Biology ofA nomuraII (A .Asa kura，e d. )，C rustac ean Resea rch，S pec ial N umber6: 15 1-166，2006

Phylogenetic relationships among western Atlantic Porcellanidae (Decapoda: Anomura)，based on partial sequences of the mitochondrial16S rRNA gene，with comments on morphology

Irene Teresa Rodriguez，Gonzalo Hernandez and Darryl L. Felder

Abstract.-We obtained 47 partial sequences as porcelain crabs. The family is considered of the mitochondrial16S rRNA gene for 20 west- awell-defined taxon within the superfamily ern Atlantic，three amphi-American，and two Galatheoidea Samouelle，1819 (Haig，1960) eastern Pacific species of porcellanids，grouped Other members ofthe superfamily are Aeglidae into eight genera.Neighbor Joining，Maximum Dana，1852 ，Chirost ylidae Ortmann，1892 ，and Parsimony，Maximum Li kelihood，and Bayesian Galatheidae Samouelle，1819 (Maロin & Davis， Analysis were the methods used to infer phy- 2001). logenetic relationships between and within the Porcelain crabs occur worldwide，primar- genera. The family is divided into two main ily in intertidal and sublittoral zones of tropi- clades，one with POかonyx and Porcellana，and cal and subtropical regions where they occupy the other with the remaining six genera treated here.Larval ，sperm ，and spermatophore mor- habitats such as depressions under stones， phology largely support this obvious dichotomy. spaces in worm tubes，cavities of sponges，and Species of Clastotoechus were grouped closely excavations in coral reefs (Haig，1960; Gore， together，which raises questions regarding the 1972，1982 ;Harvey ，1999; Hernandez，1999; validity of the monotypic genus Madarateuchus Werding etal. ，2003). Most species are free- recently proposed for Clastotoechus vanderhorsti. living ，but afew live commensally (Haig，1960 ; Intrageneric divergence in Megalobrachium Werding，1977 ，1983b; Baeza & Thiel ，2000 ; suggests the possible need for resurrection of Baeza eta l. ，2001). Porcellanids are primarily Porcellanopsis. Megalobrachium soriatum ap- filter feeders and use their highly specialized pears to contain acryptic species. Neopisosoma is setose third maxillipeds to extract minute food supported as agenus distinct from Pachycheles， particles of plankton and detritus 企omthe wa- which contained two independent lineages. ter.They can also take advantage of accumu- Petrolisthes is resolved into two clades，as sup- lated detritus by direct deposit feeding (Kropp， ported by previous morphological and molecular 1981). Their feeding habits allow them to evidence. In one clade Petrolisthes violaceus is ， occupy awide variety of habitats .Within the closely related to P. tridentatus，P. quadratus，P. family，Parap etroli sth estortug ensis (Glassell ， lewisi，and P. magdalenensis. The second clade contains three lineages: one with Petrolisthes ar- 1945) is unique in the shape of the fingers matus，P. marginatus，and P. politus as asubclade; and the third maxillipeds.Fingers are deeply another with P. rosariensis as an independent grooved along cutting edges，spooned and trun- lineage; and lastly the Petrolisthes galathinus- cate at tips (Haig，1962) .The filter setae of the complex，with P. caribensis，P. galathinus (with third maxillipeds are short and scarce ，with a cryptic species) ，and Parapetrolisthes tortugensis. brush-like setation on the dactylus that seems an adaptation to scraping material from the sub- strate (Kraus etal. ，2001). These features are INTRODUCTION shared with the galatheids and could represent Porcellanidae Haworth，1825 ，is afamily homologous，ancestral characters (Kraus etal. ， of anomuran crustaceans commonly known 2001) or convergent adaptations to their feed- ρ 7 T P H i し n o u ρV 152 - - U J u L G u u b I l r ing habits. The genus P etrolist hes Stimpson，1858 ，is Most porcellanids hatch as aprezoea and the largest in the family，containing over 100 molt through only two subsequent zoeal stages species. Based on aqualitative examination and one megalopal stage (postlarva or decapo- of morphological characters，several natural dite) .The reduced number of zoeal stages is groups，including two large groups，have been an abbreviated scheme of development，usu- distinguished. One large group consists of spe- ally considered as phylogenetically advanced， cies that have teeth or spines on the anterior in which the larval history is foreshortened in margin of the carpus of chelipeds，one or two comparison to related taxa，both in number of posterodistal spines on the merus of walk- instars and the duration of each (Gore，1985) . ing leg 1，and epibranchial ，supraorbital ，or This can result in areduced period of plank- other spines on the carapace.The other large tonic life in which larval dispersal is potentially group consists of species in which the ca叩us very limited . of the chelipeds is not armed with teeth or Porcellanids number nearly 280 species spines and spines are lacking on the carapace partitioned among 30 genera. Near1 yhalf of and walking legs (Haig，1960). The genus the species in this family have been recorded Pachycheles Stimpson，1858 ，is the second in the Americas (Gore，1982; Werding et al. ， most speciose of the family and includes over 2003). Seven species are reported to oc- 40 species (Rodriguez et al. ，2004). This ge- cur on both East and West American coasts: nus is aheterogeneous assemblage of species， Pachycheles chacei Haig，1956 ，P .monil 件r some of them previously assigned to Pisosoma (Dana，1852) ，Petrolisthes armatus (Gibbes， Stimpson，1858 ，which is currently considered 1850)，P. galathinus (Bosc，1802) ，P .robsonae asynonym of Pachycheles (Haig，1960). Glassell ，1945 ，P. tonsorius Haig，1960 ，and P. Two previous molecular-based studies of tridentatus Stimpson，1858 (Haig，1956 ，1960; Porcellanidae have been published. A mo・ Gore & Abele，1973; Scelzo，1982 ;Abele & lecular phylogeny of eastern Pacific genera Kim，1989; Rodriguez etal. ，2005). Forty- Petrolisthes and Pachycheles was based on mi- eight valid species and two undescribed species tochondrial 16S rRNA gene (Stillman & Reeb， grouped into eleven genera occur in the westem 2001) and included afew sequences of Atlantic Atlantic (Rodriguez et al .，2005). specimens for amphi-American species of Phylogenetic relationships of porcellanids P etrolis thes. In this study，the subdivision of are poorly understood. Porcellanid genera Petrolisthes into two main clades supported ear- have long warranted revision on aworldwide lier division based on morphological characters basis にhace，1942; Haig，1956 ，1960; Werding (Haig，1960). One of these clades containted et α1.，2001; Rodriguez et al .，2005) . ln the only species possesing regular serrate spines ， last 50 years，the number of described porcel- and the other clade contained species that are lanid species in the western Atlantic increased smooth or irregularly decorated (“ spiny"and from 31 to 48 with the addition of newly col- “smooth" species，respectively ，se nsu Stillman lected and described cryptic species (Werding and Reeb，2001). A second molecular study & Hiller，2002; Rodriguez etal. ，2005). The used partial sequences of the mitochondrial family includes three widely distributed and COI gene to infer phylogenetic relationships morphologically variable species complexes， between two species of the genus Neopisosoma Petrolisthes armatus，P. galathinus ，and P. Haig，1960 (Werding etal. ，2001). It was sug- tridentatus; and their taxonomic status remains gested that Neopisosoma should not be treated unclear (Werding et al. ，2003; Rodriguez et al. ， as separate from Pachycheles ，because it was 2005). found to be paraphyletic. However，a limited Morphological and molecular studies can number of taxa have been studied to date，and facilitate understanding of systematic and phy- the status of Neopisosoma remains unclear logenetic relationships within the Porcellanidae. (Rodriguez etal. ，2004 ，2005). Phylogeny of westem Atlantic Porcellanidae 153

In the present study，partial sequences of obtain apelle t. The pellet was washed in 20μ1 the mitochondrial 16S rRNA gene were used to 70% ethanol ，centrifuged at 14，000 rpm for 1 elucidate phylogenetic relationships within the min，and Iyophilized in aONAIIO SpeedVac Porcellanidae in the westem Atlantic in order (TherrnoSavant) at high drying rate for 30 min. to provide insights into classification ，detect Genomic ONAwas re-suspended in 10 -50μl cryptic species，and gain knowledge of phylo- of 10X TE Buffer (1 mMEOTA ，10 mMTris ， geographic pa口ems in this family. pH 7.5) or ddHpand stored at -20 oC. DNAamplification. - Amplification of a- 545 basepair product from the mitochon- MATERIALS AND ETHODS 九' 1 drial 16S rRNA gene was carried out by a Sample coIl ection and depositories polymerase・chain-reaction (PCR) using two Most porcelain crabs were collected in forward primers，16Sar (5'-CGCCTGTTTAT coastal Florida (J uly 2000，July -August 2001)， CAAAAACA下3') (Palumbi etal. ，1991) and along the coasts of Louisiana (June 2001)，and 16L2 (5'-TGCCTGTTTATCAAAAACAT・3') Texas (September 2001)，U.S.A.; from Veracruz (Schubart etal. ，2002) ，and the reverse primer to Quintana Roo，Mexico (J uly 2002); in Belize 1472 (デー AGATAGAAACCAACCTGG-3') (October 2002); in the vicinity of La Tortuga， (Crandall & Fitzpatrick，1996). The PCR mix Margarita，and Cubagua Islands，Venezuela for each sample contained 6.7μ1 ddHP，3.0μl

(March-June 1996，April -June，2001); and on 10 X PCR buffer，3.0μ1 MgCI2(25 mM)，5.0 the Paci白c coast ofNicaragua (September 2000， μ1 Betaine (5 M)，4.0μ1 dNTP's (1.25 mM)， November 2001). For each species，one speci- 0.5μ1 of each forward primer (20μM)，I.Oμl men per locality was used in the analyses，as of reverse primer (20μM)，0.5μ1 AmpliTaq listed in Table 1.AII specimens were preserved Gold骨 (5 U /μ1 ) ，and 1. 0μ1 ONAtemplate (-20 in 70-95% ethano l. Genetic vouchers were de- ng/μ1 ) .The PCR profile was 42 cycles (de- posited in the Zoological Collection，University naturing at 940 C I1 min，annealing at 480 C I1 of Louisiana at Lafayette，Louisiana ，U .S.A. min，and extension at 72 0 C I1 :30 min). Four (ULLZ). Some previously archived mater卜 μ1 of PCR products underwent electrophoresis als from ULLZ and the Coleccion Nacional de on a 1.5 % agarose gel and were visualized Crustaceos，Instituto de Biologia，Universidad with ethidium bromide staining. PCR prod- Nacional Autonoma de Mexico，Mexico ，O. ucts were purified with Microcon 100骨日ters. F. ，Mexico (CNCR) were also included in the DNAsequencing. -Sequencing reactants analyses (Table 1) . for each strand (forward and reverse) of ONA included 3.8μ1 ddHP，8.0μ1 2.5X Buffer (200

・ DNAextraction ，amplification ，and sequenc mMTris ，5 mM MgCI 2，pH 9.0) ，4.0μ1 ABI mg Big Oye@terrninator mix，3.2μ1 primer (2μM) Procedures were modified a仇er Kocher et (the two forward primers were used combined， al. (1989) and Schubart et al. (2002). 1. 6μ1 of each)，and 1.0μ1 purified ONA.The DNAextraction. - Genomic ONAwas profile was 28 cycles (denaturing at 960 C /l 5 obtained from muscle tissue of one or both sec ，annea I1 ng at 500 C Il 0sec ，and extension at chelipeds.Tissue was ground in 600μ1 Iysis 60oC /4:20 min).Seven μ1 of each sequencing buffer (100 mMEOTA ，10 mMTris-HCI ，pH product were visualized with ethidium bromide 7.5，1 % SOS) and incubated for 1-3hours at staining on a 1.5% electrophoretic agarose 65 0 C . ONAwas precipitated by adding 200 ge l. ONAstrands were purified in 800μ1 (8%) μ1 7.5 M NHpAc and centrifug巴d at 14，000 Sephadex columns，centrifuged at 1，500 rpm rpm for 4min. Supematant was transferred to for 5min .Sequences were obtained on an ABI atube containing 600μ1 of cold isopropanol， 3100 Genetic Analyzer (Applied Biosystems， and ce ntrifuged at 14 ，000 rpm for 1-3min to Inc.). 154 1. T. Rodriguez et al

Table 1. List of porcellanid specimens for which partial sequences of the mitochondrial 16S rRNA gene were used in this study. (P) = Eastem Pacific. ULLZ = Zoological Collection ，University of Louisiana at Lafayette ，Lafayette ，Louisiana ，USA; CNCR = Coleccion Nacional de Crustaceos，Instituto de Biologia， Universidad Nacional Autonoma de Mexico，Mexico.

Collection and GenBank Species Locality number number

Claslotoechus Haig，1960 C. nodosus (Streets ，1872) Mexico: Veracruz ULLZ 5370 DQ865311 C. nodosus (Streets ，1872) Venezuela: Isla Cubagua ULLZ 5341 DQ865312 C. vanderhorsti (Schmi口， 1924) Venezuela: Isla La Tortuga ULLZ 5342 DQ865313 Megalobrachium Stimpson，1858 M. poeyi (Guerin ，1855) Mexico: Quintana Roo CNCR 9818 DQ865326 M. poeyi (Guerin，1855) Venezuela: Isla Margarita ULLZ 5343 DQ865327 M. soriatum (Say，1818) Texas: South Padre Island ULLZ 5279 DQ865324 M. soriatum (Say，1818) Florida: Bahia Honda ULLZ 5262 DQ865325 Neopisosoma Haig，1960 八r. angustifrons (Benedict，1901) Mexico・Veracruz ULLZ 5373 DQ865336 N. angustifrons (Benedict，1901) Venezuela: Isla Cubagua ULLZ 5345 DQ865337 Pachycheles Stimpson，1858 Packleianus A. Milne-Edwards，1880 Florida :Looe Key Reef ULLZ 4824 DQ865332 P monilifer (Dana，1852) Mexico: Veracruz ULLZ 5388 DQ865330 P monilifer (Dana，1852) Venezuela: Isla Cubagua ULLZ 5348 DQ865331 Ppilosus (H. Milne Edwards，1837) Mexico: Quintana Roo ULLZ 5389 DQ865328 Ppilosus (H. Milne Edwards，1837) Venezuela: Isla La Tortuga ULLZ 5349 DQ865329 P susanae Gore &Abele ，1973 Mexico: Quintana Roo CNCR 7273 DQ865333 P susanae Gore &Abele ，1973 Venezuela: Isla Cubagua ULLZ 5352 DQ865334 Pachycheles sp. Nicaragua: Miramar (P) ULLZ 5298 DQ865335 Parapetrolisthes Haig，1962 P tortugensis (Glassell ，1945) Florida: Looe Key Reef ULLZ 4825 DQ865304 Petrolisthes Stimpson，1858 P armatus (Gibbes，1850) Florida: Fort Pierce ULLZ 5252 DQ865310 P caribensis Werding，1983 Mexico: Quintana Roo CNCR 5675 DQ865297 l宅caribensis Werding，1983 Mexico:Quintana Roo CNCR 9701 DQ865296 P caribensis Werding，1983 Belize: Twin Cays ULLZ 5428 DQ865295 Pgalathinus (Bosc，1802) Mexico: Quintana Roo CNCR 10937 DQ865303 Pgalathinus (Bosc，1802) Mexico: Quintana Roo ULLZ 5401 DQ865298 Pgalathinus (Bosc，1802) Belize: Carrie Bow Cay ULLZ 5430 DQ865299 Pgalathinus (Bosc，1802) Belize: Carrie Bow Cay ULLZ 5431 DQ865300 Pgalathinus (Bosc，1802) Venezuela: Isla Cubagua ULLZ 5355 DQ865301 Pgalathinus (Bosc，1802) Nicaragua: Estero Aserradores (P) ULLZ 5316 DQ865302 Pjugosus (Streets ，1872) Mexico: Veracruz ULLZ 5405 DQ865314 Pjugosus (Streets ，1872) Mexico: Quintana Roo ULLZ 5406 DQ865315 P lewisi (Glassell ，1936) Nicaragua: Estero Nagualapa (P) ULLZ 5321 DQ865317 P magdalenensis Werding，1978 Venezuela: Isla Margarita ULLZ 5358 DQ865316 P marginatus Stimpson，1859 Mexico: Veracruz ULLZ 5410 DQ865308 Phylogeny ofwestem Atlantic Porcellanidae 155

P marginatusStimpson ，1859 Venezuela: Isla Margarita ULLZ 5359 DQ865309 Ppolitus (Gray，1831) Mexico: Quintana Roo ULLZ 5412 DQ865306 Ppolitu s(Gray ，183 1) Venezue1a: Isla Cubagua ULLZ 5360 DQ865307 Pquadratu sBenedict ，1901 Mexico: Quintana Roo CNCR 3724 DQ865318 Pquadratu sBenedict ，1901 Venezuela :Isla La Tortuga ULLZ 5361 DQ865319 Prosari ensisWerding ，1978 Belize :Carrie Bow Cay ULLZ 5435 DQ865305 Ptrid entatusStimpson ，1859 Mexico: Quintan aRoo ULLZ 5414 DQ865322 P tridentatusStimpson ，1859 Venezue1a :Isla Margarita ULLZ 5363 DQ865323 Pt ridenlalusStimp son ，18 59 Nicaragua: Estero N ag ua1apa (P) ULLZ 5324 DQ86 532 1 P trident alu sStimpson ，18 59 Panama: Isla Naos (P ) ULLZ 5364 DQ865320 P violaceus(Guerin ，1831) Chile :Las Cruces AF260608 1 Polyonyx Stimpson，1858 Pg ibbes iH aig，1956 Florida: Fort Pierce ULLZ 5254 DQ8653 41 Porcellan aL amarck，1801 Ps ayana(Leach ，1820) Loui siana :offshor e ULLZ 45 74 DQ865338 Psayana (Leach，1820) Venezuela: Isla Margarita ULLZ 5366 DQ865339 Psi gsb eiana A. Milne- Edwards，1880 Florida: Straits of Florida ULLZ 4481 DQ865340 Outgroup Eumunida Smith ，1883 E. sternomaculata Saint Laurent & New Ca1edonia AY3512602 Macpherson，1990

1Sequence of 447 bp deposited in GenBank byStillm an& Reeb (2001). 2Sequence of 515 bp deposited in GenBank byMachordom & Macpherson (2004)

Phylogenetic analyses Starting with the default values of gap-opening Consensussequences were obtained by cut- and gap-extension penalties for pairwise and ting the primer regions and aligning the forward multiple alignments，several values were ap- and reverse DNAsequences using the program plied until 12 and 6，respectively ，were selected Sequencher™ 3.0(Gene Codes Corporation). as the values that achieved the alignment with These sequences were deposited in GenBank the minimum number of gaps. The ali gnment (accession numbers DQ865295 to DQ865341， was checked and manually improved using the Table 1) . Sequences from each related fam- program BioEdit version 7.0.4 .1 (Hall ，1999). ily within the Galatheoidea (Aeglidae: A egla MODELTEST 3.06 (Posada & Crandall， papudo Schmitt，1942 ，GenBank Accession 1998) was used to select the model of sequence No.AY050032. Chirostylidae :Eumunida st er- evolution that best fitted the data. PAUP* nomaculata de Saint Laurent & Macpherson， 4.0b 10 (Swofford ，2002) was used to anal yze 1990，AY351260. Galatheidae:Munida the data and construct phylogenetic hypotheses quadrispina Benedict，1902 ，AF436050) were by the following methods: Neighbor Joining tried as outgroups. With the exception of the (NJ)，with distances set to maximum likeli- Chirostylidae，which we used as the outgroup hood in order to apply the evolutionary model; for analyses that follow，these groups exhibited Maximum Parsimony (MP) analysis byheu- large distances from the Porcellanidae. ristic search under TBR branch swapping and ClustalX version 1.81 (Thompson etal. ， random taxon addition ，with gaps treated as a 1997) was first used to align the sequences. “自 fth base" and aconsensus of the most par・ 156 1. T.R odri guezet al. simonious trees gathered by applying the 50% provide substantive change in results .Variable Majority Rule; Maximum Likelihood (ML) characters numbered 294 (52% )and 263 char- through heuristic search by stepwise addition， acters were parsimony-informative (47%). taking into account the evolutionary mode l. Confidence values (cv) for each method were Phylogenetic relationships estimated bybootstrappin g(Felsenstein ，1985 ). The model of evolution selected was One thousand replicates were use dfor the NJ TVM+ I+ f(transversional model)(Rodriguez et tree and MPconsensus tree ，and 100 replicates al. ，1990) .Ba se frequencies were A = 0.4 213， byfast ste pwise -addition for the MLtree ，re- C =0.06 84，G =0 .1 265，T = 0.3838. The rate taining only groups with frequency > 50 %. matrix was 0.3097，7.0460 ，1. 1015，0 .8 444， Confidence values of 80 or above were consid- 7.0460. The proportion of invariable sites (1) = ered high values 0.2543. The gamma distribution shape param- Bayesian Analysis (BA) was performed us・ eter =0 .39 43 ing the program MrBayes v3.0b4 (Huelsenbeck Topology of the trees generated with four & Ronquist，200 1) .The parameters of the methods (NJ，MP ，ML ，and BA) issi milar for selected model of evolution were stated. most of the clades. ln general ，howe ver，NJ 2，000，000 generations with asa mpling fre- ismore congruent w ith MP，a nd MLwith BA. quency of 100 produced 20，001 tr ees.The first The MLph ylogram is presented to illu st rate 2，000 trees were eliminated. A consensus of genetic di vergence (Fig. 1) .The MPconsensus 18 ，001 trees was reconstructed with PAUP* us ・ of the five most parsimonious trees after boot- in gthe 50% M 司ority Rule. strapping was selected as the tree to show the reliability of the clades according to bootstrap- ping (NJ，MP ，and ML) and the posterior prob- RESULTS abilities of BA(Fig. 2) .Clades with con白dence Data set values above 50 in at 1east two methodswere Forty-seven partia1 sequences of the 16S incl uded，except for the clade grouping spe- rRNA gene of specimens belonging to 25 spe- cies of Pachycheles ，which was supported by cies in eight genera of Porcellanidae were MPonly (cv =80). Because the relationships obtained and used in the analyses along with between Pachycheles and Neopisosoma are not sequences from GenBank for two species， clear，we decidedto make anexception to our Petrolisthes vio la ceus Guerin，1 83 1(type spe- rule in order to include the information pro- cies of the genus)，and Eumunida sternomacu- vided by thi sone method. The tree was modi- lata (outgroup) (Table 1) . This species was fied to group both clades containing species found to be the clo ses tto the Porcellanidae in a of P etrolisth es ，as supported by ML(cv =55) preliminaryanalysis that included as pecimen and BA(cv =99) .P etroli sth es vio laceus was 合omeach family within the Galatheoidea; thus excluded from the group P .ju gosus (Streets ， it was used in subsequent analyses as amono- 1872)-Clastoto ec hus due to the low support for phyletic sister group. thi srelationship (cv = 65)，and it was instead Sequences ranged from 54 ト549 bp，except grouped with the clade P. tridentatus-P. qua- for Megalobrachium soriatum (Say，181 8) from dratus-P .lewisi -P .magdalenensis because of Texas (477 bp)，P etroli st hes violaceus (447 bp)， higher BAs upport (cv =80) . and Eumunida sternomaculata (515 bp) .The The followingclades have statistical sup- ali gnment of sequences of 48 porcelain crabs port by atleast two methods.C lade A ，which and the outgroup consisted of 561 characters. groups P olyonyx- Porcellana ，is supported Position s40 -50 and 235-294 were the most bya ll four methods. This clade is external variable; even so theywere included in the final to the rest of the Porcell anidae . Clade B ， alignment after analyses excluding them did not which groups the rest of the Porcellanidaein Phylogeny of westem Atlantic Porcellanidae 157

Pelr o!i slh es cari bensis BEL P elr o!i slh es caribensis MEX P elr o!i slh ιSc aribensis MEX Parapelr o/i slh es lorlugensisFL Pelr o!i slh es galalhinusMEX P elr o!i slh es galalhinus BEL Pelr o!i slh es galalhinus BEL P elr o!i slh es galalhinll sV E Pelr o!i slh es galalhinusNIC (P ) Pelr o!i slh es galalhinusMEX Pelr olis lh es rosari ensisB EL P elrolislh esp o!i lu sM EX Pelr o!i slh es polilusV EN

D C lasloloechll sva nderhors li V EN P elr o!i slh es jugosusM EX P elro!i slh esj ugosusMEX Pelr o!i slh es viola cell sC HILE Pelr o!i slh es m agdalenensis V E Pelro!i slh es lewisiN IC (P) Pelro !i slh esquadralu sM EX E Pelr o!i slh es quadralll sV EN Pelr o/i slh es IridenlalusPAN (P) P elr o!i slh es Iridenlalus NIC (P) P elr olislh esIridenlalll sMEX P elr o!i slh esIrid enlalll sV EN M egalobrachium sorialllm TX M egalobrachillm sorialum FL M egalobrachillm poeyiM EX Megalobrachillm poeyi V EN

Neopisosom aa ng川，lif ト0 附 M EX Neopisosom aa ngz 川 jトons V E Pachychelesp il oslls MEX Pα chychele spil oSl/sVE N Pachycheles ackl eωnusFL B Pachycheles m onil i戸rMEX Pachychelesmonilif erVEN Pachychelessusana eMEX PachychelesS ll sanaeVE N Pachycheles sp.NI C(P) P orcell anas ayana LA A Porcellanasaya naVE N

Eumllnida sl ernomacul ala 一- 5% di ver gence Fig.l Maximum llitelihood phylogenetic distance tree for poreElam crabs based on pamal sequences of the 16S rRNA gene(561 characters)Model of sequence evoltitlon TVM+l+r.Letters Ato Fir1dl cate the main clades. FL =Florida; LA= Louisiana ;TX = Texas ;MEX = Mexico; BEL =Belize; VEN =Venezuela ;PAN = Panama; NIC = Nicaragua;(P) = Eastern Pacific collections where indicated ，all other porcellanid collections are westem Atlantic . 158 1.T. Rodrigueze ta l

Petrolisthes coribensisA EL 100/1 00 P etrolisthes caribensis MEX 99/1 00 P etro!i sthes caribensis M EX P etro!i sth es galafhil1l1 sM EX Pefrolisfhes galafhil1l1 sB EL Pefro!i sfhes galathinlls BEL P etro!i sfhes galafhil1l1 sVEN F P etr oli sfhes galafhinllsN IC( P) 89/98 P etr olis fh es gαlafhinlls M EX Parapetr o!i sfh es fOrfllgensis FL 80/100 P efr olis fhesrosariensis BEL P efr oli sfhes poli/ll sMEX P efrolisfhes p oli /ll sVEN P etrolisthes marginaflls MEX P efro!i sthes marg inatll sVE N Petrolisth es arm alus FL Clasfofoechlls nodoslls M EX :D C/asfofoechlls nodosll sVEN 175 Clasfofoechlls vanderhorsfi VEN 55 /99 53 /56 P efrolis fhesjllgosll sMEX P efro!i sfhesjll goslI sMEX P etr olis th es νio lacell sCH I LE Petrolisfhes m agdalenensis VE P etr olis thes lelVisi N IC(P) P etr o!i sfhes qlladrall1 SMEX P etro!i sfh es quadratllsVEN Petrolisfhes fridenfαfu sPAN (P) B P efrolisfhes fridenfafllS N IC(P) Pefrolisfhesfridenfa fll SMEX 93/1 00 P efro!isfhes fridentafl1 SVEN Mega!υhrachium sorialUm TX 66/81 M egalοbrachiuln sorialutn FL 73/100 Megalοbrachillln poeyiM EX M egalobrachil1 m poeyi VEN Pachycheles piloslls MEX P achycheles pilosl1 sVEN Pachycheles ackleianlls FL Pachyche!es m oniliferMEX Pachyche!es m onilifer VE C Pachyche!出 sllsanae M EX 70/93 Pachycheles Sll sanae VEN 65 1 P achycheles sp.N IC (P) 100/100 Neupisosoma “ng usl(斤O I1 S MEX 98/1 00 Neopisosoma angllsfijrons VEN A Porcellana sayana LA Porcellana sayana V EN 97/1 00 Porcellana sigsbeiana FL 90/100 Pο Iyony. λcgihhesi FL Eutnunida slernumaζulala

Fig.2 .Ma ximum parsimonyph ylogeneti cco nsen sustr ee based onp arti alsequ ences ofth e1 6S rRNAge ne (56 1c harac ters) m odifi edat das hedlin es tos how clades support edb yo th erm eth ods. Confid ence va lu es basedo nNe ighborJ oinin g(N J)，M aximum Parsimony( MP)，M ax imum Likelih ood( ML)，an dB ayes ian A nalys is( B A) m eth ods w ith m odelof se quence evoluti onTVM +I+ r. U pperva lu es corr es pond to N J/MP (bootstr apof 1000 replicat es) and lowerva lu es corre spond toM Ll BA (bootstr apof 100 repli ca tes and posteri orpr obabiliti es ，res pectiv ely) .V alues ar es how nfo rn odes w ith support > 50%. Lett ersA toF indi cateth emain cl ades. FL =Fl orid a; LA= Loui siana ;TX =T exas; M EX =Mexic o; B EL= B eli ze; V EN =V enez uela ;PAN =Panam a; NIC =N ic ara gua; (P) =E astern Pac ific coll ecti onswher eindic ated，a ll oth er porc ell anid collection sare w es tern Atl anti c. Phylogeny of westem Atlantic Porcellanidae 159 this study，is also supported by all four meth- populations of nearly all species occurs across ods. This clade is further divided into clade their geographic ranges (Avise，2004) ，we C ，which groups Neopisosoma-Pachycheles ， sought to exemplify this difference between supported by NJ and MP(cv = 70 and 93， specimens from Mexico (Louisiana in the case respectively)，and clade D ，which groups of Porcellana sayana) and specimens from Aイegalobrachium-"smooth" Petrolisthes- Venezuela for 13 species. In most of these Cl as totoechus-Parapetrol isth es-"spiny" cases，the full range of distribution for the spe- Petrolisthes ，supported by MLand BA(cv = cies was covered or very nearly covered. We 55 and 99，respectively). Clade D is further also intentionally included specimens of the divided into clades E and F. Clade E groups trans-Floridian species Megalobrachium soria- Megalobrachium-"smooth" Petrolisthes- tum，as well as representatives of two amphト Clastotoechus，and is similarly supported by American species，Petrolisthes galathinusand the four methods. Megalobrachium is extemal P. tridentatus. An exception was made for the to the clade of “smooth" P etrolisthes ，which in- widely distributed amphi-American species P. cludes the groups of P. tridentatus-P. quadra- armatus，which was included here as asingle tus-P. lewisi-P. magdalenensis-P. vio lac eus sequence only to clari かits relationship to other and P. jugosus-Clastotoechus. Clade Fgroups species in the family; this species is beingtreat- “spiny" species of Petrolisthes，and it is also ed exhaustively in aseparate paper dealingwith supported by the four methods. Within this last population relationships over its entire range clade，three lineages are distinguished，one for (Rodriguez & Felder，in review). P. armatus-P. marginatus-P .politus ，a second lineage represented by P. rosariensis ，and a Phylogenetic relationships third lineage of species of the Petrolisthes gal- The Porcellanidae has long been postulated athinus-complex，which includes P .ca rib ensis ， to have its closest affinities with the family P. galathinus，and Parapetrolisthes tortugensis Galatheidae on the basis of morphological (Fig.2). characters (Haig，1960). Using molecular char- acters ，Morrison & Cunningham (1999) found the Lomisidae close lyrelated to the Aeglidae. DISCUSSION Spermatozoal characters support this relation- Species in the data set ship as well (Tudge & Scheltinga，2002). C . Nearly half of the porcellanid species of Morrison [pers .comm. in Martin & Davis common genera distributed in the western (2001)] suggested aseparate lineage for these Atlantic were included in thi sstudy .Onl ythe two groups，however ，Martin & Davis (2001)， genera Euceramus Stimpson，1860 ，Minyocerus following McLaughlin (1983)，decided not to Stimpson，1858 ，and Pisidia Leach，1820 in c1 ude the Lomisidae within the Galatheoidea were not included，either for lack of materi- but rather place it in an adjacent superfam・ als or because available specimens could not ily ，the Lomisoidea. The close relationship be used successfully in sequence analysis [see between Aeglidae and Lomisidae was further Rodriguez etal .(2005) for acomplete account confirmed using partial sequences of four genes ofspecies and genera in the region]. (COII，16S ，18S ，and 28S) to show gene rear- Genetic divergence and geographic distance rangements (Morrison etal. ，2002). In the were not compared analytically in the pres- same study，the Chirostylidae and Galatheidae ent work，as we did not set out to exhaustively were groupedclosely together and somewhat include multiple populations for each species. more distantly to the Porcellanidae .In amore Nonetheless，an empirical approximation for recent phylogenetic study of the reptantian the level of r巴lationship was gathered by using Decapoda with morphological characters and species representatives from distant localities molecular sequencesof two genes (COI and where possible. As genetic differentiation in 16S) (Ahyong & O'Meally，2004) ，the Aeglidae 160 1. T. Rodriguez elal is grouped with the Porcellanidae in the mor- ships of C. vanderhorsti and its congeners. phological analysis and with the Lomisidae in Megalobrachium constitutes adistinct the molecular and the combined analyses，thus group in clade E ，b asally separated from the supporting aseparate lineage for Aeglidae and species comprising C/astotoechus and as ub- Lomisidae. In the same combined analysis，the set of Petrolisthes (Figs. 1，2). Divergence Chirostylidae is c1 0ser to the Porcellanidae than within the genus Megalobrachium was very the Galatheidae (Ahyong & O'Meally，2004). high (Fig. 1) ，which could argue for the resur- In the present study，the Porcellanidae is found rection of two previously separated genera， more closely related to both the Chirostylidae Megalobrachium，represented in this study and Aeglidae，than it is to the Galatheidae， byM. poeyi，and Porcellanopsis，represented though this conclusion must be preliminary as by former P. soriata，currently ass igned only one partial sequence of one gene for each to Mega/obrachium (Chace，1942 ;Haig ， family was used in the analyses. 1956). Haig (1960) decided to synonymize The family Porcel1 anidae is divided into Porcellanopsis with Megalobrachium after con- two main clades，c1 ade A grouped the genera sidering them aseries without cJ ear subdivision Polyonyx and Porcellana，and clade B grouped based on morphological characters. The genus the rest of the genera treated in this study (Figs. Megalobrachium contains 12 species distrib- 1，2). The distinction between clades A and uted on both coasts of the Americas (Rodriguez

B is largely supported on the basis of larval et al. ，2005). Further molecular and morpho・ mo中hology (Lebour，1943 ;Hemandez ，1999) logical analyses that in cJ ude more species of and sperm and spermatophore morphology Megalobrachium (studies in progress) should (Tudge & Jamieson，1996a ，b ;Tudge ，1997) . further clari 今the status of the genus. Shared character sets of the Porcellana- Specimens of Megalobrαchium soriatum group of larvae suggest arelationship be- from Texas and the Florida Keys have larger

tween the westem Atlantic genera Euceramus， genetic divergence (branch length) than speci ・ Minyocerus，P 山 dia ，Polyonyx，and Porcellan α， mens of M. poeyi from Mexico and Venezuela， whereas larval characters of the P etr olist hes- which have a larger geographic distance group characterize the genera Clastotoechus， separating them (Fig. 1) . Trans-Floridian Megalobrachium，Neopisosoma ，Pa chycheles ， divergence has been documented for closely and Petrolisthes. Thus，molecular analyses related species of decapods within the genera suppo口 separation of the major groups of the Sesarma，Uca (both Brachyura)，Callichirus Porcellanidae (c1 ade A or Porcellana-group， (Thalassinidea)，and Pagurus(Anomura) (see and clade B or Petrolisthes-group) into dis- Felder & Staton，1994 ;Staton & Felder，1995; tinct taxa at the subfamily level as suggested Young et al. ，2002). Divergence，or at least by Tudge & Jamieson (l 996b) and Hemandez sorting of divergent haplotypes for some Gulf (1999). of Mexico endemics or eastern and western The two species of Clastotoechus are Gulf counterparts，especially those confined grouped closely and there is high support for to shallow coastal environments，may relate to their generic level clade (Figs. 1，2). In apre- periods and patterns of contact and isolation vious molecular analysis using sequences of during peak glacial advances in North America the COI gene for the same species，the genus (Felder & Staton ，1994; Felder，2001). These was also well supported (Werding et al. ，2001). glaciations were initiated between 3.1 and 2.5 The monotypic genus Madarateuchus Harvey， million years ago (Mya) (Haug & Tiedemann， 1999，proposed to receive C. vanderhorsti ，is 1998)，with the last maximum estimated to unsupported by the current molecular analy- 18 ，000 years ago (Young etal. ，2002). lf ex- sis. Further morphological and molecular pected divergence rates were applied [0 .533% studies，with the inclusion of more species of or 0.366 % per million years，estimated for C lastoto echus，may fu 口her clarifシthe relation- Petrolisthes armatus by Stillman & Reeb Phyl ogeny of westem Atlantic Porcellanidae 161

(2001) and Rodriguez & Felder (unpublished covered with granules and bear atel sonthat data) ，respectively] ，then populations of M. so- is 7-plated. The second lineage，P. monilifer riatum should have diverged at least 10 Mya. -P. ackl eianus-P. pilosus is supported byMP While this estimation of genetic divergence and BA(Fig. 2). Morphologically，P. pilosus perse mostlikely rules out the latest of glacial is unique in the western Atlantic in havi ng the maxima as the sole agent of divergence，these chelipeds thickly covered with stiff bristles . cycles of contracting and expanding temperate Pachycheles αckl eianusand P. monil件r have coastal environments in the Gulf of Mexico lon gitudinal rows of large tlattened tubercles would almost certainly have played arole on the chelae，and all three species in thi sclade in distributional sorting. Oivergence among have 5plates on the telson. A combined anal y- populations of M. soriatum is similar to that be- sis of western Atlantic (this study) and eastern tween cryptic species within the P .g alathinus Pacific (Stillman & Reeb，2001) specimens and P tridentatus clades (Fig. 1;see length of currentl yassigned to Pachycheles (in progress) the branches).Unfortunately ，in the case of M. will certainly helpto define the limits of each soriatum ，no morphological variation could be group. correlated to genetic divergence in the two sib- P arapetrolisth es tortugensis is grouped ling populations (Rodriguez etal. ，2005). within the P etrolisth es gα lathinus-complex Within the limits of our present analysis， (Figs .1 ，2) and is especially close to P .ca- Neopisosoma is supported as agenus distinct rib ensis(Fig. 1) . The presence of di st inct ， from Pachycheles. In clade C ，Ne opisosoma transverse，piliferous striations on the carapace is clearly external to the clade of species as- seems to be aunif yin gcharacter within this signable to Pachycheles (Fi g.2) .These two complex (Rodriguez etal .，2005) .The genus genera can be distinguished primarily on the Parapetro listh es Haig，1962 ，is defined based structure of the posterior portion of the lateral on the shape of the fingers and chelipeds wall of the carapace，which is membranous in Maintaining aseparate genus for this species Neopisosom αb u t consists of one or more calci- is not supported by the present molecular data， fied pieces separated by membranous regions and neither is the previously postulated close in Pachycheles (Haig，1960) . The status of relationship with the Galatheidae. Neop isosoma as agenus has been questioned Species of the genus P etroli sth es are in- since Haig (1960) establ ished it and has been c1 uded in two main clades，E and F (Figs. 1， further discussed by Gore (1977)，Werding 2) ，as occurred with eastern Pacific species (1986)，Werding & Muller (1990)，Werding et of the genus (Stillman & Reeb，2001). This al.，(2001) ，and Rodriguez et al. (2004，2005) division also has morphological support (see A worldwide revision of both genera is required Introduction). Clade E contains “smooth" in order to clari 命their relationships . (sensu Stillman & Reeb，2001) species of Oivergence between specimens of P etr olis th es grouped with M egalobrachium Neopisosoma angustφonsis relatively large and Clastotoechus (Figs. 1，勾)2 and before Haig (Fig. 1). This is probably due to geographic (υ196ω0) 臼separat匂ed the 叩species of αiC ω astωotωoe町chu附必4S distance (Mexico and Venezuela)，although it is these were ∞consl凶der閃ed “abe町rr悶an川1 叫t"species of also possibl ethat acryptic species is contained P etr oli ωf注st的hes. P etr oli st hes jugosus is weakly in thi staxon. grouped with Clastotoechus (Figs. 1，2) ，how- The monophyletic status of Pachycheles ever，these taxa are morphologically not simi- is supported by MPonly (cv = 80) (Fig. 2). lar .Haig (1960) suggested aclose relationship The genus is represented by two independent between P. jugosusand P. lewisi base don linea ges: the first containsP. susanaeand morphological characters，but this relationship Pachycheles sp.(a species similar to P .ca lculo- is not supported in the present study. sus)and is supported by NJ，MP ，and ML(Fig. Petr olisthes violaceus ，type species of the 2). Both of these species have naked chelipeds genus，is grouped with the clade of P .tridenta- 162 1. T. Rodri gue ze tal .

tus -P. quadratus-P. lewisi-P. magdalenensis 1982，1983a ;Werding & Hiller，2002). It dif- with BAsupport (cv = 80) (Fig. 2). Petrolisthes fers from the species in the complex by having quadratus and P. tridentatus are grouped two epibranchia lspines. The specimen used closely to one another，and further grouped in this study agrees with “Petrolisthes sp. 1" with the clade of P. lewisi and P. magdalenen- (Werding，(9 77) in having astrong supraocular sis，both of these groupings being supported by spine. M aterials listed ，illustrated ，and briefl y MP，ML ，and BA(Fig. 2). Morphologically，P . diagnosed by Werding (1977) were later associ - violaceus is also close to these groups，as could ated with the name P .rosariensis (Werding， be inferred by Haig (1960)，who distinguished 1978).Thus the name became valid in 19 78， alarge group of unarmed species that included well in advance of the date intended by the au- both P. violaceus and P. quadratus. On the ba- thor (Werding，(982). This later description of sis of molecular，adult ，and larval characters，it the species disagrees with previous materials is our opinion that P. lewisi，P .magdalenensis ， in lacking astrong supraocular spine and might P. tonsorius，and P .tridentatus should be in represent asecond species (see Rodriguez et this unarmed group as wel l. al. ，2 005，for discussion on the identit yof P . The clade encompassing specimens of rosariensis) Petro /i sthestridentatus from distant geographic In the third linea ge within clade F，speci- regions shows substantial intraspecific di- mensof P etrolisth es galathinus from Mexico vergence，and several distinct species should and Belize group together to the exclusion of P. almost certainly be recognizedamong them galathinusfrom Venezuela，most probablydue (Fig. 1) . Specimens from the Pacific coasts to geographic distance.Petrolisth es galathinus of Panama and Nicaragua have identical se- from the Paci 自c coast of Nicaragua and asec- quences，but the specimen from the Caribbean ond haplotype from Mexico are excluded from coast of Mexico is well diverged from the the previous clade (Figs. 1，2). Genetic diver- Pacific ones，and the specimen from Venezuela gence is noticeablylarge among specimens of is diverged from all the others (Figs. 1，2). To P .galathinus (Fig.1) ，supporting r巴cognition determine which are in fact new species，all of new species that also appear to be distinct in must be compared to the type material of P. tri- minor morphological features and coloration dentatusfrom St. Thomas and Barbados. (studies in progress). Specimens of P. cari- Clade F includes “spiny" species of bensis group together in avery well suppoロed P etrolisth es arranged into three lineages. The clade within the branch encompassing the first lineage includes P .armatus in close rela- Petrolisth es galathinus-c omplex (Fig. 2). As tion to P. marginatus，with both of these further discussed above，Parapetrolisthes tortu gensis grouping with P. politus. These relationships is also included within this complex (Figs .1 ， are supported by all four phylogenetic methods 2) . (Figs. 1，2). P etrolisthes armatus and P .mar- Polyonyx is closely related to Porcellana ginatu sare characterized by having one epi- with hi ghsuppo 口 for adi st inct clade (A) 合om branchi alspine ，while P. politus has no spines the four phylo genetic methods (Figs. 1，2).The on the carapace. The carpus of P. armatus subclade encompassing Porcellana included and P. politus is armed with 3(sometimes 4in both species analyzed with similar hi gh sup- the former) low，wide-set ，spine-tipped teeth ， po口(Fig.2). Porcellana sayana showed high while P. marginatus usually has 4broader teeth intraspeci 自c divergence (Fig. 1). This species (Haig，1956; Gore，(9 83). has awide range of distribution in the westem The second linea ge in clade F is that of Atlantic and varied host associations，so such P etroli sth es rosariensis，which was excluded alar ge genetic di vergence between specimens from the remainder of the P. galathinus ・com- could suggest the existence of distinct cryptic plex clade (Figs. 1，2) ，although it was previ- species. Detailed examination of recordsand ously considered pa口ofthi s complex (Werding， host associations from throughout the range of Phylogeny of westem Atlantic Porcellanidae 163 distribution might help in distinguishing popu- and Petrolisth es; see Haig，1960) .Additional lations within P. sayana. morphological characters of adults and larvae Phylogenetic relationships among the might help to improve the recognition of natu- Porcellanidae are more complex than previ- ral groups within those taxa. To make the dis- ouslythough t. The ability of these crabs to tinction possible，representative type specimens occupy awide variety of habitats ，coupled from around the world must be examined. In with alimited larval dispersal ，might have en- the molecular approach，additional analyses hanced diversification and regional endemism. should be focused on one or only afew c1 0sely Highly variable genera (i.e. ，M 旨galobrachium， related genera at atime to avoid ambiguities in Pachycheles ，and P etrolisthes) and species the alignments (see Stillman & Reeb，2001). (i .e. ，M egalobrachium soriatum，Neopisosoma Clearly，inclusion of the broadest possible angustifrons ，P etrolisthes galathinus，P .triden- specimen base anda larger set of genes are tatus ，and Porcellana sayana) are in need of required for definitive phylogenetic analyses. further morphological and molecular studies to Combining morphological and molecular better c1 arify their taxonomic status. phylogenies could be of great help in resolving The use of the 16S rRNA gene proved to the status of problematic taxa within the be very useful in assessing relationships among Porcellanidae and providing insights for porcelain crabs at various taxonomiclevel s， updating their c1 assification . such as family，gen us，and species.This marker has variable and conserved regions that make it Acknowledgements.-We thank Fernando suitab lefor aw ide range of applications，with Alvarez and Jose Luis Villalobos for loaning the constraints of being subject to aspecific materials from CNCR，a nd Carlos Lira，Juan secondary structure and sometimes having cop- Bolanos，Jose Cuesta，Fernando Mantelatto， ies of this gene translocated into the nuclear ge- Rafael Lemaitre，Joel Stake，Antonio Ordonez， nome (Schubart et al. ，2000c) .The use of 16S and Julieta Flor es for help in collecting sample s. and COI genes together may enhance resolution Rafael Robles helped in collecting samples and in molecular system atic studies of decapods， analyzing the data. The manuscript benefited and these analyses are currently underway with greatly from comment soffered by Raymond porcellanids .These genes are complementary Bauer，Joseph Neigel，Rafa el Lemaitre，and in that 16S has abroad range of applications Fernando Alvarez. Akira Asakura，Chr istoph er at different taxonomicle vels and COI has Tudge，and two other anonymous reviewers been useful to c1 arify relationships at terminal provided valuablesuggestions that significantly nodes (Schubart etal. ，2000c; Machordom improved this document. This study was & Macpherson，2004). Analyses of multiple supported by National Science Foundation genes，including nuclear genes，h ave also been grant nos. DEB-0315995 and EF-0 531603， used to enhance confidence in results and are U. S .Department of Energy grant no. DE- envisioned for future treatments. FG02-97ER12220，Department of the Interior A comprehensive approach to the study (Program USGS/AID-Nicaragua) cooperative of phylogenetic relationships should com- agreement award 00CRAG0009，and travel bine evidence from morphology (adult and awards from the Smithsonian In stitution CCRE larval characters) and molecular genetics (us ・ and SMSFP programs to D. L. Felder at the ing mitochondrial and nuclear genes). In the University of Louisiana at Lafayette，and Porcellanidae，substantial amounts of work awards from the Consejo de Investigacion de remain to be undertaken with both approaches laUniversidad de Oriente to G. Hernandez in Commonly used morphological character sets Venezuela. This paper is contribution number have sometimes proven insufficient to dif- 117 from the UL Laboratory for Crustacean ferentiate between even some closely re lated Research and contribution number 664 from taxa at the generic level (e.g. ，Pach ycheles the Smithsonian Marine Station ，F t. Pierce. 164 1. T. Rodri guez et al

LITERATURE CITED of Mexico and Central America (Crustacea: Decapoda: Anomura). Smithsonian Contributions Abele，L .G. ，& Kim，W .，19 89. The decapod to Zoology，363: 1-34 cru staceans of th ePanama Cana l. Smithsonian Gore，R. H. ，1983. The identit yof P etr o!i sthes Contributi on to Zoology，482 :1- 50. marginalus Stimpson，1859 ，and the description Ahyong，S. T.， & O 'Meally，0. ，2004. Phylogeny of of Petrolisthes dis simulatus n.sp. (Crustacea the Decapoda Reptantia: R eso luti onu sin gthree Decapoda: Porcellanidae) .Proceedings of the molecular loci and morphology. The Raffles Biological Societyof Washington，96: 89- 102 . Bulletin ofZoology，52: 673-693. Gore，R. H. ，1985. Molting and growth in decapod Avise，J. C. ，2004. Molecular Markers，Natural larvae .In :A. M. Wenner，(ed.) ，Larval Growth， Hi story，and Evolution . 684 pp. Sinauer Crustacean Issues 2.A. A. Balkema，Rotterdam / Associates，Inc .，Sunderland ，MA. Boston，pp. 67- 126. Baeza，J. A. ，& Thiel ，M. ，2000. Host use pattern Gore，R. H. ，& Abele，し G. ，1973. Three new and life history of Liopetro/i sth esmitra ，a crab spec ies of porcellanid crabs (Crustacea， assoc iate of the black sea urchin Tetrapygus A nomur a) from the Bay of Panama and adjacent niger. Journal of the Marine Biological Caribbean waters. Bulletin of Marine Sc ience， Association of the U nited Kingdom，80 :639 23: 559-573 645. Haig，J. ，1956 .The Galatheidea (Crustacea Baeza，J. A. ，Stotz ，W. ，& Thiel ，M. ，200 1. Life Anomura) of the Allan Hancock Atlantic history of Allopetrolisthes spinグrons，acrab Expedit ion wit ha review of the Porcellanidae associate ofthe sea anemone Phymactis c/ematis of the western North Atlantic .Allan Hancock Joumal of the Marine Biological Association of Atlantic Expedition ，8: 1--4 5. the Unit ed Kingdom，8 1: 69-76. Haig，J.， 1960.The Porcellanidae (Crustacea Chace，F. A. Jr. ，1 942. R eports on the scienti fic Anomura) of the eastem Pacitic. Allan Hancock results of the Atlant isexpeditions to the West Pacitic Expedition ，24: 1--4 40 Indi es ，under the joint ausp ic es of the University Haig，J. ，1 962.Papers from D r. Th.Mortensen ' of Havana and H arvard U niversity. The sPacific Expediti on 1914-191 6. LXXIX. anomuran Crustacea. 1. Galatheidea. Torreia， Porcellanid crabs from eastern and western (11 ): 1- 106 A merica.Videnskabelige Meddelelser 什a Dansk Crandall ，K. A. ，& Fitzpatrick ，J. E .J r. ，1996. nat urhi stor isk Forening iKjobenhaven ，124: 171 Cray fi sh m0 1 ecu 1ar syste matics: us ing a 19 2. combination of procedures to estimate Hall ，T .A. ，1999 .BioEdi t: auser-friendly biological phylogeny. Systematic Biol ogy，45: 1-26. sequence alignment ed it or and ana lys is Felder，D. し， 2001. Diversity and ecologica l program for Windows 95/98川丁目 Nucleic Acid s signi 自cance of de ep -burrowing m acrocru stacea ns Symposium Series ，41: 95-98. in coastal tropical waters of the Americas Harvey，A. W.，1 999. A review of the genus (Decapoda: Thalassinidea). Int ercie ncia ，26 :440 Claslotoechus Haig，with description sof a --4 49. new genusa nd two new species (Decapoda: Felder，D. L. ，& Stato n，J .L .，1 994. Genetic Anomura: Porcell ani dae). AmericanMuseum differentiation in trans-Floridian spec ie s Novitates ，3255: 1-32. complexes of Sesarma and Uca (Decapoda: Haug，G. H. ，& Tiedemann，R. ，1 998.Effect of the Brachyura). Journal of Crustacean Biology，14: formation of the Isthm usof Panama on Atlantic 191 -209 Ocean thermohaline circulation .Nature ，393: Fe lse nste in ，J. ，1 985. Confidence limit son 673-676 phylogenies: an approachusing the bootstrap. Hernandez，G. ，1999. Morfologia larvaria de Evoluti on，39: 783-791 cangrejos anomuros de la familia Porcellanidae Gore，R .H .，1972 .P etrolisthes armatus (Gibbes ， Hawort h，1825 (Crustacea: Decapoda)，con una 1850): The development under laboratory clave para las zoeas de los generos del Atlantico conditions of larvae from aP acific specimen occidenta l. Ciencia，7 :244 -257. (Decapoda，Porcellanidae). Crustaceana，22: 67- Huelsenbeck，J. P. ，& R onquist ，F. ，2001. 83 . MrBayes: Bayesian infer ences of phylogeny. Gore，R .H. ，19 77.Neopisosoma angustijトons Bioinformatics，17: 754-755. (Benedict，1 901): The complete larval Kocher，T. 0. ，Thomas ，W. K .，Meyer ，A. ，E dwards， de ve lopment under laboratoryconditions ，with S. v.， Paabo，S. ，Villablanca ，F. X. ，& Wilson， note son larvae of the relat ed genus Pachycheles A. C .，1989. Dynamics of mitochondrial (Decapoda，Anomura ，Porcellanidae) . DNA evoluti on in animals: Amplific ation an d Crustaceana，33: 284-300 sequencing with conserved primers. Proceedings Gore，R. H. ，1982. Porcellanid crabs 什om the coasts of the NationalAcademy of Sciences of the Phylogeny ofwestem Atlantic Porcellanidae 165

United States of America，86: 619ふる 200. Rodriguez，1. T. ，Hemandez ，G .，Magan ， 1.， Bolanos， Kraus，H. ，Navas ，G. R. ，& Werding，B. ，200 1. J. A .，& Felder，D. し， 2004. Larval development Feeding behavior in Parapetrolisthes tortugensis of Pachycheless erratus (Decapoda: Anomura: (Glassell ，1945) (Anomura，Porcellanidae) ，an Porcellanidae) under laboratory conditions， evidence for its special role in the relationship with notes on the larvae of the genus. Joumal of between the families Galatheidae and Crustacean Biology，24 :291 -308. Porcellanidae? IX Congreso Latinoamericano Rodriguez，R. ，Oliver ，1.し， Martin，A. ，& Medina， de Ciencias del Mar，San Andres Isla ，Colombia. J. R. ，1990. The general stochastic model of Abstract. nucleotide substitution. Journal of Theoretical Kropp，R. K. ，1981. Additional porcelain crab Biology，142: 485-50 1. feeding methods (Decapoda，Porcellanidae) . Scelzo，M. A.，1982. Observaciones sobre la Crustaceana，40 ・307-310. presencia y distribucion de crustaceos de la Lebour，M .Y. ，1943. The larvae of the genus familia Porcellanidae (Decapoda:Anomura) Porcellana (Crustacea Decapoda) and en aguas Yenezolanas yMar Caribe.Zoologia related forms. Journal of the Marine Biology Neotropical，2: 1129- 1146. Association of the United Kingdom，25: 721 Schubart，C. 0 .，Cuesta ，J .A .，& Felder，D .L. ，2002 . 737. Glyptograpsidae，a new brachyuran family 什om Machordom，A. ，& Macpherson，E .，2004. Rapid Central America: larval and adult morphology， radiation and cryptic speciation in squat lobsters and amolecular phylogeny of the Grapsoidea. of the genus Munida (Crustacea，Decapoda) Joumal ofCrustacean Biology，22: 28-44 and related genera in the South West Pacific: Schubart，C .0 .，Neigel ，J. E .，& Felder，D. し. ，2000. molecular and morphological evidence. The use of the 16S rRNA gene for phylogenetic Molecular Phylogenetics and Evolution，33: 259 and population studies of Crustacea. Crustacean -279. Issues，12: 817-830 Martin，J .W .，& Davis，G .E. ，200 1. An updated Staton，1.し ，& Felder，D. し， 1995.Genetic classification of the Recent Crustacea. Natural variation in populations of the ghost shrimp History Museum of Los Angeles County，Science g enus Callichirus (Crustacea: Decapoda: Series，39:1 - 124. Thalassinidea) in the westem Atlantic and Gulf

McLaughlin，P. A.，1983. A review of the of Mexico. Bulletin of Marine Science，56 目 523 phylogenetic position ofthe Lomidae (Crustacea: 536. Decapoda:Anomala) .Journal of Crustacean Stillman，J. H. ，& Reeb，C. A.，200 1. Molecular Biology，3:4 31 -437. phylogeny of eastern Pacific porcelain Morrison，C. し， & Cunningham，C. W.， crabs，genera Petrolisth esand Pachycheles， 1999. Dramatic mitochondrial DNA gene based on the m tDNA 16s rDNA sequence: rearrangements clarify relationships among phylogeographic and systematic implications anomuran crustaceans.Program and Abstracts， Molecular Phylogenetics and Evolution，19: 236 The Crustacean Society 1999 Summer Meeting， -245 Lafaye白e，Louisiana: 40. Swofford，D. L. ，2002. PAUP*: Phylogenetic Morrison，C し，Harvey，A. W.，Lavery ，S. ，Tieu ， Analysis Using Parsimony (*and other methods). K .，Huang ，Y.， & Cunningham，C. W .，2002 Yersion 4. Sinauer Associates，Inc. ，Sunderl and， M itochondrial gene rearrangements confirm MA. the parallel evolution of the crab-like form. Thompson，1. 0. ，Gibson ，T .1. ，Plewniak ，F. ， Proceedings of the Royal Society of London Jeanmougin，F. ，& Higgins，D .G. ，1997 .The Series B-Biological Sciences，269 (1489): 345 ClustalX-Windows interface: Flexible strate gies 350. for multiple sequence alignment aided by quality Palumbi，S .，Martin ，A. ，Romano ，S. ，McMillan ， analysis tools. Nucleic Acid Research，25: 4876 w. 0 .，Stice ，L .，& Grabowski，G. ，1991 .The -4882. Simple Fool 'sGuide to PCR. Yersion 2.0. 46 pp. Tudge，C. C .，1997. Phylogeny of the Anomura

University of Hawaii，Honolulu 目 (Decapoda，Crustacea): spermatozoa and Posada，0. ，& Crandall ，K .A .，199 8. MODELTEST: spermatophore morphological evidence. testing the model of DNA substitution . Contributions to Zoology，67: 125- 141. Bioinformatics，14: 817-818. Tudge，C. C. ，& Jamieson，B .G. M.，1996a. Rodriguez，1. T .，Hernandez ，G. ，& Felder，D . Spermatophore and spermatozoal morphology し， 2005. Review of the Western Atlantic in the Porcellanidae. 1. Aliaporcellana sulu ensis Porcellanidae (Crustacea:Decapoda :Anomura) and Pisidia longicornis (Decapoda: Anomura: with new records，sys tematic observations，and Porcellanidae) .Crustacean Research，25: 73-85 comments on biogeography. Caribbean Journal Tudge，C .C. ，& Jamieson，B .G .M. ，1996b. of Science，41 :544 -582. Spermatophore and spermatozoalmorpholog y 166 1. T. Rodriguez el a/

in the Porcell anidae. 1 1. The genera Petrolisthes Werding，B. ，& Hiller，A. ，2002. A new species and Polyonyx (Decapoda: Anomura). Joumal of of the Petrolisthes galathinus complex from Crustacean Biology，16: 535-546. the southern Caribbean sea ，with adi scu ssion Tudge，C. C. ，& Scheltinga ，D .M. ，2002. on the identity of P. galalhinus(Bo sc，1 802) Spermatozoal morphologyof the freshwater (Decapoda，Porcellanidae). Crustaceana ，7 5: 849 anomuran Aeg/a longiro stri Bond-Buckup & 857. Buckup，1994 (Crustacea: Decapoda: Aeglidae) Werding，B. ，& Muller，H. G. ，1990. Larval from South America. Proceedings of the development of Neopisosoma neglecfum Biological Society of Washington，115: 118 - 128 Werding，1986 (Decapoda: Anomura: Werding，B. ，1977 .Los porcelanidos (Crustacea: Porcellanidae) under laboratory conditi ons Anomura:Porcellanidae) de la re gion de Santa Hel golander Meeresuntersuchungen，44: 363- Marta，Colombia. Analesdel Instituto de 374. lnvestigaciones Marinas de Punta de Betin ，9 ・ Werding，B. ，Hiller ，A. ，& Misof，B. ，2001 .Evidence 173 -214 of paraphyly in the neotropical porcellanid Werding，B. ，1978. Los porcelanidos (Crustacea: genus Neopisosoma (Crustacea: Anomura: Anomura: Porcellanidae) de la region de Acandi Porcellanidae) based on molecular characters. (Golfo de Uraba)，con algunos encuentros Hydrobiologia，449: 105- 110. nuevos de la region de Santa Marta (Colombia) Werding，B .，Hiller ，A. ，& Lemaitre，R. ，2003. Anales del Instituto de InvestigacionesMarinas Geographic and depth distributional pattern s de Punta de Betin ，10 :213 -221 of western Atlantic Porcellanid ae(Cru stace a: Werding，B. ，1982. Porcellanid crab sof the lslas Decapoda: Anomura)，with an updated li stof del Rosario，Caribbean coastof Colombia， specie s.Memoirs of Museum Victoria ，60 ・79 with adescription of Petroli slh esrosariensis 85 new species (Crustac 回目 Anomura). Bulletin of Young，A. M .，Torres ，C. ，Mack ，J. E .，& Marine Science，32: 439-447. Cunningham，C. w.， 2002.Morphological and Werding，B .，1 983a. Der P etroli sthesgalathinus- genetic evidence for vicariance and refugium Artenkomplex im karibi schen Raum mit in Atlantic and Gulfof Mexico populations of Beschreibung von P. caribensisn. sp .und the hermit crab Pagurus longicarpus. Marine P. columbiensis n.sp. (Crustacea :Anomura: Biology，140: 1059ー 1066 Porcellanidae) .Senckenbergiana biologica，63: 407-4 18 Addresses: *ITR DLF Department of Biology Werding，B .，1983b. Kommensalische Porzel laniden ， ， ， aus der Karibik (Decapoda，Anomura). Universityof Loui siana at Lafayette，P.O. Box Crustaceana，45 ・1- 14. 42451 ，Lafayette ，Louisiana 70504，U.S.A. ; Werding，B. ，1986. Die GattungNeo p is osoma e-mails:( ITR)trodriguez99@ hotmai l. com. (DLF) Haig 1960 im tropischen Westatlantik mit der ， ， dlf4517@ louisiana.edu. GH，Universidad de Oriente， Beschreibung von Neopisosoma neglectum spec .nov. und Neopisosoma orienlale spec. Apartado Postal 074，La Asuncion，Isla Margarita， nov. (Crustacea: Anomura: Porce llanidae). Venezuela; e-mail: gonzalo@ c-com.net. ve Zoologische Mededelingen ，Leiden ，1 6: 159- *Author for co 汀 espondence 179.