Molecular Phylogenetics of Phalaenopsis (Orchidaceae) and allied Genera : Re-evaluation of Generic Concepts

2005·1.8 MB·English

by  Yukawa, Tomohisa

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ISSN 1346-7565 Acta Phytotax .GeoboL 56 (2) :14]-161 (200S)･
MolecularPhylogenetics Phalaenopsis(Orchidaceae)
of and allied
Genera: Re-evaluation Generic Concepts
of
TOMOHISA YUKAWAi, KOICHI KITA2, TAKASHI HANDA2, TOPIK HIDAYAT3
and MOTOMHTo3
i71sukuba Botanica lGarcien N,lational Sciene eMtiseum ,Amakuho, Tyuketb a3,05-OO05 .Jopan; 21hstitute of
Agricultn raendforestn)). Uhivensity qf'Tgukub af,ennodai ,71yukub a3,05-857Z Japan; 3Graduate Schoot ofArts
and Seience ,Uhivensity of7bd yK,bmoa,ba, 7bkyo ,J53-8902 ,JZu)an,
Molecular phylogeneti acnalyscs were performed using data sets derive dfrom DNA sequences ofthe
plasti gdenome (mat Kand trnK intron sa)nd the nuelear genome (rDNA ITS) in an examination ofrela-
tionships of all sections ofPhataenqpsis and closely related gcnera .The fo11owin ginsigh twsere pro-
vided: (1 )The genera Leslie aand IVbthodorit airse nested within Phalaenopsis, (2 )Phalaenopsis
subgenus Aphyila eand section EsmeJ'ald dof,ten treate das thc independen gtenera Kirrgidi uamnd Doritis
respectively, are also nested within Phalaenqpsis .(3 )Two subgenera of Phalaenqpsis, namely,
Phalaenopsi asnd 1larishianae a,re not monophyletic. (4 P)halaenopsi ssections Deliciosae ,SZautqglottis,
Amboinenses and Zehrinae are not monophyletic. (5 )lnconsistencies bctween the plasti adnd nuclear
lineage isndica tae hybrid origin ofPhalaenopsis minus and Phalaenopsi pshitmpinensi (s.6 )In ligh tof
these finding sa,nd to accommodate phylogeneti icntegri tanyd stability in nomenclature, we adopt a
broadl ydefinc dDoriti scharacterized by the possessio onf fbur pollinia ,an explicit character state.
Key words: Doritis ,introgressio nI,TS, mati(l moleculag Orchidacea eA,halaenopsi sp,hylogcncti cttst,nK
Phakzenopsis Blume is an orchid genus to which 62 tion ofthe genus has been thoroughly reviewed by
species are currently assigned (Christen s20o0n1). Sweet (1980 I)n. the present study, we fbllow the
According to Dressler' cslassification ofthe Orchid- system proposed by ChristensQ n(2001 )as this
aceae (1993 )th,e genus belongs to subfamily wotk incorporat tehse most recent data C[fa b1)l ,eIn
Epidendroidea Leindl .tribe Vlindeae Lindl. sub- his reyision, Christenso n(2001 t)reated Phalae-
tribe Aeridina ePfitzen The genus occurs from nopsis in a broad sense, sinking the genera Doritis
southern Indi aand Sri Lanka in the west to New Lindl .and Kingidium P. F. Hunt, both of which
Guinea and northern Australi ain the east, It extends have been treated as independen gtenera by other
as far north as southem China and 1faiwan .The authors (e. gSe.idenfaden 1988, Dressler 1993),
center of diversi toyf the genus is Borneo, from intoPhalaenopsis,
where 13 species are currently known, This major inconsiste namcoyng taxonomists is
Severa lmonographic treatments ofthe genus largel dyue to differe nwetighting attributed to the
have been publishe ndotably by Roife (1886 S)w,eet critical morphological character, pollinium num-
(1968- 61998,0) and Christenso n(2001 S)h.im ber .Although polliniu rnunmber has traditionally
(198 4re)vised the generi cclassification wnh empha- been censidered a heavil yweighted character in
sis on flor amolrphology, The histor yofclassifica- the classification oforchid genera i,t sstate is not uni-
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TABI,E 1 . Species of Rhalaenopsi sand allied genera used for matK-trnK intron sand ITS sequencing. Infrageneric
classificatio nfo11ow sChristenso n(2001),
'Sp
'ecles Vbucher
SllbgenusProbescidioidd(sRolfCeh)riste]son

Phalaenopsis lowii Rchb. f TBG i44316*
Subgenus (H.R. Sweet)Christenson
!{plij,tlae
Phataenopsi sbraceana (Hook f.. )Christenson 7BG 144566
Phaldenopsi sminus (Seiden CEh)ristenson 71BG 145839
PhataenopsiswilsoniiRolfe TIBG 144214
Phalaenopsi ssp. litkawa 29689
Subgenusilarishiana(eH.R.Sweet)Christenson

Phalaenopsi suppendiculat aC. E. Carr 71BG 144305
Phataenqpsi slobbi i(Rchb .£) Sweet 7BG 78865
Phalaenopsi sparishi iRchb. f. 71BG 13388J
Subgenus Ilolychile(sBredaC)hristenson

SectioInlolychilo(sBredaC)hristenson

Phalaenopsi scornu-cervi <Bred aBl)urne & Rchb. f. T:BG 145696
Phalaenopsis mannii Rchb. f 713G l18380
Phalaenopsi spantherina Rchb. f, 7)vukahara s, n.
SectionFuscataeH.R.Sweet

Phalaenopsi scochlearis Holttum .71B G14420P
Phalaenopsi skunstler Hioek. f 71BG137083
Phalaenopsi sviridis J. J, Sm. ZEIG 14J055
Phalaenqpsis sp. TBa 14223S
SectionAmboinenses H.R.Sweet

Phataenopsi samboinensis J ,J .Srn. TZIG133762
Phalaenopsi sbastian Giriuss & ROIIke 71BG140637
Phalaenopsis belli n(aRehb f.) Christenson 71BGi18533
Phataenopsi sdb",et:yt iGnarsaiys & Christenson llBG144564
Phalaenopsisfasciat Rachb. f 7BG 145 726
Phalaenopsis.fimbria Jt, aJ. Sm,
subsp, suneatrana (J J., Sm.) Christenson 71BGf45860
PhalaenopsisY7oresensisFowlie llBG145748
Phataenopsi sgigantea J. J. Sm. 71BG i3 7307
Phalaenopsi shiertrglyphi (cRaehb ,f) H. R. Sweet ZBGf45743
PhataereopsisJ'avan iJc. Ja. Sm. 71BG 14586S
Phalaenopsi sluecidemannian aRchb. f 71BG145733
Phalaenopsi slueddemanniana Rchb. t lvar. ochraeea Rchb. E 71EIGI45863
Phalaenopsi smaculata Rchb. f. 7BGJ44569
Phalaenopsi smariae Burb. ex R. Warneer & B. S. Williams 7:BGJ41056
Phalaenopsi smicholitzii Rolfe TZIG140.566
Phalaenopsih' mode,sta J, J, Sm. 71BG 140664
Phalaenopsi spallenE (Lind lRc,h)b, fi llBG145744
Phalaenopsispttlck r(aRch bf,i )H, R, Sweet 7:BG145761
Phataenopsi sreichenbachiana Rchb, f & Sander 7]BG145S75
Phalaenopsis venosa Shim & Fowlie 7BGJ45770
Pkalaen(tpsis violacea Wjtte 71BCf45785
SectionZlebrinaePfitzer
Phalaenopsis inscriptiosinen Fsoiwslie M(l i44571
Phataencrps issumatrana Korth ,& Rchb. fi 1:BG i42440
Phalaenopsi stetra,spi sRchb, f 7]BG145841
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TvLBLE1.(continued).

Speeies Vbucher
SubgenusPkalaenopsis
SectioInViataercopsis

Phalaenopsi samabitis (L. B)lumc TIIG145847
Phalaenopsis amabilis (L. B)lume
subsp. rosenstromii (F ,M. Bailey C)hristenson 71BG140316
Phalaenqpsis aphrodit eRchb, f (1) 7BC14140S
Phaiaenopsis aphrodite Rchb. £ (2) 71]G141I51
Phataenopsisphilijzpine nGseilsamco ex Fowlie & Tleing 7LBG118SS2
Phalaenopsi ssanderiana Rchb. E Atagawa s. n.
Phaiaenopsis schilleriana Rchb. f IIBG 0309
Phataenopsi sstuartiana Rchb, f, 7BG l45 758
SectionDeliciosaeChristenson
Phaiaenopsis chibae T, Yukawa 71BG115846
Phalaenopsis deticio Rscahb. f ,(1) 7BG145842
Phaiaenopsi sdltlici Rocshab. f .(2) 1;BG144594
SectionEsmeraldaRchb.f,

Phaiaenopsispttlcherrim(aLindlJ..J).Srn. 71BG1J8342

Phalaenopsi ssp, "buyssoniana" TBG l45823
Sectionstauroglot(tSicshauerB)enth.

Phaiaenopsis equestri.s (Schau eRrch)b. £ 71BG141154
Phalaenopsi scetebensis H, R. Sweet ZBG J45822
Phalaenopsi slindeni Ii.oher ZBG140568
Allied taxa
Lesliea mirabitis Seiden£ TLBG145844
IVbthodorit zihsojiangensis Z. H. Tsi TBG f3 750i
O"tgroupAmesiella
monticola J.E. Coote s&D. P, Banks 7BG 123790
*TBG series indica atcecession nurnbers in livi ncegllection databa saet Tsukuba Botanica lGarden.
fbrm within Phalaenctpyi sT.he predoinina nnumtber shed their leaves durin gthe dry season.
in the genus is two, but the subgenera Proboscidi- Phylogentic relationships in Phataenopsis
oides, Aplrylla eand Ftirishianae all have four pol- remain largel yunresolyed, Woodward (1951),
lini aas, do the sections Deliciosa aend Esmeralcla Sagawa (1962 )S,hindo & Kamemoto (1963),
foot ihn subgenus Phalaenopsis) .Immense diversity Sagawa & Sheji (1968 A)re,nds (1970 )S,hej i(1976,
in the structure Qf reproductive organs, particularly1980), Aoyama (l993 A)o,yama et al. (199 4an)d
that of the labellu mand the stipe, has also resulted Kao et at. (20 10 ) investiga tcyetdological characters,
in inconsisten tinterpretatio nosf relationships in Brandange et aL (197 11,972) examined alkaloid
the group, content and Tsai et al. (2003 c)onducted a molecu-
Phalaenopsts also exhibits rematkable diversityla rphylogenetic analysis ofthe intema ltranscribed
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 i)n subgenus Phalaenqpsi lsn. the present
tcrrestr ilailf eform .On the other hand ,plant isn sub- study, we compared and combined DNA sequences
genera Apilylla IeZ,firishia anndae Proboscidioides of a maturase-encoding gene (matK )flanke dby
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Subgenus Section

811008874 P.arnabilis Phalaenepsis Phalaeriopsis
P. amabilis subsp. rosenstromii Phalaenopsis Phalaenopsis
P. aphrodite (1) Phalaenopsis Phalaenopsis
6010016 75 P. aphrodite (2) PhalaenopsisPhalaenopsis
P. sznderlana Phalacnopsis Phalaenopsis
313 P. IilldeniiP, Phalacnopsis Stauroglottis
celebe]sis PhalacnopsisStauroglottis
lee P. equestris Phalaenopsis Stauroglottis
99 loe98 73 PP,, psthuialritipapniancnsis PPhhaallaaeennooppssiiss PPhhaulluaeennooppssiiss
100 P, schilleriana Phalacnopsis Phalaenopsis
P sp.R PolychilosPo]ychFiulsocsalt'aoelFyucshcia]toasePFoulsyccahtialeoFsuPsoclaytcaheiAlmobsoPionleynesheistosPolyehilosPolychitosPolychilosPolychilosPolychilosPetvchilosPolychilosPolvchilosPolychilosPolychilosPolychilosPolychilosPolychilosPolychilos
10064 kunstleri
100g4 P. cochlcaris
leo P. viridisP.
doweryensis
100
P, giguntea Ambeinenses
P. maculata Amboinenses
100 P. amhoinensis Amboinenses
626010096966164P, venosaP. Amboincnsc$
belhnaP. Arnboinenses
violaceaP Amboinenses
53 90 R ffilmobrreiasteanssujbssp.sumatrana AArmnbbooiinneennsseess
go 9686866065 P, inscriptiosincnsis ZebrinaeAmboinenses
P.javanicaP.
tetraspis ZebrinacZebrinaeAmboinenses
P. sumatrana
P. modestaP.bustianii
Arnboinenses
P. mariaeP. Arnboinenses
Iueddemanniana var, ochracea Po]ychilosPolAyrcnhbiolionsenses
4516 661299 PP.. IfuaesdcdieamlaanPn.iana Po]ychilos AAmrbnebioneinnseensses
hieroglyphica PolychilosPolychAirlnobsoincnscs

4349 99 P. reichenbachiuna Amboinenses
P. pallensP. Polychilos Amboinenses
pu]chraP. Polychilos Ambeinenses
928963 rnicholitzii Polychilos Amboinenses
P. pantherina Polychiles PolychilosPolychilosPolychilos
66 P. comu-cervi Polychilos
R manniiP. ?olychiles

minusLeslieumirabilis AphylLae
nappendiculata Parishianae
58100
50 45 R lobbiiP. PaTishianae
41 10064968310094 parishiiP. Parishianae
99 chibaeP. PhulaenopsLs DeiiciosaeDeHciosaeDeliciosaelkmera]da
93 delicio (s1a) Phalaenopsis
R deticiosa(2) Phalaenopsis

R sp ,''buyssoniana'' Phalaenopsis
1001005610033 R pulcherrima Phalaenopsis tsmera]da
P. bsip,aRceama AphyllacAphyllacAphyUaeProboscidioides
R wilsonii
98 R bwiiNethedoritiszhejiangensis
99
Amcsiclla
FIG.1. Stric tconsensus of24 most-parsiinenious Fitch trees based upon matK-trnK intron ssequences: length=593 ,consistency
index=O,7808 (O.63 2ex8cluding uninfbrmative characters), retentien inde xof O.875 1N.umbers above internQd eisndicate
bootstr vaaplues from 1,OOO replicatcs ofFitch parsimony analysis (maxtr leiemi tof 1,OOO per replicate), Numbers below intern-
odes indica btoeotstr avaplues from 1,OO Oreplicates ofneighborsioining distan canealysis.
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intron softhe plasti dgene imK; the trnK intron sand trees were calculated using ACCTRAN optimization
the ITS region from members of all sections of (Swoffb &r dMaddison 1987) .Distanc etrees were
Phalaenopsis and closely related genera to clarify obtained using the neighborjoining (NJ )method
relationships within the genus. (Sai t&o Nuei 1 987) with a Kimura two-parameter
correction (Kimur 1a980) ,[I bassess the relative
Materials and Methods robustness fbr branches ,the bootstra pmethod
(Felsen s19t8e5i) wnas used with 1OOO replicates.
We selected 53 representative taxa from all sub-
gener aand sections ofPhalaenopsis for inclusi oinn Results
the study. Leslie aSeidenf alnd IVbthodorit iZs. H.
Tsi were also included in the study because mor- Figure 1 shows a strict consensus of24 most parsi-
phologic aclharacters of these two genera suggest a monious (MP) trees derive dfrom the matK-trnK
close relationships with Phalaenopsi (sChristensionntron ssequences. The tree had a consistency index
2001) .Riraphalaenops iAs, D. Hawkes, a genus (CI o)f O.7808 (O.63 2ex8cluding uninfbrrnative
also regarded as being closely related to Phalae- characters) and a retention index (RI o)fO.875 1 . In
nopsis, was not included in the study becaus ean ear- this data set topologies with more than 509t 6boot-
lie rcomprehensive molecular analysis efsubtribe strap support were nearly identic abletween MP
Aeridinae had revealed wide phylogeneti cseparation and NJ analyses (Fi g1.) ,though section Stauro-
between the two (Tbpi etk al., in press )B.ased on glott ifsormed a sister group with three species of
the results ofthe same analyses, Amesietla Garay section Phataenopsi s(R philippinensi sR, schille-
was chosen as the eutgroup taxon. [[lab l1e list sthe riana and 2 stuartiana) in the NJ tree (83% boot-
materia]s used in the phylogeneti canalysis. Vbucher strap support).
specimens have been deposite adt TNS. A strict consensus of 322 MP trees on the
Experiment amelthods fbllowe dthos edescrib- basi softhe ITS data set is shown in Fig. 2. For this
ed in Yukawa et aL (1993 1,996). Sequences were tree, Cl was O.552 1(O,47 e7xc0luding uninformative
determined by amplifying matK, trnK intron sand characters) and RI was O.8169; topologies with
the ITS region (includ 5i.n8Sg rDNA and parts of more than 50% bootstra spupport were nearly iden-
1 8S and 26S rDNA) yia the polymerase chain reac- tica lbetween MP and NJ analyses (Fi g2)., although
tion (PCR )from a tota lDNA extract, PCR primers the monophyly oftwo individua olfsPhalaenopsts
were as given in Yitkawa et al. (199 9f)br the matKL uphrodit ewas well supported only in NJ analysis
trnK 3' intro nand as in Douzery et al. (1999 f)or (86% bootstrap support).
ITS, We develope dthe fbllowin gcombination of Analyse sofmatKLtrnK intron ssequences pro-
primers for the trnK 5' intron :3914FN: 5'- vided results substantially concordant with those
ATCTGGGTTGCTAACTCAATGG-3' and achieved via analyses ofITS sequences, except fbr
OMAJ]IR: 5'-CAATATGGTCAGAACGGCGTL3'. the positio nosf Phalaenopsi sphigijzpinens iasnd
DNA sequences were aligned manually. Gaps were R minus, In the plasti pdhylogenyl R phii4zpinensis
treated as missing characters, The aligned data file fbrmed a sister group with R stuartiana; in the
is available from the firs tauthor upon request. nuclear phylegeny, R philippinensis was nested
Parsimony and distanc aenalyses were conducted between the R stuartiana-schilleriana clade and
with PAUP" version 4.0b6 (Swoffb 2r0d01). The the R amabilis-aphrodite-sanderiana clade.
heurist iopction was used to perfbrT Fnitch parsi- Likewise, the plasti pdhylogcny supported place-
meny analyses (Fit c19h71) .Branch length sfor ment ofR minus in a clade comprising Leslie aand
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Subgenus Section

P. amabilis Phalaenepsis Phalaenopsis
P. amabilis subsp ,rosenstromii Phalacnopsis Phalaenopsis
R sandcriana Phalaenopsis Phalaenopsis
n aphredite (2) Phalaenopsis Phalaenopsis
E aphrodite (1) PhulaenopsisPhalacnopsis
Rphilippinensis Phalaenopsis Phalaenopsis
]schilleriana Phalaenopsi$ Phalacnopsis

P.stuartiana Phalaenopsis Pha]aenepsis
Requestris Phulacnopsis Stau[oglettis
RlindeniiP. Phataenopsis Stauroglottis
celebensLs Phalaenopsis Staurog[ottis
R sp.R PolychilosPollyfcthiislcoastPaeelFyucshcialtoasePFoutsyccahtialcoFsuseataeArnboinenses
cochlea ris
P.kunstleri
P. viridisP.
doweryensis PoLychilos
R giganteaP. Polychilos Amboinenses
maculata Polychilos Amboinenscs
P .amhoinensis ?olychilos Amboinenses
P. venosaF', Polychilos Amboinenses
bellinaR Po]ychilosPolychAim]boosiPnoe]nyscehsilosPolychilosPolychilosPolychilosPolychilosPolychilos

Arnboincnses
violaceaP.
fimbriat asubsp.suniatrana Amboinenses
P.fl{resensis Amboinense s
R livanicaP. timboinenses
inscriptiosinensis ZebrinaeZebrinaeZebrinaeAmboinenses

P, tetraspis
Rsumatrana

P,modestaRbastianii Po]ychilos
Polychiles Amboinenses

R mariacR Polychilos Ambeinenses
lueddemanniana var. ochracea PolychilosPolychAimlboosiPneelnyscehsitosPolvchilosPolvchilosPolychilosPolvchilosPolychilosPolychilosPolychiLosPolychilos
P, lueddemanniana Amboinenses

P i'asciataP. Amboinenses
hieregLyphica Arnboincnses

P. pallcnsP, Amboinenses
pulchraP.reichenbachiana Ambeinenses
Amboincnses
P.rnicho]itzii Amboinenses
P. pantherina Polychi]nsPolychilosPolychilos
P,coniu-cervi
P, mannitiR
sp.R AphyllacAphyllacAphy]laeAphyllaeProboscidioides
wilsonii
P. braceana
P, minusP.
IowiiNothodaritiszhejiangensis
PP,. aIppoenbdibeui]aitaP, PPaarriisshhiiaannaaee
paTishiiP. Parishianae
chibaeP. Phalacnopsis DeliciosacDeHciosaeDeliciosaelsmeraldalsmera]da
deliciosa(L} Phalaenopsis

1' d.eticios a(2) Phalaenopsis
Leslieamirubilis
R sp."buyssoniana'' Phalacnopsis
R pulcherrima Phalaenopsis
Amesiella
Fia2. Stric ctonsellsus ef 322 most-parsimonious Fitch trces based upon ITS sequences: length=585 ,consistency index=O.5S2 1(O.4770
excluding uninformative characters) ,retention inde xof O.8]69 N.umbers above interned iensdica tbeootstr avaplues from
1,OOO replicates ofFiteh parsimony analysis (maxtr leiemi tef 1,OOO peT Teplicate). Numbers below internode isndicat beootstrap
values from 1,OOO replieates ofneighborjoining distanc eanalysis.
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Phalaenopsi ssubgenera Ririshiana eand Phalae- Discussion
nopsis sections Deliciosae and Esmeiulcl awh,ereas
the nuclear phylogen yindicat ead nested positio nin Biogeog,up]u;
a clade formed ofIVbthodoritis and Phalaenqpsis The two major clades reeognized in this study are
subgenera Aphyllae and Proboscidioides. characterized by markedly differe pnattter nisn geo-
In such cases, analyses of combined data sets graphica dlistrjbut ioofn their constituent species.
excluding problemati ctaxa provid emore resolu- Thus species in Clade 1 are mostly distribu twietdh-
tion and intern aslupport fbr relationships than do in the limit sofMalesia, as define dby van Steenis
individu adlata sets (e. gO.lm,stead & Sweere 1995, (1950 T)h.e few exceptions are as fbllow sP:halae-
Wongsawad et al, 200], Yukawa 2001, Yukawa et nopsis aphrodit (esect iPhoanlaenopsis), which is
al. 2002), We thus conducted a combined analysis ef known from the Philippine asnd [faiwan ;R amabilis
the jnformatio dnerive dfrom matK-trnK introns (sect Pihoanlaenqpsi sw)id,ely disperse tdhroughout
and ITS sequences, In this analysis Phalaenopsis the Malesian region, though it srange extends also
philippinensi sand R minus were excluded. The into northern Australia ;R cornu-cervi (section
lengt hof the aligned sequence was 3768 base pairs, Pblychilos )a,]so very widely distribut eocdc,urring
Figure 3 shows a strict consensus of 896 MP trees from northeastern India to Java and Borneo; and R
from the combined data set. CI was O,6557 (O.5296mannii (sect iIoCbnlychilo as )cl,ose relative efR
excluding uninformative characters) and RI was cornu-cervi fbund throughout the Himalayas and in
O.8297 .In this data set, topologie swith more than Indochina, R mannii is the only species in Clade 1
50% bootstra psupport were almost identical that occurs exclusively outside the Malesia nregion,
between the MP and NJ analyses (Fi g3.) ,altheugh In contrast, species in Clade 2 are particularly
R ceiebensis fbrrne dthe basa lpositio nin the clade divers ein the Himalayas, southern China and
comprising Phaiaenopsi ssections Phalaenopsis Indochina, However, there are a few exceptions: R
and Stauroglot tiin sthe MP analysis, whilst it was puleherrima (sect iEsomneraldo), which also occurs
nested between Phalaenopsis sections Rhalaenqpsis in Sumatra and Borneo; R delicios a(section
and Stauroglotti isn the NJ analysis. Delieiosae) ,the most widely-distributed species in
In the combined analysis, the fo11owing the genus with a range that extends from Sri Lanka
insights were provided (Fi g3.): (1 )the genera and India to the Philippine sand Sulawesi ;R
Lesliea Aibthocloritis withinthe (sectiDoelniciosae),
and are nested genus aysorensis a species with an
Phalaenopsis ,(2 )Phalaenopsis subgenus ApJryllaiesolat eddistribut iino snouthern India ;and R appen-
and section Esmeraldd, often treated as the inde- diculata (subgen Putzsrishianae )a, species record-
pendent genera Kingidiu mand Doriti rsespectively, ed from the Malay Peninsul aand as such the only
are also nested within Phalaenopsis, (3 )Phalae- C`Malesian" member ofthis clade.
nopsis subgenera Phaiaenopsis and Ptirishianae This distincti oinn geographic adlistribution
are not monophyletic. (4 )Phalaenopsis sections indicate tshat speciation within Clade 1 has occurred
Dek'ciosae S,taurogibtti Asm,boinenses and Zebrinae mostly in Malesia, whilst that in Clade 2 has
are not monophyletic. C5 )The genus Phalaenopsis occurred predominantly in the Himalayan and
is apparently composed of two major clades, here Indochinese regions. In an attempt to explain this
termed Clade 1 and Clade 2. pattern ,two competing hypothese sare proposed.
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l48 APG M)1,56

Subgenus Section

Ramabilis Phalaenepsis I'halaenepsis
R amabMs subsp. resenstromii Phalaenopsis Phalaenopsis
Rsanderiana Phalaenopsis Phaluenopsis
R aphrodite (2} Phaluenopsis Phalaenopsis
E aphrodite Cl) Phalaenopsis Phalaenopsis
P.schilleriana Phalaenopsis Phalaenopsis
P. stuartiana Phalaenoptiis Phalaenopsis
P. equestris Phalaenopsig Stauroglottis
P, lindeniiP. Phalaenopsis Stauroglottis
eetebensis Phalaenopsis Stauroglottis
R sp.P. PolyehilosPolycFhUislcaotsaPeoFulsyccahtaielFousscPaottayeFcuhsiclatoasePAornLbyocihneinlsoessPolvchilosPolychilosPolychilosPolychilosPolychilosPolychilosPDIychilosPolychilosPolychilosPolychilosPo]ychilosPolychilosPe]ychilosPoiychilosPolychilosPolyehilosPolychilosPolychilosPolychilosPolychilosPe]ychilosPo]vchiLosPelvchilosPolychilosPolyehilosPolychilosAphylleeAphyllacAphyllae
cochlearis
P. kunstleri
P. viridis1'.
doweryensis

I' g.igantea1'. Amboinenses
maculata Amboinenses
P. amboinensis Amboinenses Qpapto-

R venosaRbe]TinaP. Amboinenses
Amboinenses
violttceaP. Amhoinenscs
tilnbria tsuabsp.sumatrana Amboinense s
P. fl{regensis Ambuinenses
P.juvunicaP, Amboinenses
inscriptiesinensis ZebrinaeZebrinaeZcbrinacAmboinenses
P, tetraspis
P, sumatrana
P, medestaP.
bastianii Amboincnses
R mariaeR Amboinenscs
lueddemanniana var, ochracca Amboinenscs
P. Iueddemanniama Amboinenscs
P. fasciataP. Arnboinenses
hierogtyphica Amboinenses
R pa]lensP. Amboinemses
pulchraP, Amboincnses
reichenbaehiana Amboinenses
P. rnicholjtzii Ambainenses
P. pantherina ?olychilosPolychilosPotychitos
P. cornu-cervi

P.manniiR
sp.R
wilsonii
P, braceana
Nethodoritiszhe.iiangensis
R IowiiLcslieamirabilis Proboscidioides
Qgk
R sp,''buyssoniana" Phalaenopsis EsmeralduIismeralda
R pulcherrima Phalaenopsis
R appendiculata Parishianae
RlobbiiR Parishianae
parishiiR Parishianae
chibaeR I'hu]aenopsis DcliciosaeDeticiosueDeliciosae
delicios (aI) Phalacnopsis
R delicies{a2) llialacnopsis

Amcsiella
FIG, 3. Stri ccotnsensus of896 most-parsimonious Fitch trees based upon mati<LtrnK intron sand ITS sequences/ length= 1153, con-
sistency indcx==O,6S5 (7O,S2 9ex6cluding uninformative characters), retention index of O.829 7N,umbers above interned iensdi-
cute bootstra pvalues from 1,OOO replicates ofFitch parsimony analysis (maxtre leimi tof 1,OOO per rcp]icate). Numbers below
intemodeg indicat beootstra pvalues fire m1,OOO replicates ofneighborjoining distanc canalysis.
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The fir sitnvoke scladogenesis of two precurso lrin- vegetative organs in Phalaenopsis and allies corre-
eages, with subsequent radiation occurring in each late swith pronounced environmental differences
lineag erelatively recently, Prior to cladogenesis, between the Malesia nand Indochines reegions. For
the only availal)le corridor linkin tghe two lineages instanc ei,n Clade 1, species in sections Amboinen-
in the Malesian and Indochinese regions was the ses, fileiseat aaend Zebrinae occur in evergreen
Malay Peninsula, dispersal fbrests lowlightlevels(Christe2n00s1o)n
and, consequently, under
betwee nthe two regions was rare, where the leave sdo not have to endure prolonged
The hypothesis in desiccating (K,Suzuki,
second emphasizes environ- periods a environment
menta1 differenc beestween the Malesian and Indo- persona lcommunication). In contrast, species in
chinese regions in demarcatin tghe observed distri-section Ilolychil hoasve succulent leave sadapted
butio npatter nofs Clades 1 and 2. The climate of the to bright places high in the forest canopy
Malesian is by (Christen2s00o1n,T.YUkawa, data),
region characterized rather constant, unpublished
high precipitat itohrnoughout the year ,In contrast, AccordinglM they are likel yto haye been able to
the Indochines eregion has a matked dry season. extend the distribut itoon the Indochinese region
Ancestral stocks in each major clade may have where a defini tdery season exists. In Clade 2, mem-
adapted exclusively to one or other of these envi- bers of Phalaenopsi ssubgenera Aphyllae f,urishi-
ronmental condjtions, with subsequent divergence anae and Proboscidioides and the genus IVbtho-
being reinforced throug helaboration of specjalized doriti sshed their leaves during the dry season. It is
function isn either lineage, interesti tnog note that the distributi oonfs these
There are three major reasons why the first taxa largel oyverlap with areas of subtropical mon-
hypothesi sshouldbe rejected. First ,pair-wise dis- soon climate where pine-deciduous Dipterocarpus
tances of nucleotide divergenc beetween the two fores atnd lower montane pine-oak fores dtominate
major clades and taxa in each clade are great ,For (Santi s1u98k8), As such, the deciduou shabi tin
example, the divergenc ebetween Phalaenopsis thes egroups likel yrepresents an adaptatien to a
eguestris in Clade 1 and R putcherrim ain Clade 2 seasonally severe water defic iMto.reoyer, these
in ITS sequences is 8.7%. Consequentl yi,t is not groups have develope dgreen ,flattene droots that
plausibl eto hypothesiz ae recent origin ofthe two serve as a photosyntheti corgan particularl ydur-
major clades and subsequent radiation in each clade. ing the deciduou speriod (Benzi netg al, 1983).
Second ,corridors linkin gthe Malesian and Indo- Species with vegetative organs suited to special-
chinese regjons were much wider during glacial ized ecological and physiologic caolnditions would
period sin the Pleistocen e(Morl e&y Flenley 1987). be less likel yto be able to establish in niches in
Thus geographica lbarrie rbsetween the two regions mailcedly differe cnlitmate regimes. We thus support
are negligible, if we take account the time-scale the second hypothesi tshat attaches importanc eto
required fbr speciation. Third, long-distan wciend environmental factor fsbr the advent of a bimodal
dispers aofl orchid seeds is frequent (e. gG,anda- distribut ipoantter wnithin Phalaenopsis.
wijaja & Arditti 1983), The occurrence of R
aphrodite in both the Philippine sand Taiwan pro- Mbrphotogy ofneproduc otrgianvse
vides a good example of long-distanc edispersal The enormous morphological diversificati oonf
because there is no geologi cevidence fbr a past reproductive organs in subtribe Aeridiinae is well
land cormection between Taiwan and the northern documented in studies based both on morphological
Philippin edsurin gthe Teniar y(fa n2002). (e. gS,eidenfade 1n988) and macromolecuiar char-
On the other hand, morphological evelntion of acters (Ibpi etk al. in press).
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150 APG Xio1 .S6
Pel]inium
Deciduousness
number

P.amabilis 2 evergreen
P. arnabilis subsp. rosenstromii 2 eveTgreen
P. sandcriana 2 overgreen
P. aphrodite (2 ) 2 evergreen
P. aphrodite (1 ) 2 evergreen
Rschillerian a 2
evergreen
P, stuartiana 2 evergreen
P. equesnis 2 cvcrgreen
R]indenii 2 cvcrgrcen
Rcelebensis 2 eveTgreen
Rsp. 2 cyergrcen

Rcochlcaris 2 evergreen
Rkunstle ri 2 cvcrgreen
Rviridis 2 cvergrccn
Rdoweryensis 2 evergreen
Rgigante a 2 evergreen

nmaculata 2 evergreen Qptpto-
P. ambeinensis 2 cvergrccn
P. venosa . 2 evergreen
P. bellin a 2 evergreen
P. violucea 2 evergreen
Pfimbriatasubasp.sumatrana 2 evergreen
Rflcresensi s 2 cvcrgreen
Pjovanica 2 evergreen
P. inscriptiosinen sis 2 evergreen
P. tetraspis 2 evergreen
P. suinatrana 2 evergreen
P.modesta 2 evergreen
P. bastiani i 2 evcrgreen
P. rnariae 2 evergreen
P, lueddemanniana var, oehracea 2 evergreen

P.Iueddemanniana 2 evergreen
P. fascia ta 2 evcrgrcen
P, hicroglyph ica 2 evergrecn
P. pal]e ns 2 el,ergreen
Rpulchra 2 evergreen

P.reichornbachiana 2 evergreen
Rmlcholitz li 2 evergreen
Rpuntherina 2 evergreen
Rcornu-cervi 2 evergreen
Ulnamiii 2 evergreen
P.sp. 4 deeiduo us

P.wilsonii 4 deeiduous
P. braccan a 4 deciduous
Nothodoritiszhejiangens i$ 4 deeidueus
Rlowii 4 decidueus
ge9N
Leslieamirabilis 4 evergreen
P. sp.''buyssoniana" 4 evergrecn
P. pulcherri ma 4 evergreen
P. append;culata 4 evergT'een
P. IobPi i 4 deciduous
k?reEidhuiious 4 deciduous

Rchibac 4 cvcrgrccn
P. dclieiesa(l ) 4 cvergrccn
?, deliciosa(2 ) 4 evergrecn
Amesiella 2 evergreen
FIG.4. Reconstmction ofevolution in polliniu mnumber and deciduousnes sin Phalaenopsi sa'nd allied genera under ACCTRAN epti-
mization. The trec is the str{ct censensus Fitch tree based sequences.
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