The JapaneseSocietyJapanese Society for Plant Systematics ISSN 1346-7565 Acta Phytotax. GeoboL 56 (2): 14]-161 (200S)･ Molecular Phylogenetics of Phalaenopsis (Orchidaceae)and allied Genera: Re-evaluation of Generic Concepts TOMOHISA YUKAWAi, KOICHI KITA2, TAKASHI HANDA2, TOPIK HIDAYAT3 and MOTOMHTo3 i71sukuba 21hstitute Botanical Garcien, Nlational Scienee Mtiseum, Amakuho, Tyuketba, 305-OO05. Jopan; of 3Graduate Agricultnre andforestn)). Uhivensity qf'Tgukuba, fennodai, 71yukuba, 305-857Z Japan; Schoot ofArts and Seience, Uhivensity of7bdy,o, Kbmaba, 7bkyo, J53-8902, JZu)an, Molecular phylogenetic analyscs were performed using data sets derived from DNA sequences ofthe plastid genome (matK and trnK introns) and the nuelear genome (rDNA ITS) in an examination ofrela- tionships of all sections ofPhataenqpsis and closely related gcnera. The fo11owing insights were pro- vided: (1) The genera Lesliea and IVbthodoritis are nested within Phalaenopsis, (2) Phalaenopsis subgenus Aphyilae and section EsmeJ'aldd, often treated as thc independent genera Kirrgidium and Doritis respectively, are also nested within Phalaenqpsis. (3) Two subgenera of Phalaenqpsis, namely, Phalaenopsis and 1larishianae, are not monophyletic. (4) Phalaenopsis sections Deliciosae, SZautqglottis, Amboinenses and Zehrinae are not monophyletic. (5) lnconsistencies bctween the plastid and nuclear lineages indicate a hybrid origin ofPhalaenopsis minus and Phalaenopsis phitmpinensis. (6) In light of these findings, and to accommodate phylogenetic integrity and stability in nomenclature, we adopt a broadly defincd Doritis characterized by the possession of fbur pollinia, an explicit character state. Key words: Doritis,introgression, ITS, mati(l moleculag Orchidaceae, Ahalaenopsis, phylogcnctics, tttnK Phakzenopsis Blume is an orchid genus to which 62 tion ofthe genus has been thoroughly reviewed by species are currently assigned (Christenson 2001). Sweet (1980). In the present study, we fbllow the According to Dressler's classification ofthe Orchid- system proposed by ChristensQn (2001) as this aceae (1993), the genus belongs to subfamily wotk incorporates the most recent data C[fable 1), In Epidendroideae Lindl. tribe Vlindeae Lindl. sub- his reyision, Christenson (2001) treated Phalae- tribe Aeridinae Pfitzen The genus occurs from nopsis in a broad sense, sinking the genera Doritis southern Indiaand SriLanka in the west to New Lindl. and Kingidium P. F. Hunt, both of which Guinea and northern Australia in the east, It extends have been treated as independent genera by other as far north as southem China and 1faiwan. The authors (e.g. Seidenfaden 1988, Dressler 1993), center of diversity of the genus is Borneo, from intoPhalaenopsis, where 13 species are currently known, This major inconsistency among taxonomists is Several monographic treatments ofthe genus largely due to different weighting attributed to the have been published notably by Roife (1886), Sweet critical morphological character, pollinium num- (1968-69, 1980) and Christenson (2001). Shim ber. Although polliniurn number has traditionally (1984) revised the generic classification wnh empha- been censidered a heavily weighted character in sis on floral morphology, The history ofclassifica- the classification oforchid genera, its state is not uni- NII-Electronic Library Service TheTheJapaneseSocietyforPlant Japanese Society for Plant Systematics 142 APG Vbl. S6 TABI,E 1 . Species of Rhalaenopsis and allied genera used for matK-trnK introns and ITS sequencing. Infrageneric classification fo11ows Christenson (2001), 'Sp 'ecles Vbucher Sllbgenus Probescidioidds (Rolfe) Christe]son Phalaenopsis Rchb. lowii f TBG i44316* Subgenus !{plij,tlae (H. R. Sweet) Christenson Phataenopsis braceana (Hook. f.) Christenson 7BG 144566 Phaldenopsis minus (SeidenE) Christenson 71BG 145839 PhataenopsiswilsoniiRolfe TIBG 144214 Phalaenopsis sp. litkawa 29689 Subgenus ilarishianae (H. R. Sweet) Christenson Phalaenopsis uppendiculata C. E. Carr 71BG 144305 Phataenqpsis lobbii (Rchb. £ ) Sweet 7BG 78865 Phalaenopsis parishii Rchb. f. 71BG 13388J Subgenus Ilolychiles (Breda) Christenson Section Ilolychilos (Breda) Christenson Phalaenopsis cornu-cervi <Breda) Blurne & Rchb. f. T:BG 145696 Phalaenopsis mannii Rchb. f 713G l18380 Rchb. f, 7)vukahara s, n. Phalaenopsispantherina Section Fuscatae H. R. Sweet cochlearis Phalaenopsis Holttum .71BG14420P PhalaenopsiskunstleriHoek. f 71BG137083 Phalaenopsis viridis J. J, Sm. ZEIG 14J055 Phalaenqpsis sp. TBa 14223S Section Amboinenses H. R. Sweet Phataenopsis amboinensis J, J. Srn. TZIG133762 Phalaenopsis bastianii Gruss & ROIIke 71BG140637 Phalaenopsis bellina (Rehb. f) Christenson 71BGi18533 Phataenopsis db",et:ytinsis Garay & Christenson llBG144564 Phalaenopsisfasciata Rchb. f 7BG 145 726 Phalaenopsis.fimbriata J, J. Sm, subsp, suneatrana (J. J, Sm.) Christenson 71BGf45860 PhalaenopsisY7oresensisFowlie llBG145748 Phataenopsis gigantea J. J. Sm. 71BG i3 7307 Phalaenopsis hiertrglyphica (Rehb, f) H. R. Sweet ZBGf45743 PhataereopsisJ'avanica J. J. Sm. 71BG 14586S Phalaenopsis luecidemanniana Rchb. f 71BG145733 Phalaenopsis lueddemanniana Rchb. tl var. ochraeea Rchb. E 71EIGI45863 Phalaenopsis maculata Rchb. f. 7BGJ44569 Phalaenopsis mariae Burb. ex R. Warneer & B. S. Williams 7:BGJ41056 Phalaenopsis micholitzii Rolfe TZIG140.566 Phalaenopsih' mode,sta J, J, Sm. 71BG 140664 Phalaenopsis pallenE (Lindl,) Rchb, fi llBG145744 Phalaenopsispttlckra (Rchb, fi) H, R, Sweet 7:BG145761 Phataenopsis reichenbachiana Rchb, f & Sander 7]BG145S75 Phalaenopsis venosa Shim & Fowlie 7BGJ45770 Pkalaen(tpsis violacea Wjtte 71BCf45785 SectionZlebrinaePfitzer Phalaenopsis inscriptiosinensis Fowlie M(l i44571 Phataencrpsis sumatrana Korth, & Rchb. fi 1:BG i42440 Phalaenopsis tetra,spis Rchb, f 7]BG145841 NII-Electronic Library Service The JapaneseSocietyJapanese Society forforPlant Plant Systematics August2005 YUKAXMPL et at.: Molecular phylegenctics of Phataen opsis 143 TvLBLE 1.(continued). Speeies Vbucher SubgenusPkalaenopsis Section IViataercopsis Phalaenopsis amabitis (L.) Blumc TIIG145847 Phalaenopsis amabilis (L.) Blume subsp. rosenstromii (F, M. Bailey) Christenson 71BG140316 Phalaenqpsis aphrodite Rchb, f (1) 7BC14140S Phaiaenopsis aphrodite Rchb. £ (2) 71]G141I51 Phataenopsisphilijzpinensis Gelamco ex Fowlie & Tleing 7LBG118SS2 Phalaenopsis sanderiana Rchb. E Atagawa s. n. Phaiaenopsis schilleriana Rchb. f IIBG 0309 Phataenopsis stuartiana Rchb, f, 7BG l45 758 SectionDeliciosaeChristenson Phaiaenopsis chibae T, Yukawa 71BG115846 Phalaenopsis deticiosa Rchb. f, (1) 7BG145842 Phaiaenopsis dltliciosa Rchb. f. (2) 1;BG144594 Section Esmeralda Rchb. f, Phaiaenopsispttlcherrima (Lindl.) J. J. Srn. 71BG1J8342 "buyssoniana" Phalaenopsis sp, TBG l45823 Section stauroglottis (Schauer) Benth. Phaiaenopsis equestri.s (Schauer) Rchb. £ 71BG141154 Phalaenopsis cetebensis H, R. Sweet ZBG J45822 Phalaenopsis lindenii I.oher ZBG140568 Allied taxa Lesliea mirabitis Seiden£ TLBG145844 IVbthodoritis zhojiangensis Z. H. Tsi TBG f3 750i O"tgroupAmesiella monticola J.E. Cootes &D. P, Banks 7BG 123790 *TBG series indicate accession nurnbers in living cellection database at Tsukuba Botanical Garden. fbrm within Phalaenctpyis. The predoininant number shed their leaves during the dry season. in the genus is two, but the subgenera Proboscidi- Phylogentic relationships in Phataenopsis oides, Aplryllae and Ftirishianae all have four pol- remain largely unresolyed, Woodward (1951), linia, as do the sections Deliciosae and Esmeralcla Sagawa (1962), Shindo & Kamemoto (1963), footh in subgenus Phalaenopsis). Immense diversity Sagawa & Sheji (1968), Arends (1970), Sheji (1976, in the structure Qf reproductive organs, particularly1980), Aoyama (l993), Aoyama et al. (1994) and that of the labellum and the stipe, has also resulted Kao et at. (200 1 ) investigated cytological characters, in inconsistent interpretations of relationships in Brandange et aL (1971, 1972) examined alkaloid the group, content and Tsai et al. (2003) conducted a molecu- Phalaenopsts also exhibits rematkable diversitylar phylogenetic analysis ofthe intemal transcribed in terms of ecology. Most species are epiphytes, spacer rcgions of the 18S-26S nuclear ribosomal though the species of section Esmeralda have a DNA (ITS) in subgenus Phalaenqpsis. ln the present tcrrestrial life form. On the other hand, plants in sub- study, we compared and combined DNA sequences genera Apilyllae, IZfirishianae and Proboscidioides of a maturase-encoding gene (matK) flanked by NII-Electronic Library Service TheTheJapaneseSocietyforPlantSystematics Japanese Society for Plant Systematics 1" APG Xlo1. 56 Subgenus Section 811008874 P.arnabilis Phalaenepsis Phalaeriopsis P. amabilis subsp. rosenstromii Phalaenopsis Phalaenopsis P. aphrodite (1) Phalaenopsis Phalaenopsis 6010016 P. aphrodite Phalaenopsis Phalaenopsis 75 (2) P. sznderlana Phalacnopsis Phalaenopsis 313 P. IilldeniiP, Phalacnopsis Stauroglottis celebe]sis Phalacnopsis Stauroglottis lee P. equestris Phalaenopsis Stauroglottis 99 73 P, philippincnsis Phalaenopsis Phaluenopsis loe98 P, stuartiana Phalaenopsis Phulaenopsis 100 P, schilleriana Phalacnopsis Phalaenopsis P sp.R PolychilosPo]ychilosl'olychi]osPolychilosPolychilosPolyehitosPolyehilosPolychitosPolychilosPolychilosPolychilosPetvchilosPolychilosPolvchilosPolychilosPolychilosPolychilosPolychilosPolychilosPolychilosFuscataeFuscataeFuscataeFuscataeAmboinenses 10064 kunstleri 100g4 P. cochlcaris P. viridisP. leo doweryensis 100 P, giguntea Ambeinenses P. maculata Amboinenses