动物学研究圳2,on.23(5):36l一366
Zoolo西曲IRe辩arch
cN53—1040/QISSN0254—5853
PhylogeneticRelationshipsof1lBumblebeeSpecies
(Hymenoptera:Apjdae)BasedonMjtochondrial
Cytochrome6GeneSequences
SHA0Zhi—yon91~,MAOHong—xinl,FUWen-junl,ZHANGYa—pin92,3
k6。ml。ry0,o删“山mn,掰m咖-i,,幽业矿,‰m肌”面切&&d昭y,乩。嘴^“hc“咖加目“瞎mfsc扣n懈
£kn‘榭P^唧d咖y0,&”n咄,卧。增h£200025;2Ⅲ咖Ⅲ。,^“叼“h跏fu如n口“&no删埘删ny,
KunⅡ西嘈hlA山。,zon研,m吼啪船陆唧矿&"n咄,n删M650223,∞啪)
Abst舶d:Phylogenetic—ationshipsofllburnblebee8pecies,including58ubgeⅡem:舶m6淞(59Pecles),丁k—
mr060m6m(3species).肘end吖f60m6m(18Pecies),n州协6Dr抽咄(18pecies)arldf枷6。m6m(18Pecles),were
analyzedb踮edonthe357bPmitochonddaIcytochrome6genes。q呦ees.Thereare65sindelo“polym07phlcsiksand
7lpar㈣“yi村jmlativ。PolyⅡmrphicsitesint}l诂DNAsegment,叩d45polymorphicsiteswiIhLnthetotal119tmnslated
am¨10acidss咄mentBotllNJtreeandMPt⋯howthat肌mk晶om6w(B鲫i,瑚m珊淞)isba8altootllers,fouowed
bynnid060H小珊(日pe珊一眦H拓w);P,"如m6瑚(口.En删诂m)andBom6掷aresistersub萨nera;thesubgenusof
&lm6{“ismonophyletic,inwhich占.培nmudiv。rgednrst.
Kqwords:8nr拈吣;C”0chmme6gene;DNAseq“ence;Amino卵1d88quence;Mokcula‘phyloge”y
细胞色素西基因序列与11种熊蜂的系统进化
邵志勇∽,茅红新1,符文俊1,张亚平2
巾国科学院上海生命科学院植物生理牛志研究所脚同进化实验室.上海:00025
2.中国科学脘昆明动物研究所,云南昆明650223)
摘要:通过比较一段357bp的细胞色素6的序列,
了熊蜂属5亚属11种熊蜂系统发育关系。5亚属包
括胁柚哪(5种)、7硒r删60m瑚(3种)、胁mk西Dm6哪(1种)、n兀Jfd060m瑚(1种)和胁6叩诂m(1
种)。该序列有65个单变异多态位点和71个简约信息多态位点,翻译成119个氨基酸序列后有45个氨基酸变
异位点:根据P一距离构建的邻接树(NJtree)和最大简约树(MPtr比)都显示同样的结果:胁,“妇i6叩小咐
(B.n"i袱,醇口‰)分化最早;胁dD如m琳(日.雕邶灿H面吣)次之;P呻。柚Ⅲ(口.i”删曲邶)和舶m淞形
成姊妹群;凰m6w亚属是单系群.其中曰咖血m在所研究的5个种中分化最早。
关键词:熊蜂属;细胞色素6基因;DNA序列;氨基酸序列;分子系统发育
中图分类号:Q96954;Q951.3;Q75文献标识码:A 文章编号:0254—5853(2002)05一0361一06
Thereare239bumblebeespeciesreco印izedin
lheworld(williams,1998),but山ephylogeneticre—
laIionshi∞amo“gthemarestillpoodyunderstoodbe-
causeoftheir“monotonous”moIphology(Michener,
1990),especiallywithinsubgenus,suchas肋m6w
(Williarns,1991).S0someresearchershavetumed
tom01ecularmarkers(St。phen&Cheldelin,1973;
Pekka打nen“nf.,1979;Pamiloef nf., 1987;
Pedersen,1996;Koulianos,1999;Koulianos&
Schmid—Hempel,2000).However,inBom6瑚5,
HeceIvedd小:2002一03一11;Ace唰eddⅢ8:2002—05一13
F⋯IⅢon沁m:n⋯n吐⋯仲o^dbyckPm乎狮fofK掣胁哪lad∞d5占Tc00p哪fi∞Pmjechdch啪(2001cB7
uondNmur出Sc㈣F0undmion0fCh曲a
3 CoⅡ唧ondl“g叫thor,nl:86—871—5190761,E—m越l:zh蚰g冲@publ】ckmyncn
万方数据
362 动物学研究
s打themolecularresuIts(Pamilo““..1987:Ped.
ersen,1996)didnotconcurwithmorphologyresults
(PlowrighI&st。phen,1973;w“ams,1994).The
phylogenPIicrelationshjpsofthosebee8peciesaredif_
femnIfbmauthorto author(Sakagami,1976;
Williams,1994;Pedersen,1996;Koulianos&
schm¨Hempel,2000).
MilochondrialDNA(mtDNA)is8 goodgenetic
markerbecauseofitshi曲mutationrateandhaploid
withoutrecombination(Bmwn“。f.,1979;Avise以
Ⅱ2.,1987;Modtz“口1.,1987).Thehoneybee’s
complPIemtDNAgenomewassequenced(Cmzier&
Crozier,1993),whichmakesthegeneticanalysisof
beesmucheasier.
Thereare10speciesinsubgenusofBom6瑚,and
moslofthemdismbuteinAsia(Wmi锄s,1998).
Herewesequenced。ytochmme6 genetosmdytlle
phylogeneticrelationshjpsamo“g11speciesincludi“g
subgenus舶m6淞andfourofhersubgenerawilh却如
口坤Ⅱ扣m8s山eoutgm“p.
1 Materi蚰sandMethods
1.1 Mate—alsandPCRsequencing
BumblebeeswerecollectedhDmBe玎i“g(踟m6瑚
i卵n出and口.^,pocmⅡ),Jilin(口.potn咖£珊)and
zhejiang(B.咖打础)inChina,KoreatB.函nit船)
andJapan(B.咖缸W)duing2000∞d2001.The
sarnpleswerekepti“purealcoholatmomtempera—
tu砖.T0talDNAfmm16B.i印血w,4丑.^”)Dc市口
and2口.阳£n譬缸£蛐wasextractedfrDmtheheads,
山orax(Gamerye£d1.,1991)orl。gs(Koulianos,
1999)usingthestandardphenol/chlomfo砌methods.
ParLialcytochr0珊e6 genearnpli矗edw油 the
p^merCBlandCB2(simon以口Z.,1994)c00rdi—
natesll400一11425andll859—11884asde.
sc^bedinCmzier&Cmzier(1993).Thereaction
mixtureconsistedoflO一20ngtotalDNA,2扯Lmixed
dNTP(4mmol/pI.),l pLeachkindofprimers(10
pm01/卜L),3肛LMgcl2(25mm01/fJL),5肛Lb嘣h
(10×,500mmol/LKCl,100mm01/LT^s-HCl,
pH9.O,T^tonX一100),2肛LBSA(bovinesenlm
albumin)and2UThqDNApolymem8e,thenaddi“g
ddH20tototal50肚L.Aneraninitialdenaturationat
23卷
96℃for3min,35cvcleswerepe南丌nedwi【hd明a—
Iuringa{96℃for1min,anneali“gat54℃forl min
andex慷nsionat72℃forI mi【l,fotkwedbvanend
extensionfor5minat72℃.ThePCRpmduclswere
8equencedwithBigDyel”renTlinatoroycles。quenci“g
kits(Perkin—ElmerBiosystems)usi“gABl377automat-
icsequencer(PE).Eachindividualwass。quencedin
botlldirections.Thesequenceswefecheckedbyeyes
卸deditedwilhDNAstar(DNASTARInc.1996)pack—
age.Cytochrome6 genes8quencesCenBankacces-
sionnumbersofB,^,P0c也,B.培耐fwandB.pn£-
昭缸汹areAF498397,AF498398andAF498399.
1.2 Sequ蛐c船f哪nGenB锄k
Thesequencesjn8bumblebeespeciesof5 sub—
genemand却厶,n础咖“1werecollectedfromGenBank
(Table1).
TaMel Sequen倪血tafromGenb蛆kinbumbkb畔
蛐dhoneybeesped器
subgenu9 sP州∞ No.of8。quence
助,n6l* Bf㈣
日㈣“÷
Z^0r咖bm6ⅫB fu5c“dw
B.7‘Ⅱ,训”
8 r【Ⅲk一Ⅲ
胁nd砒160m6“日“⋯如口w
jh幽抽Ⅳ*m 口Ⅲ灿∽Ⅻ
毋mb础蛐 日‘呷of如w
A咐m幽≈目。
AFD0”22
^F【K坨72l
AFol7516
AF017517
AF002723
AFl81610
AFl8161l
AFt81169
lD6178
1.3 Phyl0驴neticⅫalysis
Th8polymorPhicsitesandsyno“ymous/non一8y“·
on聊oussubsⅡtuteswerecalculatedbv sof}ware
DNASP3.O(RDz船&Rozas,1999).Weuselhe
珊inoacidsequencetoconstnJctth8phylogenictrees.
NJtreew踮constnlctedwilhMEGAversion2.1(Ku-
mar“nf.,2001)b船edonP.dislance(Nei&Ku-
m盯, 2(×)o),whilemaximumparsimo“ytreew汕
PAuP’4,08b(swofford,2001)bvheuristicsearoh
withTBRbranchsw印pjr培.Ap曲胱盯澹mwasusedas
iheoutgro“p.ForbothNJaIldMPtree,weused
2(X)0hootstr印replica“onsto esIimatethes“pport
raIe.Wereconst兀lctedNJ【reeofsub只enus日om6嬲
basedonDNAsequencelofur山eranalyzeIh。phyIog。一
“y, becausewithinsubgenusDNAsequencecanre-
necttllerelationshipbetlerthan踟inoacidsequence.
万方数据
5娴 目日志勇等:细胞色素5摹田序列与11种熊蜂的系统进化(英文
2 ResultsandDiscussion
ThP陀are65singleto“polymorphicsitesand7I
parsimo“ymformativ8polymo。phicsite8inthisDNA
s8卯lPnl:45polymorphicsiteswilhinthetotal119
translateda¨1inoacidss。gment;butnog且portenninal
codewerefound(F酿.1a,b).Themulatio“panem
0fthesegnlents“ggeststhatitis pmbablyfrommito-
(‘hondnalgenome,notfromnuclea。psuedogene.Be—
a
cause£hemeantransiIjon/“ansve巧ionjs0.6347and
therateofaminoacidI)olymorph记si忙 is high
(37.82%),weusedtheaI【ljnoacidsequencetocon—
stnlcIfhephylogenel证trees.BolhNJandMPtrees
showedthesameDattem:胁,谢6w曲om6邶(口.n口i,m-
口抽Ⅲw)isb如alloothers,f01lowedbv一0埘i如60m6淞
(日.∥邶咖dn把w);F,060m6瑚(曰.i”叩Ⅱ#搪瑚)and
口Dm6邶aresistersubgenera(F培.2a,b).Thesub—
genusof曰om占u5ismonophyletic:曰.1Dcr帆,口.p叫一
111111111111111111111111111111111111111
11 1 1222333344445555555778899011 1 12233334444455555666667777778888999
3592589124013656890134789251423802477925682457813479356890124780369258
ATTTTTT从TMTA^cATTTc^TG^^MT^TTAT^TT^GCATT从从GATMT^ATATTGTCAT从从T^
TA G C T TT CC^CTT CTCT T^^TTT T
^A T^TT T ^T CT T T ^ T^TTTTT
AC^ T^Tr T ^T CTTT T G r^T^TTr
TA^CCTT T T C T AT T T ^ TAT TT
^ T T . ^T CTTTT T AGTAT T T
ACA T T C ^T T ^ T^ TTCT
^^ TAT CTT. ^T CT TT T ^ T^TCT T
AACT T TT C AT T A T A CT^TT
T ACT C T G T ^T T T A TAT TT
T ACT T T T ^T T TCA TAT TT
C CTTATTGTAcATT^TTT.CTT^从TATTT^CTTGCCT^TTATTT
12222222222222222222222222222222222222222222233333
900∞011”1223333444445666666677777778888889900011
9123470136明901470367991245678们246892345685746902
^TCAT从T^TTT从CTCATT从T从T^cMc^TT^“T^TTT^TTT^M^
TA C T TT T T TTT MTATT
^ ^ TTT. . ^ TCM G^TT
^ TTT. T T TCMACTTT
A ..T T C T .T T T MT T^
C T^T TT T .T AT T A ^CTTT
C ^ C TcTC T T T T TC从 T^
A TTT T T TCAAT^CTTT
A TT T A C TC C^
AC TCTCCT T T T ^ATA T
^C TCTCCT T T T AAT T^
CGG^TT^TCAT TCTT^GGTTTA^CTCAT^CCT丁T
3333333333333333
111122223344S555
5789015736281457
TAT^TTAT^TTGATCA
C T^^T^C
G G鼬 ^T
A G^ ^ TT
T GAT A T
A AG^ A TT
T ^GATA T
^ GGG^ TT
A^TCATT
TCAA A T
TCAA A T
^“G^CCT
¨"”¨”¨””"仲¨垤
¨””””讣”¨”m¨堙
万方数据
动物学研究 23卷
1 111 111”1222222222233333333334444444444555555555566668666667
f2“567890123456789012345678901234567890123456789012345678901234567890
舢ITNLlSAIPYlGOFTVEWlWGGFSI洲DTLNRFYSFHFILPFIlL删V冈HLlIL⋯TGSSNP⋯SK
V I ^ T LL M L
L L M M
L L I M M
L I M
LL I W L
. L M
L. 。 L M
L ■I M
. L I M
. .. .... . L l M
T L .DTlL ... A. .FL LL.FILF^L LGN
1 11111111111111”111
7777777778888888888999999999900000口00D∞1l1f11l 1f
1234567890123456789012345678901234567890123456789
LNJYKI肝HPyFTJKDLJTJ儿TFJL朋FJ NLQ咿YlLSDPDNFK舢肿
S Ll F’L¨
■ _ SU F V.G S
M F S■ I F .G . S
H F -l L. L.F■. . .
S .哪L¨.I.F-G S
_ F.S■L L¨HG.. ..
H F Sl I F G S
M H F.S . ..■. .
- . F ■I.L . LF- , .
一 F MI..L..LF■ .
FN S S LGFYlILFl. F.F H.G . I.
Fig.1se鲫emmutaIion出t啪0fc”0chrome6DNAanda|Ilinoacids。quen钾in12bumbl幽ee
andho“eyb∞8pecies
a.357一sEF“凹Ic一·妇me6DNA咖㈣e;b¨9·辨鲫lr一衄hmme6a商肿“dsequ朗ce
1)Bi呻叫wu.2)口.删i砌蒯lm,3)丑P帆∞叫,4)B.^um以b,5)日如cⅢm,6J丑舢y厶Ⅷn埘u,7)
B枷f嗑,8)且.n‘d咖n咐,9)占枷妇Ⅱ,lO)B^m盯妇,11)口芦哪池咐,12)APbm础毋m
ng“tdbovc小。即枷qu洲莳l髑j8I】le5jkp∞j6蝴dmese印1朋ls。qlJ洲;d时罄j口出聃把山⋯d∞bd抖jd册lJ(一m
mm0fB☆HⅫ觚
Ⅱg讯螂and日.^印0c一£Ⅱarec10selyreIatedtoeach
other,and8.喀n矗瑚div。rged矗rst锄ongtllosenve
species(Fig.2a,b).MPtreeien殍his83,Consis—
tencvIndexis O.819.Retentionlndexis0.712and
RescaIedCoflsistencvIndexis0.583.SimiIarresuIts
forthesubgenusof口o,如淞wereobtainedusi“gDNA
s。quencedirectly,exceptthat曰.p武ngiⅡ£wand
口.^卿r缸dbecomethemostderivedsister8pecies.
ThephyJogenyofbumbIebeeshasnotbeenre—
solvedyetbecauseof“monotonous”morphology
(VnIIi锄s,199I).Nowbvtheana【vsisofthemito—
chonddalcytochmme6 s8quence8,w。providednew
b
¨”"””们"¨”∞¨他
”外”¨”D”∞”仲¨惶
万方数据
5期 邵志勇等:细胞色素6摹田序列与1J种熊蜂的系统进化f英文)
b
005
却bmcll≈erd
BlmpⅡ“eHs
Bh即ocrtl0
8pdlogln}us
Bl(-”es饥t
B曙"Im‘
Bpnsc"£t~s
Bh㈣lB
BⅢ拈mrlus
BpennsvlvdH“m
B㈣v*llus
占。卅6Hj
^,~60mb“‘
norⅡco^oⅢ她5
I,㈣如^Ⅲ缸,
、Mend㈣bombm
BD,"6Ⅳ5
7许洲obo卅^“s
F㈣∞¨mh“s
^{endnclbombHs
Fig.2Phylogenetictreesof11bumblebee8pecieshsedonmit∞hondnalcvt。chrome
6genes。quences
a NJl⋯tmsh咖dw汕MEcAV2l basedonP—djs伽㈣;bTopdoEyhmMP丑d!slsusl“g
PAUP‘V408b
nenumbeHmlhnod咄b00lst阳ppi091ndlcesof2000。eP№at糌《tIlelevd0f5uPp0^forl一Ⅲdu出
no(Ies,姐ddm㈣dis珊BBedwhenfh川ppo^rneisle辨50%n㈣pondl“g、uh98neraare
sh㈣mmen出.
jnfonnationonphyfogeneticreIationshipofbumb【e-
bePs
ThetopoIogyofsubgenerafromourmoleculardata
is sim订arf0 thatofWjlli锄s’morph0109icstudy
(Williams,1994),butdiffersfmmotherrecenl
mulPcuIarstudies.Pedersen(1996)showedthat
曰om6ujwasbasaltoJp们60rn6瑚andseveralother
subgenenlontheb8sisofCOIsequence.Evenbased
on【hes帅eCOIse口uences.theresultsofKoulianos
&Schmid.HemDel(2()00)difkredfromthatofPeder_
s蛐(1996).Thefonnerresuhs;ndjcatedtha{B嗍6瑚
andP,r060m6础werepar8phyIelicbetweentwo
clades,butthe1alteronesshowedthat疗om6Mswas
basaltoPm60,H6榔inamonoDhvleticclade.TherP.
sultbasedoncvt(mhrome6 sequenceswassimjlarto
thatofPedersen(1996)forIhetwosubgenera(Kou.
1ianos&Schmid—Hempel.2000).However,Ⅱ¨the
studiessharedsomesimilarpaftemsthatthesubgenus
日om6∽ismonophvletic,
Becauseof【hehighrateofpolymorphicsitesa—
mon只ouranalvsisdatd,webelievethatmutationof
珊lDNAamong5ubgener口jsalJ】】0slsalur丑Ied,solhe
万方数据
动物学研究
franslatedaminoacidsequenceisbetterthanDNAse—
quenreforth8phylogene“canalysisofbumhlebeesub一
酽“PraInlhefuture,“isnecessa‘yIosludymore
8pecies岫i“gothermo陀conservativ8genes.
Acknowledgemenb:WethankMyenong-lye01
Refbrences
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r“I’J1,:l¨n】110chondr锄bndgebct啪pop山mio“舻nrtlcsand
815⋯na【1es⋯^⋯腑nⅣf斯“.墙:489—522.
B⋯月wM.‘浦删eJR.Wjls蛐AC 】979R印】de州“,⋯fanj-
mdmlluchondn出DNA【JJ Pw№f A曲d scf【嘲,
76 1967一197I
(:州MH¨,CrozicrYC l帅3 T¨mIIochondrI出肿nome0fhonev—
l,忡^P“,耻“扣m:co“pleIe8蜘uence蛐dgenomeo‘g阴Izmm
『JJ&耻帆f,133:97一117
G岬9口【.Vau㈨lD,C⋯etJM,“一 199lPhyJ呷n“*畔l叶
m)nhhl|.sInthe胛nus^puinkⅡedhDmmJI小-hondidDNAse一
‘l“P⋯P·jala[J】.月pi撕M,22:87—92
K叫l⋯}S1999Phyl雌ene|⋯el砒lonshIpsofIh}Ⅲmbleh⋯ub一
矿“u‘,枷60,柚w(Hy⋯q’1阿a:ApldaP)inkⅡedfmmmitochon·
tln—c-yIm—hmIlM6 andcytochmmeox埘aseI 8eq㈣es。J]
^,m£hn叫S∞^m,92355—358.
Knuj⋯*s,卧h⋯l—H㈣p8lP.2000.Ph一09enetLcrdmlonshlP4一
⋯#l,urnlllHb㈣(舶柚Ⅵkf州k)l山蒯抽m如toch叩dimc卜
I∞h10mP^a11dcylwh⋯ldasPI8。q⋯”[J]^“.P^Ⅳ
£幻f.14:335—34I
K⋯rS,I'⋯raK.JakobsenIB.“d2001.MEGA2:M·】l⋯
larevolu【lonag群“e【lcsandy5lssonw。ue[J]舶Er扣r,删础.17
(1:):1244—1245
M¨-h㈣rr(:D1990cl鹊sincmlon0fI¨APld胛(Hy唧叩Iera)[J]
【mfⅣm吖‰mm5⋯&if“m.“:75一164
M¨rIIz(:.DowllnRTE,BmwnWM.1987.EvolⅢi哪of帅im出删to一
‘hondrld:Rd⋯ce如7p‘十”lmi¨nbiology肌d8yBtem“ic8l J].
^⋯I m”‰f.S"f.187:269—292
^PIM,Kurn盯S2000Mol⋯larEvDIu㈨㈨dPhylo舻ndlcs【MJ
NPwYork:OxnmlUm⋯lvPress
r⋯I¨P.P“k虮⋯A.V洲SL_1987ClL同erinzdl,umblebee
,ub妒nPrab鹊edonlnl盯5Pe。i6。矿““c商mlonshlpB(Hym蛐叩tcr-
a:、pldae:舶舶ma|IdPp雠mⅡ)【JJ ^,m“.z捌.凡肌I_
rf.24:19—27
P州⋯nBV 1996A phylo站neti㈨dvgis0fcuek00bumblebe髓
(Rrl^y兀u,姆妇w)锄dbumblebecs(舶,曲琳,胁孵龇)ln一
23卷
Lee(Koma),YUw(mg一】un,YANCMao—fa,and
YANGDa—m“gforsamplec01lePtion;PANGJIln—
fe“g,YAOYo“g—ga“g,LL0Huai—m“g,QuKai—
xin,PANGDe”g,WLCui—she“g,l川ANYun—xia.
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万方数据
细胞色素b基因序列与11种熊蜂的系统进化
作者: 邵志勇, 茅红新, 符文俊, 张亚平
作者单位: 邵志勇(中国科学院上海生命科学院植物生理生态研究所协同进化实验室,上海,200025;中国
科学院昆明动物研究所,云南,昆明,650223), 茅红新,符文俊(中国科学院上海生命科学院
植物生理生态研究所协同进化实验室,上海,200025), 张亚平(中国科学院昆明动物研究所
,云南,昆明,650223)
刊名: 动物学研究
英文刊名: ZOOLOGICAL RESEARCH
年,卷(期): 2002,23(5)
被引用次数: 10次
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版) 2009(3)
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4.吴冬霞.郝家胜.朱国萍.陈娜.苏成勇.潘鸿春.张小平 基于线粒体Cyt b基因的线蛱蝶亚科的系统发育[期刊论文]
-动物学研究 2007(1)
5.齐兴柱.尹绍武.娄甜甜.廖经球.陈国华.张本 海南产花鳗鲡细胞色素b基因的克隆及序列分析[期刊论文]-海南大
学学报(自然科学版) 2007(4)
6.代金霞.郑哲民 基于Cytb基因序列探讨蝽亚科11种昆虫的系统发育关系[期刊论文]-昆虫知识 2005(4)
7.陈艺燕.钱开诚.任岗.陈迪.章群 大黄鱼与小黄鱼细胞色素b基因全序列的比较分析[期刊论文]-生态科学
2005(2)
8.徐其放 实验动物Beagle犬微卫星研究及线粒体DNA全序列测定分析[学位论文]硕士 2005
9.芦荣胜 中国斑腿蝗科部分种类分子系统学研究[学位论文]博士 2005
10.代金霞.郑哲民 蝽科部分昆虫细胞色素b基因序列及其系统发育关系的探讨[期刊论文]-动物学研究 2004(5)
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