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Inverlchnih'SvsleDUilics, 2002. J6, 839 847

Molecular phylogeny of the mud lobsters and mud (Crustacea : : Thalassinidea) using nuclear 18S rDNA and mitochondrial 16S rDNA

C. C. Tiidge^' and C. W. Cunningham B

•^Biolog)' Department, American Universit)', 4400 Massachusetts Ave. NW, Washington, DC 20016-8007, USA and Department of Systematic Biology, National Museum of Natural History, Smithsonian Institution, Washington, DC 20560-0163, USA. '^Department of Zoology, Duke University, Durham, NC 27708-0325, USA. '^To whom correspondance should be addressed. Email: tudge.christopher(a)nmnh.si.edu

Abstract. Partial sequences of the 18S nuclear and I6S mitochondrial ribosomal genes were obtained for 14 species of thalassinidean (families Callianassidae, Laomediidae, Strahlaxiidae, Thalassinidae and Upogebiidae) and a further six species in related decapod infraorders (families Aeglidae, Astacidae, Lithodidae, Palinuridae, Raninidae and Scyllaridae). Maximum-likelihood and Bayesian analyses show equivocal support for the monophyly of the Thalassinidea, but show strong support for division of the infraorder into two major clades. This dichotomy separates representatives in the Upogebiidae, Laomediidae and Thalassinidae from those in the Strahlaxiidae and Callianassidae. The Laomediidae is shown to be paraphyletic, with the thalassinid species, sqiiamifera, being placed on a branch between Axiaiuissa and a clade comprising ./¿¿v«/ and Laomeclia. the three current laomediid genera. Fora monophyletic Laomediidae, the family Axianassidae should be resurrected for the genus Axianassa.

Introduction relationships among four thalassinidean families and the (Fig. \B). A period of 56 years elapsed before any The decapod infraorder Thalassinidea is a group of cryptic, further publications on the phylogenetic relationships within marine, burrowing shrimp-like or lobster-like this obscure, but taxonomically large, infraorder emerged. that occur worldwide (with the exception of the coldest polar Poore ( 1994: 120) published a comprehensive revision ofthe waters) in mostly shallow (<200 m) benthie habitats. Most members ofthe Thalassinidea based on morphology, including species form complex burrow systems in soft sand or mud a new classification scheme, keys to families and genera and environments, but several taxa live in stony or coral-rubble a phylogenetic tree (reproduced in Fig. ICto the family level areas and some even excavate burrows in living coral and only). This tree was the first cladistic analysis of the infraorder sponge colonies (Dworschak 2000). There are currently as a whole and separated the 22 selected genera into the 528 species in 84 genera spread across 11 recognised currently recognised 11 families and three superfamilies families of the three superfamilies: Axioidca, Thalassinoidea (despite showing a basal dichotomy with only two major and Callianassoidea (Poore 1994; Dworschak 2000; clades). He also proposed a monophyletic origin for the P. Dworschak, personal communication). infraorder, with the Anomura being the closest sister-group. Borradaile (1903; 551), in his seminal work on the Several papers have been published employing cladistic classification of the Thalassinidea, presented a 'genealogical analyses of relationships among or within genera of certain tree' of proposed relationships among 12 known genera in the thalassinidean families. These investigations cover the four families recognised at that time. His tree (reproduced in families Axiidae and Calocarididae (Kensley 1989), Fig. \A with current family designations) separates these Callianidcidae (Kensley and Fleard 1991), Callianassidae genera into five major clades corresponding with the femilies (Staton and Felder 1995; Staton et al. 2000; Tudge et al. Axiidae (Axiop.sis, Axiiis, Ciilocaris, Scytoleptits), 2000) and Ctenochelidae (Tudge et al. 2000). Laomediidae {Jaxea, Luomedia), Thalassinidae ( Thalassina) In some cases, thalassinidean representatives have been and the two subfamilies, Callianassinae (Calliunassa, used as either ingroup or outgroup taxa for larger cladistic Calliunidea, Glyplurus) and Upogebiinae {Gehicula, analyses of various Decapoda; these include studies using Upogehiu), of the Callianassidae. Later, Gurney (1938: 343) morphological characters by Martin and Abele (1986), used larval morphology to present an intuitive tree of Scholtz and Richter (1995) and Tudge (1997). Several

;CSIRO2002 10.1071/1502012 1445-5226/02/0608.^9 840 C. C. Tudgc and C. W. Cunningham

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(C) Fig. 1. (//) (iencalogicdl relalioiis between families in llic Tliaiassinidca according to Borradailc (modilied from Borradailc 1903). (B) Intuitive tree of relationships between some thalassinideans based on larval characters (inodiried from Ourncy IWS). (t) Cladograin of phylogenetic relationships between 1 1 families of the Thalassinidea based on morphological characters (modified from Poore 1994). Note: the trees of both Borradailc and Poore have been modilied to show ta.xonoraic relationships at the family-level or higher. molecular phylogenetic analyses of various decapod Spears el al. 1992; Crandall el al. 2000; Morrison el al. 2002; taxonomic groups have also included thalassinidean taxa for Pérez-Losada el al. 2002). Similarly, the mitochondrial 16S comparison with anomurans (Spears and Abele 1988; Pérez- rDNA gene has been regularly used to investigate decapod Losada el al. 2002), astacideans (Crandall el al. 2000) and relationships (Cunningham ct al. 1992; Crandall and -like decapods (Morrison e! al. 2002). Also, see the Fitzpatrick 1996; Tam el al. 1996; Ovenden ei al. 1997; synopsis of recent research into decapod phylogenelics by Kitaura et al. 1998; Tam and Kornfield 1998; Crandall el al. Schratn (2001) for a review of the different approaches and 1999, 2000; Schubart el al. 2000, 2001; Duffy el al. 2000; dala sets being applied to the field. Morrison el ul. 2002). Different regions of ribosomal genes The contribution of the small-subunil I8S ribosomal evolve at varying rales, making these useful molecular (r)DNA nuclear gene to phylogeny is well known markers across a broad taxonomic spectrum. and has been useful in investigating relationships across a This paper presents the results of a phylogenetic analysis wide variety of groups (Spears and Abele 1988, 1997, 1998; of partial sequences of mitochondrial I6S rDNA and nuclear Abele eral. 1989, 1992; Kiin and Abele 1990; Abele 1991; 18S rDNA from 14 thalassinideans in five families Molecular phylogeny of Ihe Thalassinidea 841

(Callianassidae, Laomediidae, Strahlaxiidae, Thalassinidae Materials and methods and Upogebiidae). Six otlier decapod taxa from tlie Specimen collection Astacidea, Palinura, Bracliyura and Anomura are included in ^ith the exception ofthc Raninoides specimen from 1991, for the genetic analysis were mostly collccled over a 5-ycar period tfie analysis as outgroups. Similarities and differences to (1995-1999) from a variety of subtidal and intertidal locations previous evolutional^ trees of tlie Tlialassinidea, most worldwide(Table l),btit with''an"emphasis on coastal Australia and the notably those of Borradaile (1903), Gurney (1938), Poore United States. The intertidal specimens were collected by'yabby'pump, (1994) (Figs 1/l-C) and Tudge et al. (2000), are discussed. digging or by hand and the subtidal specimens were trawled or dredged.

Table 1. Specimens, collection, voucher and GcnBank-scquence information Higher-level classification from Martin and Davis (2001). Abbreviations: AMNH, American Museum of Natural History (New York); MV, Museum Victoria; Qld, Queensland; USA, United States of America; USNM, National Museimi of Natural History (Washington DC); Vic. Victoria.

Astacidea Cambaridae Pmcamhanix cUirkii (Girard, 1852). Louisiana, USA, .Ian. 1998. Voucher: AMNH 1 7874. GenBank sequences: AF436Ü40 ( I6S), AF436001 (18S) Palinura Palinuridae Pumilims argus (Latreille, 1804). GenBank sequences: AF337966 ( 16S), U19182 ( 1SS) Seyllaridae Tlienus orientalis (Lund. 1793). Gulf of Carpentaria. Qld, Australia, 1997. Voucher: not available Brachyura Raninidae Raninoides louisianensis Rathbun, 1933. Gulf of Mexico, USA, 1991. Voucher: T. Spears personal collection. GenBank sequences: AF436()44 (16S), AF436005 (18S) Anomura Aeglidae Aegia urnguayuna Schmitt, 1942. San Antonio de Areco, Buenos Aires, Argentina, 10 Mar. 1996. Voucher: C. Cunningham personal collection. GenBank sequences: AF436051 (16S), AF436012 (18S). Lithodidae Cryplolilhodcs typiciis Brandt, 1849. British Columbia, Canada. Voucher: S. Zaklan personal collection. GenBank sequences: AF425325 (16S), AF436019(18S) Thalassinidea Superfamily Thalassinoidea Thalassinidac Thalu.ssimi sí/nanüjera de Man. 1915. Town of 1770, Qld. Australia, 29 Dec. 1997. Voucher: MV .141662 Superfamily Callianassoidea Callianassidae BiJJarins arenosns (Poorc. 1975). Dunwieh. Qld. Australia, 21 May 1997. Voucher: MV .140669 Biffaiius delieutulus Rodrigues & Manning. 1992. Fort Pierce, Florida. USA, 9 .lun. 1999. Voucher: USNM 309754 Callianassüßlholi A. Milne Edwards. 1878. Otago Harbour, New Zealand. 14 Nov. 1997. Voucher: MV J44818 Callichirns major {Say, 1818). [sles Dernières, Louisiana, USA. 27 Mar. 1996. Voucher: M V J39()44. GenBank sequences: y\F436041 ( 16S), AF436002(18S) Neocallichirus ralhhnnae (Schmitt, 1935). Fort Pierce, Florida, USA, 8 Jun. 1999. Voucher: USNM 309751 Neolrvpaea californiensis (Dana, 1854). Marina del Rey, California. USA, 20 .lun. 1997. Voucher: A. Harvey personal collection. GenBank sequences: AF436042(16S), AF436003 (18S) ^ - - Sergio mericeae Manning & Felder, 1995, Fort Pierce, Florida. USA, 8 .lun. 1999. Voucher: USNM 309755 ' - - ^ ^'- Laomcdiidac Axianassu aiislralis Rodrigues & Shimizu, 1992. Sao Sebastiáo. Sao Paulo, Brazil, 20 .ltd. 1997. Voucher: MV J44613 Jaxea nocturna Nardo, 1847. Loch Sween, Argyll, Scotland, 10 Nov. 1996. Voucher: MV .139045. GenBank sequences: AF436046 (16S). AF436006(18S) Lanmedia heaiyi Yaldwyn & Wear, 1970. Western Port Bay, Vic, Australia, 16 Apr. 1997, Voucher: MV J40697 Upogebiidae Gebiacantha plantae (Sakai, 1982). Yorkc Island, Qld. Australia, 26 Feb. 1998. Voucher: MV ,144914 Upogebia affinis (Say, 1818). Fort Pierce. Florida, USA, 15 Nov. 1995. Voucher: MV .140668. GenBank sequences: AF436047 (I6S). AF436007(18S) Supeiiamily Axioidea Strahlaxiidae Neaxius glyptocercus von Martens, 1868. Dunwieh, Qld., Australia, 2 I May 1997. Voucher: MV J39643 842 C. C. Tudiie and C. W. Cunniiiüliam

Molecular proUHob 2058 bp. Our final alignments are available from the authors Tissue samples were preserved in ')5'!^p elhanol. DNA vv;is isolated from (CWC). An ILD test (Farris el al. 1995) showed no fresh, frozen or ethanol-preservcd specimens by grindiny, small significant incongruence (P > 0.05) and the two genes were fragments of muscle tissue in a buffer (0.1 M EDTA.O.Ol viTrispH 7,5, analysed separately and together. 1 % SDS, Pakimbi cl ul. 1991 ), followed by plienol-chloroform-isoamyl For both genes, the best-fit model found using ModeiTest alcobol extraction and precipitation with 7.5 M ammonium acetate and cold isopropanol as described by Palumbi el ai (1991). Partial under maximum-likelihood was GTR + gainma + inv. sequences of mitochondrial I6S rDNA were amplified using rDNA Bootstrap consensus trees for both genes, analysed separately primers (LR-N-13398, alias 16Sar. and LR-J-12887, alias 16 Sbr. for the maximum-likel ihood bootstrap and Bayesian analyses, Simon cl al. 1994). Most of the nuclear ISS rDNA gene was amplified are presented in Fig. 2. There is weak support in the IBS using ISE-F (5'-CTGGTTGATCCT(iCCAGT-3') and 1.SSR3 (5'- analysis for a titonophylelic Thalassinidea (70% Bayes, 42% TAATGATCCTTCCGCAGGTT-3'). The amplification and sequencing of the two genes ( I6S and I8.S) was identical (except where indicated) bootstrap). Although Bayesian analysis of the I6S gene and entailed the following regime. Polynicra.se chain reaction (PCR| showed rather strong support fora paraphylctic Thalassinidea was carried out on a Perkin Elmer GeneAmp PCR System 9600 (85%), there is little ML bootstrap support fora paraphylctic (www.pecorporation.com) using 50 (.iL reactions consisting of 1 jiL Thalassinidea (33%), The conflict betweeti the 18S and 16S template DNA, 3 fiL primers. 4.5 (-iL lO^ reaction bufler. 5 |,tL Taq genes is not strong (also reflected by a non-significant ILD polymerasc, 4 ^iL deoxynucleoside triphosphates, 5 |aL MgCl, and 24,5 |.iL water. Amplification involved 20 sec denaturation al 94°C. 1.5 min test off > 0.05) and they were combined for ftirther analysis. ani\ealing at 50°C (40°C for 16S) and 2.5 mm of extension at 72°C for The ML analysis found a tree (Fig. 3) with Ln likelihood 35 cycles. The Pt'R product (3 .uL) was visualised, via electrophorcsis. = -8184,94 that weakly supports a paraphylctic on a riii agarose gel using efhidium bromide staining. The remaining Thalassinidea (76% Bayes, 56% bootstrap), A monophylelic product (45 ^iL) was purified using the Wizard'' PCR Preps Thalassinidea could not be rejected by an SH test. (www.promega.com) DNA Purification System. Applied Bio.systems. Inc. (ABI) (www.appliedbiosystems.eoni) Prism BigDyc Terminator Discussion Cycle Sequencing Ready Kits and an ABl Prism 3700 DNA Analyzer were used for sequencing. Sequences are available from the authors In these analyses, monophyly of the Thalassinidea was (CWC). equivocal, being weakly supported by the 18S dataset only Scc/iience aliginiieiit ami phylogennlic iiinilysis (Fig, 2, left side) and not by the 16S dataset (Fig, 2, right side) Sequences were aligned using Clustal X (Thompson e/ al. 1994, 1997) or the combined analysis (Fig, 3). Poore(1994; Fig, 1 C) found with gap insertion and extension costs at 10 and 5. respectively, and a monophylelic Thalassinidea based on two morphological regions of uncertain honiology removed before phylogenetic analysis. synapomorphies (reduction ofpleurobranchs on gills to seven Maximum-likelihood (ML) analyses using heuristic searches and TBR or less and presence of a setose lower margin on the propodus branch swapping were applied to the aligned sequences using PAUP* 4.0 (Swofford 2001), For both maximum-likelihood and Bayesian and carpus ofpereopod 2), but a molecularstudy by Morrison analyses, we used the general-time-reversible (GTR) model to estimate et al. (2002) foutid strong support for non-monophyly of the the proportion of invariant sites and the alpha parameter of the gamma Thalassinidea (89% ML bootstrap support). The latter study distribution from the data (the best-fit model using ModeiTest, Posada included more sequences (16S and 18S, as well as partial and Crandall 199S). Iiicongruencc testing was carried out under the sequences of mitochondrial COII and nuclear 28S ribosomal parsimony criterion using the incongruence length difference (ILD) test DNA), but fewer thalassinidean laxa than the present study. (Farris cl al. 1995). as implemented in PAUP* 4.0. The Bayesian analyses were performed using the program It is possible that the more rapidly evolving sequences used MRBAYFS 1.11 (I luel.scnbeck and Ronquisl 2001 ). MRBAYFS uses a in the Morrison el al. (2002) study give a tnore reliable metropolis-coupled Markov chain Monte Carlo (MCMCMC) algorithm indication of relationships than ju.st the two genes used in the to sample from the posterior tlislribution. Bach Markov chain was present study. started from a random tree and run for 5 million cycles. The chain was .sampled every 500 cycles in order to minimi.se the size of the output Between the Bayesian and the ML analyses there is no clear files and to ensure that Ihcse samples were independent. Four chains candidate for a sister-group to the Thalassinidea as a whole were run simultaneously with a 'temperature' of 0.2. The initial 40% of (Figs 2, 3), Interestingly though, the Bayesian analysis found cycles were discarded as burn-in and convergence was checked by strong (90"/i) support for a monophyletic sister-group to soine ensuring that the likelihood values of sampled trees had approached a taxa in Ihe Thalassinidea, including the spiny and slipper level distribution. Each analysis was performed twice to ensure that lobsters {Panulirus and Theinis), the anomurans (Aegia and convergence was rcpeatablc. Tests of alternative topologies were performed using the Cryptulilhotles) and the brachyuran (Raninoide.s). Although Shimodaira-Hascgawa (Sll) lest (Shiniodaira and Hasegawa 1999), this is intriguing, the weak support for this clade in which docs not require that the hypotheses be designated a priori. boot.slrapping (59%) suggests that further data may be Probability values were calculated in PAUP* 4.0 using 1000 RELL necessary to test this hypothesis. replicates to obtain a distribution. Within the Thalassinidea, we found two strongly supported Results major cladcs (Figs 2, 3), The first clade includes a After removing regions of uncertain alignment, our aligned monophyletic Callianassidae as the sister-group to a sequences consisted of 1731 base pairs (bp) of nuclear ]8S representative in the axioid family, Strahlaxiidae (Nea.xiiis rDNA and 327 bp of initocliondrial 16S rDNA for a total of glyplocerciis). Within the subfainily Callianassinae, the genus Molecular pliylogeny of ihe Thalassinidca Ü43

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Fig. 2. A combined figure showing the opposed consensus trees for both Bayesian and maximum-likelihood analyses of the I8S data set (left side) and the I6S data set (right side). Percentage support for Bayesian posterior probabilities (upper) and rnaximum-likelihood bootstrap values (lower) are given for each node. Best-fit model = GTR + gamma + inv.

BiffariiLs is paraphyletic, with representatives of the genus monophyletic Callianassidae in this molecular analysis (Figs apparently grouping according to biogeography, uniting the 2,3) is supported by at least 10 morphological characters (see American taxa Biffarim delicatulus and Neotrypaeu family diagnosis in Poore 1994). In addition to Tudge et al. californiensis, and uniting the antipodean taxa, Biffarius (2000), a recent synopsis of the family Callianassidae has been arenosus and Callianassufilholi. These associations, although provided by Sakai (1999). His paper indicates a polyphyletic at first appearing enigmatic, should be viewed in light of family, extensively synonymises members of the comments by Tudge et al. (2000) that generic relationships Callianassoidea and has been considered of limited value in within the family Callianassidae should be treated with some helping to elucidate the relationships in this group of caution pending a re-diagnosis of the inclusive taxa, the thalassinideans (Tudge et al. 2000; Poore 2000). subfamily Callianassinae and the nebulous genus Callianassa The large clade constituted by the Callianassidae and in particular. The apparent biogeographic associations Strahlaxiidae in this molecular analysis (Figs 2, 3) is also between the four species mentioned above are contradictory supported by six morphological synapomorphies. Three of to those proposed for the same taxa in the morphological these synapomorphies can be found in the analysis of Poore analysis (Tudge el al. 2000), where the two Biffarius species ( 1994), but are also shared by representatives in several other are included in a monophyletic clade (only 59% supported) thalassinoid families (Callianideidae, Ctenochelidae, and C. ßlholi and N. culiforniensis are in another smaller Micheleidae and Thomassiniidae) for which tissue was not monophyletic clade (100% supported). The same available for this molecular analysis. These characters are (/) morphological analysis by Tudge ct al. (2000) grouped these the presence of dense tufts of setae on abdominal somites 3-5 four cal I ianassine taxa into a monophyletic clade and Indicated (2) the absence of the appendix interna on male plcopod 1 and a comparable lack of resolution between this clade and the (J) a similar absence on male pleopod 2. Three additional callichirine taxa, Callichirus, Sergio and Neocallichirus. The morphological synapomorphies were found to support the 844 C. C. Tutlge and C. \V. Ciinninuliam

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Fig. 3. A Cüiiipositc of the consensus trees obtained (rom the Bayesian analysis and the single maximum-likeliliood tree for tlic combined 16S and I8S data sets. Percentage support for Bayesian posterior probabilities (upper) and maximuni-lilceliliood bootstrap values (lower) arc given lor each node. Bcsl-fit model = GTR + gamma - inv. Some higher taxonomic categories are provided on specific cladcs and the six outgrotip taxa are indicated. Decapod images are modified from (top to bottom): Hobbs I9SI: Rathbun l')37; Jara and Palacios 1W9; Bliss 1982; Holthuis 1991 ; Rodrigues and Shimizii 1992; Sakai 1992: Yaldwyn and Wear 1970; Kenslcy et al 2000: Felder and Felgcnhauer 1993; Hart 1982; Holthuis 1991. Note: images arc not necessarily the same species as used in the analysis, but arc representative of the genus, or a closely related genus. association of the Callianassidae with the Strahlaxiidae in an inclusion of Thalussina. This latter, alternative hypothesis, unpublished analysis by the senior author, which did not use although lacking any convincing morphological support (see the exact corresponding taxa. These characters are (4) a below), could not be rejected by an SH test of the molecular chelate pereopod 2 with the dactylus as long as the fixed finger data. The paraphyly of the laomediids, suggested in the ( .5) the absence of spiniform setae on the dactylus of pereopods present analysis, resurrects an interesting issue on the 3 and 4 and (ó) a row of lateral phirnose setae on abdominal validity of the monolypic thalassinidean family, somite 2. Axianassidac Schmitt, 1924. Ken.sley and Heard (1990) The second major clade in Fig. 2 (left side only) and Fig. 3 succinctly summarised the history of the debate on whether includes three genera currently in the Laomediidae (Jaxca, the Axianassidac is a valid family in the Thalassinidea, and Laomcdia, Axianassii), as well as representatives of the listed the supporters of the Axianassidac (Gumey 1938; Thalassinidae and Upogebiidae. Within this clade, the Wear and Yaldwyn 1966; Goy and Provenzano 1979; Poore placement of the thalassinid, Thalussinu squamiferu, makes and Griffin 1979) and those who believe the genus the Laomediidae paraphyletic, or monophyletic with the A.xianas.sa is one of five in the Laomediidae (de Man 1928; .7*^

Molecular phylogcny of Clie Tlialassinidca -- 845

Baiss 1957; Le Loeuft'and Inles 1974; Ngoc-Ho 1981). Our the I8S gene sequence data (Fig. 2, left side) and data support retention of the family Axianassidae for the unsupported by the I6S data (Fig. 2, right side) and the genus Axianussu. with Ja.xca and l.aomediu remaining in the combined analysis of both genes (Fig. 3). Borradaile ( 1903), Laomediidae. As well as the molecular evidence for a valid in his paper on thalassinidean classification, did not Axianassidae presented here (Figs 2, 3), we have identified implicitly state that the group is monophyletic. but inferred 10 morphological characters (six of them apomorphics) in this in his intuitive genealogical tree (Fig. \A) and Gurney which Axiunassa differs from Jaxeu and Laomedia. These (1938; Fig. Iß) advocates paraphyly for the Thalassinidea he apomorphic characters are: (/) the linea thalassinica studied. Although a paraphyletic Thalassinidea has some displaced dorsally (possible autapomorphy); (2) absence of support in the present molecular analysis, the question of lateral lobes on abdominal somite 1 ; (i) linear epipods on the monophyly for this enigmatic group will require more gills; (4) reduction (or absence) of an exopod on maxiUiped analysis of both morphological and molecular data sets to 3; (5) unequal chelae on percopod I; and (6) dense tufts of resolve. lateral setae on abdominal somites 3-5. A dichotomy within the Thalassinidea was previously The association of the Thalassinidae and the Laomediidae suggested by Poore (1994) based on morphological data, but (the 76% ML bootstrap supported node with Thalassina, neither of his major clades corresponds to ours. In contrast, Jaxea, and Laomedia in Fig. 3) is interesting, b\ú lacles the dichotomy described by Gurney (1938; Fig. 1B) based on strong morphological support. In fact, only two larval characteristics is reminiscent of the relationships seen synapomorphies(?), the posterior margin of the carapace in our 16S tree and combined gene tree (Figs 2 (right side), with strong lateral lobes and both anterior and posterior teeth 3). This closer relationship of the thalassinidean families on the mandibular incisor, can be gleaned from the eladistic Laomediidae and Upogebiidae with the Anomura in analysis by Poore (1994). The former is shared with Gurney s proposed classification (Fig. IS) is only weakly representatives of the Axioidca, whereas the latter character supported (under both Bayesian and ML analyses) here and is shared with members of the Thomassiniidae (once again the anomuran representatives are always part of a larger taxa missing from the current molecular analysis). Some decapod sister-clade (Figs 2, 3). Since our analysis only support, however, for this laomediid-thalassinid association, includes half of the families in the Thalassinidea, ftirther is provided by studies of larval characters (Sankolli and sampling will be necessary to determine the composition of Shenoy 1979) and gill-cleaning structures and mechanisms these major clades, which appear with regularity in eladistic (Batang and Suzuki 1999; Batang cl al. 2001 ). Although the analyses of this group. latter authors found gill-cleaning characters to be The three monophyletic superfamilics (Axioidea, conservative at the family level, there is still debate over their Thalassinoidea and Callianassoidea) indicated in the utility in phylogenetic studies (Poore 1994; Suzuki and classification of Poore (1994) are not maintained as McLay 1998). monophyletic entities in our analysis. In fact, in Poore's No unique morphological synapomorphics support the phylogeny (Fig. 1 C), these superfamilics arc not adequately major monophyletic clade containing the Upogebiidae, represented by monophyletic clades at the same level cither. Axianassidae, Thalassinidae and Laomediidae, seen in This discrepancy was noted by Martin and Davis (2001), Figs 2, 3, even though there is strong molecular support in who followed Poore's revision in their recent work. Until both Bayesian and ML analyses. In the morphological better taxonomic congruence is achieved, and more of the analysis of Poore ( 1994; Fig. 1C), Upr)i;ehia, Thalassina and 11 families are represented in molecular analyses such as the Laomedia do not form a distinct cladc, but three present study, direct comparisons will remain challenging. morphological characters variously ally these three taxa, which are part of a monophyletic cladc (with other taxa) in Acknowledgments the current molecular analysis. These are (/) the rostrum This project was initiated when the senior author (CCT) was augmented with ridges (with the exception of the an Australian Research Council Postdoctoral Research Laomediidae), (2) a cylindrical carpus and propodus on Fellow (1996-1999) at Museum Victoria, Melbourne, pereopod 1 (a symplesiomorphy shared with some axioids Australia, and the authors acknowledge the assistance of and some outgroup representatives) and (i) the loss or Gary Poore, Les Christidis, .lanette Norman and Megan reduction of the male first pleopod (except in Thalassinidae). Osborne from this institution for project support, access to This last morphological character is a possible laboratory facilities (CCT) and training in molecular synapomorphy shared with some callianassids (Poore 1994) techniques (CCT). The authors thank Cheryl Morrison, Bo or a symplesiomorphy also shared with some members of the Kong and Todd Oakley for technical support within the outgroup. Department of Zoology at Duke University. Also, Kristian In summary, the monophyly of the Thalassinidea Fauchald and Rafael Lemaitre (Department of Systematic suggested from a eladistic analysis of morphological Biology, National Museum of Natural History, Smithsonian characters (Poore 1994; Fig. I C) is only weakly supported by Institution) are thanked for supporting the Research 846 C C Tudízo and C. W. Cunniuaham

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