Biogeography 2. 103-111. Aug. 28, 2000

Molecular Phylogeny of of the Tribe (: Papilionidae) - Using All of the -

l 2 Sadaharu Morinaka , Nobuhiro Minaka , Masayuki SekiguchP, Erniwati4, Siti Nuramaliati Prijono\ Ida Ketut GinarsaS, Tadashi Miyata6 and Toshitaka Hidaka7

I Aichi Study Center, The University of the Air, Yagotohonmachi, 10 1-2, Showa-ku, Nagoya 466-0825, Japan

2 National Institute of Agro-Environmental Sciences, Kannon-dai 3-1-1, Tsukuba, Ibaraki 305-8604, Japan

J National Institute of Neuroscience, Ogawahigasimachi 4-1-1, Kodaira-shi, Tokyo 187-8502, Japan

4 Biological Division, Research and Development for Biology, Indonesian Institute of Sciences, Gedung Widyasatwaloka J1. Raya Cibinong, Cibinong Bogor, 16002

5 Pk. Jelati, Yeh Sumbul, Mendoyo Bali, 82261 Indonesia h Department of Biological Mechanisms and Functions, Graduate School of Bioagricultural Sciences, Nagoya University, Huro-cho, Chikusa-ku, Nagoya 464-8601, Japan

7 The University of Shiga Prefecture, Hassaka 2500, Hikone, Shiga, 522-8533 Japan Corresponding address: S. Morinaka, Totsukahasami-cho 11-20, Kawaguchi-shi, Saitama-ken, 333-0805 Japan (e-mail: [email protected])

Abstract. The molecular phylogeny of birdwing butterflies based on the NOS gene of mitochondrial DNA (mtDNA) was studied by the maximum-likelihood method. maximum-parsimony method and neighbor- joining method. All species of the genus Omithoptera, some species of the genus and other species representing most genera of the tribe Troidini were used. Results suggest the genus Omilhoptera forms a clade with Troides, supporting the previous results (Morin aka el al., 1999). In the case of the genus Omilhoptera, each of the subgenera Omithoptera and Schoenbergia was found to be monophyletic, however, phylogenetic relationships of O. alexandrae and O. vicloriae (the subgenus Aetheoptera) could not be clarified. In the subgenus Schoenbergia, two clades (0. paradisea + O. meridionalis, and O. goliath + O. chimaera + O. rothschildi) were found, however, the phylogenetic relationships of O. tithonus and other species remain unclarified, so as in the case of the genus in the tribe Troidini.

Key words: Aetheoptera, birdwing butterflies, mitochondrial DNA, molecular phylogeny, NOS, Omithoptera, Ripponia, Schoenbergia, Trogolloptera, Troides, Troidini.

Introduction region), Central to South America and Indo- Australian regions. This tribe includes bird wing The tribe Troidini includes about 10 genera, butterflies, which are important for mankind because which inhabit Madagascar Is. (the Afrotropical their fate, except some species adapting to the

-103- Molecular Phylogeny of Birdwing Butterflies of the Tribe Troidini countryside, is linked to the existence of tropical treated as hybrids of O. victoriae X O. priamus, and . They are considered as a symbol O. rothschildi X O. priamus, respectively, and O. exemplifying biodiversity. richmondia and O. euphorion as subspecies of O. Many taxonomic and/or phylogenetic studies on priamus. Permission was obtained for species troidine butterflies have been published (Zeuner, protected by the international contract, "Conservation 1943; Ford, 1944; Munroe & Ehrlich, 1960; Munroe, on International Trade in Endangered Species of Wild 1961; Hancock, 1983; Miller, 1987; Haugum & Low, Fauna and Flora" (C.I.T.E.S.) or other regulations 1978-1985; D' Abrera, 1975; Igarashi, 1979, 1984; when necessary. Tsukada, 1980; Ohya, 1983; Tyler et al., 1994; Fujioka et al., 1997; Parsons, 1999: Morinaka et al., Preparation of mtDNA, amplification and 1999). Their classifications are partly different from sequencing of ND5 gene each other. Morphological characters are useful for Details of DNA extraction, amplification and investigating phylogeny, whereas DNA sequences sequencing techniques for the ND5 gene of mtDNA, encode more discrete characters, determination of are described in Morinaka et ai. (1999). Nucleotide which would yield a high degree of accuracy sequences of the primers are as follows: a, 5'- (Brower, 1996; Su et al., 1998; Caterino & Sperling, CCTGTTTCTGCTTTAGTTCA-3' (Su et al., 1996); 1999; Yagi et ai., 1999, Maekawa et al., 1999). We and B3, 5'-TAACCTCTATATATYTCTCTT-3'. have previously reported that the genera of Troidini excluding the genus Bartus are well united, forming Phylogenetic Analyses one clade; Tl'Oides and Ornithoptera also form one Two or more individuals from an intrataxon were clade (Morinaka et al., 1999). However, the used for examination, except for some species which phylogeny of Trogonoptera and the intrageneric one were difficult to obtain. In the analyses, 877 of the of Ornithoptera remain unclarified. In this study, 924 nucleotide sequences were used (Fig. 1). phylogenetic analyses are carried out using Editing and aligning of the sequence data were Trogonoptera and all species of Ornithoptera. completed utilizing DANASIS (Hitachi Software Engineering Co., Hitachi) and CLUSTAL W Materials and Methods (Thompson et ai., 1994). Phylogenetic trees were constructed by the maximum-likelihood (ML) Taxa used in the study method, maximum-parsimony (MP) method and Taxa used in this study are shown in Table I, neighbor-joining (NJ) method with the evolutionary including their sampling localities and accession distances by Kimura's two-parameter method using numbers in DDBJ/EMBLlGenBank. Species name of PAUP Version 4.0b4a (Swofford, 2000). The Troides, Ornithoptera and Tl'Ogonoptera are based on statistical confidence of each clade was examined Aoki et al. (1998). O. aiotti and O. akakeae are with the bootstrap tests (Efron, 1982; Felsenstein,

a: 20 mer 83: 21 mer ... 924 bp .... His UbP I Phe 877 bp N05 Gene

Fig. I. Schematic representation of the amplified and coding regions of the NDS gene used in the study. a: 5'-CCTGTTTCTGCTTTAGTTCA-3'; B3: 5'- TAACCTCTATATATYTCTCTT-3'.

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Table I. Specimens used in the study (classified arbitrarily).

Genus (Subgenus) Species Locality DDBJI EMBU GenBank Accession Number hahneli Brasil AB027579 Par. elraces Upper HualJaga Y., Peru AB027580 Byasa alcinous Tokyo, Japan AB027583 B. polyeuctes Formosa AB044646 Atrophaneura luchti Jawa Is., Indonesia AB027584 A. nox Bali Is., Indonesia AB027586 Losaria coon Jawa Is., Indonesia AB027587 Pachliopta aristolochiae Bali Is., Indonesia AB027590 Troides (Ripponia) hypolitus Sulawesi Is., Indonesia AB027592 T. (Troides) amphrysus Jawa Is., Indonesia AB027593 T. (T. ) helena Bali Is., Indonesia AB027594 T. (T. ) haliphron Sulawesi Is ., Indonesia AB044649 T. (T. ) prattorum Buru Is., Indonesia AB044650 T. (T. ) staudingeri Lakor Is., Indonesia AB04465 I Ornithoptera (Ornithoptera) priamus Seram Is., Indonesia AB027596 O. (0. ) priamus Malaita Is., Solomon Isis AB027597 O. (0. ) priamus Timika, Irian Jaya AB044656 O. (0. ) priamus I Is., Solomon IsIs AB044657 O. (0. ) croesus Halmahera Is., Indonesia AB027595 O. (0. ) croesus Bacan Is., Indonesia AB044654 O. (0. ) aesacus Obi Is., Indonesia AB044655 O. (Aetheoptera) alexandrae Papua AB044652 O. (A. ) victoriae Malaita Is., Solomon Isis AB027598 O. (A.) victoriae Gela Is., Solomon Isis AB044658 O. ( Schoenbergia) paradisea Timika, Irian Jaya AB027603 O. (S. ) paradisea Arfak Mts, Irian Jaya AB044659 O. (S. ) meridionalis Timika, Irian Jaya AB044653 O. (S. ) goliath Seram Is., Indonesia AB027599 O. (S. ) goliath Arfak Mts, Irian Jaya AB027600 O. (S. ) rothschildi Arfak Mts, Irian Jaya AB027601 O. (S. ) chimaera Wayland Mts, Irian Jaya AB044660 O. (S. ) tithonus Arfak Mts, Irian Jaya AB027602 Padang, Sumatera Is. AB044647 Trog. brookiana Cameron Highland, AB044648 protenor Tokyo, Japan AB027604 Pieris (Pieridae, as outgroup) rapae Mie, Japan AB044594

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P. ro"'poe Ahhrcvialions: r :;llIlding.:r; AI I : A.-fak MiS, lriun Jay" Iio/iphr. (I 8m.;: Bi1(.·tm Is v' T. prof/arum Cam; Cameron Highland Gd : Gclu. Solol11on Isis T f,elello GUll: GumJ<1lcanul. Solomon Isis r gmphl)'slIs Hal: Hnlmnhclu Is T hypolilus Mal: MaJaila, Solomon [sis ,------O . o/sxol1drcre Aelileoptera SCI: S!.:ram Is Sum: SUIOi.llcra r;;:--- 0 pad Tim Tim : Tirnika, Ilian Juyu '---- 0 men 'ono/(s golio/h i\rr Sclroe"hergiaJ ,,,. 0.. 0.. o llOSer 0. chimaera ____":.- .:.._-:..--_-_-_-_-_- 0. lolhschildi Omifhopfero L O. filhonus L _"-.. pr/omu!> Tim ' ()d. JflOmU5 Sar . POOIT/US Gu. 0 ill tJ o ptiomus M:t! rJI lOp era o.. aesaCfiS I QoIJO. croesus Bac 12 0. croe5u5 Hoi o.. "it...! /i ,,001 O. vic/a fiae M .1 B !X,I /,JeI 1'-4 B oHnous L_ __ .. A luchli 1------4% ====- A. nox L ____ '" Pa r. hahne/i Poe ori5/%chioe 91 t coon '-______-"-' '',' Trag. bfLlOk/al1o :\"'" Trogonoptero Trag brookiono Cam I prolenor -- 001 subslitutions/site a

r------P ropae T, 510udinge ri f. ho/iphron T prof/arum T he/ella '------' T ompluysus T /lypo/iIV$ ,------() 01 ondtne Aetheoplera " O .... priamus Tim 67 O.pnomus Ser j II 0 priomus Gua O. priomus Mal OrllltllOptera O . oe50CUS O. croe5us Bac O. croesus Hal Omifhop/era O . porodlr!)' T.m '----- 0 me/idiono/is j 0., go/,a/lt Arl' Scl,oel/hergia .12 0 .. 9 Jlo Ser I., 0. '------0 /ilhonus L------fiWll" 0.. IIICI(;>1 /oe Gel 0.. vlc/orloe Ml.1 ------...I ::---E====-EZ-::i. B. po/yeltc/es [ B. "'CIIlOU5 1::. "-.--- -- A. "d,n fl O Par e lr'i!.ces '" r-or. hohne/i L ori5l%chioe ___ tn. L. coon L....______'""' ''I T/Of) brookiono Sum Trogonoplera Trog brookiono Cam I L....______Pap. pro/enar --- 10 changes b-1 Fig. 2 Phylogenetic trees of birdwing butterflies based on 877 nucleotide sequences of the N05 gene (mtONA), constructed by the three methods using PAUP 4.0b4a. The bootstrap values of 50% or more besed on 1000 replicates are shown at each branching point. Abbreviations are shown in Fig. 2a. a: By ML method (Stepwise addition: as-is; Branch swapping: tree-bisection-reconnection); b(1-2): By MP method (General:

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rJpoe r------T ./oudlll ali T hnl/{"Ihr q '------T p/o/torum T '------T amphrysu5 T l"!}"polfllJ5 ,------U ·, o/exandtae "D._plio!'"lS 67 a nncupvs 5tl j 1I J1J u . Gl,.la O . ptlamU5 M .lIl Or"itllOptera ,------l .. D . oesocus v 0 croesus Bac O. croesus Hal Omithoplera r::,.-- 0 porod/seo Arf "" D porodlseo Tim '------O . meridionoli5 J 6.' .. O . goJiruh Arf Sc/wel/bergia " 901/0 Sel' 0 , chlm era ., 0 ro/hscMdl '------0 '------l'W 0 viclorioe Gel ______--' o victoriae Mal B. polyeuc1es .. " B. alcrnous L____ -{... A. Iltchli A. /lOX L ___ ... elParmc&1 I1nhnell L____ orisl%chioe L coon '-______..ll"'''''I. Trag. brookinno Sum Trogonopleral Trog . brookiono Cam '------Pap. prolenor ---- 10 changes b-2 r------P. rapae 1. staud,rgeri 1. l'lCl /iphron F.:----- T. pralt . lum T he lena T '------T hYRD"lus ,------0 oIe)(o,.,d,oe Aetheoptera ".::-, ------0. poroelisea Al'f 0 pCll or/i .• eo Tim '------O . mer/clonoI l'5 '------O. lilllolwS ,"0 o goliolh Arf ,,_.J , O . go/ia Ser O . Chlmtlyl,('Id Omithoptera '-'-'------O . rothscnrl< I 'J O. victorioe Gel VI 0 vicloriae Mal £ O. pflO/IJu .• Tim 16 0 eriamu5 Ser III O . prfomvs Gua '------1'00 0 priam Mal 0 , Oe50C(JS V'r 9\10 croesus Bac (,) t. f' Hal .------'"",'l'l II'P9 Grookiono Sum j Tr g. bro kiano Cam Trogonopleral L. coonPOc:. orisloIor.;hfoe OJ B. a/croousB. po lye IJdes '- A Anox IlJc hl/ L___ ., Par hahneli '------Pap. prolenor --- 001 changes c Minimum trees only; Starting trees: Obtained by stepwise addition; Stepwise addition: 200 random reps; Branch swapping: tree-bisection-reconnection). Two most parsimonious trees (tree length: 1186, CI:0.460) were obtained; c: By NJ method with the evolutionary distances by Kimura's two-parameter method.

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1985) based on 1000 replicates in the three methods with O. alexandrae and Schoenbergia (NJ) . using the same software above. The values of 50% or Trogonoptera brookiana is not shown to form a clade more are shown at each branching point. with other birdwing butterflies in any trees. Phylogenetic trees constructed based on amino acid sequences encoded in the same region were Discussion supportively confirmed by MP and NJ (not shown). Pieris rapae (Lepidoptera: Pieridae) was used as an Previous studies suggest that birdwing butterflies outgroup species for the analyses. underwent diversification over a long period (Zeuner, 1943; Corbet, 1944); therefore, results of Results phylogenetic analysis tend to be ambiguous. We attempted to analyze as many sequence data and Only very few differences in their pairwise genera of Troidini as possible. We analyzed 6 species nucleotide sequences were found in some individuals of Troides and all species of Ornithoptera; however, of the same troidine species for the same area. The , Cressida and were not number of differences in sequences among troidine included because of the lack of some nucleotide species was I - 133/877 bp (0.1 - 15.2%). The G+C sequence data. content of the sequenced region ranged from 16 to 20 In Ornithoptera, relationships between species in %. Neither deletions nor insertions were found. This the subgenus Ornithoptera, O. tithonus and two sequenced region was considered to be useful for this clades (0. paradisea + O. meridionalis, and O. analysis. goliath + O. chimaera + O. rothschildi) in A phylogenetic tree constructed by ML, two most Schoenbergia remain unclarified. In bird wing parsimonious trees (tree length: 1186; CI: 0.460) by butterflies, the relationship of Trogonoptera with MP and a tree by NJ are shown in Fig. 2 with the other genera was also unclear. These obscure results bootstrap values of 50% or more. In these trees, the are considered to be caused by decreasing accuracy of same phylogenetic relationships of some birdwing nucleotide sequence information because of their butterflies are found. T. staudingeri and T. haliphron, rather long evolutionary period. In addition, it O. paradisea and O. meridionalis, and O. chimaera seemed that the lack of nucleotide sequence data for and O. rothschildi are shown to be sister species. O. some genera rendered the results unclear. goliath forms one clade with another clade of O. However, some interesting results were obtained. chimaera and O. rothschildi. Results suggest that each of the subgenera In Ornithoptera, each of the subgenera Ornithoptera and Schoenbergia is monophyletic. Schoenbergia and Ornithoptera forms one clade. T. Two groups (0. paradisea + O. meridionalis, and O. hypolitus (the Subgenus Ripponia) forms one clade goliath + O. chimaera + O. rothschildi) were found to with another clade of all other species of Troides and be one group, although the confidence of the clades Troides forms one group. Ornithoptera forms one was not so high. Troides was found to be one group clade with Troides based on their high bootstrap with high bootstrap values. Ornithoptera formed one values in all trees. Other relationships, however, are group as determination by MP and NJ, however, the inconsistent depending on the methods. The genus bootstrap value was considerably low (52% in MP). Ornithoptera forms one clade in MP and NJ, Moreover, taking into consideration the relationships however, it was divided into two groups by ML. In of G. victoriae with other species as obtained by ML, particular, the relationship of O. victoriae (the more studies should be carried out in determining subgenus Aetheoptera) is markedly inconsistent phylogenetic position of Ornithoptera. among trees, forming a clade with Troides and Comparing our results with Parsons, phylogenetic Ornithoptera (ML), with Ornithoptera (MP), and trees of Ornithoptera (Parsons, 1996 [Fig. 17 A)), the

-108- S. Morinaka, N. Minaka, M. Sekiguchi, Erniwati, S. N. Prijono, 1. K. Ginarsa, T. Miyata and T. Hidaka relationship between O. paradisea and O. Physical Culture Junior College), Mr Keiichi meridionalis as well as that between O. chimaera and Matsumoto, Mr Jiro Arai and Mr Ichiro Kurosaki O. rothschildi are similarly shown to be sister species. (Tokyo Metropolitan Takao Meseum of Natural These results are considered to be good examples of History) for lending precious materials. We express compatibility between molecular and morphological our thanks to Mr Shinichi Oshima (Saitama), Mr approaches. O. richmondia has been treated as Yasusuke Nishiyama (Tokyo), Mr Yasushi Sorimachi another species distinct from O. priamus (Common & (Saitama), Mr Tetsuo Miyashita (Tokyo), Mr Kaoru Waterhouse, 198). Parsons (1996) also concluded Adachi (Tokyo), Mr Akihiko Takenaka (Hyogo) and that O. richmondia is another species Mr Hiromi Detani (Bali, Indonesia) for their great phylogenetically far from O. priamus as compared to efforts to obtain permission and/or precious O. croesus or O. aesacus. This idea is very specimens for the study. We thank Dr Takashi Yagi interesting subject for our future studies of molecular (Osaka Prefecture University), Mr Katsuya Natsuhara phylogeny. (Nagoya University), Mr Noriaki Asou (Kanagawa), In conclusion, the present study confirmed the Mr Kazuaki Seta (Tokyo) and Mr Naokazu Nagahata close relationship between Ornithoptera and Troides, (Saitama) for their kind help and also Mr Yoso as well as the subgenera Ornithoptera and Bumantoro (Irian Jaya, Indonesia) for obtaining Schoenbergia to be monophyletic, respectively, as permission from C.I.T.E.S and precious specimens. reported by many previous investigators based on the We are grateful for the grant-in-aid No. 99-B3-008 morphology. The relationships of O. alexandrae and from "The Toyota Foundation". O. victoriae with other species, as in the case of Trogonoptera in Troidini, however, remain References unclarified and therefore should be the subject of future studies. Aoki, T., Uemura, Y., Otsuka, K., Nishizawa, T. & Yamaguchi, S. 1998. The World of Birdwing Acknowledgments Butterflies. In Kondo, N. & Nishida, M. (Eds), The Research Institute of Evolutionary Biology, We expres s our gratitude to Dr Sugihiko Library No.3. Shinzansha, Tokyo. (In Japanese.) Hoshizaki, Dr Yukio Ishikawa and Dr Sadahiro Brower, A. V. Z. 1996. Parallel race formation and Tatsuki (University of Tokyo) for their kind support the evolution of in Heliconius butterflies: and advice on molecular biology. We wish to A Phylogenetic hypothesis from mitochondrial acknowledge Dr Kenji Tanaka (Aichi Study Center, DNA sequeuas. Evolution, 50: 195-221. The University of the Air) for supporting our study. Calerino M. S. & Sperling F. A. H. 1999. Papilio We express our thanks to Dr Soetikno Wirjoantmodio Phylogeny Based on Mitochondrial Cytochrome and Dr Mohamad Amir (Indonesian Institute of Oxidase I and II Genes. Molec. Phylogenet. Evol., Science) for their kind support of our study since I 1: 122-137. "International Conference" in 1993, and Dr Common, 1. F . B . & Waterhouse, D. F. 1981. Hideo Mohri (National Institute for Basic Biology), Butterflies of (2nd, revised Edn). 682 Dr Tohru Nakazawa (The University of the Air), Dr pp., 49 pIs. Sydony. Tomohiro Maeyama, Dr Kiyoto Maekawa, Dr Tadao Corbet, A . S. 1944. Paleontology without fossils in Matsumoto (University of Tokyo), Mr Shigero Sugi the 'bird-wing' butterflies. Nature, 153: 32. (Tokyo), Mr Kazuhisa Otsuka (Tokyo University of D' Abrera, B. 1975. Birdwing Butterflies of the Agriculture) and Dr Osamu Kitade (Ibaraki World. Lansdawne Press. Melbourne. University) for support of our study. We express our Efron, B . 1982. The jackknife, the bootstrap and thanks to Mr Kunio Matsuda (Tokyo Women's other resampling plans. CBMS-NSF regional

-109 - Molecular Phylogeny of Birdwing Butterflies of the Tribe Troidini

conference series in applied mathematics, Ohya, T. 1983. Birdwing Butterflies. Kodansha Co. Monograph 38. SIAM, Philadelphia. Ltd. Tokyo. (In Japanese.) Felsenstein, 1. 1985. Confidence limit on Parsons, M. J. 1996. Gondwanan evolution of the phylogenies: an approach using the bootstrap. troidine swallowtails (Lepidoptera: Papilionidae): Evolution, 39: 783-791 . cladistic reappraisals using mainly immature stage Ford, E. B. 1944. Studies on the chemistry of characters, with focus on the birdwing the Lepidoptera, with reference to Omithop/era Boisduval. Bull. Kitakyushu Mus. their bearing on systematics. 4: The classification nat. His!., 15: 43-118. of the Papilionidae. Trans. R. ent. Soc. Lond., 94: Parsons, M. J. 1999. The Butteflies of Papua New 201-223. Guinea: their Systematics and Biology. Academic Fujioka, T. Tsukiyama, H. & Chiba, H. 1997. In Press., London. Fujioka, T. (Ed.), Japanese Butterflies and Their Shirozu, T. 1960. Butterflies of Formosa in Color. Relatives in the World I. Shuppangeijutusha. (In Hoikusha Publishing Co., Ltd. Osaka. Japanese.) Su, Z. -H., Ohama, T., Okada, T. S., Kajiwara, E., Hancock, D. L. 1983. Classification of the Nakamura, K., Ishikawa, R. & Osawa, S. 1996. Papilionidae (Lepidoptera): a phylogenetic Geography-linked phylogeny of the Damaster approach. Smithersia, 2: 1-48. ground beetles in ferried from mitochondrial ND5 Haugum,1. & Low, A. M. 1978-1985. A Monograph gene sequences. J. molec. Evol., 42: 130-134. of the Birdwing Butterflies, I: 1-308; 2: 1-355. Su, Z. -H ., Tominaga, 0., Okamoto, M. & Osawa, S. Scandinavian Science Press Ltd. Klampenborg. 1998. Origin and Diversification of Hindwingless Igarashi, S. 1979. Papilionidae and their early Stages. Damaster Ground Beetles Within the Japanese Kodansha, Tokyo. (In Japanese.) Islands as Deduced from Mitochondrial ND5 Gene Igarashi, S. 1984. The classification of the Sequences (Coleoptera, Carabidae): Molec. BioI. Papilionidae mainly based on the morphology of Evol., 15: 1026-1039. their immature stage. Tyo Ga, 34: 41-96. Swofford, D. L. 2000. PAUP*: Phylogenetic Analysis Maekawa, K., Kitade, O. & Matsumoto, T. 1999. Using Parsimony (*and Other Methods). Version Molecular Phylogeny of Orthopteroid 4. Sinauer Associates, Sunderland, Massachusetts. based on the Mitochondrial Cytochrome Oxidase Thompson, J. D., Higgins, D. G. & Gibson, T. J. II Gene. Zool. Sci., 16: 175-184. 1994. Clustal W: Improving the sensitivity of

(: Miller, J. S. 1987. Phylogenetic studies in the progressive multiple sequence alignment through (Lepidoptera: Papilionidae). Bull. am. sequence weighting, positions-specific gap Mus. nat. Hist., 186: 365-512. penalties and weight matrix choice. Nucleic Acids Morinaka, S., Maeyama, T., Maekawa, K., Erniwati, Res. 22: 4623-4680. D., Nuramaliati, S. P., Ginarsa, I. K., Nakazawa, Tsukada, T. & Nishiyama, Y. 1980. Papilionidae. In T. & Hidaka, T. 1999. Molecular Phylogeny of Tsukada, T. (Ed.). Butterflies of the South East Birdwing Bullerflies Based on the Representatives Asian Islands. Part I. Plapac Co., Ltd. Tokyo. (In in Most Genera of the Tribe Troidini (Lepidoptera: Japanese.) Papilionidae). En!. Sci., 2: 347-358. Tyler, H., ·Brown, Jr, K. S. & Willson, K. 1994. Munroe, E. G. 1961. The classification of the Swallowtail Butterflies of the Americas: A study Papilionidae (Lepidoptera). Can . Enl. in biological dynamics, ecological diversity, (Supplement), 17: 1-51. biosystematics, and conservation. Scientific Munroe, E. & Ehrlich, P. R. 1960. Harmonization of Publishers. Florida. concepts of higher classification of the Yagi, T., Sasaki, G. & Takebe, H., 1999. Phylogeny Papilionidae: J. Lepid. Soc., 14: 169-175. of Japanese papilionid butterflies inferred from

-110- S. Morinaka, N. Minaka, M. Sekiguchi, Erniwati, S. N. Prijono, I. K. Ginarsa, T. Miyata and T. Hidaka

nucleotide sequences of the mitochondrial ND5 Archipelago. Trans. zoo!. Soc. Lond., 25: 107- gene. 1. molec. Evo!., 48: 42-48. 184. Kurosawa, Y. (trans!., 1983). Acta Zeuner, F. E. 1943. Studies in the systematics of Rhopalocerologica, 4: 2-65. (In Japanese.) Troides HUbner (Lepidoptera Papilionidae) and its allies: Disdtribution and phylogeny in relation to (Accepted July 21,2000) the geological history of the Australian

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