Genetic distances among Oeneis norna, O. polixenes, O. bore and O. melissa (Satyridae) based on mitochondrial DNA (ND1) partial sequences

2010·0.38 MB·

TThhee LLeepipdiopdteorpoltoegircaollSoocgieitycal Society ooff JJaapapnan
uctts,kTrans.Iepid.Soc.Jlipan60 (4):293-297,March 2010

Genetjcdistances Oeneis O.polixenesO,.bore O.
among norna, and metissa
(Satyri dbaasee)d on mitochondrial DNA (NDI) partial sequences
Takao IToH*, Kazuhiko HiRANo and Kazuo WATANABE
GraduateSchool IntegrateAdrts ScienceH,iroshimaUniyersity,
of and
Higashi-Hiroshima City,739-8521 Japan

Abstract Partia ]sequences (4S2 bp) of mitochendrial DNA (mtDNA) IVDJ of Oeneis norna and
Oeneis ntetissa have been determined and compared with reperted NDI sequences of Oeneis polix-
enes, O. bore (O. taygete) and O. uhteri. I tis inferr efdrom the comparison of parti amltDNA se-
quences that O. hore, O. polixenes, O. norna and O. melissa are distinc tspecies to one another and
their ancestors were separated at almost the same age, It is mentioned also that at presen tthere is
no clear evidence showing that O. non!a is inhabjting North America including Alaska, whieh cen-
tradicts the description sgiven in a number of illustrat erdeference books on Japanese butterflies,
Key words Oeneis noma, Oeneis polixenes, Oeneis bore, Oeneis metissa, Mitochondrial DNA
(NDI) genes, geneti dcistance.
Introduction
The species of genus Oeneis Ellr efound in the north of the New and Old Worlds. All the
species are dull colored, medium in size and principa] lfyound in Arctic or alpjne habitat ass
well as in taiga fores t.In Japan two Oeneis species, O. noma and O. melissa, distribute,
both of which are regarded as high-altitu dspeecies inhabitin glimite dareas above the tim-
ber]in e.The fbrrne rspecies inhabit sthe Hjda meuntain chain (th eNorthern Japan Alps)
and Mt, Yatsugatak ein Honshu Island ,while the latte irnhabit sthe Daisets umountains and
the Hidaka mountain chain in Hoklcaid oIsland .In most of the illustrat reedference books
published after 1975 in Japan, it is describe dthat O. norna ranges both Eurasia and North
America (Kawazoe & Wakabayashi, 1976; Fukuda et at,, 1984; Matsuka, 1994; Inomata &
Matsumoto, 1995,Shir6zu,2006),Itis in foreignillustrated book
also written a reference
lhat O, noma ranges North America (Higgi n&s Riley ,1970) .However, at presen tthere is
no evidence showing that O, norna is disuibute dNorth America includin gAlaska, although
there are morphological similarities among noma, bere (tayge taned) potixenes. Although
O. taygete and O, bone were treated recently as the same species (Ople r& Wright, 1999;
Lukhtanov & Eitschberger, 2001), the relation between O. norna and O. polixene sis un-
known, It is ,therefore, of importance to investigat ethe relationship among the Oeneis
species. The molecular phylogenic analyses Df the species of genus Oeneis have been car-
ried out based on mtDNA (ND5) (Usam iet al., 2005), but only litt lienfbrmatio nis avail-
able fbr those based on mtDNA (NDI) ,We have investigat etdhe relationship among
noma, polixenes b,ore (tayge taned) melissa through mtDNA NDI (45 2bp) gene analyses.
Materials
and methods
Specimen of O. norna asamana was obtained in the vicinity of Mt Yakushidake ,Japan at al-
titude near 2,700 m under the permission of Ministr yof the Environment, Japan (cf
*Corresponding author, e-mail: [email protected] .jp
NII-Electronic Library Service
TThhee LLeepipdiopdteorpoltoegircaollSoocgieitycal Society oofJfap anJapan
294 Takao IToH, Kazuhiko HiRANo and Kazuo wrTANABE
Oeneis noma p$
Oeneispolixenes
'M
Oeneisbore -$

Fig. 1. 0utline of the distribut iToannges of Oeneis norna. O. potixenes and O, bore. The ranges at
Europe were ebtained from Higgin sand Riley (1970 )t,hose in North America were ob-
tained from Opler and Wright (1999 )a,nd those in Russia were obtained from Lukhtanov
andEitschberger(2001).
Acknowledgements) in 2001, Specimen of O. melissa l"cill ahas been obtained in the
vicinity of the Loveland pass at altitude near 3,800 m in Rocky Mountains, Colorado ,while
one of the authors (T ,Itoh )was staying in Colorado as a visiting Professo rof Universit yof
Colorado in 2002. These specimens were dropped into ethanol immediately after capturing.
MtDNA NDi sequences of O, polixenes, O. bore (O, ldygete) and e. uhleri as well as
Neohipparehia statiiinus used as the reference were obtained from DDBJ data base (Martin
2000),
et al.,
Total genomic DNA was extracted from male specimens, using proteas edigestio nfo11owed
by two phenol-chlorofor emxtractions and ethanol precipitation .Mitochondrial gene region
of A44DH dehydrogenase subunit 1 (NDI) was applied by PCR using the primers ,5'-CG-
TAAAGTCC[[IAGGT[IATATTCAGAI"TCG-3' fOr forward and 5 -ATCAAAAGGAGCTC-
NII-Electronic Library Service
TThhee LLeepipdiopdteorpoltoegircaollSoocgieitycal Society ooff JJaapapnan
Geneticdistances Oeneis 295
among species
Table 1.The mt-DNA (NDI) sequences (452 bp) of Oeneis norna, O. potixenes, O. bore and O, metissa.
1 fiO
O. 6GGTTATTACAGCCTTTTTCTGATGCTATTAAATTATTTACTAAAGAACA
nomaO.polixenes
･･T-･---･･･--･--･-,--････-.･..･..･........,.......
･･T････････････････-･･････････････.･.･･..･........
O.boreO.

･･T･-･･･････････････････････-･････.････..･.･......
melissa
51 100
O. AATTTATTTAAATTATTCAAATTATATTTTTTATTATTTTTCTCCTGTAT
norna
･G････CC･G･･･････････-････････････････････.････.･.
O. polixenes
･G････････････････.･･･････,･,･･..･.･.....,........
0.boreO.

･G･･-･C-･-･･････････････････････････････-･････････
metissa

10t I50
O. norna TAAGATTTATGTTATCTTTAATAATTTGGATAGTGATTCCATATTATTTT
･･････････A･･･････････････････････････-･･････,･.･･
O.polixenes
･-････････A･･･････････････････････,･,･･.,,...,.,..
O.boreO.
･-･-･････-A･-････-･･･････････････C･･･C････････････
melissa

151 2oo
O. AATTTAATTGTTTTTAATTTAGGGGTATTATTTTTTTTATGTT6TATAAG
noma
････････････････････････A･･････････････,･････.･.･･
O.potixenes
･･･････････C･･････A･･･････････････,,･.･..,.....,.,
O.boreO.

tnetissa
201 250
O.norna TTTAGGGGTTTATACTGTTATAATTGCTGGTTGGTCTTCTAATACTAATT
･････････････････････--･･････････A･･･.･.･･..･.･...
O. potixenes ････････････････････････････････-A･････,･････,･.･･
O,boreO.

..........,...,........･....･.･･-A･-･･-C･-･･-･････
metisffa
2Sl 300
O. norna ATTCTTTATTAGGGGGATTGCGGGCAGTGGCACAGACTATTTCTTATGAA
･･････････････････････････････････A････.･････.･.･･
O.polixenes
････････････････G･････････････････A･･･････････････
O.boreO.

･･････････････････････････････････A･････-･･A･･････
rrtelissa
301 350
O. noma GTTAGTTTAATTTTATTAATATTATCTAGTATTATTTTAATTTTGGATTT
O. pol ixenes
.......-G･･･''''''''''''''''''''''''''''''''''''''
O.boreO,
melissa
3Sl 400
O. noma T･A･A･T･T･T･A･ATT･･A･A･A･A･T･T･T･A･T･AAG･A･T･T･A･T･C･AAG･G･･A･A･T･T･A･A･T･T･T･G･A･T･T･T･A･T･A.T･T･TA
O.potixenes
･･･････T････････････････････A･･････････････････,･･
O. bo reO.
･･･--T---･･-････--･･ ･-････--･-v.･..,
metissa
401 45e
O. norna TAATATTACCTTTAAGATTTTGTTTTATGTCTTCTTTAATGGCTGAAACT
･-･････････････････A･････････.････..･.･.....･.....
O.poiixenes
O.boreO. ･--･-･･ T･-････････A･･････････････････････････････

･･･････････････････A････････････････････A･････････
melissa
45tAA
O. noma
O,potixenes
''
O.bore

''
O.melissa

Gidfir[[lkGTTTC- f3o'r reverse (Auber tet ai. 1996). Thermal cycle parameter for NDI were
2 min at 940C (1 cycle); 30 s at 94MC, 30 s at 55"C, 1 min at 72"C (30 cycles); 5'min at 720C
(1 cycle); 10 min at 4aC (1 ¢ycle) .The PCR product was purifie bdy passing through a gel
and fragment sequencing was canied out using a sequence analyzer (ABI Prism 3 1OO).
NII-Electronic Library Service
TThhee LLeepipdiopdteorpoltoegircaollSoocgieitycal Society ooff JJaapapnan
296 Takao ITeH, Kazuhiko HiRANo and Kazuo WATANABE
Table 2, Relative pairwise distance between the Oeneis taxa analyzed using
Kimura's twe-parameter rnethod, along with the standard deviation
inthe
shown parentheses.
polixenes bore melissa uhteri
O.030(O.O07) O,029(O.O08) (o.oos)o.o7s(o.e13)
nornapolixenes
O.027 (O,O08)oo..oo223 g(o,oo s)e.o63 (o,o12)
boremelissa O,027(O,O08)O.065(O,O13)

O.065(O,O13)

Results discussion
and
rlleLb]e
In 1, we show the sequences of the partia lmtDNA NDI gene of O, norna and O.
melissa, along with the reported sequences of O. polixenes and O. bone (tayget eAm)o.ng
mtDNA NDI sequences of the five Oeneis species each consisting of 452 bp, norna dilfers
from polixenes by 13 bp, diiier fsrom bore by 13 bp, and differ fsrom melissa by 13 bp,
while polixene sdiffer fsrom bone by 12 bp. The habita tof polixene sand bore overlaps in
part in North America, and these are treated as distin cstpecies (Howe, 1975). In Europe the
habita tof norna and bore also overlaps in part and there bore and noma are treated as dis-
tinct species to each other (Higgin &s Ri]ey ,1970) .Outline sof the distributi orannges of
norna, polixene sand bore are displaye din Fig. 1.
Table 2 shows relative geneti cdistances between the Oeneis taxa analyzed using Kimura's
two-parameter method. It is seen in Tbble 2 that there are no appreciable differenc eisn phy-
logenic distanc eamong norna, polixenes, bore and melissa, suggesting that ancestors ef
these four species have separated at almost the same age. Noteworthy is that the distance
between norna and melissa, and that between norna and polixenes are almost the same, al-
though norna belongs to the group differe nftrom that of metlisa or bore (Lukhtan o&v
Eitschberge r2,001). On the other hand, there is a Iarg egeneti cdistanc ebetween O. uhleri
and other four Oeneis species, which is consistent with the fact that the morphology of O.
uhleri i' smarkedly differe nitn size and in color from the four other Oeneis species,
In 1988, Oeneis philipi inhabitin Aglaska very locall ywas described ,which is morphologi-
cally almost indistinguisha blferom polixene s(Troubrid geet al., 1982; Troubridg e&
Parshal l1,988) .Further r,ecently O. taygete and O. phitip iwere treated as subspecies of O,
bore and O. rosovi, respectively, where tvsevi is an Oeneis species distributin gin the far
east edge of Eurasia (Ople &r Wright, 1999; Lukhtanov & Eitschberge r2,001). Lukhtanov
and Eitschberger wrote in a recently publishe dmonograph that the male genitali aof O.
philipi are identica lto those of O, norna. Therefbre, the conspecificity of O. nonna and O.
philipi cannQt be excluded. According to Layberry et al, (1998 O), philipi is conspecific
with O. rosovi. Thus, O. rosovi, O. norna and O. philip imay be treated as the same species
in future .However, at presen tthere is no clear evidence showing that O. noitna is distrib-
uted North America includin gAlaska. This contradicts the descriptio ngsiven in illustrated
reference books on Japanese butterfli peusblishe dup to 2006.
Acknowledgements and notes
The presen twork is based in par ton the survey carried out in 2001 in the vicinity of Mt
Yakushi-dake (2,92 6m) locate dat the Northern Japan Alps in Honshu island .The speci-
mens have been obtained under the permissio nof Ministry of the Environment, Japan with
the permissio ncode, Kan-Chubu-Kyo 280 (2001 )(T .Itoh) ,A part of the presen tresults has
been publishe das a DDBJ databas e(AB 1 82583) in 2004.
NII-Electronic Library Service
TThhee　 LLeepipdiopdteorpoltoegiroaollogical　 SSoooiceityety 　ooff　 JJaapapnan
Genetic　distance　sarlong 　Oeneis　species 297
References
Aub 呱 J，，　B，　Barascud，　H ．　Descimon 　and　F．　Miche1，1996．　 Systematiqu　emoleculaire 　des　Arygynnes
　　（Lepidopter：aNymphalidae）．　 Comptus 　Rendus　Acadeni．ie‘ies　Science　slll−Vie，　pp．319，647−651 （in
　　 French）．
Fukuda，　H．　et　al．，　1984，　The　tif　ehistorie　sqfbuぴerflies　in／apan 　4，　pp．91−94，　Hoiku−sha，　Osaka．（ln　Japanese
　　with 　English　abstracts ）
Higgins，　L．　G，　and　N ，　D，　RileyJ970 ．　A　Fielご1　Guideω the　8郡惚 rflies　of　B厂itai　nand 　Europe，　pp　150−151．
　　 Collins，　London ．
Howe ，　W ，　H，1975，　The　Butterjlie　osfNorth 　America ，　pp．80−87．　Doubleday ＆Co．　NY ．
Inomata．　T．　and 　K．　MatsumotoJ 　995．　Yama−Kei　Fieid　Books　l　i，　BuUer伍es，　p．222，　Yama −Kei　Pubs．　Co．
　　Tokyo ．（ln　Japanese）
Kawazoe ，　A．　and　M ，　WakabayashiJ976 ．　 Coloだ 4 躍配∫砌 ’∫o’z．v　of 　the　Bl’ttediies φf／4ρ‘m ．　pp　275−277．
　　 Hoikusha　Publishing，　Osaka （ln　Japanes）e．
Laybe叮y，　R．　A．，　P．　W ．　Hall　and 　J，　D ，　Lafontaine．1998．η〜θ 81廨 耀18∫ of　Canada ．　p．235．　Universit　oyf
　　 Toronto　Press，　Toronto．
Lukhtanov ，　V，　and 　U ．Eitschbergcr，200　L　Catalogue　of　the　genera　Oeneis　and 　Davidina　in　BuUerflies　of　thc
　　world ，　Suppliment　4．　pp．8−9．　Hillsid　eBooks ，　Kent．
MartinJ ．　F．，　A．　Gille　sand　H，　Decimon．　2000．　Molecular　phylogeny　and　evolution 　pattems　of　the　European
　　Satyrids（Lepidoptera：Satyrida）eas　revealed 　by　mitochondrial 　gene　sequences ．　 Molec ．　P加’ogenet ．
　　 Evo’．15：70−82． ’
Matsuka，　H ．．1994．　Shir6zu，　T．（supervised ）Jwahash」1．（Ed，）1994，　A　Guidebook　qttheノφごtnese 　Butteij7i、se，
　Op　lerp，　コP．　1A2．．　aPnHdP　A　I，　nBst．．　　IWnc肖．tgTokhyoし199．9（．　lAn　　JFaipeatnde　sGeu）i．de　to　We∬e厂n　Butteijlie（s2nd　Edn）．　pp．367−369 （Peterson
　　Field　Guides　Series）．　Houghton　MiMin 　Co．．　New 　York．
Shir6z，u　T．，2006．　The　Standard　of　Butterflie　sin　Japan，　p，249．　Gakushukenkyusha，　Tokyo，（In　Japanes）e．
TroubridgeJ ．　T．　and　D．　K，　Parshal」1988，　 A　review 　of　the　Oe’leis ρo’ixenes（Fabricius）（Lepidopter：a
　　 Satyridac）complex 　in　North　America．　Can．Ent，120：679−696．
T　ro　ub丘riomdg　et，h　Je　，　NTor．，　しhK　．A　mWcr．　iPch乱ilAirp，c　Jt．　iA．c　．　CSαc’to．　tEt　’aπnd．1　1J4．　：H8．8　S1−h8e8p9a．rd，1982，　A　new 　species　of　Oeneis （Satyridae）
Usa賦 S．，　S．　Ueda ，　T．　NakataniJ ．　ltoh　and 　Y、　IwE置moto ，2005．　Molecular　phylogenic　analyses 　of　the　species
　　of　genes　Oene ’∫．52nd　Meeting　of　Lepid．　Soc　Jpn，　p．11．
摘 要
ミ トコ ン ドリア DNA （1＞Dl ）の 部分塩基配列に基づ い たOeneis　norna ，0 ．　polixenes，0．　bore　and
O．melissa の種間の遺伝的距離 （伊藤隆夫 ・平野和比古 ・渡辺一雄）
互 い に形態的に酷似 して い る4種の Oeneis　．K 種 （Oeneis’norna 、　Oeneis　pθhLxene，　sOeneis　bore，　Oeneis
melissa ）の m の NA （〈の 1）の 452 塩基対配列か ら，これ ら4種は互 い に明瞭な独立種で ほぼ同時代 に分
岐 したこ とが示唆された．また，現在の 所，種 タカネヒカゲ（Oeneis　norrta ）が新大陸に分布してい る明
確な根拠はな く，この こ とは冂本で出版 された多数の図鑑の記述 と矛盾する ことを言及 した．
（Accepted　Scptember　30，2009）
一
NNI工I工-EElleoetcrotniroonic　 LLiibrbarryary 　Service