto,1lopte'u'sspeciesusedinthisstudy.Nume「alS
correspondIothoseshowninTabletandFig.1.●,lineage1;▲,lineage2;■,linea9e3.
noseContinent(seefig.32ofIMuRAeta1., l998).Figure2andtheND5treesindicate
thatthedjstributionrangesofthethreelineagesarewideandoverlapconSide「ably.No
definitedistributionboundariesbetween themcanbedrawn, Thissuggeststhat the
Apotomopterus ancestor was divided into at least three isolates upon gee9raPhiC
changesofthecontinentca37MYA.Theneachisolateexpandeditsdistributionto
occupythepresenthabitat.
Patter morp ofogz'ca evoftefzon
Foursauterisubspecies,dayaoshanicus,yunkaicus,sauteriandssp.(population
fromMt.JinggangShan)differonlyslightlyfromoneanother,eventhoughtheyhave
beenclassifiedassubspecieswithinthesamespecies(IMuRA, l994,1995&1999).
However,theirseparationoccurredlongtimeago,ca22MYA(Fig.1).Carabus(A)
clermontjanusandC.(A)sp arealsosimilar inmorphology,buttheyshowaremote
phylogeneticrelationship,separatedca26MYA.Thus,evenifmorethantwotaxaa「e
treatedasthesameoralliedspeciesbecauseofsimilarmorphology,their timeofdi-
EvolutionofApotomopte,・usGroundBeetles 647
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648 Cheong-GenKIMetal
vergenceisnotnecessarilyrecent.Sucha“silentmorphologicalevolution”canberec-
ognizedinothercarabidssuchasDamasterblaptoides,Phricocarabusglabratus,To(-
mocarabusspp.,etc.(SUetal.,1999).
Asnotedabove,C.(4.)hupeensissplit fromoneoftheC.(A)sauteristemsin-
cludingC.(A)sdayaoshanlcusandC.(A)s.ssp.ThissuggeststhatC.、(A)hupeen-
sisemergedwithaconsiderablemorphologicalchange fromC. (A)sauteri whose
morphologyhasremainedalmostunchanged foralong time.Thespeciationofthis
typeresemblesthebranchingofC.(Euleptocarabus)porrectico11is fromtheC. (Lim-
nocarabus)clathratusstem(IMURAeta1.,1998;KIM,SUeta1.,1999;SUeta1.,1999).
Thedifferenceinmorphology betweenC.、(A)cyanlpennisandC. (A) laoshanlcus,
andbetweenC.(A)arrowiandC.(A) irisareconsiderable,andyettheirdiversifica-
tionoccurredrecently.Thus,evolutionofApotomopterushasbeenbroughtaboutby
havinga silent phaseandaphaseaccompaniedby a considerablemorphological
change,ashasbeenwidelyrecognizedintheothercarabidgroups(Suetal.,1999)
Acknowledgements
Wearegrateful tothe followingcolleagueswhokindlysuppliedwithvaluable
specimens for thepresentstudy:Prof ]) CAwAzzUTl (Pagno, Italy), Dr. S.MURzIN
(Moskow),Dr.ShuheiNoMURA(NationalScienceMuseum,Tokyo),Mr.A.SHAMAEv
(Moskow),Mr.J.TuRNA(Czech)andDr.H.-Z.ZHou(AcademiaSinica,Beijing).This
study issupportedinpartbytheGrant-in-aidNo.09041167forOverseasResearchof
theInternationalScientificResearchProgramfromtheMinistryofEducation,Science,
Sports andCulture, Japan, to the fieldwork in Vietnamby Dr.Shuhei NoMuRA.
ThanksarealsoduetoHidekoTANAKA forskillful technicalassistance.
要 約
金 衝;坤・蘇 智慧・井村有希・大澤省三: ミトコンドリアND5造伝子によるトゲオサムシ
IIE属(Apoto″1opterus)の系統解析. - Apoto,;,opterusは, 雄の交尾器内袋基部に基棘と呼ばれる
硬化片を持つトゲオサムシ群の唯一の構成貝で, 中国南東部を中心に分布し, 100種近くを含
む大亜属である. 本研究では, IMuRAら(l998)の報告に加え, 中国とその周辺地域からえられ
たトゲオサムシ亜属19分類単位 (27個体) のミトコンドリアND5遺伝子の塩基配列から系統
関係を検討した結果, 3系統の存在が認められた. これらの3系続は, オサムシの一斉放散
(約5~4千万年前) 直後にほぼ同時に分岐し, さらに各系統の中で2~3回にわたって分岐が起
きたと推定された. Ca,abus(11りotomopte,・us)saute,iは, 中国南東部と台湾にぃくっかの亜種を
産するが, 形態的にはたがいによく似てぃるため, その分化は比較的最近に起つたものと想像
されがちである. しかし, mtDNAの結果ではsaute,・iは3つの系統に分かれ, それぞれ2200万年
前の古い時代に分岐したことが示唆された. また, c (d)sp. (ブロイニングトゲオサムシに近
縁な未記載種) とC.(A)clermontianusも基本形態はたがいによく似ているが, mtDNAから両者
は非常に古い時代に分化していることが示された. 一方, C.(A)sauteriの一つの枝からC.(A)
EvolutionofApotomopte,-ttsGroundBeetles 649
hupeensisが分岐しているが, この事実は, 長いあいだ形態変化がほとんどなかったsauteriから,
形態的に異なる種が出現したことを示唆している. さらに, C. (A)cyampennisとC.(A)
1aoshanicus,およびC.(A)a,,-owiとC.(A),risとのあいだでは, 顕著な形態的差異にもかかわら
ず, 分子系統樹からみるとそれぞれの種の分岐した時期は比較的新しいことが判明した. 以上
の結果から, トゲオサムシ亜属の進化には, 形態が長いあいだほとんど変わらなかった時期と,
短いあいだに変化した時期のあることが示唆された.
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