aqua, International Journal of Ichthyology

On the validity of the cirrhitid genus Itycirrhitus

Michelle R. Gaither1-2 and John E. Randall3

1) California Academy of Sciences, 55 Music Concourse Dr., San Francisco, CA 94118, USA. E-mail: [email protected] 2) Hawai‘i Institute of Marine Biology, P.O. Box 1346, Kaneohe, HI 96744, USA. 3) Bishop Museum, 1525 Bernice St., Honolulu, HI 96817-2704, USA. E-mail: [email protected].

Received: 6 June 2012 – Accepted: 17 September 2012

Abstract der Familie (COI, d = 8-18%; cyt b, d = 12-20%). Dieser The hawkfish wilhelmi Lavenberg & Yañez was Grad der Divergenz ist typisch für andere exemplarische described from Easter Island in 1972, reclassified in Vergleiche innerhalb der Cirrhitidae (COI, d = 15-20%; cyt by Pequeño (1989), and the range extended b, d = 15-21%). to the Pitcairn Islands by Randall (1999). Randall (2001) described the new genus Itycirrhitus for this species. The Résumé similarity in general morphology and color to the Hawai- Le poisson-faucon Cirrhitus wilhelmi Ravenberg & Yañez ian fasciatus and C. mascarenensis, type locality a été décrit en provenance de l’île de Pâques, en 1972, et Mauritius, prompted the present molecular study to deter- reclassé comme Amblycirrhitus par Pequeño (1989), et la mine the validity of Itycirrhitus. Combining mitochondrial distribution étendue jusqu’aux îles Pitcairn par Randall cytochrome oxidase I (COI) and cytochrome b (cyt b) (1999). Randall (2001) a décrit le nouveau genre Itycirrhi- sequence data from specimens of I. wilhelmi with compa- tus pour cette espèce. L’analogie en morphologie générale rable data from the two species of Cirrhitops, we found et en couleur avec Cirrhitops fasciatus d’Hawaï et C. mas- high levels of divergence between the two genera (COI, d carenensis, localité-type l’île Maurice, a déterminé la = 15%; cyt b, d = 20-21%) that are similar to comparisons présente étude moléculaire pour tester la validité d’Itycir- between Itycirrhitus and other genera of the family (COI, rhitus. En combinant les données des séquences de la d = 8-18%; cyt b, d = 12-20%). This level of divergence is cytochrome oxydase I (COI) et du cytochrome b (cyt b) de typical of other generic comparisons within Cirrhitidae spécimens de I. wilhelmi avec des données comparables de (COI, d = 15-20%; cyt b, d = 15-21%). deux espèces de Cirrhitops, nous avons trouvé des niveaux élevés de divergences entre les deux genres (COI, d = 15%; Zusammenfassung cyt b, d = 20-21%) similaires à des comparaisons entre Ity- Der Büschelbarsch (Korallenwächter) Cirrhitus wilhelmi cirrhitus et d’autres genres de la famille (COI, d = 8-18%; Lavenberg & Yañez wurde 1972 von den Osterinseln cyt b, d = 12-0%). Ce niveau de divergence est typique beschrieben und später von Pequeño (1989) in die Gattung d’autres comparaisons génétiques parmi les Cirrhitidae Amblycirrhitus eingeordnet, hinzu kam eine Erweiterung des (COI, d = 15-20%; cyt b, d = 15-21%). Verbreitungsgebietes bis zu den Pitcairn-Inseln von Randall (1999). Bald danach wurde von Randall (2001) für diese Art Sommario die neue Gattung Itycirrhitus beschrieben. Die Ähnlichkeit Il pesce falco Cirrhitus wilhelmi Lavenberg & Yañez è nach Färbung und allgemeiner Morphologie zu den hawai- stato descritto dall’Isola di Pasqua nel 1972, riclassificato ianischen Cirrhitops fasciatus und C. mascarenensis, Typus- in Amblycirrhitus da Pequeño (1989) e la sua area di pre- lokalität Mauritius, gab den Anlass zu der vorliegenden senza estesa alle isole Pitcairn da Randall (1999). Randall molekularen Studie mit dem Ziel, die Gültigkeit von Itycir- (2001) ha descritto il nuovo genere Itycirrhitus per questa rhitus zu prüfen. Auf der Grundlage der Sequenzdaten der specie. La somiglianza nella morfologia generale e nel colo- mitochondrialen Cytochromoxidase I (COI) sowie des re alla specie hawaiana Cirrhitops fasciatus e a C. mascare- Cytochrom b (cyt b) wurden Exemplare von I. wilhelmi mit nensis, località tipo Mauritius, ha incoraggiato il presente solchen der beiden Cirrhitops-Arten verglichen; es zeigte sich studio molecolare per determinare la validità di Itycirrhitus. ein hoher Grad an Divergenz zwischen den beiden Gattun- Combinando i dati di sequenza della citocromo ossidasi I gen (COI, d = 15%; cyt b, d = 20-21%), ähnliche Werte wie (COI) e del citocromo b (cit b) mitocondriali da campioni beim Vergleich zwischen Itycirrhitus und anderen Gattungen di I. wilhelmi con dati comparabili delle due specie di

219 aqua vol. 18 no. 4 - 15 October 2012 On the validity of the cirrhitid fish genus Itycirrhitus

Cirrhitops, abbiamo trovato alti livelli di divergenza tra i Key to the Genera of Cirrhitidae due generi (COI, d = 15% , cit b, d = 20-21%), simili a 1a. Snout not elongate, its length about 2.8-4.1 confronti tra Itycirrhitus e altri generi della famiglia (COI, in head length; body not slender, the depth d = 8-18%, cit b, d = 12-20%). Questo livello di diver- 2.0-3.4 in SL; canine teeth in jaws markedly genza è tipica di altri confronti generici all'interno Cirrhi- tidae (COI, d = 15-20%; cyt b, d = 15-21%). longer than inner villiform teeth, those at front of upper jaw and side of lower jaw enlarged ...... 2 INTRODUCTION 1b. Snout elongate, its length 1.85-2.0 in head The 33 species of of the perciform family length; body slender, the depth 4.4-4.6 in SL; Cirrhitidae, popularly known as hawkfishes, are canine teeth in jaws only slightly longer than found in the tropical and subtropical Indo-Pacific inner villiform teeth and nearly uniform in region, except for three in the Atlantic and three in size ...... Oxycirrhites the eastern Pacific (two of which, 2a. Caudal fin rounded, truncate, or slightly oxycephalus and , are range exten- emarginate; dorsal soft rays 11-15; snout not sions from the Indo-Pacific). Cirrhitid fishes all short, its length 2.7-3.SL ...... 3 have X dorsal spines, III anal spines, 14 pectoral 2b. Caudal fin lunate; dorsal soft rays 16 or 17; rays (the lower five to seven rays unbranched and snout short, its length 3.6-4.1 in head length thickened), two flat opercular spines, a serrate pre- ...... Cyprinocirrhites opercle, cycloid scales, one to several cirri at the tip 3a. Small scales on cheek in more than 12 rows.. of each membrane of the dorsal spines, and no ...... 4 swim bladder. Most species occur in shallow water 3a. Rows of large scales on cheek 4-6 (small scales on reefs or rocky substrata, often in areas also usually present) ...... 8 exposed to wave action. When in a surge zone, they 4a. Lower 7 pectoral rays unbranched and thick- use their thickened lower pectoral rays to wedge ened; first 2 supraneural bones in space before themselves in cracks in the reef or within branches second neural spine; more than 40 cirri in 2 of coral. Hawkfishes feed mainly on benthic crus- series on posterior flap of anterior nostril ...... taceans and occasionally on small fishes. Excep- ...... 5 tions are Cyprinocirrhites polyactis that feeds well 4b. Lower 6 pectoral rays unbranched and thick- above the substratum on zooplankton, and O. ened; first 3 supraneural bones in space before typus that often makes short forays from the bot- second neuralspine; fewer than 15 cirri on tom to prey on the larger of the demersal posterior flap of anterior nostril ...... 6 plankton. At least some of the species of the family 5a. Supraorbital ridge high, continuing more are protogynous (Sadovy & Don- than half eye diameter posterior to orbit; aldson 1995). lower opercular spine acute, forming an angle The generic classification of the Cirrhitidae has a of 45° or less; no scales in interorbital space; long and confused history, Günther (1860) recog- pectoral fins reaching slightly beyond a verti- nized eight genera in the family. A century and 11 cal at tips of pelvic fins; body depth 3.1-3.35 cirrhitid publications later, Schultz in Schultz & in SL...... Cristacirrhitus collaborators (1960) listed 13 genera, including 5b. Supraorbital ridge low and not continuing Iso bu na and Serranocirrhitus, now known to be ser- posterior to eye; lower opercular spine form- ranids. Randall (1963) included 10 genera and 34 ing an angle of 90°; a V-shaped band of scales species in his revision of the family. Randall (2001) in posterior half of interorbital space; pectoral again revised the genera of cirrhitids, adding three fins short, not reaching a vertical at tips of new monotypic genera from species formerly clas- pelvic fins; body depth 2.6-3.1 in SL ...... sified in Cirrhitus. One of these, Itycirrhitus, was ...... Cirrhitus described for a small species, Cirrhitus wilhelmi, 6a. Dorsal soft rays 13; three-fourths or more of from Easter Island. This species was first described preopercular margin coarsely serrate; palatine by Lavenberg & Yañez (1972) and later reclassified teeth absent; body very deep, the depth 2.0- in Amblycirrhitus by Pequeño (1989). Randall 2.4 in SL, and very compressed, the width (1999) extended the range to the Pitcairn Islands. 2.9-3.1 in depth; longest pectoral rays not The key to genera of Randall (2001) is reproduced extending beyond a vertical at pelvic-fin tips. here...... Neocirrhites aqua vol. 18 no. 4 - 15 October 2012 220 Michelle R. Gaither and John E. Randall

6b. Dorsal soft rays 11-12; about upper half of 9b. Palatine teeth present; maxilla not reaching a preopercular margin finely or coarsely serrate; vertical through middle of eye; dorsal spines palatine teeth present; body not very deep, 1.7-3.0 in head length; dorsal profile of head the depth 2.6-3.3 in SL, and not very com- straight to slightly convex; lower 5 to 7 pec- pressed, the width 1.9-2.3 in depth; longest toral rays unbranched; interorbital scaled or pectoral rays extending beyond a vertical at naked; snout pointed or not pointed; preor- pelvic-fin tips ...... 7 bital with or without a free posterior margin. 7a. Upper margin of preopercle finely serrate (25 ...... 10 or more serrae); exposed end of posttemporal 10a. Dorsal soft rays 14-15 (rarely 15); first 2 pec- finely serrate; first (most medial) branchioste- toral rays unbranched; snout not pointed, the gal ray strongly curved and nearly parallel dorsal profile from interorbital to upper lip with second ray; gill membrane across throat convex; interorbital naked...... Cirrhitops naked; scales on cheek separated from serrate 10b. Dorsal rays 11-13 (rarely 13); first pectoral edge of preopercle by a broad naked zone ray unbranched, second branched; snout crossed by irregular sensory channels (may pointed, the dorsal profile straight; interor- show as ridges); no scales on snout; scales ven- bital scaled or naked...... 11 trally on chest extremely small; 3 rows of large 11a. Preopercular margin finely serrate; first 2 scales above lateral line in middle of body; supraneural bones in space before second dorsal soft rays 12...... Notocirrhitus neural spine; preorbital without a free hind 7b. Upper margin of preopercle coarsely serrate margin; interorbital scaled; first dorsal soft ray (fewer than 13 serrae); exposed end of post- not produced into a filament; lower 5 (rarely temporal with 3-5 serrae; first (most medial) 6) pectoral rays unbranched ...... branchiostegal ray nearly straight and not par- ...... Amblycirrhitus allel to second ray; gill membrane across 11b. Preopercular margin coarsely serrate; all 3 throat scaled; scales on cheek extending to supraneural bones in space before second base of preopercular serrae; scales on snout neural spine; preorbital with a free hind mar- extending to below anterior nostrils; scales gin for one-fourth to one-half distance from ventrally on chest one-half or more size of lower edge to eye; interorbital not scaled; first scales on side of body; 4 rows of large scales dorsal soft ray usually produced into a fila- above lateral line in middle of body; dorsal ment; lower 6 or 7 pectoral rays unbranched soft rays 12-14 (rarely 12 or 14) ...... Cirrhitichthys ...... Itycirrhitus 8a. Rows of large scales above lateral line to base After the separation of Oxycirrhites in the key, of spinous portion of dorsal fin 5; a single cir- mainly by its snout length, and Cyprinocirrhites by rus from membrane near tip of each spine of its lunate caudal fin, the characters leading to the dorsal fin; membranes between longest dorsal other genera are less incisive, in particular those spines incised at most one-fifth spine length; leading to Itycirrhitus. This study was initiated to palatine teeth absent...... determine if an analysis of DNA sequence data 8b. Rows of large scales above lateral line to base from species of cirrhitid fishes will support the of spinous portion of dorsal fin 3 or 4; a tuft generic classification. Specifically we collected of cirri from membrane near tip of each spine mitochondrial sequence data to compare Itycirrhi- of dorsal fin; membranes between longest tus wilhelmi (Fig. 1) with the similarly colored dorsal spines incised one-third or more of species of Cirrhitops, C. fasciatus (Fig. 2) and C. spine length; palatine teeth present or absent mascarenensis. Additionally, we investigated the ...... 9 level of genetic divergence between the two mono- 9a. Palatine teeth absent; maxilla reaching to or typic genera Notocirrhitus and Neocirrhites which beyond a vertical through middle of eye; dor- are closely grouped in the key. sal spines short, the longest 2.9-3.2 in head length; dorsal profile of head convex; lower 5 MATERIALS AND METHODS pectoral rays unbranched; interorbital fully For genetic analysis a 1cm fin clip was obtained scaled; snout not pointed; preorbital without from three specimens of Itycirrhitus wilhelmi from a free posterior margin...... Isocirrhitus Easter Island. DNA was isolated using the modi-

221 aqua vol. 18 no. 4 - 15 October 2012 On the validity of the cirrhitid fish genus Itycirrhitus

Fig. 1. Itycirrhitus wilhelmi, Easter Island. Photo by J. E. Randall.

Fig. 2. Cirrhitops fasciatus, Oah‘u, Hawaiian Islands. Photo by J. E. Randall. aqua vol. 18 no. 4 - 15 October 2012 222 Michelle R. Gaither and John E. Randall

Fig. 3. Maximum Likelihood (ML) phylogenetic tree of the cytochrome oxidase I gene for 10 species (7 genera) of hawkfish (Family Cirrhitidae). Numbers above branches reflect bootstrap support for ML tree and Bayesian posterior probabilities, respectively. Only major nodes with bootstrap support of >60 are labeled.

223 aqua vol. 18 no. 4 - 15 October 2012 On the validity of the cirrhitid fish genus Itycirrhitus

Table I. Mitochondrial cytochrome oxidase I (COI) and cytochrome b (cyt b) sequences used in this study. Species name, Barcode of Life Data systems (http://www.boldsystems.org/views/login.php, Ratnasingham & Hebert 2007) sequence iden- tification numbers and GenBank accession numbers (in italics; http://www.ncbi.nlm.nih.gov/genbank/), and museum voucher IDs (in parentheses) are listed. Sequences generated in this study are in bold. Number of individuals (if >1) in which haplotype was detected is listed in parentheses.

Species COI cyt b Amblycirrhitus pinos TZAIB036-06 (HLC-12147)

A. bimacula JX645648 EU684136, JX645657

Cirrhitops fasciatus EU684132, EU684133 (BPBM 40485, 40888) EU684137, EU684138 (BPBM 40485, 40888) TZAIB307-06 (HLC-12324) GBGC10204-09, GBGC10205-09

C. mascarenensis EU684134, EU684135 (BPBM 40889-90) EU684139, EU684140 (BPBM 40889-90)

Cirrhitus pinnulatus JX645649, JX645650 (2) JX645661, JX645662, JX645663

Itycirrhitus wilhelmi JX645651, JX645652 (2) JX645658, JX645659, JX645660

Neocirrhites armatus TZAIC178-05 (HLC-10878) JX645664, JX645665, JX645666 TZAIB505-06 (HLC-13128) TZAIC695-06 (HLC-11757) JX645653, JX645654, JX645655

Notocirrhitus splendens JX645656(2) (MA655017, MA655018)

Paracirrhites arcatus TZAIB143-06 (HLC-12066) TZAIB144-06 (HLC-12067) TZAIB145-06 (HLC-12068) TZAIB147-06 (HLC-12070) TZAIB867-07 (HLC-15197)

P. forsteri DSFSG451-11(ADC11_214.7 #2) DSFSG566-11(ADC11_214.7 #4)

Epinephelus lanceolatus HQ174825, HQ174826 HQ174838, HQ174839

Plectropomus leopardus GBGC1814-06, GBGC1208-06, FSCS615-07 AY963555, AY963556 fied HotSHOT method (Meeker et al. 2007; ume. After an initial 7 min denaturation at 95°C, Truett et al. 2000). We amplified approximately each of 35 cycles consisted of denaturation for 30 s 660 bp the mitochondrial cytochrome c oxidase at 94°C, annealing at 56°C for COI and 58°C for subunit I (COI) gene and approximately 750 base cyt b for 30 s, and extension at 72°C for 45 s with pairs of cytochrome b (cyt b) gene using the a final 10 min extension at 72°C. Amplification primers employed in Randall and Schultz (2009). products were purified using 0.75 units of Exonu- Polymerase chain reactions (PCR) were carried out clease I: 0.5 units of Shrimp Alkaline Phosphatase in a 20 µl volume containing 5-20 ng of template (ExoSAP, USB, Cleveland, OH, USA) per 7.5 µl DNA, 0.4 µM of each primer, 10 µl of the pre- PCR products at 37°C for 60 minutes, followed by mixed PCR solution BioMix Red (Bioline Inc., deactivation at 80°C for 15 minutes. DNA Springfield, NJ, USA), and deionized water to vol- sequencing was performed with fluorescently- aqua vol. 18 no. 4 - 15 October 2012 224 Michelle R. Gaither and John E. Randall labeled dideoxy terminators on an ABI 3730XL the MRBAYES manual. Average percent divergence Genetic Analyzer (Applied Biosystems, Foster City, (d) between genera was calculated in ARLEQUIN 3.5 CA, USA) at the University of Hawai‘i Advanced (Excoffier & Lischer 2010). Studies of Genomics, Proteomics and Bioinformat- ics sequencing facility. Sequences for each locus RESULTS were aligned, edited, and trimmed to a common Our comparisons of Itycirrhitus wilhelmi and the length using the DNA sequence assembly and two species of the genus Cirrhitops (C. fasciatus and analysis software Geneious Pro 5.0 (Biomatters, C. mascarenensis) at two mitochondrial genes con- LTD, Auckland, NZ). In all cases, alignment was firmed the distinction of these two genera. Across unambiguous with no indels or frameshift muta- all COI sequences analyzed we detected 197 vari- tions. Unique sequences were deposited in Gen- able nucleotide sites. There were two haplotypes in Bank (http://www.ncbi.nlm.nih.gov/genbank/; see three I. wilhelmi specimens. The average pairwise Table I for accession numbers). difference between C. fasciatus and C. mascarenen- Maximum Likelihood (ML) trees were con- sis was 39 bp (d = 7%), while I. wilhelmi differs structed using the default settings implemented in from the two species of Cirrhitops by an average of the program MEGA 5.05 (Tamura et al. 2007). 85 bp (d = 15%). All generic level comparisons COI and cyt b sequences of Cirrhitops fasciatus (N resulted in average base pair differences that range = 5, Hawai‘i) and Cirrhitops mascarenensis (N = 2, between 82 bp (d = 15%; Cirrhitops vs. Neocir- Mauritius), were obtained from GenBank (Randall rhites) and 113 (d = 20%; Neocirrhites vs. Ambly- & Schultz 2009, Table I). For inclusion in the phy- cirrhitus). The exception is I. wilhelmi and Neocir- logenetic tree COI sequences from representatives rhites armatus which differ by an average of only 47 of three genera in the family Cirrhitidae were bp (d = 8%). downloaded from the Barcode of Life Data Analyses of cyt b sequences were similarly robust. (BOLD) Systems 2.5 website (www.barcod- We detected 256 variable nucleotide sites. There inglife.org) (Table I). Corresponding cyt b were three haplotypes in three I. wilhelmi speci- sequences were available from GenBank for only mens. The average pairwise difference between C. Amblycirrhitus bimacula. Additionally, tissue sam- fasciatus and C. mascarenensis was 75 bp (d = 11%). ples of Amblycirrhitus bimacula (N = 1, Hawai‘i), In contrast, the average pairwise differences Cirrhitus pinnulatus (N = 3, Guam), Neocirrhites between I. wilhelmi and the two species of Cirrhi- armatus (N = 3, Guam), and Notocirrhitus splen- tops were 140 and 132 respectively (d = 21% and dens (N = 2, Kermadec Islands, New Zealand) were 20%, respectively). Generic level comparisons obtained and sequenced at both loci and deposited resulted in average base pair differences that ranged in GenBank (Table I). Available voucher specimens between 84 bp (d = 13%; Neocirrhites vs. Itycirrhi- and tissues (Table I) are deposited in the: Biodiver- tus) and 138 bp (d = 21%; Amblycirrhitus vs. Cir- sity Institute of Ontario (HLC), Guelph, Canada; rhitops). Bernice P. Bishop Museum (BPBM), Honolulu, After repeated attempts, we were able to amplify USA; South African Institute of Aquatic Biodiver- the Notocirrhitus splendens specimens at COI but sity (ADC), Grahamstown, South Africa; and the not cyt b. When comparing N. splendens with Neo- Auckland Museum (MA), Auckland, New Zealand. cirrhites armatus, which are closely grouped in the Phylogenetic trees were rooted with two species key, we found an average pairwise difference from the family Serranidae (Table I, Epinephelus between these monotypic genera of 84 bp (d = lanceolatus and Plectropomus leopardus). For analy- 15%). A similar level of divergence was detected sis, all COI and cyt b sequences were trimmed to between N. splendens and I. wilhelmi (87 bp, d = 557 bp and 671 bp, respectively. Bootstrap support 16%), which are also closely grouped in the key. values were calculated using default settings with Phylogenetic analyses revealed a single tree topol- 1000 replicates. For comparison we ran a Bayesian ogy for both mitochondrial markers with only the Markov Chain Monte Carlo (MCMC) analysis as relative positions of the outgroups varying (Fig. 3). implemented in the program MRBAYES 3.1.1 Both Maximum Likelihood and Bayesian analyses (Huelsenbeck & Ronquist 2001). We employed indicated that N. armatus is more closed related to default settings and ran simulations for 1,000,000 I. wilhelmi than I. wilhelmi is to the members of generations until the standard deviation of split the genus Cirrhitops. frequencies were below 0.01 as recommended in

225 aqua vol. 18 no. 4 - 15 October 2012 On the validity of the cirrhitid fish genus Itycirrhitus

DISCUSSION REFERENCES Phylogenetic analyses indicate strong concor- EXCOFFIER, L. & LISCHER, H. E. L. 2010. ARLEQUIN suite dance between our molecular data and generic ver 3.5: a new series of programs to perform population level classifications within the family Cirrhitidae genetics analyses under Linux and Windows. Molecular (see key above). Species within the same genus Ecology Resources 10: 564-567. GÜNTHER, A. 1860. Catalogue of the Acanthopterygian always grouped together with intragenetic diver- Fishes in the Collection of the British Museum. British gences ranging from 7-14% at COI. The genus Ity- Museum, London, 548 pp. cirrhitus was described by Randall (2001) based on HUELSENBECK, J. P. & RONQUIST, F. 2001. MRBAYES: seemingly small morphological differences. Here Bayesian inference of phylogeny. Bioinformatics 17: 754- we report sequence data that support the generic 755. level designation for Itycirrhitus wilhelmi. This LAVENBERG, R. J. & YAÑEZ L. A. 1972. A new species of species is 15% divergent at COI from either of the Cirrhitus from Easter Island (Pisces, Cirrhitidae), species of the genus Cirrhitops; a level of divergence Gayana, Zoologia, 21: 1-11. MEEKER, N. D., HUTCHINSON, S. A., HO, L., & TREDE, common among the other genera of the family (8 N. S. 2007. Method for isolation of PCR-ready genomic to 20%). Our finding of a similar level of diver- DNA from zebrafish tissues. BioTechniques 43: 610-614. gence between and Notocirrhi- PEQUEÑO, G. 1989. Peces de Chile. Lista sistemática tus splendens supports the designation of these revisada y comentada. Revista de Biología Marina, Val- monotypic genera. paraíso 24: 1-132. Interestingly, I. wilhelmi did not cluster in the RANDALL, J. E. 1963. Review of the hawkfishes (family phylogenetic tree with the morphologically similar Cirrhitidae). Proceedings of the United States National Cirrhitops (Figs 1-2). Instead, despite significant Museum 114: 389-451. RANDALL, J. E. 1999. Report on fish collections from the morphological differences, Neocirrhites armatus Pitcairn Islands. Atoll Research Bulletin, 461, 53 pp. was found to be only 8% divergent from I. wil- RANDALL, J. E. 2001. Revision of the generic classification helmi, a similar level of divergence between the two of the hawkfishes (Cirrhitidae), with descriptions of three species of Cirrhitops (7%). In this case we found new genera. Zootaxa 12: 1-12. surprising incongruence between the morphologi- RANDALL, J. E. & SCHULTZ, J. K. 2009. Cirrhitops mas- cal characters used to delineate these species and carenensis, a new species of hawkfish from the Mascarene the level of molecular divergence detected. Islands, southwestern Indian Ocean. Smithiana Bulletin 9: 15-20. RATNASINGHAM, S. & HEBERT, P. D. N. 2007. BOLD: ACKNOWLEDGEMENTS The Barcode of Life Data System We thank the following for providing tissue sam- (www.barcodinglife.org). Molecular Ecology Notes 7: 355 ples for this study: Dr. Gerald R. Allen, Dr. Alfredo 364. Cea, Dr. Mark V. Erdmann, Daniel Pelicier, and SADOVY, Y. & DONALDSON, T. J. 1995. Sexual pattern of Dr. Gordon W. Tribble. Support for the first Neocirrhites armatus (Cirrhitidae) with notes on other author was provided by a grant/cooperative agree- hawkfish species. Environmental Biology of Fishes 42: ment from the National Oceanic and Atmospheric 143-150. Administration, Project R/HE-1, which is spon- SCHULTZ, L. P., CHAPMAN, W. M., LACHNER, E. A. & WOODS, L. P. 1960. Fishes of the Marshall and Marianas sored by the University of Hawaii Sea Grant Col- Islands. Smithsonian Institution United States National lege Program, SOEST, under Institutional Grant Museum Bulletin, 202 (2), 438pp. No. NA09OAR4170060. The views expressed TAMURA, K., DUDLEY, J., NEI, M. & KAMAR, S. 2007. herein are those of the authors and do not neces- MEGA4: molecular evolutionary genetics analysis sarily reflect the views of NOAA or any of its sub- (MEGA) software version 4.0. Molecular Biology and agencies. UNIHI-SEAGRANT-JC-09-48. This is Evolution 24: 1596-1599. contribution #1516 from the Hawai‘i Institute of TRUETT, G. E., MYNATT, R. L., TRUETT, A. A., WALKER, Marine Biology and #8739 from the School of J. A. & WARMAN, M. L. 2000. Preparation of PCR-qual- ity mouse genomic DNA with hot sodium hydroxide and Ocean and Earth Science and Technology. Tris (HotSHOT). BioTechniques 29: 52-54.

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