Evolutionary relationships of the genus Caloglossa (Delesseriaceae, Rhodophyta) inferred from large-subunit ribosomal RNA gene sequences, morphological evidence and reproductive compatibility,

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Evolutionary relationships of the genus Caloglossa Delesseriaceae, Rhodophyt...Mitsunobu Kamiya; Giuseppe C Zuccarello; John A WestPhycologia; Sep 2003; 42, 5; Academic Research Librarypg. 478


Fig. I. Drawing or a Ca[ox[v.,·.w thallul> at the node. Axial celb aI"~h,1l1ed <lnd ~econdary pit connections arc omilled. B. aba i<ll side:

D. adjacent side to lhe laleral branch: X, <ldaxi,t1 side: FLA. first<lxial cell or the lateral axis: FMA, firsl a ial cell or the main axis:LA, lateral a is: LP • laten" pcricel1lr<l1 cell: M . main axis; A.nodal <lxial cell: Of'. opposite sid", to lh'" I<lleral branch: TP . transvcrs"l pcriccmral c II; W . wing cell. Scale b<lr = 100 1'.111.

(West 1991; Tanaka & Karniya 1(93), but West (1991) reported that the dirrerences in blade shape between C. bcc('urii

and C. slipi!Uta appear in culture to be dependent on growthconditions. Purthermore, C. slipitatu had been known to haveonly adventitious branches, but Pedroehe el al. (1995) collected a putative C. stipitata specimen from Guatemala withendogenous branches. This charaetcr is generally so stable thaIthe Guatemalan specimen required a more detailed comparison with C. slipilalu.

QUI' previous studies indicated that C. Icpricllrii can be divided into two types based on the number of cell rows (singleor multiple) formed from the first axial celIoI' the main axis(FMA) opposite the lateral branch (Fig. I; Kamiya cl u/. 1995,1(98). Although these morphological entities arc reproductively isolated from each other, their phylogenetic relationshipwas not fully resolved by a molecular analysis using theRuRisCo spacer and its fianking rcgions (Kamiya el al. 1998,2(00). Asexual specimens or C. leprieurii from Pacific Mexico were described as a distinct species, C. uporneiolica West& Zuccarello (West cl u/. 1(94), but West el al. subsequentlyconcluded that asexuality arose several times within C. le

prieurii hee'Hls,=, the asexual strains isol:lted from gcogmphically distant populations were resolved as polyphyletic (Westct al. 200 I). Nevertheless, they dcrerred making a taxonomicdecision about these asexual strains until further molecularanalyses could be perrormed.

We have continued to collect algae from mangroves aroundthe world and successrully obtained most Calog!ossa taxa,including those mentioned above. Furthermore, we have established and maintained hundreds of strains, which makes itpossible to investigate morphological plasticity, reproductivecompatibi lity and physiological characteristics. Figures 2-13show a selection of Caloglossa species to demonstrate the

Kwuiya et al.: Systematics of Caloglossa 479

range or variation in overall and blade morphology: C. ad

IU/erells R.J. King & Pullock (Fig. 2) and C. COlllilll/O (Fig.II) are characteri/.ed by continuous blades without constriction; blades of C. !J('«('({rii (Fig. 5) <lnd C. .1/illiIUIU (I,'ig. (l)

arc strongly constricted at each node; C. ogu.luH'umellsi.l' (Fig.4), C. .\'({igonellsis (Fig. 9) and C. jJosliu(' (Fig. 10) have muchnarrower blades than the other species. The aim 01' this studyis to resolve the systematic problems in Culoglos,\'({ and revealthe evolutionary pattern in this genus. Nodal morphology(Fig. I), blade shape (Figs 2-13), rhi/,oidal organi/alion (Fig.14) and branching pattern were examined in both field andcultured specimens to veril'y their morphological status andtaxonomic utility. Crossing tests were undertaken to c1aril'ythe reproductive compatibility among the Illl;mbcrs or the C.conlinuu complex.

The RuBisCo spacer and its flanking regions have beenshown to be useful for inl'erring the relationship between morphologically related species of Culoglos.lu, and between populations or C. !cprieurii (Kamiya cl al. 19lJX; Zucclrello el al.

20(0), but the rate of hase substitution is Loo high to align thesequences of all the species with confidcnce (Kamiya 1'1 al.

2(00). Recent studies have suggested the potential 01' lhe nuclear-encoded large-subunit ribosomal RNA (LSU rDNI\ =

265 rDN 1\) gene sequences for phylogenetic analyses or algae(Freshwater el al. 1999; Lin ct al. 200 I) and vascular plants(Ku/.off el 01. 19(8). Zuccarello and West (2002) showed thaIthe LSU rONA had a slower mutation rale than the Ru8isCospacer in f]o.l'frvchiu species, which should make LSU rDNI\more suitable than the Rul3isCo spacer for studying the evolutionary relalionships between and wilhin species or Culog

fossa. Therefore, we initiated an analysis 01' LSU rDNA sequences in Caloglossa.

In this study, we describe the Guatemalan alga as a newspecies, C. rofundalu sp. nov., based on morphology and molecular phylogeny. We also propose thai the name C. Fieillur

dii (Ki.it/.ing) Setchell should be restored for one or the twomorphotypes of C. lepriellrii and that C. ('olllillllU ssp. uxil

loris and C. ujJo/lleiolicu should be placed into synonymy withC. /'i1ollo.l'liclw and C. leprieurii, respectively.

MATERIAL AND METHODS

The methodologies for collecting tile algae and conducting thecrossing experiments arc described in Kamiya cl al. (!lJ97)and West & Zuccarello ( 1(99). Collections or Caloglos.\'{{ species were made at the localities indicated in 'I;lble I. Livingand fixed materials were mounted in 40% sugar syrup in distilled waleI' with 0.004% collon blue, 3'Yr I'orm,din and 3'YrHel. 1\11 specimens without herbarium voucher numbers arcmaintained in culture in Kobe University Research Center forInland Seas. The isolates were incubated in Provasoli enrichedseawater medium (PES; Starr & Zeikus IlJ(3) ,It 20°C at aphotoperiod of 14: 10 h light-dark under 4-40 [Lmol photonsm-'} s I from cool-white Huorescent lUbes. Each isolate wascultured in a 100 ml clean cup 90B (Risu Pack, Tokyo. Japan)containing 5U III I PES medium, occasionally supplementedwith I mg I' Ge02 to suppress diatom contaminants. Theculture medium or each strain was changed once a month, and50-2DD [LI of an antihiotic mixturc (10,000 units pcnieillin Gpotassium, 25 mg streptomycin sulfate, D.I mg chloramphen-


4XO Phyc%gia. Vol. 42 (5), 20m

Figs 2-13. Ve ctativc thalli of Ca/og/o.\·.\·a ~peeies. Scale bar = 2mm.Fig. 2. n/og/os.Wl ndhaerens from Isobe River. Ishigaki lsland. Japan.Fig. 3. a/og/ossa henga/ensis from Daintrcc River. Queensland. Australia.Fig. 4. a/og/o.uo ogasall'arael/si.~from Tone River. hiba. Japan.Fig. S. n/og/ossa he earii from 'ungai Kolok. Thailand.Fig. 6. a/oli/ossa .~Iipi/(I/a from Amb n. )ndonesia.Fig. 7. a/og/os.1'lI 1lI00IOsliclw fr m Teluk Awang. Lombok, Indonesia.Fig. S. a/og/ossa /l/O//Ostielw from ao Paulo. Bra~il.

Fig. 9. a/og/o.\·.\·a saigo/H'I/si. from andakan. Sabah, Malaysia.Fig. 10. Ca/og/o.Ha postiat! from Georges River, N W, Auslr:llia.Fig. J J. (i1og/ossa COlllil/l/a from hiotsuka River, Fukuoka. Japan.Fig. 12. a/og/ossa /epriel/rii from mlala7.i, alaI. South frica.Fig. l3. Ca/og/ossa il/tel'll/edia from James I. lanel. South nrolina. A.

9


Kamiva et al.: Systematics of Caloglosso 4X I

"C. rotundata

Fig. 14. Comparison or rhizoidal losllHm around the nude in lIfoJ;.fos.w species. Shaded cells indicate Ihe position 1'J'()111 which rhiwid:i1filum nls originalc. Axi,i1 cells ar drawn by dolled lincs. ClIfogfo.l'.w lepriellrii, ,Iricf(uf{/ and C. vil'ilfardii do 1I()l have lhe adaxial eel I (blm;kcell in lype F) 1'1'0111 lhe FLA.

icol, and 2500 units polymyxin B sulphate pCI' millilitre ofdistilled water: Tatewaki el al. 19X9) was added to conlrolbactcrial growth. Male and femalc gametophytic strains wereusually established by isolating tctraspores released from [hetetrasporophytcs, this generation being thc dominant one innature.

DNA cxtractions followed Zuccarello et al. (1999a), andamplification and sequencing were performed as described inZuccarello & Wcst (2002). Approximately the last IWO thirdsof the gene were sequenced and analysed using the primercombinations given in Harper & Saunders (200 I) (corresponding Lo the Y and Z fragments). Taeniol11a pefpusillu/'/1 (1.

Agardh) J. Agardh, which is assumed to be from the genusmost closely related to Caloglossa (Papenfuss 1944, 1961),was lIsed as an outgroup. The sequences obtained werealigned for phylogenetic analyses using thc Clustal X computer program (Thompson et al. 1997) and refined by eye.Ambiguously aligned regions were removed beforc analysis.Phylogenetic relationships were inferred using PAUP* 4.0b6-10 (Swofford 2002).

Maximum parsimony trees (MP) were constructed using theheuristic search option, 500 random sequence additions, treebisection-reconnection (TBR) branch swapping, unorderedunweighted characters, and gaps Lreated as missing data. Theprogram Modeltest vcrs ion 3.06 (Posada & Crandall 199R)was used to find the model of sequence cvolution that bestfits the data set by a hierarchical likelihood ratio test (or. =0.(5). When the best sequence cvolution model had been determined, maximum likelihood (ML) and distance searcheswere performed in PAUP* using the cstimated parameters(suhstitution model, gamma distrihution, proportion of invariable sites). Distance trees were constructed using neighbourjoining reconstruction (NJ) (Saitou & Nci 19R7). Maximumlikelihood was also used to construct the most likely tree fromthe daLa scL (fivc random additions).

For bootstrap analysis (Felsenstein 1985), 1000 bootstrapdata sets were generated from resampled data (fivc randomsequence additions), for both M P and NJ analyses, as implemented in PAUP*, and 100 resamptings for ML analysis usingPhylip 3.6 (Felsenstein 2(02). Decay indices (Bremer 198X),on a strict consensus of the most parsimonious trees, werecalculatcd with AutoDecay version 4.0.2 (Eriksson 1998).

The 'one degree of freedom' (I D) and 'two degrees of freedom' (2D) methods were used to cxamine variation in substitution rates between lineages of the CaloM/ossa speciesstudicd (Tajima 1993; Miyashita et al. 199X). These methodsare based on thc cxpectation that under a uniform ratc of substitution, the numbcr of sites at which the nucleotide slale isshared by thc outgroup and only one of thc two ingroupsshould bc equal for both ingroups. The 2D method, especially,is applicable even when thc pattern of substitution rates isunknown and the substitution rate varies among different sites(Tajima 1993).

RESULTS

Morphological analyses of C. heccarii, C. stipitata and C.rotundata sp. nov.

We distinguished C. beccarii from C. s/ipi/ala on the basis ofits more elliptical or lanceolate blades. and identified the twospecies from fivc and six localities, respectively. A detailedexamination of blade morphology was pcrformed on fieldcollected or cultured spccimens Cfable 2). Among the morphological charactcrs examincd, the blade width in [he upperparts of thc plant was the best characLer to disLinguish bctweclIthese two species (Fig. 15). This was dcmonsLrated in [hestrains of C. beccarii and C. slipi/afa incubatcd under identicalculture conditions (Table 2; Fig. 16). Il was also apparent [hatthe formation of nodal rhizoids was more extensive towardsthe blade margins in C. ,I/ipi/({/(f than in C. heccarii (data notsbown), although this di l'f"rencc was not :dW:1YS cvidC'nt inunderdeveloped thalli. King & Pullock (1994) demonstrateddifferences in branching angles beL ween C. heccurii and C.slipitata, cspecially the angle of divergence of Ihe lateralbranch from Lhc main axis and Lhe angle of the blade definedby branching. We examined these angles in many wild andcultured specimens of C. beccarii and C. s/ipita/(f but couldnot confirm significant difference in either charactcr (data no[shown).

Detailed phenetic examination was performed on C. W/III/

data sp. nov. (Figs 17-20). The blade of this specimcn isstrongly constricted at the nodes and elliptical to rotundate in

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Table 1. Source of Caloglossa species examined in this stud.. peci s names in parcnthe e are newl, assigned on the ba. is of thi ·lUdy. New aeee ,i n numbers arc from the GenBankdatabase.

pecies

C {Idlwerell.r· adlwerell.

C adhaerens· opomeiOlica (C leprieurii)

C apomeiotica (C leprieurii)· apomeimica C leprieurii)

C apomeiotica (C leprieudi)C apomeioticn (C "ieil/ardii)C beccoriiC. beccariiC beccariiC beccariiC beccariiC. be,'coriiC. beccarii

bet/golellsisC bel/golellsisC. cOlllillllGC. cOlltllllla

· cOntillllaC. cOlllinuaC. ('0111111110 ~ p. a.ril/oris (c. monostic/w)C cOlltillua ssp. oxil/oris (c. 1II0llosticlw)C. illlermediaC. illlermediaC. leprieurii (C "ieillardii)C. leprieurii (c. "ieil/ardii)C. leprieurii (C I'ieillardii)C leprieurii (c. I'ieil/ardii)C. leprieurii (C vieil/ardii)C. leprieuriiC leprieurii

· leprieuriiC. leprieudiC leprieurii

· leprieuriiC. leprieuriiC. I/lollosticha

· llIonostic/1({C. mOIlO tiell({

· mOllostichaC. monastic/IllC. mOl1osticlwC. ogasawC/I'oensisC. ogasaworaellsisC. ogosall'araellsisC oga oll'ortlensisC. postiaeC. postiaeC. postiaeC. saigonellsis

Location

lobe River. 1 'higaki 1 land. 1 panDaintree River ro ing. Queensland. u:tralia

im Chu Kang. ingaporel. Espiritu amo. RC.S.. MexicoImercoastab Waterway. Ft Pierce. FI ridaisle of Palm. oUlh ami ina. lSADaintree River. Cookl \\In. Queen. land. u.lraliaGarden I land. delaide. Auslralia

ungei herok. Perak. MalaysiaDainlree River. Queen. land. u,traliaKranji. ingaporePasir Ris Park. ingaporeLim Chu Kang. ingapore

ungai Kolok. ThailandH.o Chi Minh. VictnamKranji. SingaporeHo Chi Minh. VietnamWenzhou. ChinaKuriyama River. Chiba. JapanTaiho Pef.. T:liwanGiao Thuy. Vietnam

oUlh Iligator River. Northern Territory. AustraliaEast Alligator River. Northern Territory. Au traliaJames Island. Soulh Carolina. U

apelo I land. Georgia. SAPort nhur. Ta mania. AustraliaPicnic Pl.. Georges Ri er. N W. u lralia

himajiri. Miyako Island. JapanNu. a Lembongan. Indone~ia

Tomkinson Ri er. onhern Territ ry. Au~tralia

Mayagucye . La Parguera. Pu rto RicoFara 'an I land. Red ea. audi rabialame [land. outh Carolina. ACobham. Wharf. Virginia. A1 la Morada. Florida Keys.Bali. Indone iaShimajiri. Miyako Island. JapanMonke Mia. Western u ·Iralia. Au traliaTeluk wang. Lombok. Ind nesiaR. Pereque. I. do ardoso. .P. BrazilPlantation, lorida Key . Florida. UBarll Puteh. Changi. SingaporeSandakan, abah. Malay iaGeorges Hall. Georges River. NSW. AustraliaGeorge Hall. George River. NSW, Au rraliaWilson Promolllor . VictOria. Au traliaSakuta River. Chiba. JapanShiira River, [riornote 1 land. JapanGeorge River. N W. AustraliaLiverpool River. Northern Territory. AustraliaPan Dick. on. Selangor. alay in

Date

23 lan. 19928 Dec. 1999131un.19919 ~ar. 1992191un.199413 luI. 19914 a t. 1995

22 lan. 19934 May 1971

ep. 199112 lan. 199317 Dec. 1991161un.1989

I ar.20002 - Jul. 200012 Jun. 198925 Jul. _00022 Mar. 199619 Dec. 199112 Mar. 2000I luI. 2000

I Oct. 199130 ug. 199714 Oct. 199524 Oct. 199027 Feb. 199316 Feb. 1999291an. 199225 pro 199922 ug. 1999

6 Mar. 1994luI. 2000

171un. 1994"!15 Jun. 199413 pro 1999I May 1991

31 Aug. 19927 pro I 99

4 pro 19901- lun. 199413 Mar. 199416 Aug. 200030 Apr. I 9830 Apr. 19981I Oct. 199722 Dec. 1991

2 lar. 199530 pl'. 199823 Aug. 199914 May 1998

Strain no.

MKI13240472996327634213173r383328

2992E 67MKI282995MKI290MKI073MK639MKI271MKI289

MK112()35353092

360DB 1MK739~970

F442340240703407GOllillgen 101.793431MKI20MK465D567397930553426MK9874094GZ81GZ613783MK620MKI061C889F406, 46

Acce.ssion no.

F52219AF522199AF-22200

F522201AF522202AF522203AF522204AF522205

AF522206AF522_07A 522208AF522209AF522210AF522211AF522212AF522232

F522233

AF512213AF522214AF522215AF522216AF522217

F522218F522219

522220AF522221

F522_22F522223

A 522224AF522_25AF-22226

F522227A -2222'AF522229AF522230AF-22231AF-22234AF52223A 522236AF522r7AF522238AF522239AF52224()AF52224IAF522242AF522243

ColleclOr

Yl. Kamiy,t1.A. West1.A. c t

1.A. e~l

J.A. We t1. . We t

J.A. estJ.A. eSI

. KumanoKing ,PullockM. KamiyaJ. anakaJ.. WestH. kiyama1. TanakaJ.A. WCSI

1. TanakaT. KitaymnaM. KamiyaA. Yokoyama1. TanakaM. KamiyaR.1. KingI.A. We I

M. Darley1. . W~tG. . ZuccarelloM. Kamiya1.A. e I

1. . WestJ. eSI

M.1. Hli ain1. . WeslF. all1. e.1M. KawachiM. KamiyaM. KrtitzenJ.. We I

1.A. We. I

1.A. WestM. Kamiya1.A. e tG. . ZuccarelloG. . Zuccarello1.A. We rM. KamiyaM. KamiyaG.C. Zuccarello1. . We I

J.A. Wesl

<


Kal1liyu et '11.: Systematics or Cu/og/ossa 41'n

I I I I

shape (Figs 17, I X). Secondary endogenous branches rrequently arise rrom the nodal cell or the first axial cell abovethe node and out or the pl<lne or the Iha/lus (Fig. Jt). Inaddition, adventitious branches arc rormed in the nodal regions of the blade by interstitial prolikration arising laterallyrrom marginal wing cells (arrow in Fig. 19), but these occurmuch less frequently than endogenous branches. Rhi/oidal filaments are derived from the nodal pericentral cells, or the lirstto third pericentraf cells immediately above and below thenoclc, or sometimes from the wing cells adjacenl to these pericentral cells (Fig. 20). The positions or the rhiwids rrcquentlyextend towards the base (type C in Fig. 14). whereas those orC. l7eccurii and C. slipiluiu extend to the blacle margins (typeJ) in Fig. 14).

Morphology and el'ossability of the C. continI/a complex

Thallus morphology was examined in both field-collected andcultured specimens of the C. continuu complex (c. conlinua

ssp. continl/lI, C. continl/O ssp. ar;//lIl';S, C. nW!losliclw, C./wsl;lIe and C. su;gonensis; Table :\; Figs 21. 22). ClI/og/os.\'((

conl;nuu ssp. uxi//uris, which is characleri/.ed by wide bladesand monosiphonous hyphae along lhe axis. was collected fromthe East Alligator River, Australi<l, close to the type localityor this subspecies. We also collected spccimens lhat werc verymuch narrower than C. lI1onosl;c!tu. from Sandakan on BorneoIsland and Selangor on the west coast or Peninsular lalaysia.ami these two specimens were identified as C. suigollensis.

The field-collected specimens or C. cont;nuu ssp. u.ri//"ris

h<ld comparatively wider blades than other species in lhe C.conlinuu complex (Fig. 21), but lhe cultured strains or thissubspecies were indistinguishable rrom c. nwnost;c/lil inblade width (Fig. 22). The field specimen or C. .lu;g{}Jwns;s

from Sandakan shows similar gross morphology 10 the typespecimen or C. suigonensis rrom Vietnam (Table :\). Theblades of both specimens are mostly less than 0.5 mm wideand significantly more slender (han any others (I' < I 'lr) except for C. jJo.lliae, which is distinguishable from C. s,,;go

nens;s by the number of cell rows from the nodal axial cell(NA in Fig. I). Two Malaysian strains of C..migo/lc/lsis maintained their slender form in culture (Fig. 22). Most specimensof C. /lWnosl;c!ta have blades that arc not or only slightlyconstricled al the nodes (Fig. 7). but the strains from Bra/.il(:\055) and the United States (]426) show pronounced constriclion at every node (!"ig. Xl.

The number of cell rows rrom the NA was comparedamong specimcns of thc C. COIII;III.f({ complex Crable :\; I:igs2:\,24). Cu/og/ossu cont;/lua and C. !)o.I/;lIe from Japan produce multiple-cell rows at every node in the field condition(Fig. 2:\), and this reature was maintained in culture (Fig. 24).Multiple-cell rows were more abundant than single-cell rowsin C. cOllt;lIua rrom Taiwan and Vietnanl. By contrast, singkcell rows were more abundant than multiple-cell rows in C.mO!1oslichu from Singapore and C. UJIIli1ll1£l ssp. aril/oris.

Only single-cell rows were seen in C. II/OIf()sticlf({ from Australia, Rnv.il, Indonesia, Malaysia and the United Stales amiin C..wigo/lcf7sis, although l11u/tiple-ccll rows appeared in thecultured material of C. saigonells;s from Sandakan (Fig. 24).

Monosiphonous hyphae were carel'ully examined ill C. COII

t;nuu ssp. ax;l/ar;s (Figs 25-27). Initiaf cells of the hyphaeusually derive I'rom transversc peri central cells (Fig. 25), but

ci.v.

,:oc

I I I I

.,j

":>cCou


4X4 Phyc%gia, Vo!. 42 (5), 2003

0-40.3

1--.,-----,\---,,--.,---,..--,---'

0.1 0.2

Likin. Guatemala (20)

Cebu, Philippines (12)

Selangor, Malaysia (5)

16

Ambon, Indonesia (12)

Barambang, Brunei (6)

0.1 0.2 0.3 0-4upper blade width (mm)

Figs 15,16. omparison of the blade width ,II Ihe upper parI betweenC. heccarii (dosed circles), . mUll/data (open triangles) and C. slipiww (open circles). umbers of specimens examined are given inparentheses. and the error bars represent ~ 1'* standard error of themean.

Fig. 15. Field-collecled specimens.Fig. ]6. ultured specimens.

Cape York, Australia (4)

Sungai Kolok, Thailand (14)

Pasir Ris Park, Singapore (13)

Kranji, Singapore (18) •

Pasir Ris Park, Singapore (20)

Kranji, Singapore (17) •

Perak. Malaysia (7)

Daintree R., Australia (8) •

15 Likin, Guatemala (8)

Cebu, Philippines (4)


sometimes from lateral peri central cells or wing cells adjacentto the lateral pericentral cells. The hyphae mostly grow towards the base, occasionally towards the apex, beginning toappear at the fifth to seventh internodes and lying betweenthe axis and the peri central cells (Fig. 25). These cells are 5~

15 f1m wide and up to 120 f1m long. They form secondarypit connections with pericentral cells or adjacenl wing cellsand repeatedly divide along the axes. The hyphae frequentlybranch off at the lower part of the thallus and make a networkover the midrib ncar thc basc (Pig. 26). Cultured spccimensof this subspecies produced far fewer hyphae than field material, and they occurred only at the lower part of well-developed thalli (Fig. 27). Similar hyphae, although less abundant,were found in some field specimens of C. monosticha fromAustralia and Indonesia, but rarely in cultured material.

Crossing tests between members of the C. continua complex samples were carried out to determine the sexual compatibility and fertility of the hybrids (Fig. 28). No cystocarpswere formed on isolated female gametophytes used as negative controls. Successful crossings between male and femalestrains that originated from the same tetrasporophyte were regarded as positive controls, although the gametophyte pairwas not always obtained in the strains examined.

Ca[o!?[ossa fJostiae and C. l71on.osticha from Florida (maleonly strain) were completely intersterile with other strains.Viable F I gametophytes were produced between C. saigonen-

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'""0e.::B'0g'0<;IJ::

C

>.co0"0.<=e-oEOJ ,,;'0

'" C-0.D ._

'- d

° >c

...OJo v:

v:.D'c C

"''-l="0c '"c .....,

.D

N E:>.. c

:0'"~ '"


Kamiya et al.: Systematics of Ca/og/ossa 4S5

17

Figs .17-20. B lade morphology of . rowllda/a sp. no .Fig. l7. Hololype specimen. Scalc har = 5mm. TNS-AL-151390.Fig. 18. Upper lhallus of' a cullured specimcn. Scale b,lr '" 2111111. TN$-AL-15/39 r.Fig:. 19. mlal pan or the thallus. Both an adventitious secondary branch (arrow) and all endogenous one (arrowhead) have dcvclopcd fromIhis node. Scale btu' = 200 f.lol11. TNS-AI.-151:l91.Fig. 20. Monosiphonolls rhi'l.oids derived from periccmral cells and wing cclls around til nodc. Scale bar'" 2001-\0111. T S-AL-151:W I.

sis from Sandakan and Selangor, the latter yielding manypseudoeystocarps with males of C. mo//oslicha from Singapore, Australia and Indonesia. The Australian and Indonesianstrains of C. //w//oslicha, which were entirely compatible witheach other, produced fertile F , tetrasporophytes in the presenceof a male C. co//ti//ua ssp. axil/aris, whereas only pseudocystoearps were formed in the reciprocal crossings. The Singaporean strain of C. mo//osticha did not make cystocarps hutmany pseudoeystoearps with some other strains. The Japanesestrain of C. conlinua was interfertile with the strains fromTaiwan and Vietnam.

Molecular phylogeny based on LSU rDNA sequences

For each or the 4S strains examined, 1461-1662 hp of thesequence from the middle region of the LS U gene were ohtained. This fragment starts at bp 1300 of the complete LSUsequence for Orvza sotiva (GenBank Accession No. M I ISSS).These sequences were easily aligned hy eye, except for thesegments around bp 445-467, 690-725, I 163-1 IS2 and12SS-1304, where the lengths as well as sequences were extremely variable between strains. An alignment of 142i1 siteswas constructed which excluded these highly variable segments. These sequences had a mean G + C content of 51.0%and included 435 variahle sites (excluding gaps); 2SS of thesewere potentially phylogenetically informative.

The MP analysis resulted in five equally parsimonious trees(length = 1131 steps, consistency index = 0.55, retention index = 0.S2), and the strict consensus tree was generated fromthese trees (Fig. 29). The NJ tree was similar to the MP treeexcept for the relationships within C. /cpricurii (data notshown). Two maximum-likelihood trees (log-likelihood =

7922.75) were constructed using the estimated evolution model hy Tamura & Nei (1993) with the following conditions:equal base rrequencies; rate matrix A ---i> C = J .00, A ---i> G

3.30, A ---i> T = 1.00, C ---i> G = 1.00, C ---i> T = 4.33, G ---i>

T = I,C)O; gamma distribution = 0.49; and proportion of in-

variable site = 0.45 (rig. 30). The position of C. il/lcl'Il/cilia

varied in some reconstruction methods, although it was alwaysclosely related to C. leprieurii.

The molecular phylogeny divided the memhers of Ca/og

/ossa into three major clades, which correspond to the distribution of types of secondary branching system (Fig. 29). Thefirst clade consists of C. rollll/da/({ that produces both adventitious and endogenous hranches. Thc second clade (the adventitiously hranched group) is composed of the species thatproduce only adventitious branches, and the third clade (theendogcnously branched group) consists of the C. (,ol/Iil/lla

complex, C. illiamcilia and C. /cjJricllrii (including C. ({POIl/

ciOli('(1) that produce only endogenous branches). Among theadventitiously branchcd group, the relationship hetweenstrains C. bcc('arii from three localities was ambiguous. Ca/

og/ossa Icpricllrii is known to be di vided into two morphotypes, based on the numher of rows of cells produced fromthe FMA (sec Fig. I): eithcr single or multiple rows. Thesemorphotypes were phylogenetically different, and C. il/ler

mcilia was more closely related to thc type with multiple-ccllrows than to the single type. The two asexual strains fromAustralia were split, and one was included in each 0[· the twoC. /cpricurii lineages. The asexual strains from Atlantic Florida and Pacific Mexico showed a close relationship to eachother, but did not make a clade with either of the Austr,ilianasexual strains. The monophyly of the C. ('oll/inlla complexwas supported by all three analyses, but C. Il/ol/oslie//({ wasseparated into several lineages.

The molecular clock hypothesis was tested using the I Dand 2D methods. The numher oj· sites by which each of theingroup specimens differed from the outgroup was comparedin all possihle pairs of ingroup specimens (total lOS 1 combinations), and the null hypothesis that the substitution rate isequal for both ingroup specimens was tested for each pair.The substitution rate in C. il/lcrll/cilia was significantly faskrthan that of all the other species (/' <: I fir') in both methods.

Reproduced w



ithout permission.

Table 3. Comparison of blade morphology in field and cullUred specimens of the C. cOl/lil1l1G complex. Numbers represent ranges: mean values <lre given in parentheses. Il = number ofobscrv!llions.

Cell no. inCell row no. second~order Cell row no. Bhtde width (mm)

from internodal ro\v at from nodal Blade lengthSpecies Locality II axial cell internode axial cell Upper Middle Lower (0101)

ield specimens'. COIl/il/lla Chiba, Japan 20 4-9 (7) 12-18 (15) 2-6 0.5-1.0 (0.7) 0.5-0.9 (0.7) 0.4-0.8 (0.6) 0.9-2.4 (1.6)'. cOlI/il/lla Taipei. Taiwan 10 4-6 (5) 14-19 (17) 1-4 0.6-1.2 (0.9) 0.6-1. I (0.8) 0.S-o.9 (0.6) I. 1-2.S (1.7)'. COlI/illllO Giao Thuy. Vietnam 13 3-S (4) 9-19 (16) 1-4 O.S-13 (0.8) 0.4-1.2 (0.8) 0.3-0.9 (0.6) 0.7-1.7 (1.2)

C. mOf/os/icha Bali. Indonesia 10 S-6 (6) 16.....25 (21) I 1.0-1.9 (1.4) 1.2-2.0 ( 1.5) ().7- '-3 (0.9) 1.6-3.3 (2.4)'. c. ssp. axil/oris East Alligator River. Northern 10 5-7 (6) 23-33 (27) 1-4 1.0-2.5 (1.7) 1.0-2.2 (1.7) 0.9-1.6 (1.3) 11-2.6 (1.9)

Territory. Australia· mOllo.wicha Derby. Western Australia. Australia 20 4-5 (4) 18-28 (23) I 0.8-1.8 (1.2) 1.0-1.7 (1.3) 0.6-1.0 (0.8) 1.6-3.3 (2.3)'. 1Il0f/()s/iclw Pulau Ubin. Singapore 20 5-7 (6) 12-23 (17) 1-4 0.6-1.3 (0.9) 0.6-1.2 (0.9) 0.4-0.9 (0,7) 0.7-1.9 (1.3)'. lIlol1Qs/icha Bali. Indonesia 10 5-6 (6) 16-25 (21) I 1.0-1,9 (104) 1.2-2.0 ( 1.5) 0.7-1.3 (0.9) 1.6-3.3 (2.4)'. mOllos/icha Florida. USA 3 5-6 (6) 18-20 (19) I O.4-o.S (0.5) 1.0-1.3 (1.2) 0.5 (0.5) 2.4.....2.S (2.4)'. I/Iollos/icha Sandakan. Malaysia 14 4-6 (S) 15-20 (18) I 0.4-1.8 (1.3) 0.9-1.6 (1.3) 0.5-1.4 (0.9) 1.2-2.5 (1.7)'. pos/ill!! Ishigaki Island. Japan 20 3-S (4) 3-9 (S) 2.....4 0.2-0.3 (0.2) 0.2-0.3 (0.2) 0.1-0.3 (0.2) 0.6-1.6 (1.0)· saigol1ellsis Sandakan. Malaysia 8 3.....5 (4) 6-8 (7) 1-2 0.2-0.4 (0.3) 0.2-0.4 (0.3) 0.1-0.2 (0.1) 1.1-1.4 (1.2)'. saigolli!lIsis (type specimen) Cau Chu Y. Vietnam 16 3-6 (S) 3-18 (9) 1 0.2-0.6 (0.4) 0.3-0.6 (0.5) 0.2-o.S (0.3) 1.4-3.2 (2.3)

Cultured specimens'. cOll/illl/CI (MK639) Chiba. Japan 10 4-5 (4) 10-20 (15) 1-5 0.8-1.2 (1.0) 0.6-1.1 (0.9) 0.4-0.9 (0.6) 1.0-3.2 (2.4)'. cOlI/illl/n (MK1271) Taipei. Taiwan 10 3.....4 (3) 14-19 (17) 1-3 0.7-1.6 (1.0) 0.7-1.3 (0.9) 0.6-0.9 (0.8) 0.8-2.S (1.3)'. cOlllil1lw (MKI289) Giao Thuy. Vietnam 7 4-5 (5) 10-14 (13) 1-3 0.4-0.6 (0.5) 0.5-0.7 (0.6) 0.2-o.S (0.4) 0.7-1.3 (0.9)'. c. ssp. axillaris (M KI 120) East Alligator River. Nonhern 10 2-5 (4) 17-27 (22) 1-3 0.7-1.5 (1.0) 0.5-1.3 (0.9) 0.5-0.9 (0.7) 0.8-2.3 (1.4)

Territory, Australia· llIol/os/icha (MK892) Derby. Western Australia. Australia 20 3.....5 (4) 12-26 (18) 1 0.4-1.3 (0.8) O.S-1.3 (0.8) 0.2-0.9 (O.S) 0.8-3.3 (1.7)'. /1Iollos1icha (MK987) Changi. Singapore 10 3-S (4) 11 ..... 16(13) 1-2 0.4-0.8 (06) 0.4-0.9 (0.6) 0.3-0.5 (OA) 0.5-2.0 (1.4)'. lIlol1os/ic!w (3979) Lombok. Indonesia 8 2-5 (4) 9-18 (15) I 0.5-1.0 (0.7) 0.5-1.1 (0.7) 0.2-0.8 (0.5) 0.7-1.5 (1.0)· IIIQllos/idw (3426) Florida. USA 9 5-6 (S) 14-20 (17) 1 0.3-0.9 (0.4) 0.8-2.0 (1.2) 0.2-0.4 (0.2) 2.1-4.3 (3.4)· 1Il0llOS1icllll (3055) Sao Paulo. Brazil 15 6-8 (7) 13-24 (18) J 0.5-0.9 (0.6) 1.0-2.0 (1.4) 0.3-0.6 (0.4) 2.1-4.7 (3.1)'. pos/iae (MK96 I) Ishigaki Island. Japan 20 2-6 (4) 2-7 (5) 1-4 0.2-0.3 (0.2) 0.2-0.3 (0.2) 0.1-0.2 (0.2) 0.7-1.8 (1.2)

C. saigonel/sis (3846) Pon Dickson. rVlalaysia 13 3-5 (3) 3-10 (6) 1 0.1-0.2 (0. I) 0.1-0.2 (0.2) 0.1-0.2 (0.1) 0.6-4.2 (1.6)· saigonensis (4092) Sandakan. Malaysia 10 4-5 (4) 5-11 (8) 1-2 0.1-0.4 (0.2) 0.1-0.6 (0.3) 0.1-0.2 (0.1) 0.9-3.3 (2.0)

+>:x:~

'1;;::::-.

';S-~.

<2.+>N.--.Vl~

~8


/{«III/I'" L't ~\I.: SV,ll'1I1:tlil', "I' (·1110,1:10.1.11/ .+K7

23 JIJpflJl (0) <>Taiwan (13) ~

Vietnam (10) 1--{)--l

Australia (14) I-iSi

Australia (20) 0

Singapore (13) K)l

IndOnesia (8) 0

Florida (3) 0

Sandakan. Malaysia (14) 0

Japan (20)

Sandakan. Malaysia (8) •Vietnam (16)

r0 20 40 60 80 100

24 Japan (15) 1(>1

1Taiwan (9) t-{>-i

Vietnam (8) ~

Australia (12) I-fr-l

Australia (20) 0

Singapore (12) 0

IndOnesia (8) 0

Florida (9) 0

Brazil (15) 0

Japan (20) •Selangor. Malaysia (9) •

Sandakan. Malaysia (5) ~

I0 20 40 60 80 100

ra!io of mul!iple cell rows (%)

Figs 23, 24. OlllparisOIl or lhe mean ratio or lhe multiple-cell n)wswilh the single-cell row from thc nod'li axial cell in a U)alllls. Numl CI'S

of specimens cxamined arc given in parentheses. and the error bal'srepresent ::t 1% Slandard erwr or lh . mc:ul. Sec lh legend in Figs21, 22 r I' xplamnioll r the symhols.

Fig. 23. Field-eo!lccled spccimcns.Fig. 24. ulwrcd spccimcns.

1.6

o

••

o

Japan (10)

Taiwan (10)

Vietnam (7)

Australia (10)

Australia (20)

Singapore (10)

Indonesia (8)

Florida (9)

BraZil (15)

Japan (20)


Sandakan, Malaysia (10)

22

21 Jllpon (20) <>Taiwan (10) 0

Vietnam (13) 1(>1

Ausfralia (10) I---6rl

Australia (20) 0

Singapore (20) 0

Indonesia (8) I-O!

Florida (3) ~

Sandakan, Malaysia (14) IOi

Japan (20) ~

Sandakan, Malaysia (8) •Vietnam (16) •

0 0.4 0.8 1.2 1.6 2.0

0.4 0.8 1.2middle blade width (mm)

Figs 21, 22. omparison of the widlh at the middle pan of the bl"dbetween C. COII/illl/tl (open lo:t. 'nges), C. ('Olllilll/(/ ssp. axillaris (opentriangles). . 1II001(J,I'ric!w (open circles), . po.'liae (closed triangles)and . xaiXollel7xix (closed circles). Numbers of specimens c.xwn;lledare given in p,lI'entheses, and the error bars represent ± 1% standarderror of the 1lle<1Il.

Fig. 21. Field-collected sl ecimclls.Ii'ig. 22. ulwred specimens.

The molecular evolutionary clock hypothesis was also rejected at the I % level in the following combinations: C. leprieuriifrom Japan (MK739) and Australia (3328. 083 land F442);C. bengalensis (2995 and M K 1290) and C. rotundata (3375);and C. mDllOsticha from Singapore (MK987) and C. slipitata(3599. 3837 and 3844). The substitution rate of C. continua(MK639 and MKl073) and C. Inonosticha (MK127l andMK 1289) was significantly slower than that of C. s/ipi/ata(3599, 3837 and 3844) and C. bengalensis (2995 andMKI290).

DISCUSSION

Taxonomic conclusions

The Guatemalan alga is distinct from other species on thebasis of its unique arrangement of nodal rhizoids and the pro-

duction of both endogenous and adventitious secondarybranches. ea!og!ossa tric/ada (Post) R.J. King & Puttock isalso characterized by possessing both branching systems. butis obviously different from the Guatemalan alga by its lack ofthe adaxial cell row from the first axial celioI' the lalend axis(FLA) and its coalescent rhi/oid system. Thercl'ore we decided to distinguish this Guatemalan alga as a new species, C.ro/unda/a sp. nov.

Asexual specimens of C. !epriellrii had been recogni/.ed asa distinct species, C. aj7otl1eio/ica, but it is apparent that asexual reproduction arose independently more than once (Figs 29,30). This suggests that asexuality as a character is inadequateto delimit species and that C. apomeio/ica should be reducedas a synonym of C. !epriellrii. The present study strongly suggests that the two morpholypes of C. !cpricurii are phylogenetically different, so there is no reason for them to remaineonspeei1k. We observed multiple-cell rows from the FMA inthe type specimen of C. !cprieurii and a single-cell row in


---------------------------------- -- ---

4KK P!I.\'cologio, Vo!. 42 (5), 2003

Figs 25-27, Ilyphal cells or C. cOrllilll/ll ssp, axillari,I', Scale bar (illFig. 25) :; IOOJ.lI11,

\'ig. 25. Initial hyphal cells (arruwhC<lds) derived 1'1'0111 the transversal perieclltnll cells (TP ,,'rrowed) IUwards the b"se in a fieldcollccted sP' 'il1lcn.Fig. 26. Network or hypha I cells <'/eveloped ,,1011(' the axis ill a fieldcollcclcd specimen.Fig. 27, nelerdevclOI eel h phac in ;1 cullUred lhallus.

those of C. leprieurii var. !Iookeri Post, C. IlIllioides Harveyex.l. Agardh and C. vicif!ardii (Klitzing) Setchell, all of whichKing & Pu(lock (I (94) placed in synonymy with C. !cpricurii.ThercJ'ore, C. I'ieiffardii (Figs 31, 32), which was describedearlier than C. leJ}f'icurii var. !Ioo!<eri and C. //lnioidcs, shouldbe used for the single-cell row type of C. Icpricurii.

Thc close relationship between C. continua ssp. axif!arisand C. /I1ollostic!la from northwest Australia (the type locality)and Indonesia was strongly suggested by the production ofhyphal cells, reproductive fertility and close genetic distance,lL is therefore reasonable to combine C. contilllf({ ssp, axilloriswith C. /I101l0.I'tic!lo, Although the heterogeneity of C. /IlOIl

osticfw is apparent in this study, the phylogenetic relationships

within the species arc still ambiguous. Further cxamination isnecessary to resolve the systematics of C. Ilionostic!lo, especially of the Atlantic and Malaysian populations. The keymorphological characters for distinguishing Caloglossa species are listcd in Tablc 4,

Morphological evolution in the Caloglossa lineage

This study demonstratcs the uscfulness of LSD rDNA in infCITing the evolutionary relationships between and within species of Caloglossa, although there are some segments of thephylogenetic tree that cannot be aligned with confidence, Percent LSU rDNA sequcncc divergence valucs ranged from 0,4to 12,0 between species and from 9,3 to 12.1 between genera,excluding C. intcr/l1cdio, which showed extremely high genetic distance values with other species (discussed below).These ranges are much higher than those found for LSDrONA sequences from gelidialean taxa (0.1-2,4 between species, 4.0-6.R between genera) but similar to those for rhcLsequences (3.1-1 1.5 betwcen species, I 1.0-15.5 between genera) (Freshwater & Bai ley I (98). The tribe Cafoglosseae,which ineludes CaloKlossa and 7'acnioma .I. Agardh, is assumed to he ancestral within the family Delesseriaceae; all itsmembers possess simple blades and form exogenous apicalinitials, all second- and third-order initials reach the thallusmargin, and the primary eell rows lack intercalary divisions(Papenfuss 1944, 1961). It is reasonable to assume that manybase substitutions and length mutations in the LS U have accumulated in this cvolutionarily old group, The evolutionaryposition of the genus Caloglossa was unresolved in a recentphylogenetic study on the Delesscriaccae (Lin el al. 200 I),although it could still be ancestral within the subfamily Delesserioideae.

The present molecular phylogenetic results suggcst that C.rolundata, which has mostly endogenous secondary branches(rarely adventitious), is morc closely related to the adventitiously hranched group than to the endogenously branchedgroup. This relationship was demonstrated in the Ml~ ML andN.I analyses, but the bootstrap values were not always high

~C. continua C, monosticha

C, c. ssp. C,C, postiae

axillaris saigonensis

MK728 MK1271 MK987 MK892 3979 MK1120 3846 MK1122Japan Taiwan Singapore Australia Indonesia Australia Malaysia Australia

C, continua MK728 Japan + + -* - - - - -

C, continua MK1289 Vietnam + + - - - - - -C, monosticha 3426 USA - - - - - - - -C, monosticha MK986 Singapore -* -* + - - - -* -C. monosticha 890 Australia - - -* + + -* -* -C. monosticha 4092 Indonesia - - -* + + -* -* -

C, c. ssp. axillaris MK1120 Australia - - -* +0 +0 + - -

C, saigonensis 4092 Malaysia - - - - - - + -

C, postiae MK1122 Australia - - - - - - - +

Fig. 211. Rcsulls or l.hc l1ybridiz<llioll experiments, +, 1', spol'lJph les and subscqu III F, gHmClophyles wcr vil,ble (shaded); +0, F, spomphyleswcr I'cnil bUI subsequenl F, gamelophyles W'I' not viahl : _ .., pseudoeyslocllrpS were pmclu(;ecJ: -. no reproduclion occurred,


Kwniyu et al.: Systematics of C((!og!os,1'({ 4i\9

C. apomeiotica (G. vieillardii) Australia

~1001100C. leprieurii (G. vieillardii) AustraliaC.~prkum(C. weillard0Japan +

§16 +G. leprieurii (C. vieillardii) Indonesia + +-+

~J::

C. leprieurii (C. vieil/ard;;) Australia £5 Gl ~C. leprieurii USA "0 "8 '- eG. leprieurii USA

.~ c u.. ~OJ OJ <I) BC. leprleurii USA '0 :6 ! "0

991100 ro :§ ro38119 G. lepneurii Puerto Rico ::c iii .~8 c "'0 -E1 C. leprieurii Saudi Arabia 0 '0 "0

C. leprieurii Indonesia ~ ~Ql ro0.. '0

C, apomeiotica (C. leprieuril) Australia :s ""69100 I/) "'5 8 g'3 G. reprieurii Japan

c E+ 8 c i5c. apomeiolica (C. leprieuri/) USA 5!97198 .c c11 C. apomeiotica (C. leprieuri/) Mexico + ro l!!

C. apomeiolica (c. leprieuri/) USA + .0

~C. inlermedia USA + ::>C. posliae Japan 0c

98100 1001100 C. postiae Australia + Ql

8 27 C. posliae Austalia ~'0

42121 C. monosl/cha Australia + c:Ql

2 C. mOlloslicha Indonesia

5317C. C. ssp. axillaris (C. monoslicha) Australia

3 C. monosl/cha SingaporeC. conlinua China + (.!)C. conlinua Japan I-3416 1001100 + :)

2 14 C. conlinua Taiwan + u..G. conlinua Vietnam + E

651100 1001100 C. monosticha Brazil B ,g+ 8- Ol

3 10 C. monosticha USA + :: ce :c

C. monoslicha Malaysia "0 ()40/28 '0

~c:

3 C. saigonensis Malaysia + .!::l roC. saigonensis Malaysia

... -E ..aro >-

C. slipilala Philippines .SI 'x Ui1001100 + Ol CIl ::>

G. slipilala Malaysia + + 0 c:: "0 025

... + 'iii ro c::C. slipilala Malaysia '0

Ol Q)1001100 c:: Ol

15 C. adhaerens Japan Q) :s 0... Q "0C. adhaerens Australia +. « c c:: c

+ OJ ~Q)

75193 C, adhaerens Singapore () .c "02! .2' c::

3 C. ogasawaraensis Australia 0- ro+ ... UJ I/) >-

C. ogasawaraensis Japan 5 Uiwoo c. beccarii Thailand :0 :J

E 0+

..,5 C. beccarii Singapore + 0 %! ~

C. beccarii Vietnam ... "0 <ll64198 ro :>

100/100 C. bengalensis Singapore ... '01 + CO ro

34 C. bengalensis VietnamC. rolundala Guatemala ()

Taen/oma perpusillum Japan

Fig. 29. Suict consensus topology of five 'qually most parSiml)niolis trecs infcrrcd from I.SU rDNA. The numbcrs at each inlernal branchindic:lle bootstrap proportions (in %) for MP lln,i1y~is (len, above line) and .I <Ina I 'sis (right, above line) and decay values (below linc). Specicsnames in parenthcses llrc ncwly assigncd on the basis of (his study. Bhldc width: +, slender: + . inlermedi"le; + ++. bro;.Iu. Constriction at.the node: -, hardly cOllslricled: +. slightly constricted: + -1-. modcrately conslricted: +++. slrongly constricted. Sec Table 4 for d'tails ofmorphology.

(64, 72 and 9i\, respectively). CaloglosSCI !ridada is also char

acterized by having both branching systems, but this species

is morphologically similar to C. leprieurii and C. vieillardii(discussed below). Although Taenioma has neither branching

system, our results suggest that endogenous branching is ancestral to adventitious branching and that C. rotundata was

the first taxon to acquire adventitious branching in Calog!ossa.Compared with the branching pattern, the organization of

rhizoids around nodes is more variable between species of

Calog!ossa (Fig. 14). The rhizoids of C. adhaerens are scat

tered throughout the blade, except for portions near the marginand apex (type A in Fig. 14); all the other species form rhi

zoidal clusters associated with the blade nodes. Caloglossuhengalensis is distinguishable from the rest by the absence of

rhizoidal filaments from the transverse peri central cells (lype

B in Fig. 14), and this character is autapomorphic (Fig. 29).

All members of the C. continua complex arc characterized bythe development of rhizoids from thc lirst and second cell

rows within the main and lateral axes (type G in Fig. l4),whereas all the other species produce rhizoids from the centrc

of the nodes, although the arrangement is diverse among these

species. In C. heccarii. C. ogasmvaraellsis, C. s!ipiw!CI and C.rotundata, the rhizoids arc frequently derived from the peri

central cells of the axial cell just below the NA (types C, Dand E in Fig. l4), whereas the pericentral cells never produce

rhizoids in C. interm.ediCl, C. leprieurii or C. !ridada (type F

in Fig. 14). The latter group is also characterized by producingmore than one rhizoidal filament per blade cell and coalescent


490 PIJ.I'('%gio, Vo!. 42 (5), 20m

cell row numberfrom NA

cell row numberfrom FMA

single>multiple

single

single>mulliple

-- 0.01 SUbstitutions/site

single

single

multiple

multiple

mUltiple>single

single

multiple

L..... C. intermedia 3535 USA

C. apomeiotica (C. vieillardii) Australia

C. (eprieurii (C. vieillardi;) Australia

96 C. leprieurii (C. vieil/ardii) Japan100 C. leprieurii (C. vieillardii) Indonesia

C. leprieurii (C. vieillardii) Australia

C. (eprieuri; USA

C. leprieurii USA

C. leprieurii USA100 C. leprieurii Puerto Rico

C. leprieurii Saudi Arabia

76 C. leprieurii Indonesia

C. apomeiolica (c. leprieuril) Australia

C. leprieurii Japan

89 C. apomeiotica (c. lepr;euril) USA

C. apomeiotica (G. leprieuril) Mexico

C. apomeiotica (C. leprieuril) USA

100

100

44

100

52

73

87

100

100

100

96C. postiae Japan

C. postiae AustraliaC. postiae Austalia

C. monosticha Australia

C. monosticha Indonesia

C. c. ssp. axil/ads (C. monosticha) AustraliaL..... C. monostic/la Singapore

C. continua China

C. continua Japan

C. continua Taiwan

C. continua Vietnam

C. monoslieha Brazil

C. monostieha USA

,------ C. monos/icha MalaysiaC. saigonensis Malaysia

C. saigonensis MalaysiaC. stipitata Philippines

r---,lc::OO::"'-_-1 C. slipitata Malaysia

C. slipitata Malaysia

C. adhaerens Japan

C. adhaerens Australia

C. adhaerens Singapore

C.ogasawaraensisAustraliaC. ogasawaraensis Japan

C. beeearii Singapore

C. beccarii Vietnam

C. beccarii Thailand

100 C. bengalensis SingaporeC. benga/ensis Vietnam

'------- C. rotundata GuatemalaTaenioma perpllsil/um Japan

72

Fig. 30. Onl; of Ih' IWo maximum-likelihood lrces (log-likelihood = - 7922.75) from L U rONA. The numbers ill each internal branch indicatcb(lOlSlrap proportions (in %). p'cic.~ n;lJl1e, in flarclllhescs are ncwly assigned on the bnsis of thi. study.

rhizoidal clusters (King & Puttock 1994; Kamiya ef al. 2(00).1'{/eniom{/ produces only single rhizoidal lilaments from intercalary pcricentral cells without forming any rhizoid'll clusters (Papenfuss 1944: Stegenga et a/. 19(7), and such a simplerhizoid'll system is not seen in Calog/o,lsa. Most other algaelhat are mainly distributed in mangrove regions, such as Rostr.l'('hia, Calene/la Greville, Murrctyella Schmitz, Sficfosiphonia J. Hooker & Harvey and Rhi~oclol/iurn Kutzing, usuallyproduce well-developed intercalary rhizoids at segments, butthe pattern of' rhi/.oidal arrangcment and organi/.ation is notas variable as in Clilnglossa (Post 1936; Tscng 1942; King &PUllock 1(89).

Blade shape in the group with adventitious brancl1ing isdiverse and appears to change drastically in this lineage. Although C. heccarii (Fig. 5) and C. sfipilci((l (Fig. 6) showsimilar gross morphology, with moderate blade width and

conspicuous blade constriction, they make clades with C. ogasawaraensis (which has narrow and slightly constrictedblades, Fig. 4) and C. adhaerens (which has continuous broadblades, Fig. 2), respectively. It is apparent that blade widthand constriction has changed scveral times in both adventitiously and endogenously branched groups (Fig. 29).

Relationships within the endogenously branched group

Our molecular data suggest that asexuality has occurred morethan once (at least three times) within the endogenouslybranching group, and this is consistent with thc results basedon the plastid encoded RllBisCo spacer (West el a/. 200 I).Apom.ixis, which results in the repeated production of asexualspores, is not a rare phenomenon in red algae (West et al.200 J). Rosfrychia hisporct West & Zuecarello, which is now


31

synonymous with B. rnorilziml({ (Sonder) J. Agardh, was characterized by its apomictic life cycle (West et al. 1(92). Recently, asexual populations have been found throughout Austndia, New Caledonia, Japan and Micronesia, and the molecular analyses have revealed the genetic dissimilarity of theseasexual entities (West and Zueearello 1999; Zueearello et al.

1999b). In Californian populations of the geniculate corallinealga Lilholhrix aspergillwn Gray, more than 70% of the larger,loosely branched subtidal specimens bore bisporangial eoneeptaeles indicative of asexual reproduction (Pearson & Murray 1(97). In Baja California and Adelaide, the majority ofC. leprieurii and C. vieillardii specimens, respectively, areasexual (West 1'1 al. 1994, 200 I). Because both populationsare located at the edge of the distrioution for either species,asexualization seems to contribute to the acquisition of a newniche.

Caloglossa inlermedia is similar to C. leprieurii and C.vieillardii on the basis of the position and organization of therhizoids at the nodes, but differs by the presence of adaxialcell rows developed from the FLA (Pig. 1; Kamiya 1'1 al.2(00). Caloglossa inlermedia made a sister clade with C. 11'prieurii in the MP and NJ trees (Fig. 29), supported with highbootstrap values, although their relationship was unresolvedin the ML analysis (Fig. 30). Strangely, C. inlermedia has anextremely high pairwise distance value compared with other

ill!~J'(l\I!, s!,('('il's (O.IO.O.17!. 11(':lrly l''1II:J1llllll,' v:IIIII' h,'IIV"('11

till' ingroup :md olilgroliP ((J.()1J-O.17). Till' I~lll\i,( '0 SP:ll'l'rof this species is also distinct from other Caloglossa species,with 24-34 op mutations (Kamiya 1'101. 2(00). The relaliveratio test of LSU rDNA indicated that suostitution rate in C.iJ1lerl7ledia is significantly faster than in the other ingroup taxa(P < 1'Ir'). In contrast, the rbcL gene sequences showed nosignificant difference in the substitution rate oetween C. interl7ledia and C. leprieurii-C. vieillardii, and its phylogenetictree supported the close relationship oetween C. il/lcl'Il/cdiaand C. Icprieurii (Kamiya 1'1 al. 2(03). It is not known whether the difference in evolutionary rate is caused by the dillerence between nuclear and chloroplast genes, or bet ween multigene family and single copy genes, so further examinationsarc necessary to reveal the evolutionary background of thisspecies.

Caloglosso Iridada is distinguished from C. hpriellrii allliC. I'iei/lordii by the production of both endogenous and adventitious branches. This feature is also seen in C. rollll/dala,but C. rolundo!U is clearly distinguishable from C. Iric!w/o bythe distribution and organi/.ation of nodal rhi/.oids and by thenumber of adaxial cell rows from the FLA (Table 4). Cologlosso Iridodo was collected only twice from Mauritius in1868 and 1869 and has not been found since, either from thetype locality or elsewhere. Endogenous branches arc muchmore abundant than adventitious ones in the 1868 specimen(lectotype: 8M61 (478), whereas there arc only endogenousbranches in the 1869 specimen (BM531240) (data not shown).Endogenously branched species occasionally produce adventitious initials from internodal blade margins, especially underinadequate culture conditions or due to cell damage (Kamiya,unpublished observations). However, the adventitious branches of C. tric/ada certainly occur at the nodes in the 1868specimen and seem to be formed under normal conditions (notdue to damage). The taxonomic distinctiveness of C. Iric!odois still uncertain, but at least there is no doubt that it is mostclosely related to C. lepriellrii Crable 4).

Four species of the C. cOl/tinuo complex (C. cOl/lil/lla, C.mOlw.llicho, C. poslioe and C. soigol/ensis) were classified onthe basis of differences in blade width and cell row numberfrom the NA. Algal morphology is occasionally subject toenvironmental change and it makes algal taxonomy difficult(Chen & Taylor 1980; Blomster 1'1 al. 1998; Kalvas & Kautsky 1(98). However, the morphological features of the C. conlil/ua complex were maintained well under our culture conditions (Figs 22, 24), and each species was reproductivelyisolated (Fig. 28). These results support the usefulness ofblade width and number of cell rows from the NA for theelassi fieation of Caloglosso, although additional in formationis necessary for understanding C Il/ol/ostic!w, which is reproductively as well as genetically heterogeneous.

Phytogeography

Of the 12 species described in the genus Caloglos.la, eight arcendemic to the 'old' world, whereas only C. interllledia andC. rotundata arc restricted to the 'new' world. This unequaldiversity is recognized, not only in other mangrove red algae(Post 1936, 1943; King & Puttock 1(89), but also in mangrove trees (Hadac 1976; Chapman 1(84). More than 40 species of mangrove plants have been recognized in the old world

Reproduced w



ithout permission.

.j;.-t;IV

'r;~

;::~~

~ ..~Table 4. Comparison of morphological characters of CaloglossCI species. Numbers in parelllhese, are occasional observations. <c

.j;.IV

C.C. C. C. saigo- e. V1

°8C1sa \l'CI- ~

adhoae/lS beccClrii bengalensis continua i"termedia leprieurii /IIo/lostiella mensis C. po.l·tiae rOlllndCIIO /lensis e. stipiwta e. triclada vieillardii~

ndogenous branch- absent absem absent present presem presem pre,ent absem present present present absent present present 8iog

Advemitious bnll1ch- present prcsel1l presem absem absem absent absent presel1l absent rarely absent presenl mrely absel1ling pres- pres-

eot entNumber of rhizoids single single single single lllultiple multiple single single single single single single llluitiple multiple

per cellRhizoid dislribution l type A type D type B type G type r- type F lype G type E type G type C type G lype D lype F Lype FAdaxial cell row present present present present present absel1l present present presel1l presel1l presel1l present absent absent

from FLA'Number of cell rows 2-3 I (2) (I) 2-4 (I) 2-6 1 (2) 1-3 I (2-4) I 2-5 I 1-2 I (2) 1-5 1-3

from NAJNumber of cell rows 2-4 1-2 1-4 2-6 1-4 2-6 l-7 I 1-5 1-2 2-4 1-2 3-5 I (2

from FMA"Constriction at the hardly moder- moder- hardly or slightly moder- moder- slightly slightly strongly slightly Strongly !>Iightly moder-

node ately mely slight- ately mely atelyor Iy or or orslighl- slight- slight- slight-ly ly Iy Iy

Upper blade width 1.0-3.0 0.1-0.3 0.3-1.7 0.4-1.8 1.1-2.0 0.1-1.7 0.4-2.5 0.1-0.4 0.1-0.3 0.2-0.4 0.2-0.6 0.2-0.6 0.3-0.8 0.1-1.0(mm)

Middle blade width 1.0-3.0 0.4-1.4 0.8-1.8 0.4-1.5 1.2-2.2 0.2-1.9 0.4-2.2 0.1-0.5 0.1-0.3 0.9-1.9 0.2-0.6 0.6-2.0 0.6-1.0 0.4-1.9mm)

Lower blade width 10-2.8 0.1-0.3 0.3-1.3 0.3-1.3 0.8-1.5 0.1-1.3 0.3-1.6 0.1-0.2 0.1-0.3 0.1-0.3 0.1-0.5 0.2-0.4 0.2-0.5 0.1-0.8(mm)

Blade length (mm) 0.9-2.3 1.1-6.0 1.0-4.5 0.7-3.7 2.6--4.6 0.7-5.2 0.7-3.3 1.1-5.0 0.5-1.8 1.9-3.8 IA-3.8 1.1-5.1 1.2-4.0 0.8-6.6

I Each rhizoid distribution type is indicated in Fig. 14., Adaxial cell row from the first axial cell at the lateral axis..I Number of cell rows from the nodal axial cell opposite the lateral branch.4 Number of cell rows from lhe firsl axial cell at the main axis Opposile the laleral branch.


but only 10 species are known in the new world (Chapman1984). The palaeontological data suggest that many mangrovespecies occurred along the Tethys coasts from the end of theCretaceous to the beginning of the Tertiary period and thattheir distribution expanded secondarily to the Atlantic and theEast Pacitic (McCoy & Heck 1976; Mepham I9~3; Duke1995; EllisolJ el al. 1999). The difkrcncc in spccics divcrsitybetween the two regions is possibly related to the closure ofthe Tethys route (Chapman I9~4), although the reasons remain debatable (Hadac 1976; Saenger I99~).

The biogeographic relationship between the old and newworlds can be considered in C. leprieurii and C. vieillardii.CaloKlossa vieillardii is found only in the Indo-Pacific regions, whereas C. leprieurii is distributed worldwide. BecauseC. leprieurii is closely related to C. intennedia (that is endemic to the Atlantic), C. leprieurii may have originated aroundthe Atlantic rcgion and subsequently dispersed worldwide. Ifthe ancestor of C. leprieurii and C. vieillardii was distributedalong the Tethys sea coasts where mangrove plants were abundant, the closure of the Tethys sea was possibly related to thedi fferentiation of these species.

This vicariant hypothesis may be indirectly verified by theestimation of the date when these species diverged. The molecular clock hypothesis can be applied to the C. leprieuriiC. vieillardii lineage, except for the Japanese specimen of C.vieillardii (MK739) that indicated a significantly fastcr cvolutionary rate than other specimens in the same clade. In thecase of marine organisms with no fossil record, it is conventional to estimate the divergence rate of populations across theIsthmus of Panama (Lessios 1979; Bermingham & Lessios1993; Jackson et al. 1993; Knowlton el al. 1993; Knowlton& Weigt 1998; Zuccarello & West 2(02). Marine organismsliving in the pantropiC'll shallow sea around Central Americaarc presumed to have maintained gene exchange across thePanama Strait, and genetic divergence began to accumulateafter the closure of the Panama Strait, at 3.5-3.0 Ma (Coates& Obando 1996). If C. leprieurii were distributed on bothsides of the Panama Strait, the genetic difference of the Atlantic and Pacific samples would have been established afterthe closure of this strait. In this study, the specimens fromPacific Mexico (3276) and Atlantic Florida (3421) show aclose evolutionary relationship, and can be used to estimatedivergence rate. The pairwise percent divergence of LSUrONA sequence between them is 0.20%, and this value is notfar from that between an Atlantic and a Paeific isolate oJ' Bostrvchia calliplem (Montagne) Montagne (0.22%; Zuccarello& West 20(2). The estimation of the divergence rate, therefore, is 0.033-0.029% per million years (the figure for percentdivergence per million years in Zuccarello & West 2002 is amiscalculation and should be divided by 2).

The mean genetic distance between C. leprieurii and C.vieillardii, excluding the Japanese specimen (MK739), is4.21 %, so the divergenee date of these species is estimated at73.7-63.2 Ma. Many fossils of mangrove trees as well as theirpollen were recorded from the Tethys Sea coasts during theearly Eocene (60-50 Ma) (Ellison el al. 1999), so the diversityand dispersal of CaloKlossa may have been associated withthe appearance of mangrove species. However, this divergencerate is mueh earlier than the closure of the Tethys sea (15-12Ma; Ragl & Steininger 1984), and it is essential to improvethe precision of this estimate by increasing the number of

Kwniya lOt al.: Systematics of Caloglossa 493

individuals, populations and gencs cxamincd (Knowlton &Weigt 1998).

It is interesting that the number of cell rows from the NAin the C. conlinua complex corresponds roughly to their latitudinal distribution. Thc specimens producing only or mainlythe single-cell row are distributed around the southern hemisphcrc, whercas the ratio of the multiple-cell rows in a thallusincreases with increasing latitude in the northern hemispherc.Caloglossa I/Wl/ostic!1({ from Singapore (ncar the cquator) ismorphologically intermediatc between the northcrn and southern hemispheres' entities and produces many pseudocystocarps with both of them. Considering thc abundancc and diversity of the C. continua complex, it is possible that theyevolved around the equatorial Indo-Pacific and then werc dispersed towards higher latitudes in both hemispheres. It is, ofcourse, necessary to produce a well-supported phylogenywithin the C. ('ollIinua complex before we can confidentlyunderstand this alga's phylogeography.

Two distinct lineages showed a similar phylogeographicpattern: in C. leprieurii, the northern Australian isolate (F442)is more closely related to the Japanese isolate (MK739) thanthe isolate from southeastern Australia; in C. posliae, the isolate from northern Australia (F406) is phylogenetically closerto the Japanese isolate (M K 106 I) than to the southeasternAustralian isolate (C889). These relationships were supportedby crossing studies and Rul3isCo spacer DNA sequence data(Kamiya el al. 1998; Kamiya, unpublished observations). Furthermore, the pairwise genetic distance between the northernand southeastern Australian populations is almost the same inthe two species (2.6%), and this means that the interruptionof genetic exchange between these populations possibly happened at a similar time. There are several possible reasons forthere being similar phylogeographic patterns in difrerent lineages: it could be an indication of ancient coastlines, or itcould be due to collective migration on the same pieces ofmangrove trees, or to a universal migration route, as was suggested for a common mangrove species of Kandelia ('ande!(Linnaeus) Druce in East Asia (Chiang el al. 200 I). Dispersalevents in the mangrove ecosystem seem to be more frequentand complicated than expected, and detailed biogeographicalinformation should be extended to other mangrove organisms,as well as mangrove plants themselves, to reveal their patternsand mechanisms for dispersal.

Caloglossa rotundata Kamiya, sp. nov.

Figs 17-20

Ramuli endogeni ad nodos ex cellulis axialibus oriundi: ramuli advcntitii ad nodos ex ccllulis in 111arginc laillinanllll oricntcs. Lalnillae ad nodos valde constrictae, 1.9-3.S mm longae, 0.9-1.9 mmlatae. Cellulae pericentrales circum nodum et secus axcs principalcsrhi/'oidea incohaerentia effcrcntes. Cetlula prima axiatis axis lateralis cellulas adaxiales fecit. In latere axe Iaterali opposito nodi seriem unicam cellularum (cellulas monostichas) felTns. Dillcr! a C.beccurii et C. .I'tifJilaiu praesentia ramificationum et endogellarumct ct advcntitiarUlll in thallis singulis.

Endogenous branches produced from axial cells at the nodes andadventitious branches produced by cells at the margin of the bladesat the node. Blades strongly constricted at the nodes, 1.9-3.S mmlong and 0.9-1.9 ml11 wide; divergent rhizoids produced from pericentral cells around thc nodcs and along the main axes. Thc lirstaxial cell of the lateral axis forms adaxial cclls. A single-ccll rowis derived from the nodal cell on the opposite side to the tatcral


404 Phym!ogi{/, Vo!. 42 (5), 2003

axis. Dirfering rrom C. heccarii and C. slil,il{/la by thc prcscnce orboth endogenous <ll1d adventitious branching Oil individual thalli.

HIIHIII-:J, RI-:J'I·:RI:NCI': Pcdrochc 1'1 al. IlJlJ5, p. I 15, ligs 10-12.

IIOUHYI'I·:: TNS-AL-I 5 13lJO (Fig. 17).

TYI'I: 1.0CAI.ITY: Bucna Vista, Likin, Guatemala (13°53'N, lJoo42'W),22 March IlJlJ3, John A. Wcst.

SI'H'IMI:NS I:XAMINI':Il: Rucna Vista, Likin, Guatcmala (culturcd spccimcns: TNS-AI.-I 5 13lJ I, TNS-AL-I 5 13()2).

IIAlmAI: Plants wcrc round adhcring to prop roots and trunks orNhi::,oj);'orn IJ/(fngle I .inn<lclIs and intcnl1ingling with C. !clJrieur;{.

LTYMOI.OCiY: The speeilic cpithct rcrcrs to the roundish blade shape.

VH;J:TATIVI' STRlICTlIRI': Thc thalli arc lear-like, Ilat, dark brown,subdichotomously branchcd, with a total length or I cm or morc(Fig. 17). Each bladc (l.lJ-4.3 mm long and 0.5-·2.4 mm widc) isstrongly constricted at thc nodcs, clliptical to rotundate in shape,and consists or a midrib Ilankcd on cithcr sidc by monostromaticwings (Fig. IX). The midrib is conspicuous and composed or axialcells, cach with two transvcrsc and two lateral pcricentral cells, sothc thallus rcmains thrcc cclls thick in the midrib region. The axialcells arc up to 160 fLm long and 25-35 fLill in diameter, so longerand narrower than the pericentral ce\ls (up to 150 fLm long and 3545 fLm in diamcter). Thc middlc or thc internode is X-24 cells widcon both sidcs or thc midrib. Ce\ls I'orm a monostromatic bladc and,Ire irregular or subquadratc in shape (30-70 fLm in diameter), andrapidly decrease in si/e rrom midrib to margin. The first to ninthaxial ce\ls abovc thc nodc rrcqucntly produce no-wing cc\ls androrm a stipe at the lower part or the blade. Thc apical ccll is dOIlleshapcd (10-13 fLm high, 13-15 fLm in diamctcr), cutting off a fewdisc-shapcd scgmcnts postcriorly that are 3-5 fLm high and 18-20/-LIn ill dianlctcr. Exogenous lateral branches originate fro111 the thirdor rourth axial cell rrom thc apcx. A few secondary endogenousbranches arisc rrom thc nodal celioI' the first axial cell abovc thenodc, and out or thc plane or the thallus (Fig. IlJ), usually on thedorsal sidc. Advcntitious branches are sometimes (but much lessrrequcntly than endogenous ones) formcd in thc nodal region of thebladc by intcrstiti,d prolifcration arising laterally rrom marginalwing cclls (Fig. IlJ). Rhi/oidal lilaments arc regularly formed onthc vcntral sidc around the nodes (Fig. 20). Monosiphonous rhi/oids(up to 220 fLill long and X-20 fLm in diametcr) arc formcd from thcnodal pcriccntral cclls (the two-threc periccntral cells immcdiatelyabovc and below the node), or somctimcs from thc wing cclls adjaccnt to thcsc pericentral cells. These cells produce only onc rhi/oidal filamcnt per ccll. Thc rhi/oids arc discrctc and do not coaIcscc to rorm thc stipe which commonly develops in C. leprieuri;.This Guatemalan alga posscsscs onc adaxial cc\l row from the FLA.Thc nodal axial cell produces a single cell, and the first axial cellsjust above the node also rorm a single-ce\l row (occasiona\ly two)at both main and lateral axes.

RJ:I'ROllliCTIVI: STRlICTlIRL: Female reproductive structures areusually formed on the ventral sides or the apex, anc! each procarpis composed or a rour-cellcd carpogonial branch bornc on thc supporting cell. Thc carpogonium forms an clongatcd trichogyne (upto 300 fLm long and 3-5 fLm in diameter). Other reproductive structurcs wcrc not I·ound.

Caloglossa vieillardii (Kiitzing) Setchell (Setchell 1924,p. 161)

tIl'TJ:I{()TYI'IC SYNONYMS: Caloglossa leprieur;; val'. hookeri E. Post(Post IlJ36, pp. 53-60, X3, figs 2, 3), type locality: Bay or Islands,Ncw Zealand (BM000562231, RM00077005X): Post 1943, p. 127,figs 3-5, 24, 32, 37: Post IlJ63, p. lJlJ: Tanaka and Chihara In5,p. 42, figs 3, 4, IlJ88a, p. 9X, IlJ88b, p. 32: Silva 1'1 al. 1996, p.452. Caloglossa IJ/l1ioides Harvey ex J. Agardh (Agardh 1876, p.5(0), typc locality: Fricndly Islands, Tonga (L00557I 5).

l'lIRTIII:R Rl:I-I'RI:NCl:S: Kraft & Woclkcrling IlJlJO, p. 50, fig. 3.4E (asC. leprieurii): Kamiya 1'1 al. IlJ95, p. 81, figs 3-10, 16, 19 (as C.!eprieur;;), IlJlJX, p. 365, figs lJ, 10 (as C. leprieurii).

SI'I'CIMI:NS 1·:XAMINI'Il: Japan: Shimajiri. Miyako Island (M. KalJ/iva,

3 April IlJ91, TNS-AL-151393, 151394); Oura Rivcr, Nago, Okinawa Island (M. KalJ/iya, X Deccmbcr IlJ91, TNS-AI r I5 13lJ5):Wanya Rivcr, Tokuno Island (.I. Twwka, 10 March IlJXlJ, 'INS-AI.slide-565lJ); Miyara Rivcr, Ishigaki Island (.I. 'I'IJ/aka, 26 April1982, TNS-AL-slide-60(0). Austnllia: Port Fairy, Victoria (W.I I.Harvey, IXX4, I3M000770064, BMOOO7700(5): Nielscn Park, Sydney (M. Kwuiya, 17 October IlJ91. TNS-AL-I 5 13lJ6): Oak Beach,ncar Mossman, Queensland (M. Kmuiya, 20 Scptember 19lJ I, TNSAL-15 13lJ7). New Zealand: Ray or Islands (W.1f. lIar!'ey, 1867,BMOOO77(057). Indonesia: Poka, Ambon (.I. '!'lIli1ka, 12 AugustIlJX6, TNS-AL-slide-7(42). New Caledonia: Wagap (/0'. Vieillard,18h3, L0(55716).

;\I)J)ITIONAL COI.IHTIONS: Japan: Minato River, Okinawa Island (.I.'I'lIIuka, I I March IlJ88); S~lIl.ja R., Na!l0, Okinawa Island (M. KalIIiya, 8 Decembcr IlJlJ I): Oho River, Ogimi, Okinawa Island (M.Kal1liya, 31 March 19lJ I): Genga River, Nago, Okinawa Island (M.Kallliya, 7 Deccmbcr IlJlJ I); Nakama River, Iriomotc Island (M.Kallliya, 30 October 1993): Urauchi Rivcr, Iriomote Island (M. KalIIiva, 30 Octobcr IlJl(3). Austmlia: Cardcn Island, Adclaide (,J.Wesl, 10 February IlJlJlJ): Tooradin. Victoria (,J. Wesl, 25 OctobcrIlJX6): Hopkins Rivcr Falls, Victoria (.I. Wesl, 25 March IlJ,)5):Coopcrnook, New South Wales (NSW) (.I. Wesl, 22 October 1995);Brisbane River, Hrisbane (.I. West, 12 Decembcr IlJl(4): BurleighHcads, Gold Coast (,J. Wesl, 13 Decembcr IlJl(4): Jctty, Cairns (,J.West, 4 December 19lJ4); Arnhcm Land, Northern Territory (.I.WI'SI, 23 August IlJl(9): Southport Boat Ramp, Darwin (.I. Wesl,21June IlJl(7): Mangrovc Ray, Exmouth (M. Kallliya, X Octobel' 1991).New Zealand: Linkwater, South Island (.I. Wesl, 7 July IlJlJX). NewCaledonia: PI age de Fouc (,J. We.I I, 2 February 199X). Indonesia:Serangan Island, Benoa Bay, Hali (.I. Ta/1aka, 27 August 200 I);Nusa Lcmbongan (,J. Wesl, 25 April 1999). Phillippines: Tolcdo,Cebu (.I. Wesl, 3 March IlJl(6). Vietnam: Dong Nai River, Ho ChiMinh (.I. Ta/1aka, 2X July 2(00). Fiji: Viti Levu, Lau Cala Bay (,J.Wesl, 4 Junc IlJl(7). Samoa: A'll1apa Resort, Upolu Island (I'. Skel10/1, 13 February 1998).

Caloglossa leprieurii (Montagne) G. Martens (Martens1869, p. 234)

HI,TEROTYI'IC' SYNONYM: Caloglossa apol1le;olico Wcst & Zuccarelloi/1 West 1'1 a!. IlJlJ4, p. 381, figs 1-15 (typc locality: San Carlos,Bahia Magdalena, Baja Calirornia Sur, Mexico): Kamiya el al.1998, p. 361.

FURTIIER REFERI':NCI,S: Papcnfuss IlJ61, p. X, figs 1-30: Yarish &Edwards IlJX2, p. IIX, fig. 15: Lambert 1'1 al. 1987, p. 355, fig. 3G:Tanaka & Shamccl 1992, p. X4, fig. 24: Kamiya el al. IlJlJ5, p. 81,figs II, 13-15.

SPECIMI:NS I:XAMINI:Il: United States: Tcnncsscc Rivcr, Shcflield, Alabama (P. Dav;so/1, 20 April IlJlJ7, UNAF-3lJ86). French Guiana:near Caycnne (Icctotype of C'. leprieurii, L00557 17). Pakistan:Sandspit, Karachi (.I. Ta/1alw, 12 August 1990, TNS-AL-sIide7375). Indonesia: Bay or Islands (W.H. !larvey, I X67,BM000770055, BM000770056, BMOOO7700(0). Japan: TaharaRiver, Yonaguni Island (,J. TO/1olw, 9 Fcbruary 1989, TNS-AL-slide58(5).

;\IlIlITION;\L, COLLECTIONS: United States: James River, Virginia (1.Wesl, 20 May IlJ93); Stamrord, Connecticut (asexual: .I. Wesl, 13August IlJlJ 1); James Island, South Carolina (,J. Wesl, 17 JuneIlJl(4): Isla Morada, Florida Keys (1. Wesl, 15 June IlJ94). PuertoRico: Mayagueyes I., La Pat'guera (.I. Wesl, 6 March 19lJ4). Pel'll:Puerto Pi/arro, Tumbcs (.I. Wesl, 10 Fcbruary 1990). Venezuela:Laguna dcs Restingas, Isla Margarita (.I. Wesl, 13 April IlJlJ I): Punta Uricaro, Edo. Sucre (.I. Wesl, 10 April 1991). Mexico: BocaRiver San Juan, Chiapas (.I. We.ll, 25 March 19lJ3); Espiritu SantoIsland (asexual: M. Kal1l;ya, IlJ May IlJl(2): Balandra, La Paz (asexual; M. Kal1liya, 18 May 1992). South Afdca: Umlahl/i, Natal (A.erilcMey, 4 October IlJlJ I). Sd Lanka: Ncgombo Lagoon (R. Cordover, 26 August 2(02). Saudi Arabia: Farasan Island, Red Sea(M. lIu.lsail1, 8 July 20(0). Indonesia: Benoa East, Bali (slenderrorm: .I. Wesl, 10 April IlJl(9): Nusa Dua, Bali (M. Kawachi, 13April 19lJlJ). Singapore: Batu Puteh, Changi (M. Kal1liya, 13 Jan-


uary 11)1)1): Kg. Melayu. Pulau Ubin (slender rorm; M. Kallliva, 11January 1(91). Japan: Genga River, Okinawa Island (slender form;M. KWI/;ya, 7 lJL:cL:lllbL:r I ')') I); SiJilllajiri, liyaKo /slalld (sklldL:rform; M. Kalll;ya, 2 April 1')91).

Caloglossa 1110110.\·ticha Kamiya (Kamiya 1997, p. 97)

IIETUWTVI'IC SYNONYM: Ca/og/ossa con/inlla ssp. axi//aris R.J. King& Putloek (King & Pulloek 1994, p. 117, lig. 6b), type locality:Arllham Highway bridge, South Alligator River, Northern Tnritory.Australia.

I'URTI·II,R RI'I'lcl<I.'NCI:S: Kamiya el a!. 1997, p. 99, Jigs 5-~, 1012;Kamiya 1')99, p. :156, lig. I I, 2000, p. 41 I. lig. Rg. h; Wynne andDe Clerck 1999. p. 1R I.

SI'I'CIME S I'XA liNE]): Australia: Derby (M. Kl/llli"'l. 2 October190 I, TNS-AI.-421(5): Sn1<111 I.agoon, Denham (M. KWl/iYI/, I IOctober 1991, TNS-AL-23(6). Siugaporc: Kg. Melayu. Pulau Ubin(!VI. Kalll;ya, 11 January 1001, TNS-AI.-42:J(7).

ADDITIONAl. COI.I.ECTIONS: Australia: Soulh Alligator River, Northerll Territory (M. Kallliya, I October 1001); East Alligator River,Northern Tnritory (I<. King, 30 August 1097); Mandorah, Darwin(J. Wes/, 21 June 1(97); Broome (M. Kom;yo, 4 October 1(91);Cape Keraudren. Port Headland (M. Komiya, 5 October 100 I );Da11lpieer (M. Kalll;yl/, 6 October 199 I); Waper Creek, Carnarvon(M. Kalil iVll , 0 October 199 I). Indonesia: West Sawang, North Sulawesi (J. Wesl. 16 November 19(4): Nusa Dua, Bali (J. Wes/, 10April 19(9); Teluk Awallg, Lombok (.I. Wesl,27 April 10(9). Malaysia: Morib, Selanger (.I. West, 12 May 199H); Sandakall, Sabah(J. Wes/, 16 August 20(0). Bnlzil: R. Perellue. I. do Cardoso, SaoPaulo (J. Wesl, 4 April 1(90). United States: Plantation, FloridaKeys, Florida (J. West, IS JUlle 19(4). Pen" Pucrto Pi/,a1To, Tumbes (J. West, 12 July 1(94).

ACKNOWLEDGEMENTS

The authors thank Drs H. Akiyama, D. Ballantinc, R. Cor

dover, A. Critchley, P.G. Davison, Y. Ham, M.1. Hussain, U.Karsten, M. Kawachi, R.J. King, T. Kitayama, M. KrUtzcn,

D.M. McBride, 13. Sandercock, H. Sato, P. Skelton, J. Tanaka

and A. Yokoyama 1'01' collections of Ca/og/ossa material. We

arc also indcbted to thc directors of thc following herbaria for

thc loan of specimens: The Natural History Museum, I,ondon,

UK (8M), Nationaal Herbarium Nederland, Leiden, The

Nethcrlands (L), Royal Botanic Gardcns, Sydney, Australia

( SW), Hokkaido University, Hokkaido, Japan (SAP), a

tional Science Muscum, Tokyo, Japan (TNS), and University

of California, Berkeley, USA (UC). We thank Drs H. Kawai

and D. Honda for providing helpful suggestions and Dr N.

M iyashita for providing the computer program. This research

was partially supported by the Syowa Seitoku Memorial

roundation and Grant-in-Aid for Scientific Rescarches

(12740469) from the Ministry of Education, Sciencc and Cul

ture, Japan, to M.K. and the Australian Rcsearch Council toG.C.Z. and J.A.W.

REFERENCES

A(;;\RDII J.C'. I K76. S!Jec;es generu eI ordines a/ganulI, vol. 1, deF/orideis cun.le !H}s/er;ore.\'. EI)icri,'lis sysletnatis F/orideorurn. Weigel, LL'ip/,ig. 724 pp.

BERMI (,IIAM E. & LESSIOS H.A. 1093. Rate variation of protein andmitochondrial DNA evolution as revealed by sea urchins separat'edby the Isthmus of Panama. I'roceedillgs or 1/11' Na/i(JJ"'/ Academyor Sciences (d' the Uniled Stmes of Americu 90: 2734-·-273R.

K{/rniy{/ et al.: Systematics or CI//og/OSSI/ 495

131.0MSTI'R J.. MM;(jS C.A. & ,)TANIIOI'I' M..I. 1'J'JH. Mo!l'clliar alldmorphological an,liysis or 1:'lIlerOlllo/l/!'" inlesl;lwlis alld 10'. COIII!Jressu (Chlorophyta) ill thc Ilritish Islcs . .Iollmll!oF 1'!nnJ!og.'· .14:

310 ·340.I3l<1cMER K. 19KK. The Ii mils or amino acid seqllellce data ill '1I1gio

sperm phylogcllctic rcconstruction. !o'\'(}/ut;oll 42: 795· ·R03.CiIJlI'MJlN v..I. 19R4, Mangrovc biogeogl'aphy. In: Ilrdro!l;o!og\' I~Ft!«'

IlIlu/ga! (Ed. by ED. POI' & I. Dor), pp. 15-24. Junk, The Haguc.CHEN L.C.-M. & Ti\YI.OR A.R.I\. I')HO. Invcstigaliolls or distincl

straills of Cholldms crisplls Stackh. II. Cullure sludie.s. !lolall;ul

MorillO 21: 441--44R.CHII\N(; TY.. OIIAN(; Y.e.. Ollci' Y.J.. Dim: C'.1l .. HAVANO\!J) S ..

HONC; TN. & HUANC; S. 200 I. Phylogeogr'1J1hy 01' Kmll/elill ('({lIdeiin hast Asiatic mangroves based on nucleotide variation of chloroplast and mitochondrial DNAs. Mo!cut/ar I,'('ologl' 10: '2697'2710.

COJ\TI'S A.G. & OllJ\NllO .I.A. 109(,. The gL'ological evolution of theCentral Amcrican Islhmns. In: 1·.·'·O/lIlioll 111/11 ('III'iI'OIlIlI('{1/ ;11 II'O!'iCit/ AII/er;m (Fd. by .I.I).c. .hlckson. A.I': (:ludd & A.CJ Coates).pp. 21-.'i6. Univcrsity or Chicago Press, Chicago.

DUKI' N.C. 1905. Gellelic diversity. distributiollal harriers and raftillgcontinents - nlorc thoughts 011 the evolution 01' 1l1allgrnves. I!.n!rohi%gia 295: 167-IRI.

EI.I.lSON A.M .. I'ARNSWORTII E..J. & MI·.I{KT R.E. 1999. Origins ol'mallgrove ecosystems and thL' mallgmve hiodiversily anomaly. G!ol"t/F;co!ogy 1I1/{! 8;ogeogm!,hv g: 9'1-1 15.

EI<IKSSON T 199R. Alllof)eca.\', version 4.0.2 (availahle rmm the authorHt: hllp:/lwww.hotan.su.se/Systelllatikll:olkl.llll.stcn.html). Department or Botany. Stockholm University. Stockholm.

I-'LI.SENSTUN J. 19R5. Confidence limits on phylogenies: an approachusing bootstrap. r:"ollll;oll 19: 7Rl-70 I.

!'lcLSLNSTUN J. 2002. PHYUP (!lhv/ogellv ;II/,'rellce !'l/dllge), vel'sioll3.6. Departmenl of Gelletics, Univcrsity of Washington, SL:atlle.

FRI'SIlW,\ IHl D.W. & BAII.EY .I.e. 199X. A multigene phylogcny of thcGelidiales including nuclear large-subunil rR A sL:lJuellce data..Iolll'lla! or A/I!llied Pliyc%gl' 10: 229-23(,.

I:RESIIWATLR D.W., FRI'I)J:RICQ S. & 13,\II.I'Y .I.e 190'} Charactcristicsand utility of nuclear-encoded large-suhunit ribosomal gL:lle selJuences in phylogenetic studies 01' I'cd algae. PlivCO/Og;ClI/ N('selll'c11

47: 13-3R.HJ\DAC E. 1076. Species diversity or mangrove and continental dril'l.

Folia C;eo!Jolal1;ca ('I fJ/n·/o/a.wllolllica I I: 213-216.HAl<l'Ul .IT & SAUNDERS G. W. 200 I. The application of scquenccs of

the ribosomal cistmn to the systematics and classilicatioll of theIlorideophyte red algae (f'lorideophycL:ae. Rhodophyta). ('lIh;er.l di'

lJiolog;(' Mar;lIe 42: 25-1g.JACKSON .I.B.C., .IIIN(; P, COJ\I'I':s A.Ci. & COI.I.lNS I..S. 1993. Diversity

and extinction or tropical !\Illcrican In()llllSC~ and ~l1lCrgcllcc or theIsthmus of Panama. Sciellce 260: 1624-·1626.

KALVAS A. & KAUTSKY I.. 199H. Morphological variatioll in !-'tICIiS\Jesicu/osus populations along tCI11pcrature and salinity gradiellts inIceland. .Iolln",/ or the Mal';ne 8;%g;ca/ ASSO";UI;OIl 01/111' UII;led

K;lIgdol/l 7R: 9R5-1001.KAV1IYA M .. TANAKA .I. & 1-IAllI\ Y. 1')l)'i. A Illorphological study and

hybridinllion analysis of ('a/og/ossa /el'riellri; (Ceramiales. Rhodophyta) from Japan, Singapore alld Australi,l. Pln,(,(l!ogint/ Ne

selll'ch 41: R1--9 I.K/\MIYJ\ M .. T,\NAKI\ .I. & 1-1,.\1(1\ y. Ilj,)7. Compar'ltivc morphology.

crossability, and taxollomy within thc ('alog!os.\'{{ cOlililillO (Dclesseriaceae, Rhodophyta) complex from Ihe Western Pacific . .Iollma/

of Phyca/ogy 33: 97-·1 D5.KAMIYA M., WI",r .I.A., KIN(; R..J., ZUCCAI<I'I.I.0 G.e.. TANi\K/\ .I. &

Hi\RA Y 199R. Evolutionary divcl'gellce in rhe red alg"e CII!Og!OSSIl

!<'I'r;elll';; alld C. 1I!'(}Il/e;ot;cII . .I0UI'IiIl! of 1'/lI'co!ogv 34: 3(, 1-370.KI\MIYA M., TI\N/\KA .I., KIN(; R..J., WI'ST .I.A .. ZU('('/\\(I'I.I.o (j.e. &

KAWAI H. 1999. Reproductive and genetic dislinclion betweenbroad and narrow entities of eltlog!OSslI COllt;1/11II (Delessel'iacL:ae,Rhodophyta). PllIH/logill 1H: 356-167.

K""'lIY,\ M., WI'ST J.A., ZUCCAREI.I.() G.e. & KA"'J\I H. 2000. ('II!

og/OSSII ;l/tel'll/ed;1I sp. nov. (Rhodophyt") I'mlll tile Western Atlalltic


496 f>hyco!ogia, Vol. 42 (5),2003

coast: molecular and morphological analyscs with spccial reference(0 C. leprieurii and C. /lwnosticha . .!ournal of Phycologv 36: 41 1420.

KAMIY,\ M., 7.UCCAREI.LO G.c. & WEST l.A. 2003. Phylogeographyof Caloglossa It'prieurii and related species (Dclesseriaeeae, Rhodophyta) based on the rhcL gcne sequences . .!opa/lese .!oul'l1al oj'Phvcologr (S(lrui). In press.

KING R..I. & PlirrocK C.I' 1989. Morphology and taxonomy of HosIrvchia and SticIosipho/iia (Rhodoll1elaeeae/Rhodophyla). Australian S-"stenUilic Hotanl' 2: 1-73.

KING R..I. & PlirrocK C.F 1994. Morphology and taxonomy of CaloKlos.l·a (Delesseriacc<lc, Rhodophyta). Australia/l Svslcll1wic BolII/n' 7: 89-124.

KNOWLTON N. & WEIGr L.A. 1998. New dates <lnd new r<ltes fordi vergence <lcross the 1sthmus of Panama. Proceedings or the RoyalSociell' or London, Series B 265: 2257-2263.

KNOWLTON N .. WEIGl' L.A., SOJ.()RZANO L.A., Mtl.LS O.K. & BERMINeiHAM E. 1993. Divergence in proteins, mitochondrial DNA, andreproductive eOll1patibility across the Isthmus of Pan<lma. Science260: 1629·-1632.

KRAFT GT & WOU.KERI.tNG W..I. 1990. Rhodophyta. In: Riologyoj'/I1arine 1,Ianls (Ed. by M.N. Clayton & RJ. King), pp. 41-85. Longm<ln Chcshire. Melbourne.

KliMANO S. 2002. Freshwaler red algl/(' or the world. Biopress, BristoL UK. 375 pp.

KlIZOFF R.K .. SWI;ERE J.A .. SOI.TIS D.E., SOLTIS PS. & ZIMMER B.A.1998. The phylogenetic potenti<ll of entire 26S rONA sequences inpl<lnts. Molecular Biology al1d t-'volwiol1 15: 251-263.

LAMBERT G., STEI KE TD. & AIDOO Y. 1987. Algae associated withmangroves in southern African estuaries, I. Rhodophyceae. SOLtlhAji'ical/ .101.11'1101 "j' !Jolony 53: 349-361.

Lr,ssios H.A. 1979. Use of P<ln<lm<lni<ln sea urchins to test the molecular clock. Nature 280: 599-60 I.

LIN S.-M., FREDERICQ S. & HOMMERSANO M.H. 200 I. Systematics ofthe Delesseriaceae (Ceramiales, Rhodophyta) b<lsed on large subunitrONA and rhcL sequences, including the Phycodryoideae, subfam.nov. .101.11'11,,1 oj' Ph-"cology 37: R81-899.

MARTENS G. VON 1869. Beitriige WI' Algen-Flora Indiens. Flom 52:233-238.

McCoy R.D. & HECK K.L. JIC 1976. Biogeography of corals, seagrasses. <Ind mangroves: an alternative to the center of origin COIl

cept. Svsle/l/alic Zoolo!?" 25: 201-210.MEPI-IAM R.H. 1983, Mangrove floras of the southern continents. Part

I. The geographical origin of Indo-Pacific mangrove genera and thedevelopment and present status of the Australian mangroves. SoulhAli-icol/ .Iou I'lla I or !JO/{/lIV 2: 1-8.

MtYASHITA NT, KAWAIlE A .. INNAN H. & TERAlJCIII R. 1998. Intraand interspecific DNA variation and codon bias of the alcohol dehydrogenase (Adh) locus in Arabis and Arabidopsis species. Molent/or Hiology olld I:volulioll 15: 1420-1429.

PAPf'Nl'lJSS G.I~ 1944. Structure and taxonomy of Toellio/l/o, includinga discussion on thc phylogcny 01' the Ceramiales. MlldrOllo 7: 193232,

PAPENI'"lJSS G.F 196 I. The structure and reproduction of Caloglossa"prieurii. I'hvcologio I: 8-31.

PEARSON E.A. & MURRAY S.N. 1997. Patterns of reproduction, geneticdivcrsity, and genetic dilTcrentiation in California populations of thegeniculate coralline alga Lilhothrix ospergillu!11 (Rhodophyta)..!o/ll'/wl or Phycologv 33: 753-763.

Pt:DROClIt'" FE, WI'"ST J.A., ZlJCCAllt:L1.0 G.c., SENTII;S G.A. & KARSTEN U. J995. Marine red algae of thc mangroves in soutllern PacificMexico and Pacific Guatemala. fJ(}fanica Marina 38: I I J-I 19.

POSADA D. & CRANDALL K.A. 199R. Modeltest: testing the model ofDNA substitution. fJioin{fJrlllalics 14: 817-R 18.

POST E. 1936. Systematische und pflanzengeographische Notizen wrfJoslr"chio-Caloglos.m-Assoz;ation. Revue algologique 9: 1-84.

POST E. 1943. Zur Morphologie und Oko!ogie von Caloglossa. Ergebnisse der Sunda-Expedition der Notgemeinschaft der deutschenWissenschaft 1929/30. IInhiv Ii:ir Pmtistellkw/{Ie 96: 299-308.

POST E. 1963. Zur Verbreitung und (Jkologie der l3oslrychill-Colog-

losso Asso;t.iation. Inlerl1aliona!l?ev/./e der Geslllntell Hvdrobiologie48: 47-152.

R()GL E & STEI 'INGt:R EF. 1984. eogene Para tethys, Mediterraneanand Indo-Pacific seaways. Implications for the paleobiogeographyof marine and terrestrial biotas. In: Fossils and climwe (Eel. by PJ.Brenchley), pp. 171-200. John Wiley and Sons, Chiehcster, UK.

SAI;NGER P. 1998. Vlangrove vegetation: an evolutionary perspective.Marine and Fre,I'hwOler Research 49: 277-286,

SAITOU N. & NEI 1. 1987. The neighbor-joining mcthod: a new method for reconstructing pbylogenetie trees. Molecular fJiology andI,'!'olulioll 4: 406-425.

SETCHI'L1. W.A. 1924. A/l/ericall Samoa. Part I. Vegclolir", or'liuuiloIslalld. Pan II. Ethlloholal1Y 01' Ihe S0!1100I1s. Pan III. Vegefotion oj'Rose Aloll. Publications of the Carnegie Institution of Washington,no. 341. 275 pp.

SILVA pc., BAssoN PW. & MOE R.L. 1996. Catalogue or the benthicmarille algae "I'lhe Indian Ocean, University of California Press,Berkeley. CA. 1259 pp.

STARR R.C. & ZElKlIS J. 1993. UTHX - the culture of algae at theUniversity of Texas al Austin, .!ournal or Phycology 29, supplement: 1-106.

STEGENGA H., BOLTO ' .1..1. & A IlF.RSO RJ. 1997. Seaweeds "r IheSoulh IIlikan west coasl (Ed. by A.V. Hall), vol. 18, Bolus Herbarium, Cape Town. 655 pp ..

SWOFFORD D.L. 2002, PA UP"'. Phylogel1etic analysis usillg parsimollY(al1d olher melhods). Version 4. Simllier Associates, Sunderland.MA.

TAJl 1A E 1993. Simple methods for testing the molecular evolutionaryclock hypothesis. Genetics 135: 599-607.

TAMURA K. & NEt M. 1993. Estimation of the numher or nucleotidesubstitutions in the control region or mitochondrial DNA in humansand chimpanzees. Molecular 8iology and F:,'olulion 10: 5 I 2-526.

TANAKA J. & CtIlI-iARA M. 19R5. Taxonomic studies or Japanese mangrove maeroalgae II. Two taxa of Caloglossa (Ceriuniales, Rhodophyceae). Bulleli" oj'lhe Natiol1ol Science Museul1l, Serie.l· 8 I I:41-50.

TANAKA J. & CtIIHARA M. 1988a. Maeroalgae in Indonesian mangroveforests. Bullelin (d'ihe National Science Museum, Series B 14: 93106.

TANAKA J. & CtIlHARA M. 1988b. Macroalgal flora in mangrove brackish areas of East Indonesia. In: Biological system oj' n1(/l1groves, Areport 01' east Indonesian 11U1l1groFe expedition (Ed. by K. Ogino &M. Chihara), pp. 41-50. Ehime University, Ehime, Japan.

TANAKA J. & KAMIYA M. 1993. Reproductive structure of Calog!o.\'soogasawaruensis Okamura (Ceramiales, Rhodophyceae) in natureand culture. .!apanese ./oul"llol or Phycology 4 I: I 13-12 I .

TA AKA J. & SIIA 1EEI. M. 1992. Maeroalgae in mangrove forests orPakistan. 1n: CiyplOg{llnic .fIom oj' Pakisloll (Ed. by T Nakaike &S. Malik), vol. I, pp. 75-85. National Science Museum, Tokyo.

TATEWAKI M., XIAO-YANG W. & WAKANA I. 1989. A simple methodof red seaweed axenic culture by spore-washing . ./ajJane.l·c .Iou1"110 I(il' Phycology 37: 150-152.

THOMPSON J.D., GIBSON T..I.. PI.EWNIAK F, JI'ANMOlJCilN F & HIGGtNSD.G. 1997. The ClustaJ X windows imerface: flexihle strategies I'ormultiple sequence alignment aided by quality analysis tools. NucleicAcids Research 24: 4876-48R2.

T.~I'N(; C.K. 1942. Mal'ine algae of Hong Kong, II: the genus Colenel/o. .!oul'IJol (>t'lhe Washil1glon Acade/l'lv 01' Sciences 32: 142146.

WEST l,A. 1991. New records of marine algae I'rom Peru. BotanicaMarina 34: 459-464,

Wl.sr J.A. & ZUCCAllEI.I.O G.c. 1999. Biogeography or sexual andasexual populations in !Jostr)'chio nwrilzi{llUl (Rhodomelaeeae,Rhodophyta). Phycological Research 47: I 15-123.

Wisr l.A .. 7.un:ARu.1.0 G.c. & CAl.l1MPONG 1-1.1'. 1992. !Jostrychiahispom sp. nov. (Rhodomelaceae, Rhodophyta), an apomictic species from Darwin, Australia: reproduction and development in cullure. Phycologio 31: 37-52.

WEST J.A., ZUCCAREI.I,O G.C., PEIlROCI-IE FF & KARSTEN U. 1994.


CU{OX!O.'i.')'(( ufJ()fUeiOlicu sp. nov. (Cc.raillialc:-;, Rhodophyta) fnJlll

Pacific Mexico. !Jo/(ll1ic{/ Morino 37: 3R 1-390.

WEST .1.1\ .. ZllCCAREI.I.O c..C'. &. KAMIYA M. 2001. Repro<1l1clive p'll

terns or Ca/o!?lossa species (DeJesseriaceae, Rhodophyta) fromAustralia and New Zealand: multiple origins of asexuality in C.!eprieurii. Literature review on apomixis, mixed-phase, bisexuality

and sexual compatibility. Phyco!cl!?ico! Research 49: 1X3-200.

WYNNE M..J. & DE CLERCK O. 1999. Taxonomic notes on Co!og!ossa/J1ol1oslicha and Ca!og!ossa saigol1el1sis (Delesseriaceae, Rhodo

phyta). Conlrihu/ions from the UniversilV or Michigan Herbarium22: 209-213.

Y ARISH c. & EDWARDS P 19R2. A field and cultural investigation of

the horizontal and seasonal distribution of estuarine red algae of

New Jersey. Phyco!o!?ia 21: I 12-124.

ZUCCARELLO C.c. & WEST .I.A. 2002. Phylogeography or the Hoslrychill ca!!ipleru-IJ. pinnala complex (Rhodomelaceae, Rhodophyta)

KOllliyo ci al.: Systcmatics or Ca/og/ossa 407

and divcrgcnce rates based 011 IlUCIe"I!", Illilochondrial and plastid

DNA markers. I'hvco!ogio 41: 49-60.7.1ICCAI'I-:I.I.o c..c. WISr .1.1\ .. KAMIYA M. & KINC; ILl. 1()()9a. 1\ rapid

method to score plastid haplotypes in red seaweeds ,md its usc ind~teJ'll1ining parent.al inheritance or plHstids ill the I't.:.:d alga !3ostry

chia (Ceramiales). Hydrohio!oKia 40 I: 207-214.ZUCCARFJ.J.O C.c.. WEST 1.1\. & KINC; R..I. 1999h. Evolutionary di

vergence in the Roslrychia lIlorilz.iul1u/H. rudicul1s complex (Rhodomelaceae, Rhodophyta): molecular and hybridization data. !'h\'-c%gio 3R: 234-244. .

ZUCCAREI.I.O G.c., BARTI.I·IT J. & YEATES I'll. 2000. Dillercllliationor Co!og!osso !e/Jrieurii (Rhodophyta) populations ill northern andeastern Australia using plastid haplotypes. [';uro/IN/II .10u!"!la/ oj"Phyco!of{y 35: 357-:16:1.

Accep/ed 21 FehruUly 2003

Documents

Evolutionary relationships of the genus Caloglossa (Delesseriaceae, Rhodophyta) inferred from large-subunit ribosomal RNA gene sequences, morphological evidence and reproductive compatibility,