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astva Trans. Iepid. Soc. Jdpan 58 (4): 387404, September 2007

Molecular phylogenetic analysis of the Erebia aethiops group (Lepidoptera, Nymphalidae)

Takatoshi NAKATANi'), Shin-ichi UsAMi2' and fateo IToHj'

i) Graduate sehool of Science and Technology, Shinshu University,

3-1-1 Asahi, Matsumoto, Nagano, 390-8621 Japan 2' Department of Otorhinolaryngology, Shinshu University School of Mcdicine,

3-1-1 Asahi, Matsumoto, Nagano, 390-8621 Japan 3' Department of BioEogy, Shinshu University, 3-1-1 Asahi, Matsumoto, Nagano, 390-8621 Japan

Abstract Efebia niphonica on the Japanese islands belongs to the aethiops group (sensu Wurren, 1936), We conducted molecular phylogenet{cal analyses of the major subspecies and also of the geographical variations, which are supposed to belong to this group. [[Ihe resu]t implied that each of "niphonica]' E. niphonica from Honshu, and E, scoparia from Sakhalin and HokkaidQ has evolvcd

to a species level comparabie with E, aethiops, E. alcmena, and E. neriene. We also inferred the phylogeography of the aethiQps group. Southern Sakhalin is considered to have been conncctcd to northern Hoklcaido during the last glacial period, and many animals and plants were considered to have come over to the Japanese islands through this landbridge. In fact, we detected the identical "niphonica" haplotypes from populations in southeni Sakhalin and northern Hekkaido. This sup- ported thc hypothesis that genetic exchanges took place. Erebia vidleri in Nearctic region with an extemal appearance similar to those of the aethitrps group, showed a little phylogenetic relation- ship.

Key words Biogeography, Erebia, Erebia niphonica, Erebia vidteri, Nymphalidae, haplotype,

sw5. COL

1. Introduction

In his monograph, Warren (1936) divided the species-group taxa of the genus Erebia into 15 groups based on morphological characters such as male genitatia. He placed niphonica from the Japanese Islands ("niphonica" hereafter) in the aethiops group together with aethiops, alcmena, and neriene, There has however hitherto been no agreement on the sys- `Lniphonicd'. tematic position of Some treated it as a separate species firom neriene, like Warren, whereas others treated it as a subspecies of neriene. In this paper we discuss its systematic position and eyolutionary process based on molecular information and habitat,

In 1877, Janson described Erebia niphonica as an independent species based on specimens from Mt Asamayama in Nagano Pref,, central Honshu. The Hoklcaido population was de- scribed as a separate species, Erebia scoparia (type locality, Kuromatsunai, Shiribeshi, Hokkaido), by Butler in 1881, But Leech (1892-1894) and Eifinger (1906) treated these taxa, which are currcntly classified as alcmena and neriene, as conspecific under the narne sedakovii, distributed from Altai to China and Far Eastern Russia, and the Japanese popula- tion as its subspecies. Go]tz (1932) treated alcmena as a separate species, treating doii from the Kuri1 Islands (Kunashiri) as a subspecies of sedakovii, Goltz (1934) also described a new taxon septorientalis from Kamtschatka as a subspecies of sedakovii, but it has not been found again tltere since then (Korshnov and Gorbunov, I995). It is considered that the pop- ulations belonging to the aethiops group do not inhabit such a northern cold region. Warren (1936) treated sedakovii as a synonym of neriene, and at the same time, each of alcmena, neriene and niphonica as a separate species, and scoparia and doii as subspecies of niphon- ica. Esaki and Shir6zu (1951) pointed out that there was some room for doubt about treat-

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388 Takatoshi NAKtxT,xNi, Shin-ichi UsAMi and Tatco I'roH

ing niphonic;a as a separate species, as the Japanese specimens that W}uren studied were limited in numbers, and some from the Korean Peninsula showed transitional appewrances, It has been, however, a usuat practice to classify the populations from Hokkaido and Honshu as niphonica among the major illustrated reference books published in Japan there- "ni- after (Shir6zu, 1965; 1975; Kawazo6 and Wakabayashi, I976). Fojioka (1975) treated phonica" from the Japanese islands as a separate species in the first edition, but classified as a subspecies of neriene in the revised edition (1997). Asahi et al. (1999), who have studied butterfiies from Sakhalin energetically, also classified it as a subspecjes of neriene from the standpoint of adopting the wider concept of species. As for the classifications by foreign taxonomists who are mostly Russian, Kurentzov (1970) treated the pepulations from Sakhalin, the Kuri] lslands, Hokl(aido, and Honshu as niphoniea, and classified the popula- tion that is somewhat in isolation in the northern highlands of the Korean Peninsula as ner- iene, by examining the male genitalia of specimens from Mt Changbaishan (on the borders of China and northern Korea). He pointed out, however, that the difference in the male gen- italic structure ameng local populations of the sedokovii complex was very small, and could not be the basis for separating species. Thereafter Tuzov (1993), Korshnov and Gorbunov (1995), and Tuzov et aL (1997) followed in his footsteps, and recently Gorbunov (2001) also classified niphonica as a separate species based on the difference in genitalia. D'Abrera (1990) also treated them in the same way based on specimens in the British Museum. All the above taxonomists treated all the populations from Sakhalin, the Kuril Islands, Hokkaido and Honshu as a single taxonomic group, either an independent species separate from neriene or a subspecies of the continental species neriene.

On the other hand, Ono (1973) divided the population of the Japanese islands into two sepa- rate species for the first time, and pointed out the possibility of treating the Sakhalin and Hokkaido populations as a subspecies of neriene and the Honshu population as a separate species niphonica, Kogure (1975, t979) and Murayama (1975) also put forward the view that the populations from Sakhalin and Hokkaido were a subspecies of neriene, and that that from Honshu was a separate species niphonica. Ono and Murayama showed no de- tailed grounds for classifying the Sakhalin and Hokkaido populations as a subspecies of ne- riene, but Kogure pointed out the existence of the scent patches on the forewings of the males (Kogure, 1975, 1979), and also a possibi]ity of difference in the larval fbod prants (Kogure, 1981). Regarding the larval food plants the djfferences in the species of Gramineae and Cyperaceae, however, are not so significant as a basis for differentiating their parasite species, and there is a counter argument that some scent scales although only a few can be recognized in the Honshu population (Watanabe, 1986). In fact, about 10 species of Gramineae and Cyperaceae are recorded as larval food plants fOr the Honshu population (Fukuda et al., 1984). In addition larvae have been found eating the grass of im- ported species in Hoklcaido, and some cases of distribution expansion have been observed along trails where the particular grass has been cultivated (Kawata and Kitahara, 1990). Therefore, it is certain that these popu]ations are generalists for larval food plants. Kawazo6 and Wakabayashi (1976) treated those from the Japanese islands as a separate species niphonica based on some ditferences in the male genitalia, and at the same time pointed out the possibility that the populations from the Korean Peninsula may be the same species as that from the Japanese islands.

In order to eyaluate the independence of species, crossing experiments between populations are the most effective means, According to such experiments between the populations from Hokkaido and from Honshu using individuals that were bred (Kawata and Nagaoka, l990), 46 eggs were obtained after coupling males from Hokkaido (Maruyama, Sapporo) and fe- males from Yamagata Pref, (Mt Gassan). Eighteen 1arvae hatched (hatching rate 39%), only

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389 Erebia aethicrps Group

Fig. 1. Geographical locations of the aethiops group and the referenced spccics sampLing sitcs. Shadowed area dcnotcs the distribution range of each species belonging to the aethiq?s group. blue: E. aethiops, green: E. neriene, pink: E. alcmena, brown: E, niphoni.ca. Sampling sites, red cjrcle: E. aethiops, triangle: E. neriene, inverted triangle: E. al(/mena, star: E. callias and E. theano;. rectangle: E. vidteri, yellow circie: E, niphonica, For detail sainp]ing sites of E. niphonica refer to Fig. 3,

2 1arvae grew nermally, and finally 2 male adults emerged, It was also recorded that copu- lation did not succeed in combinations where the females were from Hoklcaido, although the total number of trials was not given. But if it was confirmed with many cases, then there is a possibility that a premating isolation mechanism was functioning for Hokkaido females involving the dctcction of some chemicals from the male scent scales, which exist in the males of the Hokkaido population. As only rnale adults emerged in F,, there might also be a possibility of hybrid inviability,

Sekiguchi et al, (2000, 2002) have shown results which suggest that the aethiops group is molecular-phylogenically monophyletic by analyzing 30 species of the genus Erehia based on the combined fragments (735 bp in total) ofapart of 16s rRNA (356 bp) and apart of NDI (379 bp) of mitochondrial DNA (mtDNA). Even within the same genus Erebia, how- ever, there is a great ditference in the rate of speciation between the theano species group (comprised of theano, pawlo}vskii, stubbendoi:t7i and maurisius) and the aethiops group, and they could not effectively dis¢ uss the speciatjon of the aethiops group because of insuracjent materials of the populations with geographic variations.

We have studied the genetic diversity of Erebia niphonica from the Japanese islands, in- ferred a process of distribution changes in the Japanese islands during the Quaternary, and pointed out the existence of multiple refugia in Hokhaido and Honshu (Nakatani et al., 2007), In this paper, we study the moleculur phylogeny of the aethiops group, and discuss espeeially speciation based on the results of molecular marker analysis of the major geo- graphic populations of the aethiops group.

2. Materials and methods

We have detected 24 haplotypes from Hoklcaido and 26 from Honshu as a result of studying a total of 289 populations and 423 individuals (Nakatani et at,, 2007). In this paper, we used 20 individuals from the major regions for each of these haplotypes as geographical variation samples from Holcl[aido and Honshu. In addition, a total of 31 butterflies includ- ing all species of the aethiqps group were sampled from the Continent and Sakhalin ([rbble 1, Fig. 1).

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Table 1. Samples used.

Accession No. Taxa Haplotype nLoca]ity M)5 COI E.niphonicascoparia HBOOI 310477195102Walckanai, Hokkaido AB306406 AB306454 E. niphonicascql)aria HAOOO Kaminokuni,Hokkaido AB306407 AB306455

E.niphonicascoparia HCOOI Takayarna, Rebun, Hokkaido AB306411 AB306459

E.niphonicascoparia HCOIO Kutsugata, Rishiri, Holtkaido AB306412 AB306460 E. niphonicascoparia HDOOO Kamikawa, Hokkaido AB306413 AB306461 E. niphonicascoparia HDO03 Nissho Pass, Hidaka, Hokkaido AB306414 AB306462

E.niphonicascoparia HFOOI Toyouchi, Hamatonbetsu, Hokkaido AB306415 AB306463

E niphoniea scoparia HFO02 Kamisawaki, Ohmu, Hoklcaido AB306416 AB306464

E niphonica scoparia SAOOO Russia: Khrebtovyi Riv., C. Sakhalin Reg. AB324830 AB324849 E. niphonicascoparia HBOOI 161l7610425614112742222221112122111111Russia: Yuzhno-Sakhalinsk, S. Sakhalin Reg. AB3e6406 AB306454

E. niphonica niphonica NAIOO Mt Tangoyama, Joetsu Mts range AB306419 AB306467 E. niphonica niphonica NS130 Mt Kabutoyama, Waga Mts range AB306420 AB306468

E niphonica niphonica NS150 Mt MLnamjhonnaidake, Yakeishi Mts range AB306421 AB306469

E. niphonica niphonica NS 1 70 Mt Ryumonzan, Asahi Mts AB306422 AB306470 E. niphonica niphonica NS 1 1 1 Mt Tekaridake, Akaishi Mts AB306423 AB306471

E, niphenica niphonica CHI10 Mt Myokosan, Myoko Mts range AB306425 AB306473

E. niphonica niphonica CH120 Happo-one, Hida Mts AB306426 AB306474

E, niphonica niphonica CH130 Mt Tanigawadake, Joetsu Mts range AB306428 AB306476 E. niphonica niphonica CH140 Mt Iwasugeyama, Joshin Mts range AB306430 AB306478

E. niphottica niphonica CH150 Mt Komagatake, Kiso Mts AB306431 AB306479 E. niphonica niphonica CH160 Mt Makariseji, Hakusan Mts range AB306433 AB306481

E.aethiopsaethiops AAOOO Switzerland: Goppenstein, Valais AB324812 AB324831 E. aethiops goltzi AUOOO Russia: Serga Riv., Middle Urals AB324813 AB324832 Eaethiqpsmelusina AMOOO Russia: Teberda, Caucasus AB324814 AB324833

E. aethiops rubrina AROOO Russia: Muika vil., AIan Range, Sayan AB324S15 AB324834 E.alcmenctminshani CMOOO China: Taibai Shan, Qin Ling Mts, Shaamxi AB324816 AB324835 E.alcmenaminshani CMOIO China: Datong Shan, Qinghai AB324817 AB324836 E.nerienealcmenides NAOOO Russia: Khrebet Khekhtsir, Khabarovsk AB324818 AB324837

E.nerieneatcmenides NAOOI Russia: Khrcbet Khekhtsir, Khabarovsk AB324819 AB324838 E.nerieneatcmenides NAOIO Russia: Gorin, N. Komsomol'sk na Amurie AB324820 AB324839

E.nerienealcmenides NAOIO Russia: Gornyy, N. Komsomol'sk na Amurie AB324820 AB324839

E. neriene neriene NNOOONNOIONN020VDOIOLTOOOCAOOOTHOOOBPOOOPHOOORussia: Buryatia, Mondy vi1., Sayan AB324821 AB324840 E. neriene neriene Mongo]ia:Uiaanbaatar,Terelj AB324822 AB324841 E. neriene neriene Mongolia: Tsenkher Jiguur, Hangay Mts AB324823 AB324842

E. i,idleri Canada: Mt Apex, BC AB324824 AB324g43

E, ligea takanonis Japan: Jonendake, Hida Mts AB324825 AB324844

E. cattias alttu'ana Mongolia: Chkherteyn, Bayan Ulgii AB324826 AB324845

E. theano Mongolia: Chkherteyn, Bayan Ulgii AB324827 AB324846

Boerebiapannenio Mongolia: Bayanchandman, Toy AB324828 AB324847 Paratasa herse China: HuasiBei Shan, in hai AB324829 AB324848

n, number of{ndividuals.

Locality, where OUT was collected.

For genetic markers, we used a part of the NADH dehydrogenase subunit 5 (AiD5) and cy- tochrome oxidase subunit I (COI) genes in mitochondrial DNA. DNA was extracted from the body and legs by DNeasy Tissue Extraction Kit (Qiagen). Details on PCR running con- ditions are described in Nakatani et at. (2007). As primers for PCR and direct sequencing, we used Vl, C2 (Yagi et al., 1999) for IV[D5, and mtD6, Nancy (Simon et al., 1994) for COL We aligned the nucleotide sequence data with Clustal X (Thompson et at,, 1997), and conducted the fragment comparison of432 bp fbr ND5, 51O bp fbr COI, and 942 bp in total,

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391 Erebia aethiops' Group

ff. niphonica (Honshu)

1 E aethiops

E. neriene

L-mrm NN020 94 CMOIO E･ aicmena CMCOO l Z H,,sFHoF,o,oi SI,EI,g E. niphonica CHokkaiao, 4H,A,Og,O,Sakha]in) 1OO LrHDO03 l･HDOOOSAOOOVDOIOLTOOOTHOOO

E, w'dleri

E. fi9eaE.

theano

CAOOOPHooO E. cafEas

Paralasa herse BPOOO O,O05substitvtionsisite Boesebia patTnenio

Fig, 2. Neighbourjoining diagrain of the Erebia aethiops groupbased on genetic distances (Kimura' s 2 parameter methods).

3. Results

The phylogenetic analyses were worked out with the neighborjoining method and with the maximum parsimony method. The topologies of the subspecies rank are identical between the two methods, so the neighbor-joining tree is shown (Fig. 2). For convenience we will call tihe Sakhalin and Hoklcaido population scoparia, the Honshu population niphonica, and "niphonica" the whole populations from the Japanese islands hereafter.

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Ettt. .

3.1 Diyergence in the species rank

It is mferred that scqparta, alcmena, nenene, aethiop s, and nrphonica diverged from a uom- mon ancegtral species in bpecies iank, and the divergence of these dugters is supported by high bootstrap values Populations that are considered to be geographical variations of each single gpecies form a clugter, the populations from Sakhahn and Hokkaido forrn the cluster scoparta, the populations that are considered as nertene from the dred ranging from Mongolia to Russidn Sayan and the Amur region fbrm the clugter nenene, the populations from Switzerland, Middle Uralg, Sayan and Caucasus fonn the cluster aethtops, the popula- tiong from Shaanxi and Qmghdi form the cluster alcmena, and all the populations fiom Honshu torrn the tluster nrphon"a The conventional claggification of gpecJes rank based on external characterg ig supported at the molecular phylogenetic ]evel except for the popu- lations fiom the Japanese islands bemg grouped mto two clusters (gcoj)ana and nrphonica) `"nophonica", E vidten has a similar external appearance to but does not belong to the aethtoj)s group

3.2 Diyergence in the subspecies rank The populatiens of the cluster aethiops diverged into two groupg one is ssp metustna in Caucasus and the other consist of the remamder It is mterred that the Caucagus population diverged earlier than the other populations o ± the aethiops cluster The populdtions ot the cluster nenene diverged mto two groups one is ssp atcmenideg m the Amur region and the other is sbp neriene in Sayan and Mongoha Thug the geographiL distanceg are refiected in the genetic relationships We cannot infer the relationships betxNeen the geographical posi-

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393 Erebia aethtops Group

' tf tNt-p Northern Lineage Lr . 'E 1- nyt- ']LIf1t t)T -c

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big 3 Dibtribution of the hdplotypLs, white sohd Lirdes dcnote thL loLal hdplot)pes Pale green dot denotes the distr]butien rdngc Indicdtaon unit its 75' lengitude× 5O latrtudc (a) Hokltdlde, (b) Honshu

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394 Takatoshi NAKA'rArgt, Shin-ichi UsAMi and Tateo IToH

tion and the genetic relationships fbr alcmena, because only two populations were studied.

3.3 Divergence within the local populations

"niphonica" Regarding of the Japanese islands, we mainly collected samples from the pe- ripheries of the distribution areas for the purpose of studying distribution limits of the ge- netic variations. As a result, we detected 24 haplotypes from Hokkaido and 26 haplotypes from Honshu as mentioned abeve, The genetic distance (Kimura's 2 parameter method) be- tween each pair of haplotypes was in the range of O.1-O.3%, and the interrelations were not shown accurately by the phylogenetic tree, as the bootstrap value was low. At this level of genetic distances the network tree represents lineage relations better than the phylogenetic tree. According to the statistical parsimony network (Nakatani et al., 2007), 3 lineages in Hokkaido and 2 lineages in Honshu were recognized (Fig. 3 (a), (b)), Regarding the Sakhalin population, we analyzed 3 samples frem 2 regions and the haplotype from YUzhno-Sakhalinsk in southern Sakhalin was identical to that of the population with a limit- ed distribution around W2aklcanai, in nonhernmost Hoklcaido. The haplotype from Khrebtovyi Riv. in Central Sakhalin was closely related to the haplotypes of the populations distrjbuted in Rishiri lsland and Rebun Island, northern Holtkaido.

4. Discussion

4.1 Sakhalin and Hokkaido population

The depth of the Soya Strait, which separates Sakhalin and Hokkaido, js about 60 m, and the strait was formed later than 11,OOO years ago (Ohshima, 1990, 2000). The depth of the Tartar Strait (the Mamiya Strait), which separates the Continent and Sakhalin, is even shal- lower, and it is said to have been formed later than 4,OOO years ago (Ohshima, 2000). Sakhalin and Hoklcaido had been connected to the continent almost continuously during the period of 65,OOO years from about 75,OOO years ago up to about 10,OOO years ago (Ono, 1990). They were parts of the continent for longer periods of time than the periods when they were isolated isiands. Thus, during the glacial period geographic conditions allowed the organisms to migrate southward from the continent through Sakhalin to Hokkaido, The more important factor was the existence of environmental conditions which allowed organ- isms to inhabit Holtkaido. Reconstruction studies of ancient flora by pollen analyses (Sakaguchi, 1989; Igarashi et al., 1993; Yamada, 1998; Hoshino, 1998) suggest that the low a] tj tude areas in Holckaido were the southern limit of the taiga during the last glacial period, and that there were forests richer in Larix gmelinii than those in present northern Sakhalin, In the eastern and northern parts of Hokkaido, the distribution of the taiga with Larix gmetinii as its main plant was divided in such a way that each of coniferous forests, grass- ]ands, wet vegetation areas and alpine tundra constituted segregated niches according to the altitude andlor the geographical features. Thus the areas were functioning as refugia for many organisms in the Far East (Ono, 1990).

It is inferred that such a landscape closely resembles the vegetation that is commonly seen where scoparia buttediies currently occur (Kogure, 1981; Takeuchi, 2003), Therefore, it was suggested that genetic exchanges occurred between the southern Sakhalin popu]ation and the northernmost Hokkaido population during the last glacial period. This was support- ed by the fact that the southern Sakhalin population is genetica"y identical to the northern- most Hokkaido population. It was also supported genetically by the fact that the Centrat Sakhalin population is quite closely related to the northern Hokkaido population in Rishiri and Rebun. In other words, these results supported the notion that there were genetic ex- changes between the population of Saklialin and that of Hoklcaido before the Soya Strait

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Erehia aethiops Group 395

"'t,/

(a) x/

(b)

Fig.4. Erehia aethiops ssp. (a) ssp. aethiops; Goppenstein, Valais, Switzerland. 18 July 2003. (b) ssp. goltzi; Middle Urals, Russia, 14 July 2001. (c) ssp. melttsina; Dagestan, E. Caucasus, Russia IO July 2000. (d) ssp. n{blina; Abaza Khakassiya, W. Sayan, Russia, 25 July-8 Aug. 2003.

>x-ix.

(c)

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(b) (d)

Fig,5. Erebia neriene neriene. (a) Tgenkher-Jiguur, S. Tsetserleg, Arkhangay, Mongolia. 25 July 2000. (b) Tov, Terelji, Khentey Mts, Mongolia. 20 July 1997. (c) Upper Mondy, Sayan, Russia. 9 July 2000. (d) BuJljatia, Transbaikal, Russia. 28 July 1999.

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396 Takatoshi NAKATANi, Shin-ichi UsApttt and Tateo IToH

t, -. t"t "

(a) (c) ix ,tt.tt

(b)

1M.t tttt

(e)

Fig. 6. Erebia neriene atcmenides. (a) Gorin, S. Gorin rtver, Amur region, Russia. 16 Aug. 2002. (b) Gornyy, Amur region, Russia. 17 Aug. 2002, (c) Khrebet Khekhtsir, Khabarovsk, Russia. 19 Aug. 2002. (d) Female; Gornyy, Amur region, Russia, 17 Aug. 2002, (e) Female, Tsimmermanoyka, Khabarovsk, Russia, 23 Aug, 1986.

was formed. On the other hand, the depth of the Rishiri Channel which separates Rishiri Island and Rebun Island from Holckaido is about 80-85 m (Japan coastal guard, 2005a, b, c), It is possible that Rishiri Channel was formed earlier than the Soya Strait. Thig is con- sistent with the result of the molecular phylogenic analysis, which is that the Rishiri and Rebun population diverged from the mainland Helckaido population earlier than the south- ern Sakhalin population did from the mainland Holdcaido population.

When we look at the distribution of scoparia in Sakhalin (Asahi et al,, 1999; Asahi and Kohara, 2004), the habitat was not observed in the area north of N51-520, It is inferred that sc'oparia did not adapt to such cold areas where permafrost existed. A landbridge was formed across the [Ihrtar Strait (the Mamiya Strait) during the last glacial periods colder than the present day, But the environment there was less favourable for scoparia to inhabit. Theref6re, it is strongly suggested that the scqparia population in Sakhalin and Hokkaido or in the Continent did not migrate back and forth between the Continent and SakhalinlHokkaido during the last glacial period. This is consistent with the result shown above that the populatjons of Saldlalin and Hokkaido are molecular-phylogenically quite far away from the Erebia population in the Amur Region, which is on the opposite shore across

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397 Erebiaaethiops Group

x

(a)

(b)

"niphonica]' Fig. 7. Erebia ssp. (a) ssp. scoparia; Upper Veba West Yasnoye vill., Tymovskiy re- gion, C, Sakhalin, Russia. 1 Aug. 2002. (b) ssp. scoparia; Daisetsu Lake, Kamikawa, Hokkaido. 24 July 2002. (c) ssp. niphonica; Mt Tangoyarna, Niigata, Honshu. 16 Aug. 1997. (d) ssp. ntphoniea; Mt Karaniatsudake, Nagano, Honshu. 17 Aug. 1999.

(a) (b)

Fig. 8,Erebia atctnena, (a) Xiahe (Lagrang), Gansu, Ch ina (h=3,OOO-3,400 m).(b) Taibai-Shan, Qin Ling Mts, China.

Fig,9, Erehia vidt (a) Mt Apex, Penticton, B. Canada. 24 July 20e4,Cb) Marining Park,

Penticton.Beri,,C,,Canada, 25 July 2004,C.,

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398 Takatoshi NAKA'i'ANi, Shin-ichi UsAMi and Tateo IToH in' "''.""".',,･ fva,`le.geii.'-...gew, ,."･,eegee..,lef.ag",i.･'../,.vatfi"va'-',･l/ew, .,

Fig. IO. Habitats. (a) Erebia aethiops, Goppenstein, Valais, Switzcrland (h=1,300 m). E. aethiops dominated with E. montana, E. ettnyaie and E. Iigea fiy together. (b) Erebia neriene, Tgenkher Jiguur SPA, Arkhangai, Mongolia. (c) Erebia alcmena, Xiahe (Lagrang), Gansu, China (photo courtesy of Y. Watanabe). (d) Erebia vidleri, Mt Apex, Penticton, British Columbia, Canada (photo by S. Asai),

the Japan Sea, Some have pointed out that the population belonging to scoparia is distrib- uted in the Continental region which is opposite to Sakhalin (Churkin, 2005), We had an opportunity to examine 3 males and 1 female (Fig. 6e) which Mr Novomodnyi collected on 23 Aug. 1986 at Tsimmermanovka (E139020', N51020') on the lower Amur River. The wing patterns of these specimens show a high degree of homogeneity and are characteristic of neriene. Hence, the possibility of the populations belonging to scoparia being distrib- uted on the Continental side seems to be less probable, One of us, Nakatani together with Mr Kogure guided by Mr Novomodnyi, had a chance to visit the particular place on 14 Aug. 2002, but we were unable to find any material although the weather was good. The flight- period of the species seems to be quite late in that region. Therefbre it scems to be probable that more new habitats will be discovered in the lower Amur River.

4.2 Speciation

Sekiguchi et at, (2002) discussed speciation of the aethiops group by comparing the genetic distance of each pair of taxa within the aethiops group with the genetic di stances among the theano species group, but were unable to make a definitive argument, because the geograph- ic variations of the aethiops group were not examined. And it has been suggested that the rate of speciation ditfers greatly depending on the species group even within the same genus Emebia,

In this paper we have analyzed molecular phylogenic relations by selecting specimens from the major geographical populations of each taxon belonging to the aethiops group, First of

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399 Erehia aethiops Group

all, we will discuss the species relationships of E. aethiops and E. neriene, It has been pointed out since long ago that these two taxa have a sympatric distribution from southern Siberia to Altai (Eiranger, 1906), although ali the other taxa within the aethiops group have allopatricdistributions,

Kogure (1997) made great efforts to study the distribution of neriene, but seems to have fOund no regions where both E, aethiops and E. neriene are distributed together, However, according to Lukhtanov and Lukhtanov (1994), the distribution areas of these two species overlap each other at around E86-870N500 in the Altai Mountains. According to Tshikolobets (2003), they also overlap each other in Yablonovyi and the Kentei mountain range around EI07eN500, southwest of Lake Baikal, Although Tshikolobets (2003) did not mention the altitude of habitat, according to Lukhtanov and Lukhtanov (1994), aethiops was found from the lowlands up to 1,OOO m, and neriene at 800-1,700 m in the Altai Mountains, It is considered that there are regions where the two species occur together ina microscopic view. Therefbre, it is safe to conclude that these two species are separate species in light of the biological species ¢ oncept. When we look at the phylogenetic tree with this fact in mind, it is suggested that each of the foilowing 5 taxa hag differentiated up

to the species rank: aethiops, alcmena, neriene, niphonica, and scqparia, We understand

that it is widely accepted that we cannot discuss classification of species simply based on molecular phylogeny alone (see, for example, Osawa et al., 2002). In the case of the line- age relationship of the aethiqps group (Fig, 2), however, it seems acceptable to cenclude that differentiation between scoparia and niphonica has developed to the species rank, al- though their distribution ranges are geographically located closely but not overlapping. DNA barcoding has been proposed in which the classification of species is approached by using relatively short fragments of the COI gene of mitochondrial DNA (Hebert et al,, 2004). It is worth mentioning that it has received a certain recognition as a simple means for detecting cryptic species (Consortium fbr the Barcode of Life (CBOL), Smithsonian Institution). In addition, although there is only one case with a few individuals, the result of the crossing experiment by Kawata and Nagaoka (1990) seems to be consistent with the view that the populations from Holckaido and Honshu have evolved to the species rank in respect of each other.

4.3 Molecular phylogeny of Erebia vidleri in North America

Ejebia vidleri is distributed locally from British Columbia, Canada to Wyoming, USA. It has been suggested that the external appearance is very similar te E, niphonica, although the genitalia are diiferent (Layberry et al., 1998). Warren (1936) designated the vidleri group as one species, and placed it next to the aethiops group. Its habitat is open grassland at the margin of coniferous forests (Fig, 1 Od), and is quite similar to the habitat environment of aethiops (Fig. 10a), neriene (Fig. 10b), and niphonica (Nakatani and Kitagawa, 2000; Nakatani and Hosoya, 2003). But it was suggested that E, vidleri is molecular-phylogeni- cally far away from the aethiops group.

4.4 Phylogeography of the aethiops group

The major babitats of the aethiops group such as aethiops (Fig. 10a), neriene (Fig, 10b), scoparia ([fakeuchi, 2003), and niphonica (Nakatani and Kitagawa, 2000; Nakatani and Hosoya, 2003) are open grasslands within or at the margin of coniferous forests. But at- cmena seems to inhabit the dry shrub zone (Fig. 10c) (Watanabe, 1993; Watanabe, personal communication).

When we Iook at the habitat environment and the distribution of the aethiops group (Fig. 1), it is inferred that the common ancestral species was distributed in a region where the conifl

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400 Takatoshi NAKATANi, Shin-ichi UsAMi and Tateo IToH

erous fbrests and the grasslands spread in a mosaic pattern around Lake Baikal. When the environment became unsuitable (warm or dry), the habitats were fragmented and isolated from each other, E. scqparia diverged around Sakhalin and Hokkaido, which is the eastem periphery of the distribution of the ancestral species, and atcmena, which is adapted to a dry regien, diverged in the southern periphery of distribution in China. E. neriene diverged in the nonh of Sayan and Transbaikal, When the environment became more suitable (cold or wet), aethiops expanded its distribution from Altai to the west to Caucasus, Urals and Central Europe including the British islands, and niphonica to the Japanese js]ands, E. ner- iene and aethiops expanded their distributions, resulting in an overlap in the area from Sayan to [[)ransbaikal. The population that ranges from northeastern China to the northern part of the Korean Peninsula is considered as neriene. There js a yiew, however, that the 'from population of the northern part of the Korean Peninsula is identical to niphonica the Japanese islands (Kawazo6 and Wakabayashi, 1976). It has also been pointed out that the transitional fbrm has been observed there (Esaki and Shir6zu, 1951), Therefore, we hope a future study of materials in this region will clarify the relationship of neriene and niphonica,

As has been mentioned already, except for alcmena each of all taxa belonging to the aethiops group mainly inhabits open grasslands in the coniferous forests and does not in- habit drier expansive grasslands without surrounding forests. Considering such an ecoiogj- cal requirement of the subject species, the important prerequisite for refugia in warm inter- glaciat periods is not only adequate temperature but also adequate humidity to allow conif- erous fbrests and grasslands to grow together in patched patterns, Isolation may have also been taking place in quite dry glacial periods, This situation is apparently similar to the case of Erebia epiphron in Europe (Schmitt et al,, 2006). Estimation of the precise diver- gence time needs to be discussed in the whole evolutionary processes of the entire genus Eiehia.

Acknowledgments

Regarding collection of Erebia species in Valais, Switzerland, permission was granted by Dienststelle fur Wa]d und Landschaf, and the support by Dr T. Althaus is appreciated. As for the samples in Japan, permissions were granted by Ministry of the Environment, by Agency of Cultural Affbirs, by local governments, and by other related organizations. We thank the fo11owing people who helped us for collecting samples in the Continent: J, Asahi, N. Aso, K. Sei, E. Tsilj'i, Yl Ni'shiyama, T. Nemoto, M. Hukui, A. Hojo, K. Matsumoto, E. Vl Novomodnyi (Khabarovsk Regional Lore Museum). We are also grateful to many col- leagues for collecting in Japan. Y Watanabe gave us infbrmation about the habitat of Erebia alcmena and permission to use his photos. Y Ohshima and M. Kogure planned the field study on the Continent, C, S, Guppy and S. Asai cooperated in obtaining Eiehia vid-

leri specimens,

Referenees

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Macrolqpidoptera ofthe IVbrld 1/ 94-1 14, pls 35-37. Fritz Lehmann Verlag, Stuttgart. Esaki,T, andT. Shir6zu, 1951, ButtetYTies of.lapan. Hokuryukan,Tokyo. (In Japanese). Fukuda, H,, Hama, E,, Kuzuya, T,, Takahashi, A., Takahashi, M,, Tanuka, B,, Tanaka, H., Wakabayashi, M, and Y. Watanabe, 1984. 7he LCfe Histo,:}, ofButteijlies in Japan 4. Hoikusha, Osaka. (In Japanese). Filjioka, T., 1975. Buttenyvlies ofJapan. Kodansha, Tokyo, (In Japanese). Edn). Kodansha, Tokyo. Japanese). , 1981.ButtenyfliesqfJapanCrep.second (In Goltz, V. D,, 1932. Genus Erebia DaLm, in Seitz, A (Ed.), 71eie Mac,rotqpidoptera qfthe Wbrtd (Suppl,) 1: 132-152. Alfred Kernen, Publisher, Sttutgart, , 1934. Ueber Erehia sedukoviiEv. und alcinena GT,-Grsh, Dl. ent, Z. Iris 48: 54-59. Gorbunov, P, Y,, 2001, 7:Pte Butteslaies qf Russia: Classijicarion, Genitatia, Key for Identijication "Thesis", (Lepidoptera: Hesperioidea andPapilionoidea), Ekaterinburg. Hebert, P. D, N., Penton, E, H., Burns, J. M., Janzen, D.H. and W, Hallwachs, 2004, Ten species in one: DNA barcoding reveals cryptic species in the Neotropical skipper butterfly Astmptes falgerator. Proc natn.Acad. Sci. 101: 14812-14817. Hoshino, F,, i998, Historic flora ofSouthern Holckaido. in Yasuda, Y, and N. Miyosh] (Eds), Historic Flora qfJapanese Jsls:51-61. Asakura, Tokyo, (lnJapanese). Igarashi, Y., Igarashi, T., Daimaru, H., Yamada, O., Miyagi, T,, Matsushita, K. and K. Hiramatsu,. 1993. Vegetation history of Kenbuchi basin and Furano basin in Hokl(aido, North Japan, since 32,OOO yrs BP. Quaternar:y' Res. Tbk>'o 32: 89-1OS (in Jupanese). Janson, O. E,, 1877, Notes on Japanese Rhopaloccra, with descriptions of new species. Cistuta Ent. 2: IS3- 160. Japan coastal guard, 2005a. IVautical Chart, So.va strait (1 :200,OOO).

, 2005b. NLiuticalChart,RishiriisL (1:50,OOO).

2005c. Ndutical Chart, Rebttn isl. , (1:50,OOO). Kawazo6, A. and M. Wakabayashi, 1976. Coloured Illustfutions qf the Butteijlies of Japan, Hoikusha, Osaka, (InJapanese). Kawata, M, and H, Kitahara, 1990. Food p]ants and range expansion of Erebia niphonica in Hoklcaido. Choken Field 5 (8): 28-3 1 (in Japanese). "Blaldston Kawata, M. and H. Nagaoka, 1990. Breed of Benihikage": on hyhrids of the Hokkaido and Honshu population of E, niphonica, Choken Fieid 5 (6): 6-8 (in Japanese), [`niphonica" Kogure, M., 1975. Erebia should be separated into two species (a tentative study). Usubaki (5): 1-2 (in Japanese). Ndt"re insect 14 1S-19, , !979.Erebia niphonica and EFebia neriene. (9): [`Benihikage". Tokyo. Japanese), , 1981. Butteijt}i migratedf}'omAltai, Tsukijishokan, (In ofErehia neriene Lepidoptera) in Asia. Pap. Ttikao , l997,Distributjona]sphere (SatyTidae, [Colln Seminar] (2): 123-160 (in Japanese), Takao Seminar, Chiba, Korshnov, P. Y. and P. Y. Gorbunov, 1995. Buttenyflies oj'the Asiatic Part ofRuffsia, a Handbook. Ural University Press, Ekaterinburg. (In Russian). Kurentzov, A. I., 1970. 71he Btttteijlies oj'the Far East USSR (Japanese translation Edition by M. Abe). Bun- ichi, Tokyo. Layberry, R., Hall, P. W. and J, D. Lafontuinc, 1998. 7ke B"ttecflie,s of Canadd, University of Toronto Press, Toronto.

Leech, J. H., 1 892-1894. Buttei:fiiesfrom China, Japan and Corea. R, H. Porter, London. Lukhtanov, V, and A. Lukhtanov, 1994, Die 7laghlter IVbrdwestasiens. Dr, UlfEitschberger, Marktleuthen. Murayama, S., 1975. A general view of the genus Erebia in the world. Gekhan-MLtshi (54): 9-14

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402 Takatoshi NAKATANJ， Shin−ichi UsAMI and Tateo IToH

− Ono， H ．」 973 ． Butterflies of Siberia（7）， Nature Jnsect 7 （ll）： 2 16 （in Japanese）， ！ − Ono ， Y ．，1990、 The northern landbridge of Japan． guaternar＞ Res ． Tokyo 29 ： 183 192 〔in Japanese）． − ， ， Osawa， S．， S叫 Z ．H ． aud Y ． Imura，2002， Molecular Ph）logen｝ az ！d Evotution｛ガ伽 carabid Beetles（f， the World ． Tetsugakushobo， Tokyo．（in Japanese）． Sakaguchi， Y ，，1989． Some p〔，11en rocords f士om Hokkaido and Sakhalin． Bull． Dep． Geogr．σniv ． Todyo 21 ： 1−17，

Schmitt， T ．， Hewitt， G ． M and P ． M しlller，2006． Di匍unct distributions during glacial and iIlterglacial periods ・ in mountain butterflies：E ’ebia epiphrotz as an example ．ノ． Evol． Biol．19 ： 108−113． Sekiguchi， M ．， Nakatani， T ．， Shinkawa， T ． d M ． Kogure ，2002 ． Molecular phylugenetic analysis of the Erebia 「 − genus 〔LepidopIera， Nymphalidae ）．7 rans ． lqフid． Soc． Japan 53：1 11（in Japanese）． Sekiguchi， M ，， Shinkawa， T ．， Nakatani， T ， and M ． Kogurc，2000． Molecular phylogelletic analysis of tlle − genus Erebia （Satyridae）of Japaエ1． B μ tterj7ies（26）： 14 21 （｛n Japanese）． Shir6zu， T り1965． ButteCtlies OfJapan． Hokuryukan ， Tokyo ．（ln Japanese）． 1975．Illustrated ， BookforStudents1．Gakushukenkyusha ，Tokyo ．（ln Japanese）． − Simon， C ．， Frati， F りBeckenbach ， A ．， Crespi， B 、，Liu ， H ， and P ， Flo〔｝k ，1994． EvolutiQn， weighting ， and phy − 10genetic utility of mitochondrial gene sequences and a compilation of conserved polymerase chai n reac − tion pri111ers． Ann ． ent ． Soc， Am ．87 ； 651 701 ． Takcuchi， N ，，2003 ． On the dis面 bution， variation and ecological study o 皿 Erebiα ni ρhonica in Hokkaido， − Northern Japan（1）． Choken Field 18 （7）19 17 （in Japanese）． ThonpsonJ ， D ，， Gibson ， T ．工， Plewniak ， F ， Jeallmougin， F ． and D ． G ． Higgins，1997． The C］ustal X win − dows interface ： flexible strategies f（〕r multiple sequencc aligllment aided by quality analysis tools．1＞uct ． Acids Res ．24 ：4876 −4882 ． Tshiko 】ovets ， V ． V ．，2003． The Butteifliesげ Transbaikal Siberia． BmQ −Kyiv． ’ ， ， − Tuzov ， V ． K ．，1993， The s｝non ｝mic ムist qfButtenyfliesプhom the ex U ∬ R ． Rosagroservice， Moscow ． Tuzov， V ． K ， Bogdanov， P． V 、， Devyatldn ， A ．， Kaabak ，1，， Korolev， V ．， Murzln， V ，， Sa odurov ， G ． and E ．

Tarasov 1997、 Guide ro the Butterflies of Ru ．∬ どα α層 A c 翩 7セ广厂itories 1 Pensoft Sona −Moskow ， 吻 ． ， ． Warren B （， the ， ，C ．S．，1936．Monograph f GenttsErehia，BMNH ，London ． ・ Watanabe Y り1986． Alpine ButtefLtlies：Mountain ， Butterfl｝ and me ． Tsu 跏 shQkan ， Tokyo ．（In Japanese）， ，1993．Btitteffties and Scenery勿 China． Private pub 】icadon．（ln Japanese）． Yagi T ． Sasaki G ． and H ． Takebe 1999． Phylogeny − ， ， ， ， of Japanesepapilionidbutterfliesinferredfrom nu cleotide sequences of rnitochondrial ハの 5 gene，」．〃 lotec ． E レol ，48142 −48．

Yamada ， G ．，1998． HistoricfiorainNorthernHokkaido． InYasuda， Y ． and N ． Miyoshi （Eds ）， Historic Flora − qプ加 anese lsls：39 50． Asakura， Tokyo ．（In Japanese）．

摘 要

ー Erebia aethiOPS グ ル プ の 分子系統解析 （中谷貴 壽 ・宇 佐美 真 一 ・伊 藤 建 夫）

ベ ニ ー 日本 列 島 産 ヒ カゲ の 種内系統 関係 を調 べ る た ．め ，aethiops グ ル プ の 主 要な地 理 的変異 と，外群 と て Erebia ． し 属 お よ び近縁属 合計 6 種 を用 い て 分 ゴ系 統 解析 を行 っ た，種 ラ ン ク に つ い て は 系統樹 ー （近 隣結合法 と 最 大節 約 法 に よ る解析 で は亜 種 ラ ン ク ま で の トポ ロ ジ は 同 じた め 近 隣 結合 法 に よ る を ン つ い ・ ー 樹 形 Fig2 に示 す），地域 集団 ラ ク に て は ネッ 1 ワ ク樹 （統計的節 約法 ）に よ っ て 解析 し た ．

大 陸産 近 縁種 を含 む系統関係

種 ラ ン ク で は 共通 祖 先種 か ら scoparia atcmena neriene aethiops niphonica ， ， ， ， ， が ほ ぼ 同 時期 に 分 岐 して い い い ー る事が 示唆 され， ずれ も高 ブ トス トラ ッ プ値 で 支持 され る ． Scoρaria と niphonica が 分割 され て い る 点 を除 けば ，従 来 の 形 質 に 基 づ く亜 種 レ ベ ル ま で の 分類が 分 子 系統 的 に も支 持 され る ．Fig．2 に 示 す よ うに sc ・ と nii ）honica は が い ン ， paria 分布域 地理 的 に近 接 して る もの の ， 種 ラ ク の 分化 が 進 ん で い る 事が示 唆 され る ．

Erebia vidleri − は カ ナ ダ BritishColurnbiaか ら ア メ リ カ Wy 〔〕ming に か け て 局 地 的 に 分 布 す る 種 で ， E ． ni ー ー phonicaと似 て い る と の 指摘が あ る ．Warren （1936）は 1種 で 1 グ ル プ と し aethiops グ ル プ の 隣 に 置 い て い ， の ・ る そ 生 息地 は針葉樹 林縁 の 明 る い 草 原 で あ り（Fig，5d）， aethiops （Fig．5a），’neriene （Fig．5b ）， ni ー phonica の 生 息環境 と よ く似 て い る．しか し分子系統的 に は aethiOlps グ ル プ と は 遠 い 位置 に あ る 事 が 示唆 さ れ た ，

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403 Er 召 わどα aethi （ノps Group

亜 種 ラ ン ク で の 分岐 に 関 し て は，E ． aethiops で は Caucasus 集 団 （ssp ． melusina ）の 分 岐 が 深 く，そ の 他 の ・ ・ ス イ ス （ssp． aeliziops ） Middle Ural （ssp ． gottzi） Sayan（ssp ． rubrina ）の 集 団 は あ ま り分 化 して い な い ． ・ E ．neriene で は Amur 地 域 集 団 （ssp ．‘alcmenides ）と Sayan モ ン ゴ ル 集団 （ssp ． neriene ）に 分 岐 して い る、

ベ ニ ヒ つ い 2 24 か ら 26 の ハ プ ロ 口本列 島産 カ ゲ 集団 に て は ，Sakhalinか ら 種，北海道か ら 種 ， 本州 種 ー タ イプ を検出 した．λ ッ トワ ク 樹 （Nakatani et al ．，2007）に よ る と北 海 道 で 3 系統，本州 で 2 系統 が 認 め られ た，北海道 産集 団 （Fig，3a）は，西 部系統が稚内 付 近 か ら日本 海岸 沿 い に 渡 島半島 ま で 分布 し ，中部 ・ Sakhalin，利尻 礼文両 島 の 集団 も含まれ る ，広域 分布 の ハ プ ロ タ イブ HAooo か ら狭 分 布型 の 多 くの ー ・ ハ プ ロ タ イプが 派生 して い る ．東部系統 は オホ ッ ク海沿岸 か ら 道央 日高山脈 ま で 広 い 範囲 に 分 布す る ．日 高山脈 北 部 に 狭 分布型 の ハ プ ロ タ イ プが産 す る 他 は ，す べ て ハ プ ロ タ イ プHDOOO で あ り遺 一 ー 伝 的 に 均 な集団 で あ る ．北部系統 は ネ ッ トワ ク 樹 で は北部 系統 に結 合 さ れ る が，他 の 系統 と は 遺伝

的 距 離が 離 れ て お り，北部 の 狭 い 範 囲 に分 布 して い る ．本州産集団 （Fig，3b）で は ， 北部系統 が東北 地 方 か ら上 越 山系 の 朿半分 と 赤 石 山 脈 に 分 布 す る ．ま た 南部系統 は 上 越 山系 の 西 半分 と赤 石 山脈 以 外 の 中 部 山 岳に 広 く分 布す る ，

Sa alin 集 団 で は，南 部 Sakhalin， Yuzhno −Sakhalinsk産 は 北海道北端 の 稚 内市周辺 に 限 っ て 分 布す る も ・ の と同 じ ハ プ ロ タ イ プ で あ り．また 中部 Sakhalin， Khrebtovyj Riv ．産 は 利 尻 島 礼文島 に 分布 す る ハ

プ ロ タ イ プ に 近縁 な タ イ プ で あ っ た ，Sakhalinと 北 海道 を分 け る 宗 谷 海 峽 の 水深 は 約 60 m で あ り，最 終 氷 河 期 の 約 Ll 万 年 前 以 降 に 海 峡 が 形 成 され た と さ れ る （大嶋，1990，2000）． Sakhalin南 部集 団は北 海 ・ 道北端集団 と遺伝子的 に 同 じ集団 で あ る こ とが確認 され ，ま た Sakhalin中部 の 集 団 も利尻 礼 文な ど こ れ た わ が さ れ る ま で は 北 海道 北部 集団 と 近縁 で あ る と が 遣伝子 的 に 確 認 さ ．す な ち ， 宗谷 海峡 形成 一 Sakhalin と北 海道 の べ ニ ヒ カ ゲ 集 団 の 問 に は 遺伝的交流 が あ っ た こ とが裏付 け られた、 方利 尻 島 と 礼 − 文 島を北 海道 本島 と分 ける利尻 水 道 の 水 深 は 海 図 に よ る と 約 8〔〕85m で あ っ て ，宗谷海峡 よ りも古 い ・ 時代 か ら海 峡 と な っ て い た可 能性が あ る ．北 海道 本 島 の 集 団 と 比 較 し て ，利 尻 礼 文集団 の 方 が ， Sakhalin南部 産 の 集 団 よ り分 予系統的 に 分 岐 が 古 い とい う結 果 と整 合 性 が あ る ． 4 大陸 とSakhalinを分 け る間宮 海 峡 の 水深 は宗 谷 海 峡 よ りもさ ら に 浅 く，海峡 の 形 成は約 ，000 年前以 ［ 1999 ・ 2004 よ る と に お る 降 と さ れ る （大 1鳥，2000）．朝 日 ら の 調査 （朝 日 ほ か ， ；朝 日 小 原 ， ）に Sakhalln け ° ベ ニ ヒ カ ゲ の 分 布 は 北 緯 51−52 よ り北 で は 生 息 が 確認 さ れ て お らず ，永 久凍 土が 存在す る ほ ど 寒冷 な気候 に は 適 応 で きな い も の と 推測 され る ．陸化 さ れ て い て も そ の 環境が ベ ニ ヒ カゲ の 生 息 に 適 さな い もの で あれ ば往来は不可能 で あ る ．現在 よ りも寒冷 な氷 河期 に は 間宮 海 峡 は 陸化 し て い た も の の ， ・ ベ ニ ヒ カ ゲが大 陸 と の 問 を行 き来 で きる 環 境 で は な く，Sakhalin 北海道 の べ ニ ヒ カ ゲ集 団 は 最終 氷 河 ・ 期 に 大陸 と の 間 を往来す る こ と は なか っ た と 考え ら れ る ，こ れ は Sakhalin 北 海道集団 が 対 岸 の Amur 地域 の neriene と さ れ る 集団 とは分于系統的 1に 非 常 に 離 れ た 存在 で あ る と い う解析結果か ら示唆 さ れ る ．

AethiOPS グ ル ープ の 系統地 理 ー 一 Aethiopsグ ル プ の 生息環境 を み る と，aethiol ，s （Fig．5a）， neriene （Fig．5b）， seoparia （竹内，2003），η 置 ・ rl l ・ い 1 phonica （中谷 北 川 ，2000； 谷 細谷，2003）は ，い ずれ も針葉樹林内 や そ の 林 縁 の 明 る 草原 が 主 な 生 息環境 と な っ て い る ．ま た alcmena の 生 息 環 境 は 渡 辺 （1993；私信 ）に よ る と，中 国甘 粛 省 夏 河 ー （Xiahc ， Gallsu， China）で は乾燥 した潅木帯 （Fig．5c）で あ る．Aethi・ps グ ル プ の 生息環境 と現在 の 分布 域 （Fig．1）か ら み る と ，共 通祖先種 は Baikal湖付近 の 針葉樹 林 と草原 が モ ザ イ ク状 に 拡 が る地域 を中心 に 分布 し て い た と推定 され る．そ して 環境 の 悪化 （温 暖化 また は 乾燥化）に よ っ て い くつ か の 集団 に ・ 隔 離分 布 し た も の の うち ，東部 の 分 布 辺 縁 部 に あ た る Sakhalin 北海道付近 で ．sc ・ipariaが分岐 し，辺 ・ 縁部 南 部 の 中 国 で 乾燥地 に 適応 し た alcmena が 分岐 し た、また Sayan Baika1地 域 の 北 部 で neriene が 分岐 し た．Niphonica は大 陸 の 東南 方 で 分 岐 し た 口∫能性 が 考 え ら れ る ．こ れ ら の 分岐年代 は ほ ぼ 同時期 と み ら れ る ．そ の 後 の 環境改善 （寒冷化 ま た は 湿潤化）期 に α etitivp ，〜 は Baikal， Sayanか ら 西 へ Caucasus， ー Ural を 経 て ヨ ロ ッ パ ま で 分 布 を 拡 大 し，niphoniea は 日 木列 島 へ 進 出 した ．Neriene と aethiops は Sayan ， Altaiで も分 布 を拡 大 し， Baikal地域 か らSayanに か け て 混生 状態 を生 じた．中 国東北 部 か ら朝 鮮 半 島 北部 に 分布す る 集団 は現在 neriene とされ るが，朝鮮 半 島北 部 の 集団 は 本州産 吻 海傭 c α と 同 じ と す る 見 解 （川 副 ・若林，1976）P ，移行型 が み られ る と の 指摘 （江崎 ・白水，1951）も あ り，今後 の 調査 が 期待 され る ．

べ ル ー は い れ の を 主な す で に 述 た よ うに aethiops グ プ の 内 ， alcmena を除 く タ クサ ず も針葉樹林 林縁 生 息 域 と して お り，乾燥 が 強 い 広大 な草原 に は 生 息 し て い な い ．こ の よ うな生 態 か ら 推 定す る と，ヨ

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404 Takatoshj NAKArAN1 ， Shin−ichi Us ，xMi and Tateo IToH 一 ロ ッ パ に お け る Erebia epiphr ・ J・．（Schmitt 8 ∫ 砿 ，2006 ） と 同 じ よ う に ，分布 を分 断 され孤 立化 した集 の は の 団 隔 離 ，問 氷期 温暖 期 だ け で は な く氷 河 期 の 乾 燥 が 強 い 環境 下 で も起 こ っ て い た 可 能性が 考 え られ る．分 岐 の 年代推定 は Erebia属全体 の 進 化 プ ロ セ ス の 中 で 検討 し て い く必要 が あ る ．

（Accepted January 5，2007 ）

Published by the Lepidopterological Society of Japalコ， − − 5 20，Motoyokoyama 2， Hachioji， Toky ，192 0063 Japan

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