Pseudojuloides labyrinthus, a new labrid ish (Teleostei: Labridae) from the western Indian Ocean BENJAMIN C. VICTOR Ocean Science Foundation, 4051 Glenwood, Irvine, CA 92604, USA and Guy Harvey Research Institute, Nova Southeastern University, 8000 North Ocean Drive, Dania Beach, FL 33004, USA E-mail: ben@coralreefish.com JASON M.B. EDWARD Greenwich Aquaria, 1064 E Putnam Ave, Riverside, CT 06878, USA E-mail: jason@greenwichaquaria.com Abstract The new labrid ish species, Pseudojuloides labyrinthus n. sp., is described from three specimens obtained via the aquarium trade from Kenya, in the western Indian Ocean. The species is similar in appearance to other IndoPaciic Pseudojuloides in the P. severnsi complex, distinguished mainly by the markings of the terminal-phase male, which includes a maze of lines on the head and three thicker blue stripes along the rear body. Despite the similarity in appearance, the new species is 9.66% divergent in the sequence of the mtDNA barcode marker COI (minimum interspeciic divergence, pairwise; 10.54% K2P distance) from its nearest relative, P. edwardi, also found in Kenya. A neighbor-joining tree and genetic distance matrix is presented for 12 of the 14 known species in the genus Pseudojuloides. Key words: coral reef ishes, ichthyology, new species, taxonomy, systematics, Kenya, Africa, DNA barcoding. Citation: Victor, B.C. & Edward, J.M.B. (2016) Pseudojuloides labyrinthus, a new labrid ish (Teleostei: Labridae) from the western Indian Ocean. Journal of the Ocean Science Foundation, 21, 58–70. doi: http://dx.doi.org/10.5281/zenodo.55594 urn:lsid:zoobank.org:pub:B9C0D2C5-DBBD-42E9-BF57-3D7B52491C04 Date of publication of this version of record: 15 June, 2016 Introduction The labrid genus Pseudojuloides Fowler was revised by Randall & Randall (1981), who recognized eight species in the genus, ive of which were new. Since then, ive additional new species have been described from various locations in the Indo-Paciic Ocean, including P. kaleidos by Kuiter & Randall (1995) from the Maldives and Indonesia; P. severnsi by Bellwood & Randall (2000), from the Maldives to the W. Paciic; P. edwardi and 58 Journal of the Ocean Science Foundation, 21, 58–70 (2016) P. polackorum from the southwest Indian Ocean (Victor & Randall 2014, Connell et al. 2015); and the deep-reef species P. zeus from Micronesia (Victor & Edward 2015). The genus comprises a set of small fast-swimming wrasses, typically found on deeper slopes and in habitats dominated by rubble rather than live coral. They are distinguished morphologically by having chisel-like incisiform side teeth (unusual among the labrids) and fusiform bodies with relatively large scales. Terminal-phase (TP) males are large, brightly colored individuals, while smaller initial-phase (IP) ish, usually females, are reddish orange. We describe here a new species from the African coast in the western Indian Ocean, the fourth species of the genus from that location, and compare its barcode mtDNA COI sequence to 11 of the 13 described species in the genus (all except for P. argyreogaster and P. erythrops). Materials and Methods Specimens have been examined from the Bernice P. Bishop Museum, Honolulu (BPBM). In addition, ethanolpreserved specimens of comparison species were collected for DNA sequencing from Bali (Indonesia), Moorea and the Marquesas Islands (French Polynesia), Cook Islands, New Caledonia, and Hawai‘i in the Paciic Ocean, and South Africa in the Indian Ocean, as well as obtained via the aquarium trade from the Philippines, Indonesia, Vanuatu, and Micronesia in the Paciic Ocean and Kenya and Mauritius in the Indian Ocean (Appendix 1). DNA extractions were performed with the NucleoSpin96 (Machery-Nagel) kit according to manufacturer speciications under automation with a Biomek NX liquid-handling station (Beckman-Coulter) equipped with a iltration manifold. A 652-bp segment was ampliied from the 5′ region of the mitochondrial COI gene using a variety of primers (Ivanova et al. 2007). PCR ampliications were performed in 12.5 µl volume including 6.25 µl of 10% trehalose, 2 µl of ultra pure water, 1.25 µl of 10× PCR buffer (10mM KCl, 10mM (NH4)2SO4, 20mM TrisHCl (pH8.8), 2mM MgSO4, 0.1% Triton X-100), 0.625 µl of MgCl2 (50mM), 0.125 µl of each primer (0.01mM), 0.0625 µl of each dNTP (10mM), 0.0625 µl of Taq DNA polymerase (New England Biolabs), and 2 µl of template DNA. The PCR conditions consisted of 94°C for 2 min., 35 cycles of 94°C for 30 sec., 52°C for 40 sec., and 72°C for 1 min., with a inal extension at 72°C for 10 min. Specimen information and barcode sequence data from this study were compiled using the Barcode of Life Data Systems (Ratnasingham & Hebert 2007). The sequence data is publicly accessible on BOLD and GenBank. Sequence divergences were calculated using BOLD with the Kimura 2-parameter (K2P) model generating a mid-point rooted neighbor-joining (NJ) phenogram to provide a graphic representation of the species’ sequence divergence. Genetic distances were calculated by the BOLD algorithm, both as uncorrected p-distances and as K2P distances. The length of specimens is given as standard length (SL), measured from the median anterior end of the upper lip to the base of the caudal in (posterior end of the hypural plate); body depth is the greatest depth from the base of the dorsal-in spines to the ventral edge of the abdomen (correcting for any malformation of preservation); body width is measured just posterior to the gill opening; head length from the front of the upper lip or anterior upper teeth (whichever is most anterior) to the posterior end of the opercular lap; orbit diameter is the greatest leshy diameter of the orbital rim, and interorbital width the least bony width; snout length is measured from the median anterior point of the upper lip to the nearest leshy rim of the orbit; caudal-peduncle depth is the least depth, and caudal-peduncle length the horizontal distance between verticals at the rear base of the anal in and the caudal-in base; predorsal, prepelvic and preanal lengths are angular measurements; lengths of spines and rays are measured to their extreme bases; caudal-in and pectoral-in lengths are the length of the longest ray; pelvic-in length is measured from the base of the pelvic spine to the tip of the longest soft ray. Morphometric data are presented as percentages of the standard length. Proportional measurements in the text are rounded to the nearest 0.05. The upper rudimentary pectoral-in ray is included in the count. Lateral-line scale counts include the last pored scale that overlaps the end of the hypural plate as +1; scales above the lateral line are counted in an oblique row from the pored scales under the mid-spinous dorsal in, the much smaller scale abutting the base of the in is counted as 0.5 scales. The count of gill rakers is made on the irst gill arch and includes all rudiments. The counts and measurements for the larger paratype (>50 mm SL) is shown in parentheses following data for the holotype (not listed if damaged). Proportional morphological measurements are presented in Table 1. 59 Figure 1. Pseudojuloides labyrinthus, BPBM 41257, TP male holotype, 65 mm SL, Kenya via aquarium trade (B.C. Victor). Pseudojuloides labyrinthus, n. sp. Labyrinth Pencil Wrasse urn:lsid:zoobank.org:act:A14ED652-FFF9-474D-9AA4-D4C281EDF3BC Figures 1–6, Table 1. Holotype. BPBM 41257, 65.0 mm SL, TP male, Mombasa region, Kenya, aquarium-trade, about April 9, 2015. Paratypes. BPBM 41258, (2) 43.0 & 60.6 mm SL, IP females, same collection data, about May 8, 2014. 60 Figure 2. Pseudojuloides labyrinthus, BPBM 41257, TP male holotype, 65 mm SL, Kenya via aquarium trade (M. Stern). Figure 3. Pseudojuloides labyrinthus, BPBM 41258, IP paratype, 43 mm SL, Kenya via aquarium trade (V. Altamirano). Diagnosis. Dorsal-in rays IX,11; anal-in rays III,12; pectoral-in rays 13; lateral-line scales 27 (+1 on caudalin base); no scales on head; gill rakers 14; a single pair of large, projecting, and slightly recurved canine teeth anteriorly in each jaw, the upper pair slightly out-laring, the lowers curving forward and itting between uppers when mouth closed; a short irregular row of 3–7 chisel-like incisiform teeth on each side of upper and lower jaws, no canine posteriorly at corner of mouth; elongate body, body depth 5.0–5.4 in SL; only slightly compressed, body width 1.7 in depth; caudal in slightly rounded in initial phase, truncate in terminal-phase male; initial phase reddish orange to pink, often with more yellow tint anteriorly and grading to white ventrally on the head and abdomen, a band of bright relective white running from tip of upper jaw back to under posterior orbit; terminalphase male in life greenish yellow with three bright blue stripes along posterior half of body, head and anterior body often abruptly darker, with maze of lines, thinner and ranging from bright blue to reddish on head, wider (about one scale high) and bright blue on anterior body; dorsal and anal ins with broad yellow bands bordered above and below with blue stripes, upper and lower margin of caudal in banded with yellow; iris red. Description. Dorsal-in rays IX,11; anal-in rays III,12, all soft dorsal and anal-in segmented rays branched, last split to base; pectoral-in rays 13, the irst rudimentary, the second unbranched; pelvic-in rays I,5; principal caudal-in rays 14, the upper and lower unbranched; upper and lower procurrent caudal-in rays 6; pored lateralline scales 27 (+1 on caudal-in base); scales above lateral line to origin of dorsal in 4.5; scales below lateral line to origin of anal in 8; median predorsal scales about 7–10; gill rakers 14. Body elongate, the depth 5.0 (5.4) in SL, and only slightly compressed, the width 1.7 (1.7) in depth; head length 3.1 (3.0) in SL; dorsal proile of head nearly straight on snout, forming low angle of about 20° to horizontal axis of body, and slightly convex on nape; snout sharply pointed, its length 3.7 (3.8) in HL; orbit relatively small, diameter 5.1 (4.7) in HL; interorbital space broadly convex, the least bony width 4.2 (4.8) in HL; caudal peduncle short and narrow, the least depth 3.5 in HL, caudal-peduncle length 3.0 in HL. Figure 4. Pseudojuloides labyrinthus, BPBM 41258, IP paratype, 60.6 mm SL, Kenya via aquarium trade (B.C. Victor). 61 TABLE 1 Proportional measurements of type specimens of Pseudojuloides labyrinthus, n. sp. as percentages of the standard length holotype 62 paratypes BPBM BPBM BPBM 41257 41258 41258 TP IP IP Standard length (mm) 65.0 60.6 43.0 Body depth Body width Head length Snout length Orbit diameter Interorbital width Caudal-peduncle depth Caudal-peduncle length Predorsal length Preanal length Prepelvic length Base of dorsal in First dorsal-in spine Ninth dorsal-in spine Longest dorsal-in ray Base of anal in First anal-in spine Second anal-in spine Third anal-in spine Longest anal-in ray Caudal-in length Pectoral-in length Pelvic-spine length Pelvic-in length 19.8 11.8 32.0 8.8 6.3 7.7 9.2 10.6 29.4 56.6 35.5 56.0 6.2 8.6 11.1 34.0 2.6 5.7 7.2 9.8 16.5 15.4 9.4 14.9 18.5 11.1 33.2 8.7 7.1 6.9 29.9 58.4 36.0 6.6 3.1 6.9 7.9 15.8 10.1 13.5 19.5 11.9 33.7 8.8 8.1 7.0 10.0 9.8 33.0 58.1 35.6 55.1 7.0 9.8 12.8 31.2 3.3 4.9 7.7 11.6 21.9 18.1 11.2 15.6 Mouth very small, terminal, the corner of gape with closed jaws well anterior to anterior nostril; end of maxilla buried, even when jaws gape. Lips moderately thick, the upper puffed with striations on the underside, the lower lip with prominent ventral-projecting lap along side of jaw. A pair of large, moderately projecting, and slightly recurved canine teeth anteriorly in each jaw, the upper pair slightly out-laring, the lowers curving forward and itting between uppers when mouth closed; a short row of 3–7 irregularly placed chisel-like incisiform teeth along each side of upper and lower jaw; no canine tooth posteriorly on upper jaw. Upper preopercular margin free nearly to level of lower edge of orbit; lower margin free anterior to a vertical through anterior nostril. Gill rakers short, the longest on irst arch (at angle) about one-ifth to one-tenth length of longest gill ilament. Nostrils small, in front of upper edge of orbit, the anterior in a short membranous tube elevated posteriorly, the posterior in advance of a vertical through front of orbit by a distance slightly less than internarial space. Pores on lower half of head comprise one over rear maxilla, then two anterior to orbit, followed by a curving suborbital series (counting up to rear mid-eye level) numbering 5–7 in single series; preopercular pores in a curved series after start of free edge near mandible, numbering 9 or 10 along free margin of preopercle, plus 1 or 2 more up to rear mid-eye level, in a single series at distal tips of canals. Scales thin and cycloid; scales on side of thorax less than half as high as largest scales on side of body, becoming still smaller ventroanteriorly; head naked except for small partially embedded scales on nape in irregular rows; median predorsal scales extending forward to slightly posterior to a vertical through upper free end of preopercular margin; ins naked except for several progressively smaller scales on basal region of caudal in and mid-ventral scale projecting posteriorly from base of pelvic ins. Lateral line continuous, nearly following contour of back to 18th pored scale, below base of eighth dorsal soft ray, where delected sharply ventrally to straight peduncular portion, single small pore per scale, last pored scale on caudal-in base. Origin of dorsal in above anterior edge of second lateral-line scale; dorsal-in spines progressively longer, the irst 5.2 (5.0) and the ninth 3.7 in HL; longest dorsal-in soft ray 2.9 in HL; origin of anal in below base of last dorsal-in spine; irst anal-in spine very short, 12.2 (10.6) in HL; second anal-in spine 5.6 (4.8) in HL; third anal-in spine 4.4 (4.2) in HL; longest anal-in soft ray 3.3 in HL; caudal in with slightly extended upper and lower lobes in terminal-phase males, caudal-in length 1.9 in HL; third pectoral-in ray longest, 2.1 (2.1) in HL; pelvic ins short, 2.1 (2.5) in HL. Color in life. Based on two TP male individuals (holotype and one living non-type)(Figs. 1,2 & 5 top), head and body greenish yellow grading to white ventrally, one specimen with anterior body and head abruptly darker, sparing ventral abdomen and thorax; head with reticulated pattern of thin lines, colored bright blue to red (in same male holotype) to purplish; maze of lines on head continuing onto anterior body and partially breaking up into spots, then transitioning at mid-body to form three distinct stripes ending near base of caudal in, body spots and stripes bright blue and wider than head stripes, about one scale wide. Dorsal and anal ins with a broad yellow band over a narrow blue stripe near base of in, in edged with a thin blue margin; caudal in with thick yellow bands along the upper and lower margins also edged with a thin bluish line, central portion of in translucent; pelvic and pectoral ins translucent. Iris reddish orange. IP individuals reddish to pinkish or orange (Figs. 3,4 & 6), lighter ventrally, white on lower half of head and thorax; a continuous band of brighter, relective, pearly white from tip of upper jaw back to below rear orbital margin. Fins translucent. Iris orange to bright red. The living holotype had a scattering of black pinpoint spots over the head and body; it is unclear if these are pathological or parasitic. They do not persist on the preserved specimen. Color in alcohol. TP male uniform grey brown dorsally grading to white ventrally, except for some residual blue in stripes. Fins are translucent. IP ish are uniform yellowish with no markings. Etymology. Named for the maze-like pattern of lines on the head and body. The speciic epithet is a noun in apposition. Distribution. The new species is described from specimens from the coast of Kenya. A record of “P. erythrops” from Seychelles (Randall & van Egmond 1994) is a 57-mm SL initial-phase specimen that is indistinguishable from P. labyrinthus: given the distance of Seychelles from the type location of P. erythrops in Mauritius, the record is more likely P. labyrinthus. Interestingly, the east African coast now has more sympatric species of Pseudojuloides than any other location, i.e. four species: the new species plus P. edwardi, P. polackorum, and the elusive P. argyreogaster. 63 Figure 5. TP male colors in aquaria– top: Pseudojuloides labyrinthus, non-type, Kenya (T.J. Engels); upper middle: P. erythrops, Mauritius (J.E. Randall); lower middle: P. severnsi, Japan (K. Nishiyama); bottom: P. edwardi, Kenya (V. Altamirano). 64 Barcode DNA sequence. A 652-nucleotide sequence of the segment of the mitochondrial COI gene used for barcoding by the BOLD informatics database (Ratnasingham & Hebert 2007) was obtained for the holotype. Following the database management recommendation of the BOLD, the sequence of the holotype (GenBank accession number KT352046) is presented here as well: CCTCTATCTAGTATTCGGTGCCTGAGCTGGGATGGTGGGCACAGCCCTAAGCCTGCTCATTCGGGCT GAACTTAGCCAGCCCGGTGCTCTCCTCGGAGACGACCAAATTTATAACGTAATCGTTACGGCCCAC GCCTTCGTAATAATCTTTTTTATAGTAATGCCAATTATGATTGGCGGGTTCGGAAACTGACTAATTCC TCTGATGATTGGGGCCCCTGATATGGCCTTCCCTCGAATGAACAACATGAGCTTCTGACTCCCATCT TTCCTTCTCCTCCTTGCCTCATCTGGTGTAGAAGCGGGAGCTGGAACTGGCTGAACAGTCTACCCC CCTCTGGCTGGCAACCTCGCCCACGCAGGGGCCTCTGTAGACTTAACTATCTTCTCCCTCCACTTAG CCGGCATCTCATCGATCCTAGGGGCAATCAACTTTATTACAACTATTGTAAATATGAAGCCCCCTGCT ATTTC TCAATACCAAACACCTCTCTTTGTTTGAGCCGTCTTAATTACAGCAGTCCTACTTCTTCTCTC ACTACCCGTGCTTGCTGCGGGCATCACAATGCTGCTAACTGATCGTAACCTCAATACCACCTTCTTT GACCCTGCAGGGGGAGGAGATCCCATCCTTTACCAACACCTC Comparisons. Among the Pseudojuloides, P. labyrinthus most closely resembles the P. severnsi species complex in basic marking patterns on the TP male, i.e. an abruptly darker head and anterior body, reticulated lines on the head, and blue stripes along the body (Fig. 5). The new species differs by having a third blue stripe along the posterior half of the body (vs. two), the third running well below the lateral midline. The TP male of P. erythrops also differs from P. labyrinthus in having the light ventrum ending abruptly at mid-abdomen, blue spots and reticulations against a dark background on the anterior abdomen, and only a single stripe across the upper head (vs. a maze). TP males of both P. edwardi and P. severnsi are missing the maze of lines on the upper head and upper anterior body. The initial-phase specimens of the entire genus are very similar and mostly non-descript and reddish orange; however, IP P. labyrinthus have the prominent relective white stripe below the eye extending in a complete wide band from the tip of the upper jaw to past the orbit, while those of the other species have the relective white more limited to the maxilla, or maxilla and separately under the eye, not forming a continuous streak (Fig. 6; and see Victor & Edward 2015). Figure 6. Pseudojuloides edwardi vs. labyrinthus, initial phase, (identiications by DNA barcode): P. edwardi at left, P. labyrinthus paratype, 43 mm SL, at right; Kenya via aquarium trade (J.M.B. Edward). 65 Morphometrics (>50 mm SL) are similar to other congeners, although the new species is more elongate than most, with body depth 18.5–19.8% SL, less than the P. cerasinus complex and P. atavai (23 to 25% SL; Randall & Randall 1981, Connell et al. 2015) and P. edwardi and P. severnsi (19% to 23% SL; Victor & Randall 2014); about the same as P. erythrops (18.6% to 19.6% SL; Randall & Randall 1981); and slightly wider than P. zeus and P. mesostigma (less than 18% from Victor & Edward [2015]). DNA Comparisons. The neighbor-joining phenetic tree based on the COI mtDNA sequences of 12 of the 14 known Pseudojuloides species, following the Kimura two-parameter model (K2P) generated by BOLD (Barcode of Life Database), shows deep divergences between species and relatively small differences within species, except for the P. edwardi and P. severnsi sequences, which are very close (Fig. 7). As a broad generality, among most reef TABLE 2 K2P distances for mtDNA COI sequences of 12 species of Pseudojuloides Minimum Interspeciic and Maximum Intraspeciic Distances (%) ata cer edw elo kal lab mes pol pyr sev P. atavai P. cerasinus P. edwardi P. elongatus P. kaleidos P. labyrinthus P. mesostigma P. polackorum P. pyrius P. severnsi P. xanthomos P. zeus 0 19.32 17.97 17.78 16.61 20.19 6.89 14.83 18.57 17.76 16.22 18.2 0.18 16.06 18.8 10.4 16.63 17.64 9.47 3.5 16.34 11.34 17.49 0.31 15.81 15.9 10.54 9.07 14.58 15.27 0.46 15.51 9.29 0.62 19.6 17.54 17.5 18.69 20.38 16.14 19.16 17.94 NA 15.9 16.49 8.42 10.71 15.83 4.48 17.51 xan 0.31 11.78 0.93 16.37 14.83 0.31 16.89 16.76 9.47 NA 10.75 9.3 14.34 15.61 0.93 16.52 15.08 7.5 11.23 15.45 NA 12.0 5.31 17.09 17.23 8.61 16.28 zeu NA P-distances (uncorrected pairwise) for mtDNA COI sequences of 12 species of Pseudojuloides Minimum Interspeciic and Maximum Intraspeciic Distances (%) ata cer edw elo kal lab mes pol pyr sev 66 P. atavai P. cerasinus P. edwardi P. elongatus P. kaleidos P. labyrinthus P. mesostigma P. polackorum P. pyrius P. severnsi P. xanthomos P. zeus 0 16.5 15.67 15.67 14.59 17.36 14.9 13.21 15.98 15.51 14.29 15.82 0.18 14.12 16.41 9.55 14.68 15.37 8.74 3.4 14.34 10.36 15.2 0.31 14.17 14.02 9.66 8.41 12.93 13.55 0.46 13.71 8.57 0.61 17.05 15.51 15.51 16.28 17.51 14.44 16.74 15.82 NA 14.13 14.44 7.83 9.83 13.98 4.3 15.21 xan 10.75 0.92 14.44 13.21 0.31 14.9 14.75 8.76 NA 9.83 8.6 12.75 13.82 0.92 14.59 13.36 7.07 10.29 13.67 NA 10.91 5.07 14.9 15.05 7.99 14.29 zeu NA Kenya } Kenya 0.02 Kenya P. labyrinthus, n. sp. P. zeus Micronesia Vanuatu } Vanuatu Kenya } Kenya Kenya Kenya P. mesostigma P. edwardi Philippines New Caledonia Philippines New Caledonia Indonesia New Caledonia Philippines } P. severnsi P. kaleidos P. xanthomos aqua Mauritius South Africa } South Africa Kenya Kenya South Africa P. polackorum P. pyrius Marquesas Islands Hawai‘i Hawai‘i Hawai‘i Hawai‘i New Caledonia New Caledonia New Caledonia Cook Islands Fr. Polynesia Fr. Polynesia Fr. Polynesia } P. cerasinus } P. elongatus } P. atavai Bali, Indonesia Leptojulis cyanopleura Figure 7. The neighbor-joining phenetic tree of Pseudojuloides following the Kimura two-parameter model (K2P) generated by BOLD (Barcode of Life Database). The scale bar at left represents a 2% sequence difference. Collection locations for specimens are indicated, and Leptojulis cyanopleura is used as an outgroup. The “aqua” label indicates an aquarium-trade specimen of unknown provenance. GenBank accession numbers and collection data for the sequences in the tree are listed in Appendix 1. 67 ishes the minimum interspeciic distance between close congeners is often up to an order of magnitude greater than the maximum intraspeciic distance, which is precisely what makes the barcode database particularly useful. It appears that the majority of reef ish species (with many exceptions) differ by more than 2% from their nearest relatives (Steinke et al. 2009, Ward et al. 2009, Victor 2015). Our genetic results show that P. labyrinthus falls in a broad clade made up of P. severnsi/edwardi and P. zeus and P. mesostigma. The nearest-neighbor sequence to P. labyrinthus is P. edwardi, which differs by 9.66% in COI sequence (uncorrected pairwise distance; 10.54% by K2P). The divergence is on a similar scale to that from the remaining species in the complex, i.e. P. severnsi, P. zeus, and P. mesostigma. It is somewhat unexpected that P. labyrinthus is more distant from P. severnsi than are P. zeus and P. mesostigma, since the TP males of P. labyrinthus share the basic marking patterns of P. severnsi and P. edwardi and appear quite different from P. zeus and P. mesostigma. Interestingly, in this case, the initial-phase marking differences (although slight) may be more relective of the genetic relationship than the TP male pattern. Genetic divergences within the genus Pseudojuloides vary widely (Table 2). For all but one pair, minimum interspeciic distances range from 3.4% to 17.51% (uncorrected pairwise; 3.5% to 20.38% by K2P). The maximum intraspeciic distances range from 0 to 0.92% (uncorrected pairwise; 0 to 0.93% by K2P), showing a clear “barcode gap” between species. The exception is the species pair of P. edwardi and P. severnsi, which diverge by only 0.46% (three nucleotides of the 652-bp barcode segment), and may be an example of phenotypic differences outpacing the rate of neutral substitutions in the mitochondrial COI DNA sequence early in the process of speciation (Victor & Randall 2014, Allen et al. 2015). Acknowledgments We thank Loreen R. O’Hara and Arnold Suzumoto of the Bishop Museum for curatorial assistance, and Vincent Altamirano, T.J. Engels, Kazuhiko Nishiyama (Kazu), and Michael Stern for graciously providing photographs. Comparison sequences on the Barcode of Life Database (BOLD) were provided by Serge Planes of the Centre National de la Recherche Scientiique and Jeff Williams of the U.S. National Museum of Natural History, via CRIOBE (Centre de Recherches Insulaires et Observatoire de l’Environnement CNRS-EPHE), BIOCODE (Moore Foundation), CORALSPOT (MEDDE, ANR, Polynésie), and the LABEX “CORAIL”; as well as by David Carlon of the Bowdoin Marine Laboratory, Brunswick, Maine and the University of Hawaiʻi and Anuschka Faucci of the University of Hawaiʻi. The assistance of Michael Stern of NY Aquatics is appreciated. Comparison specimens/ tissues were provided by David Bellwood, Alonso Gonzalez Cabello, the late Allan Connell, Arie deJong of De Jong Marinelife of the Netherlands, and Antoine Teitelbaum. George Walsh and Walsh Paper Distribution, Inc. of Westminster, CA sponsored preparation and publication of the project. 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Journal of Fish Biology, 74, 329–356. 69 Appendix 1. Specimen data and GenBank accession numbers for the mtDNA COI barcode sequences used to generate the phenogram in Fig. 7, following the order in the tree. Holotype in bold. 70 Genus species Collection site Voucher GenBank # Collector/Source Pseudojuloides Pseudojuloides Pseudojuloides Pseudojuloides Pseudojuloides Pseudojuloides Pseudojuloides Pseudojuloides Pseudojuloides Pseudojuloides Pseudojuloides Pseudojuloides Pseudojuloides Pseudojuloides Pseudojuloides Pseudojuloides Pseudojuloides Pseudojuloides Pseudojuloides Pseudojuloides Pseudojuloides Pseudojuloides Pseudojuloides Pseudojuloides Pseudojuloides Pseudojuloides Pseudojuloides Pseudojuloides Pseudojuloides Pseudojuloides Pseudojuloides Pseudojuloides Pseudojuloides Pseudojuloides Pseudojuloides Pseudojuloides Leptojulis labyrinthus, n. sp. labyrinthus, n. sp. labyrinthus, n. sp. zeus mesostigma mesostigma edwardi edwardi edwardi edwardi severnsi severnsi severnsi severnsi severnsi severnsi severnsi kaleidos xanthomos polackorum polackorum polackorum polackorum polackorum pyrius cerasinus cerasinus cerasinus cerasinus elongatus elongatus elongatus atavai atavai atavai atavai cyanopleura Mombasa, Kenya Mombasa, Kenya Mombasa, Kenya Majuro, Marshall Islands Vanuatu Vanuatu Mombasa, Kenya Mombasa, Kenya Mombasa, Kenya Mombasa, Kenya Philippines New Caledonia Philippines New Caledonia Indonesia New Caledonia Philippines aquarium trade Mauritius South Africa South Africa Kenya Kenya South Africa Marquesas Islands Hawai‘i Hawai‘i Hawai‘i Hawai‘i New Caledonia New Caledonia New Caledonia Rarotonga, Cook Islands Moorea, French Polynesia Moorea, French Polynesia Moorea, French Polynesia Bali, Indonesia BPBM 41258 (43.0) BPBM 41257 BPBM 41258 (60.6) BPBM 41215 BPBM 41216 60.6 BPBM 41216 60.9 BPBM 41172 je14pe1 BPBM 41173 63.4 BPBM 41173 70.6 BPBM 41174 72.3 BPBM 41175 55.4 M1496 BPBM 41175 70.1 je13ps qm14ps2 BPBM 41174 79.8 je14pk610 dej13px360 DSFSG592-11 DSFSG925-13 BPBM 41207 BPBM 41208 ac13pc MARQ-424 FLHI398-09 h83pc370 FLHI318-09 h83pc260 jr14pe3 jr14pe2 jr14pe1 ck98425pa210 MBIO1549 M106 MBIO1289 bal11700px124 KP975975 KT352046 KT352050 KJ591656 KP975989 KP975968 KJ591643 KP975964 KJ591642 KJ591644 KJ591652 KJ591651 JQ839573 KJ591655 KJ591653 KJ591654 JQ839574 KP975974 KJ591657 KF489719 KP975998 KP975967 KP975996 KP975978 KJ591650 KJ591646 JQ839570 KJ591645 JQ839571 KJ591647 KJ591649 KJ591648 JQ839568 JF435150 JQ839569 JF435151 JQ839546 J. Edward/aq. trade J. Edward/aq. trade J. Edward/aq. trade J. Edward/aq. trade J. Edward/aq. trade J. Edward/aq. trade J. Edward/aq. trade J. Edward/aq. trade A. DeJong/aq. trade J. Edward/aq. trade J. Edward/aq. trade A. Teitelbaum D. Bellwood, JCU A. Teitelbaum J. Edward/aq. trade A. Teitelbaum J. Edward/aq. trade J. Edward/aq. trade A. DeJong/aq. trade A. Connell/ SAIAB A. Connell/ SAIAB J. Edward/aq. trade J. Edward/aq. trade A. Connell/ SAIAB J. Williams/S. Planes D. Carlon/A. Faucci B. Victor D. Carlon/A. Faucci B. Victor A. Teitelbaum A. Teitelbaum A. Teitelbaum B. Victor S. Planes/J. Williams D. Bellwood, JCU S. Planes/J. Williams B. Victor