Two Species of Entada in Japan as Evidenced by cpDNA Phylogeny

2008·0.95 MB·English

by  Wakita, Norihisa

TThhee JJaapapnaesneSeosciee tySociety ffoorr PPllanatnt SSyystsetmaetmicastics
ISSN1346-756S ActaPhytotax.Gcobot. 59(3):183-1(923008)

Two Species inJapan by Phylogeny
ofEntada asEvidenced cpDNA

NORIHISA WAKITAi, YOICHI TATEISHI2, TETSUO OHI-TOMA3, JIN MURATA3
and TADASHI KAJITAi'
JDEzpartment ofBiolo Ggrayduate Sbhool ofScien ceeni,ba U)iiversi t1y-33 }21yoi-ch ion,age-ku, (:hiba 263-8522,
Jdpan. *tkciji@bculcychiba-u,ip (Ziutho rcofrroesprondencel,' ?DE:partment qf'Sciene Eeducation ,haculty of
Education ,Uhivensit oyfth Reyukvu sOk,inawa 903-O129 ,Jinpan; 3Botanical Gatdlens ,Graduate School ofScience,
711 iUeniversit oyf7lol 1y1o2-OOOL .lapan
The phylogeneti crelationships ef Entadu (Fabace aweere) examined to determine if two distinct
species occur in Japan. Nucleotide sequences of the trnK/ and trnL intron sand the intergenic
spacer of trnL-trnF of cpDNA resolved 19 haplotype asmong 39 widely collected samples of
E. phaseoloicies-1 ipkleants ,In the resultant tree ,two distin ccltades corresponded to two groups
alse distinguish ebdy diagnosti cseed characters. The clades agree with the expected chorology of
samples and with the diagnosti mcorphology from previou staxonomic work. One clade corresponds
to Entacl aphaseoloides and the other to E. tonkinensis. A distincti vcoencentration of haplotypes
was observed. Although seeds of Entadu phaseoioide sare renowned as sea drif tseeds, our results do
not suggest frequen tgene flow caused by long distanc eseed dispersal,
Key words: Entacla koshunensis,Entadophaseoloicles,Entad tloankinensis, match-box bean ,seed
dispersal
Entada Adans. (Leguminosa Mei:mosoi- tropica land subtropical areas of the Pacific ,from
deae) consists of about 30 species (Niel s1e98n1, eastern to southeastern continental Asia, the south
1992, ILDIS 2005) distribu tien dthe tropics and Asian island sO,ceania, and Polynesia. The north-
subtropics. While more than half of the species eastern limi tof the species' range is Japan (Nielsen
in Entacla are shrubs or subshrubs with small 1992, Huang & Ohashi 1993). The wide distribu-
pods, some species are liana swith giganti cpods tion of Entacl aphaseoloides is thought to have
that can be more than 1 m long ,The larg ewoody been achieved by long distan cdeispers aofl the
seeds ofEntatla are also known fbr drifti nign sea- seeds, Although Entado phaseotoide shas some
water. These remarkable seeds are called "match- distincti fveeature isn it sfirui atnds seeds, consid-
box beans" (th ceommon name fbr Entacl aphase- erable taxonomic confusion exists as to it sclas-
oloides (L. M)err. and E, rheedii Spreng. )or "sea sification in Japan and adjacent areas. While most
hearts "(E .gigas (L. F)awc, & Rendle), and are studies treat the species of Entacl ain Japan as E.
frequentl fyound on beaches far from their origin, phaseoloides s,ome works have variously treated
Entacl aphaseotoides is widely distribut eidn it as E, rheedii Spreng, ,E. parvijiol Miearr., or E.
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koshunensis Hayata & Kaneh. Some authors rec- presumed type localit yof Entacl aphaseotoides
ognize two distin cstpecies (Ho 1985, Huang & in Indonesia, Key seed shape characters used in
Ohashi 1993, Ohashi 2001), but the recegnition of Wakita et al. (2005 w)ere also examined to deter-
E. rheedii and E. parvijbli ian the Ryukyu Islands mine the correspondence between seed character
(Okinaw aJ)a,pan ,is apparently misleading and E. groups and molecular phylogeneti cgroups .The
koshunensi scannot readily be separated from E. goal of this study was to reveal the phylogenetic
phaseoloides according to a detaile dtaxonomic relationships of Entadu phaseoloide sand closely
study that fbcused on this taxonomic confusion related species, and to determine if indeed two
(fatei est haii. 2008), species ofEntacla occur in Japan.
RecentlM Wakita et aL (2005 s)urveyed the
number of lcafle tpairs on the second pinnae, Materials and Methods
morphelogical characters of the seeds, and single
strand confomiation polymorphism (SSCP )band Entado phaseoloides-like plant swere col-
pattern sof the icps16 intro nin chloroplast DNA leete dfrom eastern to southeastern Asia and the
(cpDNA )fbr species of Entadu in the Ryukyu South Pacific (Tabl 1e) .Samples from Indone-
Islands A,lthough the number of pairs of leaflet sia and Vanuatu were obtained from individu-
on the second pinnae did not clearly divide the als cultivated in the Bogor Botanical Garden
samples into two groups t,he seeds and molecu- (Bogo rI,ndonesia )and the Tsukuba Botanical
lar markers clearly indicated the two groups of Garden (Ibara kJaip,an), respectiyely (Tabl 1e).
Entacla .The firs gtroup has a sma]1 (`S' c)on,vex Sincc plant sof Entada are generally very large,
seed type, 29.4ul6. 9mm wide by 32,4-52. 7mm spreading over the fores tcanopy, distinguishing
long with an angular margin. The second group individua lwsas difficu lWte, collected more than
has a larg e(`L' c)om,pressed seed type, 39.5-59.1 one sample per populatio nonly when individu-
mm wide by 45.8-64.7 mm long with a rounded als could be clearly distinguishe Sde.ed samples
margin (Waki teta al. 2005) .Differen StSCP band were not obtained from all the population sK.ey
pattern sc,aused by a nucleotide substitution and seed shape characters of all avai]able seeds were
some jndels in the ips16 intron w,ere also c]early observed, and seed type (S or L) was determined
observed in the two groups .Wakita et al. (2005)according to Wakita et al. (2005).
considered the firs gtroup as corresponding Enta- Leaf samples of the species of Entada
tla koshunensi sand the second to E. phaseoloides obtained from eastern and southeastern Asia,
based on seed characteristics, fbllowin gthe cri- Oceania, and the South Pacjfic were used fbr a
teria of Ho (1985 f)or distinguishi nspgecies of molecular phylogenetic study ([lab 1i)e ,Entado
Entacla in [laiwan .Wakita et at, (2005 )ho,wever, gigas (L. )Fawc. & Rendle, E. rheedii, and E.
did not resolve the taxonomic status of these two spiral iRsidley ,which belong to the same subsec-
groups due to insufficie snatmpling limite donly tion (Ehta cslubasect. Entadu) as E. phaseoloides,
to the Ryukyu Islands A. molecular phylogenetic were also include din the phylogeneti canalysis.
study using additional samples from a wider geo- Outgroups were Entacla ato?ssinica Steud .(ftom
graphi carea is necessary to clarify the presenc eof Tanzania )and E, polystachya DC. (fro mMexico).
two species in Japan. Those species were confirmed as not being in the
In this pqper ,we report the results ofmolecu- ingroir pby our preliminar aynalysis (N ,Wakita et
la rphylogenctic analyses using samples obtained al., unpublished data )using publishe dtrnL-trnF
fror na broad geographica lrange, includin gthe sequences of Mimosoideae (Lucko wet al, 2000,
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November 2008 WAKIIIA & AL.: Entada in Japan by cpDNA Phylogeny l85
2003). A tota lof47 samples, includin goutgroups, CA, USA). Purifie DdNA was directl syequenced
was used for the sequence analysis. Vbucher using an ABI PRISM Big Dye Terminator ver-
specimens of the samples used in this study were sion 3.1 Cycle Sequencing Ready Reaction kit
deposit eidn QCNE, TI, TNS, or URO ([fa b1)l.e and an ABI PRISM 377 DNA sequencer (Applied
Prior to genomic DNA extraction, crushed Biosystems, Foster City ,CA, USA). Multiple
leaf tissue was washed using HEPES buffe rfbl- sequence alignment was perfbrrned with ClustalX
lowing the methods of Setoguch i& Ohba (1995).ver, 1,83 (Thompso ent al. 1994) using the default
Total genomic DNA was extracted from washed setting, and the obtained alignrnent was manually
leaf pellet ussing the methods of Doyle & Doyle modified using Se-Al Sequence Alignment Editor
(1987 )T.he two regions of cpDNA, the trnL version 2.0al1 (Rambau 2t002). Combined nucle-
intro nincludin gthe trnL-trnF intergen iscpacer otide sequences were used fbr furth earnalyses
(IGS ;hereafte rreferred to as trnL-trnF) and the because the incongruenc elengt hdifferen c(eILD)
trnK intro n(inclu dmaitKn;g hereaft reefrerred to test implemented in PAUP* ver4.0. detecte dno
as trn?O, were amp]ified by PCR. The fo11owing significant incongruen bceetwee nthe two genetic
oligonucleotide primers were used fbr PCR ampli- regions fbr phylogeneti acnalyses,
ficatio nand sequencing: c, d, e, and f for trnL- Maximum-parsimony (MP) and neighbor-
trnF, designe bdy [faberl eett aL (1991 t)r;nK-F joini n(gNJ) analyses were conducted using
(forwar5d'-:TGGGTTGCTAACTCAACGG-3') PAUP' 4,Obeta10(Swoffbr2d002),
version
and trnK-R (rever s5e'-:GGAACTAGTCGGPCIr- Branch support was assessed with 1,000 rep-
GGAGT-3') designe adccording to the complete licat efsbr MP and 10,OOO fbr NJ. A Bayesian
cpDNA sequence data of tobacco (GenBank phy]ogeneti acnalysis was perfbrmed in MrBayes
accession numbers ZOO044 and S54304) ,matK- 1O version 3.1.2 (Ronqui s&t Huelsenbeck 2003)
(Kat oet aL 1998), matK-AF (Takayam eat al. with the best-f imtodel (GTR+I+G )for both of
2005), matK-1777L, and matK-1932R (Hu et al. trnL-trnF and trnK selected by Akaike's infor-
2000) for trnK. The PCR reaction mixture con- mation criterion in MrModettest 2.2 (Nylander
tained 2,O pL of O,2 mmol!L dNTP solution, 1.0 et al, 2004). Metropolis-couple dMarkov chain
pL of 5 mmollL of each primer ,O.5 unit of ExTaq Monte Carlo sampling was perfbrmed with one
DNA polymerase (TaKaR aBie, Ohtsu, Shiga, cold chain and three heated chains that were run
Japan) ,2.5 uL of 1Ox ExTaq buffer a,nd 1O-30 ng fbr 1,OOO,OOO generation sT.rees were sampled
ofgenomic DNA in a total volume of25 pL, PCR every 100 generation sI,n total ,1O,OOO trees were
amplification was performed in a PCR Thermal obtained and the firs t2,500 samples from each
cycler Dice (TaKaR Baio). The PCR cycle fbr run were discarde das burn-i nto ensure the chains
trnL-trnF was 950C for 1 min; fbllowed by 25 had become stationary. A majority rule consensus
cycles at 950C for 45 s, 51OC for 1 ,5 min, and 720 tree was generate dto compute posterio prrobabili-
C for 1.5 min; and a fina lextension at 720C for 10 ties .Insertion!delet ievoennts (inde lwesre) not
min. For trnK, the settings were 960C fbr 1 min; include din the NJ and Bayesian analyses, Indels
fbllowed by 35 cycles at 960C fo r45 s, 480C fbr ofmore than two base pair swere treated as binary
1 min, and 720C for 1.5 min; and a fina lextension (O! 1c)haracters for the MP analysis ifnot caused
at 72eC for 15 min. The extension time in both by a simple sequence repeat.
PCR program swas increase dby 2 s every cycle.
Amplified DNA was purifie dusing a GeneClean
III DNA purificati oKnit (BIO 101, Carlsbad,
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186 ActaPhvtotax．Geobot． v 1，59
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188 Acta Phytotax .Geobot. Nlt) 15.9
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and
FIG.1, Geographi dcisnibutio nof Entada haplotypes .Haplotypes are designate dby name as they appear in Table 1 and Fig. 2.
Circle sand niangles indicat ethe clade to which each haplotype be]ongs (se eFig, 2). Numbers in parcnthcsc safter haplotype
names indicat enumbers of individual sS.eed typ ei sshown as S or L according to Wakita et al, (2005 )(.a)-( cDi)s:nibution
rnaps. (d) P:icture sshowing seed types .Scal ebar =: 1cm.
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E. Tanzania
aby
E. pol.ustacl t- yMaexico
POI:Japan;Ibu(2)-Okinawalsl.K,ohamaIsL,

Uganzaki-lshigakiIsl.

Taiwan; Chiopen g(3)
P02:Japan;YUtsun-IriomoteIsl.

gRgetw\.R
P03: Taiwan; Naajen-shan
P04:Japan;Ota-IshigakiIsl.

P05: Japan;Motonagura -IshigakiIsl.

P06:Japan;Nagura,Sokohara-IshigakIisl,

P07:Japan;Nakama -InomoteIsl,

P08:Japan;Funaura-IriemoteIsl. om-am

P09:Japan;YbnaguniIsl

PIO: Vanuatu ;Efate Isl.
Pll:IndonesiaJ;avaIslB8

P12:Indonesiaf'ernateIsl SulawesiIsl.
tsgg"Q"N"ts
P13:IndonesiaA;mbon IsL

P14: Indonesia;JavaIsl.E43a,KaiIsls.

P15:AustraiiaQ;ueenslan3d01

P16: Samoa; Safat a605 -Upolu Isl,
P17: AustraliaQ;ueensland303

Samoa;Safata80I -UpoluIsl

TOI: Japan ;Yaku Isl .A,mami-Oshima Isl (.2) gygNoNx-aRo s･
Taiwan;Juntou(2)1lahtung(2)

China;Hong Kong

ps
Vinh Yen
rheedii Thailand ;Chiag Mai
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rheedli
E. -AustraliaQ;ueensland
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- 5 changes
FiG. 2. Stric tconsensus tree of 1 23 rnost parsimonious trees of E. phaseoloides-lik seamples and their allies obtained from trnL-
trnF and trnK sequences (leng =t h202 steps. CI = O.861, Rl == O.8607, RC = O.741) .Measures of support are given at each
node: NJ bootstra psupporttMP bootstra psupportfBayesian posterio rprobability.
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well supported (BS of99, 99, and 1.00 for Clade
Results I and 98, 99, and 1,OO for Clade II) .Clade I con-
sisted ofsamples from southern rfaiwan, Vanuatu,
Seedmoilphology Samoa, Australia, and Indonesia, where S-type
Using the key character of seed shape report- seeds were obtained, Clade II consisted of sam-
ed in Wakita et al. (2005 )th,e seeds were sorted ples from northem Taiwan, Hong Kong, and Viet-
into one of two groups, The seed type assigned nam, where L-type seeds were obtained. Samples
rlbrpe
to all available seeds is shown in Table 1, collected from the type locali toyf Entacl aphase-
S from Japan(Okinawa), (AmbonIsland,Indonesia) included
seeds were obtained oloides were
Taiwan,Indonesia,AustraliaV,anuatu, inCladeI.
southern
and Samoa. Type L seeds were obtained from
Japan(Kagoshima), [laiwan,China, Discussion
northern and
Vietnam. The geographic distributi oonf seed
types is mapped in Fig. 1. Pnesence qftwo species qf Entada in .Jdpan
The two well-defined clades ( Iand II) in
Mblecular analyses our phylogenetic tree are clearly consistent with
The trnL-trnF sequence was 1,OSO-1,057 the two seed typ egroups (S or L) recognized by
bp long and the trnK sequence was 2,513-2,537 Wakita et al. (2005 O)u.r results clearly indicate
bp long, Using the combined sequences, 19 the presence of two species ofEntado in Japan.
haplotype swere resolved among the 39 Entacla Entacl pahaseoloide wsas originally described
phaseoloides-lik seamples. Twenty-four indels by Merrill (1917 )Th,e type specimen (draw ans
were obtained across the matrix of all haplotypes fuba marina in tab 4, Herbarium Amboinense 5:
and three wcre coded as potentiall yphylogeneti- 5-8, Rumphius 1747) was obtained from Ambon,
cally informativ efor parsimony analyses. The Indonesia A.ccording to some floris twiorcks that
phylogeneti canalysis using Entacla abyssinica treated this species (Walk e1r976, Nielse n1992),
and E. polystachya as outgroups produced 164 the diagnosti ccharacters of Entacl aphaseoloides
most parsimoniou strees of 226 steps, with a con- are 1-3(or 4) pairs of asymmetric elliptic leafiets
sistency inde xof O,876 ,retention inde xof O.877, and seeds 3-6 cm long .Entado phaseoloides is
and rescaled consistency index of O.769 (Fi g2.), distribut tehdroughout subtrop{cal to tropica lareas
The NJ and Bayesian tree topologies were nearly from Southeas tAsia to the Pacifi cislands T.hjs
consistcnt with the MP tree presente dhere (Fi g2),, genera lcharacterization is applicable to all sam-
Entacl agigas was sister to another clade, which ples considered to be Entadn phaseoloides-like in
was furth edrivided into two major clades. One this study, The wide distributi roenported thus far
of those clades was composed of Entadd rheedii may represent the sum of the two species reyealed
and E, spirali s(bran csuhpport (BS )by bootstrap here .Our study, however, clearly suggests distinc-
or posterior probability of 71 (MP analysis), 85 tive distribut airoeans fbr the two species, Sam-
(NJ analysis), and O.90 (Bayesi aananlysis); BS is ples include din Clade I were from Southeas tAsia
presente din this order in the fo11owi ntgext )T,he to the South Pacifi cw,hereas samples in Clade II
second clade was composed of all Entado pha- was collected only in areas extending from Viet-
seoloides-like samples (BS of 96, 96, and 1.00). nam to Japan (Fi g1.) .Since the sample obtained
This latte crlade was furth esrubdivided into two from the type localit yof Entada phaseoloides
clades (Clad eI sand II) ,both of which were very (Ambon Island ,Indonesia )appears in Clade I,
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we conclude that this clade corresponds to E. the Henchun Peninsula ,southern Taiwan, Some
phaseoloides A.ll available seeds obtained from studies distinguis hthis species from Entada
samples in CladeIwere of the S type, Thus, the phaseoloides based on it ssmaller seeds (ca ,3
diagnostic seed character of Entacla phaseoloides cm) and greate rnumber of leafle tpair s(Hayata
is a convex seed with an angular margin, and the 1921, Ho 1985, Huang & 0hashi 1993) ,but other
range of E. phaseoloides encompasses the islands studies treat it as E. phaseoloici ebsecause of its
of eastern and southeastern Asia, a part of Oce- morphological resemblance (Walk e1r976, Huang
ania, and Polynesi a(Fi g1.). & Ohashi l977) .In this study, four samples col-
The second clade (Clad IeI )was composed lecte dfrom the type localit oyf Entado koshun-
of samples from Japan (Amami-Oshi manad Ykiku ensis (Hench uPenninsula, southern Taiwan) had
islands), Taiwan,Hong Kong, Viet- hapletypesPOI P03.Thesehaplotypes
northern and and were
nam, All avai]able seeds from these samples were included in Clade I and formed a clade having
type L (Tab l1e, Fig. 1) ,The samples compris- weak support with samples from the Ryukyu
ing Clade II have generall ybeen misidentified in Islands (Fi g2.), but no clear rnorphological or
Japan and Taiwan as Entacl aphaseoloide sA,fter biogeographic aalssociatien with this clade was
surveying various taxonomic works from Japan, observed, Furthe rtaxonomic study also supports
Taiwan, China, and Vietnam, the seed morphol- the dithcult yin distinguishi nEgntadZ ]koshunen-
ogy of Entacl atonkinensi sGagnep. appears to be sis as a separate species (Tatei est haLi 2008),
censistent with the samples in Clade II .Entado
tonkinensis was originally reported from Vietnam Geographic distributi oonfE .phaseoloide asnd E.
(Gagnepa i19n11) ,and it scharacteristically large tonkinensishaplompes
seeds were recorded as 50 mm wide and 60 mm This study revealed the approximate distri-
long ,Clade II contained samples fror nVietnam bution sof haplotype sof Entacl aphaseoloides and
and the seeds obtained from this clade were 39.5 E. tonkinensis (Fi g1.) .Both species have very
-59.1 mm wide and 45.8-64. 7mm long, which larg eseeds that can drif tin seawater, which might
is consistent with the seed size reported for E. contribute to their populatio nstructure. Other sea-
tonkinensis. Althoug hthe species has not been disperse dplant shave recentiy been studied using
Vietnam, distance
reported outside our study suggests a molecular markers and evjdence oflong
wider distributi oenxtending to China, Taiwan, dispers ailn haplotyp estructure has been reported
and Japan. (Takaya meta ai. 2006, Nettle & Dodd 2007).
Unlike the above discussion W,akita et aL For example, Hibiscus tiliaceu sL. (Malvaceae)
(2005 a)ssigned E}itad uphaseoloide sto the group has a wide distribut tihoronughout the Old World
with type L seeds and E. koshunensi sto the group tropics. Common haplotypes were distributed
with type S seeds, based on samples obtained throughou tthe Pacifi acnd Indian ocean areas, and
from the Ryukyu Islands ,Their assignment of geneti cdifferentiati aomnong population swas
Entado phaseoloicies to the type L seed group is very low (Takayam eat ai. 2006). However, eur
not likel ycorrect, as discusse dabove. Regarding results from the study of Entado in Asia and the
their assignment of Entada koshunensis to the Pacifi cregion are somewhat contradictory. Based
type S seed group ,our phylogeneti tcree does not on the distributi oofnhaplotypes (Fi g1,) ,few
show clear evidence to distingu itshhi sspecies, widespread haplotype sin Entada phaseotoides
Entado koshunensi swas originally describe dby are observed and a geographic concentration of
Hayata (1921 )based on a sample collected in haplotypes appears to exist, Haplotypes POI-
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P09 occur in Japan and Taiwan, haplotype sPl1- very large ,individu aglrowth and seed production
P14 in Indonesia ,and haplotypes PIO and P15- over many years might be one explanation for the
P17 in Australi aand Samoa (Fig .1) ,Although the colonization of an inlan dhabitat,
number of samples surveyed was 1imite dn,o com- In this study, a phylogeneti canalysis using
mon haplotype swere fbund among these regions. cpDNA sequences revealed that two distinct
The only case of long distanc edistributi oofn a phylogenetic groups of Entacla are present in
common haplotype is in Australi aand Samoa, Japan, Morpho]ogically ,ene clade corresponds
separated by about 5,300 km, for haplotyp eP17. to Entado phaseoloide sand the other to E. tonk-
The seeds ofEntacla phaseoloici earse viable inensi sA,lthough seeds of Entada phaseoloides
for at leas t1 year in seawater (Nakani s1h9i94); are remarkable examples of sea-drifted seeds, evi-
thus, seeds can be disperse dby ocean currents dence of frequen tlong distanc seeed dispers awlas
over very long distance sT,he distributi oofn each not obtained. Even if the seeds can be dispersed
haplotype ,however, was geographicall lyimited by ocean currents across very long distances
(Fi g1.). One possibl eexplanation for this is the (Ridl e1y930) ,the geographical concentration of
low ability of seeds to establish in new localitieshaplotype ssuggests that another facter ,such as
after successfu1 dispersa tE.ntacl aphaseotoide sis diMculty in establishment, prevents greater gene
rarely seen in littor aarleas and is more common flow among regions,
in the canopy of inland forest A.lthough seeds
frequentl ryeach open sandy beaches ,subsequent We thank Drs B. Adje, K. Chayamarit ,C. Nyomdham, T.
colonization may be dirncul tO.ther factor ssuch Yikawa, Josph Yip, L. K. C. Chau, N, N. Thin, S. Mat-
D. Neil, Bogor BotanicalGarden,MEXU,
as the directio onfocean currents may also result sumura, and
in the geographical llyimite dhaplotyp edistribu-QthCaNnkED rfsbrY .thWeairt ahneol,p Ti.nD cobntadinai,ngT.A ssaamkplaews.a,K .WTeak aalysoa-

tion. Further studies using more samples from rna for their valuablc cornments and uscfu1 suggestions
various localiti easre necessary to clarify this during this study, We also express our great thanks to
issue. Drs G. Keniccr and D. Boufford fbr critically reading
For Entadu tonkinensis, Clade II included the manuscript. This work was mainly supported by
only two haplotypes w,hereas Cladc I of E, pha- JSPS K.AKENHI No. 16370043 (t oT.K.) and 14405015
17haplotypesfrom23 (t oY/T.) P.art of the study and sampling were supportcd
seoloides contained sam- by JSPS KAKENHI forforeign 14405015
expeditions
ples. This perhaps reflects the population size and and 12575011 (t oY,T. )and 17255004 to (J.M. )T.his
distribut iroannges of these two species, In Clade work is part ofthe PhD dissertati oofnN.W.
II, haplotype(TOI) fbundin
a common was eight
infive (.Fi2g),.The distribu-
populations regions References
tion ofhaplotype TO1 might have been due to dis-
persa lof seeds from continental Asia northward Doyle, J. J .& J, L. Doyle. 1987. A rapid DNA iso]ation
to Taiwan and Japan along the islan cdhain, as the procedure for small quantiti eosf fresh leaf tissue.
habita tof Entacl atonkinensis in Japan and Hong Phytochmem. Bull ,9: 11-15.
Kong is on slopes exposed to the sea at 20-150 Gagnepain, F. 1911. Mimosees nouvelles. Jn: H.
m. The occurrence in Taiwan, however, is inland Lecomte, Notulae Systematica c2: 56-61,
Hayata,B, 1921. Entada. Icones Plantarum Formo-
about 15-30 km from the seashore, How this spe-
sanarum 10: 3-5.
cies has successfu11y colonized an inland forest Ho, F.-C, 1985. Notes on the genus thtado of faiwan. J.
from the seashore on which seeds washed ashore Taiwan Museum 38 (1) 7:S-SO.
is still unknown. As the vines ofthis species grow Hu, J,-M, ,M. Lavin, M, F. Wojciechowski & M, J.
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