J. Ocean Univ. (Oceanic and Coastal Sea Research) DOI 10.1007/s11802-014-2609-x ISSN 1672-5182, 2015 14 (1) http://www.ouc.edu.cn/xbywb/ E-mail:[email protected]

The Taxonomic Status of Japanese japonicus (Bloch, 1791) (: Nemipteridae) with a Redescription of this From the South China Sea Based on Morphology and DNA Barcodes

NING Ping1), *, SHA Zhongli1), Paul D. N. HEBERT2), and Barry RUSSELL3)

1) Laboratory of Marine Organism and Phylogeny, Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266071, P. R. China 2) Biodiversity Institute of Ontario, University of Guelph, Guelph ON N1G 2WI, Canada 3) Museum and Art Gallery of the Northern Territory, Darwin NT 0801, Australia

(Received May 9, 2013; revised December 20, 2013; accepted January 4, 2014) © Ocean University of China, Science Press and Springer-Verlag Berlin Heidelberg 2014

Abstract Because of its importance as a food source, (Bloch, 1791) (Nemipteridae) or Japanese threadfin bream is the best studied of these taxa, and numerous investigations have examined its , its biology and biochemistry. De- spite such intensive work, the taxonomic status of N. japonicus has never been seriously questioned and it is regarded as a common species, widely distributed throughout the Indo-Western Pacific Ocean. In fact, Bloch’s description of the type specimen of N. ja- ponicus has ambiguous collection data and lacks a designation for the type locality, though it is probably Java. In this paper, DNA barcode results based on COI gene support the existence of two geographically separated lineages of the Japanese threadfin bream, both being an Indian Ocean and western Pacific lineage, with 2.7% sequence divergence, and the results indicate a possible existing of some cryptic species. The two lineages also possess a diagnostic difference in their belly color, with specimens in the South China Sea having a silver belly, while those from the Indian Ocean isolate specimen have a yellow coloration. Based upon new collections from the South China Sea, this species from the western Pacific is morphologically redescribed and its details of DNA barcode diver- sity are shown for the future investigations.

Key words South China Sea; Indian Sea; Indo-Pacific Barrier; COI gene

details DNA barcode diversity in N. japonicus on the ba- sis of specimens from the South China Sea. 1 Introduction Nemipterus Swainson, 1839, the largest in the family Nemipteridae, includes about 26 species (Russell, 2 Materials and Methods 1990) which are distributed in the Indo-Western Pacific 2.1 Sample Collection and Examination Ocean. Because of its importance as a food source, Ne- There are 15 examined specimens collected from the mipterus japonicus (Bloch, 1791) is the best studied of southern coastal waters of China from 1950 to 2011 and these taxa and numerous investigations have examined its deposited at the Marine Biological Museum (MBM), In- fisheries, its biology and biochemistry (Krishnamoorthi, stitute of Oceanology, Chinese Academy of Sciences (IO- 1971; Russell, 1990; Chawla et al., 1996; Joshi, 2010; CAS). The other 22 specimens examined are in: Academy Naqasha and Nazeera, 2010). Despite this intensive work, of Natural Sciences (ANSP) Philadelphia, USA; Aca- the taxonomic status of N. japonicus has never been seri- demica Sinica (ASIZ), Taipei, Taiwan, China; the Natural ously questioned and it is regarded as a common species, History Museum (BMNH), London, UK; California Aca- widely distributed throughout the Indo-Western Pacific demy of Sciences (CAS), San Francisco, USA; Museum Ocean (Froese and Pauly, 2013). and Art Gallery of the Northern Territory (NTM), Darwin, While carrying out morphological and DNA barcode Australia; Senckenberg Museum (SMF), Frankfurt, Ger- studies, we discovered evidence for genetic divergence many; Smithsonian Institution (USNM), Washington DC, within N. japonicus that is linked to collection locality. USA; Zoological Museum (ZMB), Berlin, Germany. This paper provides a morphological redescription and 2.2 Morphological Description * Corresponding author. E-mail: [email protected] Color and pigmentation patterns were examined in

2 NING et al. / J. Ocean Univ. China (Oceanic and Coastal Sea Research) 2015 14 digital images of freshly collected specimens, and de- “HKY+G+I” for ML was chosen based on results from tailed measurements were made on specimens preserved the option of “Find Best DNA Models”: heuristic method in 95% alcohol or 10% formalin. The technical terms and is Nearest-Neighbor-Interchange (NNI), bootstrap values measurements followed Russell (1990): SL means stan- are computed with 1000 replications. NJ bootstrap values dard length. HL means head length. Distributional data were estimated using 1000 replicates with Kimura’s derived from Russell (1990), FishBase (Froese and Pauly, two-parameter model of substitution (K2P distance) evo- 2013) and the Taiwan Database (Shao, 2013). lution model. The genetic distances were estimated based on Kimura’s two-parameter model using MEGA 5.0 2.3 DNA Extraction, PCR, and Sequencing software. Genomic DNA was extracted from three individuals (IOCAS 135FB00144-1, IOCAS 135FB00144-2, IOCAS 3 Results 135FB00144-3) using Tiangen® Marine Tissue DNA Extraction Kit and following the manufacturer’s 3.1 Synonym instructions. Polymerase chain amplification was con- Nemipterus japonicus (Bloch, 1791) ducted using standard fish primers (Ward et al., 2005) for the barcode region of the COI gene:

COI F1 5′- TCAACCAACCACAAAGACATTGGCAC-3′

COI R1 5′- TAGACTTCTGGGTGGCCAAAGAATCA-3′ Each reaction had a total volume of 25μL containing 1μL of template, 2.5μL of 10× PCR buffer, 1.5μL of Mg2+, 1μL of dNTP mix, 1μL of each primer, 0.3μL of Taq DNA polymerase, and 17.7 μL of dH2O. The thermocycler re- gime was 3 min at 95°C, followed by 35 cycles, each at Fig.1 Nemipterus japonicus (IOCAS 135FB00144-1), 94 °C for 30 s, at 55 °C for 30 s, at 72 °C for 1 min, and 151.5mm SL, City, Island, China. Scale final extension at 72 °C for 10 min. The reaction products bar=1.0 cm. were separated by electrophoresis on 1.0% agarose gel, Sparus japonicas – Bloch, 1791: 110 (original type local- stained with ethidium bromide, and visualized under ul- ity: lacks a designation for the type locality; the type traviolet light. Sequencing reactions were performed at specimen has ambiguous collection data, but probably the Key Laboratory of Experimental Marine Biology Java) (KLEMB) of IOCAS. Three COI sequences had been ?Coryphaena lutea – Schneider in Bloch and Schneider, deposited into the NCBI database with the access num- 1801: 297, fig. 58 (Tranquebar, India) bers from KF134003 to KF134005, corresponding to Cantharus filamentosus – Rüppell, 1828–30 (1829): 50, voucher ID from 135FB00144-1 to 135FB00144-3. pl. 12, fig. 3 (Massaua, Red Sea) Dentex tambulus – Valenciennes in Cuvier and 2.4 COI Sequences Downloaded from GenBank and Valenciennes, 1830: 249, 558 ('Rade de Pondicherry', Checked by BOLD India) Dentex luteus – Valenciennes in Cuvier and Valenciennes, COI sequences for additional specimens were obtained 1830: 250 ( Pondicherry, India) from GenBank: N. japonicus from the Indian Ocean ?Dentex striatus – Valenciennes in Cuvier and (GenBank accession numbers: EF609553-EF609555) Valenciennes, 1830: 252 (Tranquebar, India) (Lakra et al., 2011); N. mesoprion (GenBank accession Spondyliosoma guliminda – (non Valenciennes) Cantor, numbers: EF609557-EF609559); N. virgatus (GenBank 1849: 1032 (Penang) accession numbers: FJ237833-FJ237835); N. hexodon Dentex Blochii – Bleeker, 1851: 176 (Batavia =Jakarta) (GenBank accession number: EF609414); N. peronii Synagris japonicus – Günther, 1859: 378 (Batavia (GenBank accession number: EF609415); N. furcosus =Jakarta) (GenBank accession number: EF609413). A sequence Synagris filamentosus – Günther , 1859: 378 (Red Sea; from eriomma (GenBank accession num- of Pondicherry, India) ber: HQ945943) was used as an outgroup. The COI se- Dentex japonicus – Bleeker, 1865: 173 (Siam) quences from GenBank and those obtained in this study Synagris grammicus – Day, 1865: 14 (Cochin, Malabar were validated on BOLD (Ratnasingham and Hebert, coast of India) 2007) to ensure that specimens were properly identified. Heterognathodon fiaviventris – Steindachner, 1866: 778, p1. 13, fig. 6 (Zanzibar) 2.5 Sequence Alignment and Phylogenetic Analysis Dentex (Heterognathodon) filamentosus – Steindachner, All sequences were aligned using the CLUSTAL W, 1868: 976 (Mauritius) and then truncated to a uniform length by MEGA 5.0. The Nemipterus japonicus – Jordan and Seale, 1907: 21 aligned sequences were subjected to both maximum- (Cavite, Philippines); Akazaki, 1962: 91 (Tonkin Bay, likelihood (ML), and neighbour-joining (NJ) analyses Vietnam; Iran); Cheng, 1962: 505–510 (South China using MEGA 5.0 (Tamura et al., 2011). The model of Sea); Wu, 1985: 187–193 (East China Sea); Lee, 1986:

NING et al. / J. Ocean Univ. China (Oceanic and Coastal Sea Research) 2015 14 3

168, pl. 3, fig. 13 (Kaohsiung, Taiwan); Russell, 1986: Synagris flavolinea) (ANSP 53454), Saukiwan, Hong 19–35 (species catalog, Indian Sea); Li, 1987: 336–337 Kong, China; 1 ex. (110.0 mm SL, paratype of S. fla- (China sea); Russell, 1990: 40, pl. II, b, fig. 72 (Indian volinea) (ANSP 53455), Saukiwan, Hong Kong, China; 1 Ocean and West Pacific); Lee, 1993: 369–371 (Tai- ex. (156.0 mm SL) (ASIZ P055432), Kao- hsiung, Tai- wan); Russell, 1993: 295–310 (eastern Pacific) wan, China . Synagris flavolinea – Fowler, 1931: 299, fig. 8 (Saukiwan, Hong Kong, China) 3.3 Diagnosis Synagris flaviventris – Fowler, 1933: 94 (Philippines; Pectoral fins very long, reaching to or just beyond level Hong Kong, China) of origin of anal fin; pelvic fins moderately long, reaching Euthyopteroma blochi – Herre, 1934: 55 (Manilla, Phil- to or just beyond anus; upper lobe of the caudal fin is ippines) produced into a short or moderately long filament (this character is not obvious in small specimens). Upper part 3.2 Specimens Examined of body pinkish, becoming silvery below; belly silvery Holotype – 1 ex. (146.0 mm SL) (ZMB 8147), lacks a from lower jaw to base of pelvic fin, rest yellow; top of designation for the type locality. head behind eye with a golden sheen; 11 to 12 pale Material examined from the South China Sea – 1 ex. golden-yellow stripes along body from behind head to (191.5 mm SL) (IOCAS 55-1758), Changhua [=Town], base of caudal fin; a red blotch below origin of lateral line; Hainan Island, China; 1 ex. (228.5 mm SL) (IOCAS a pale lemon stripe near base of , this stripe 56-9188), Beibu Gulf (Gulf of Tonkin); 1 ex. (171.5 mm narrows anteriorly and widens on posterior part of fin; SL) (IOCAS 76-018), Sanya [=City], Hainan Island, anal fin whitish with pale lemon broken lines or scribbles China; 1 ex. (147.5 mm SL) (IOCAS 76-333), over most of fin; caudal fin pink, upper tip and filament [=Town], Hainan Island, China; 1 ex. (191.5 mm SL) (IO- yellow. CAS 83-370), Guangzhou [=City], Guangdong Province, China; 1 ex. (142.5 mm SL) (IOCAS JXY-10- 0038), 3.4 Description Beihai [=City], Guangxi Zhuang Autonomous Region Dorsal fin X, 9; anal fin III, 7; pectoral fin 17; pelvic [=Province], China; 1 ex. (144.5 mm SL) (IOCAS JXY- fin i, 5; caudal fin 17; lateral-line scales 43–47; transverse 10-0039), Beihai, Guangxi Zhuang Autonomous Region, scale rows 3/10; gill rakers 14–17. China; 1 ex. (140.5 mm SL) (IOCAS JXY-10-0040), Body depth 2.7–3.4 in SL, head length 3.1–3.5 in SL, Beihai, Guangxi Zhuang Autonomous Region, China; 1 caudal peduncle length 6.1–7.3 in SL, pectoral length ex. (111.5 mm SL) (IOCAS 20101023001), Zhanjiang 2.8–3.3 in SL, pelvic length 3.3–4.1 in SL, dorsal fin base [=City], Guangdong Province, China; 1 ex. (126.5 mm length 1.8–2.0 in SL, anal fin base length 4.9–5.8 in SL, SL) (IOCAS 201010221614), Zhanjiang[=City], Guang- predorsal length 2.5–2.8 in SL, length from the origin of dong Province, China; 1 ex. (97.5 mm SL) (IOCAS pelvic fin to the origin of the anal fin 3.0–3.6 in SL. 201010231005), Beihai, Guangxi Zhuang Autonomous Snout length 3.1–3.9 in HL, eye diameter 3.1–4.0 in Region, China; 1 ex. (151.5 mm) (IOCAS 135FB00144- HL, length of upper jaw 2.7–3.2 in HL, depth of subor- 1), Sanya, Hainan Island, China; 1 ex. (159.5 mm) (IO- bital bone 3.7–5.7 in HL. CAS 135FB00144-2), Sanya, Hainan Island, China; 1 ex. (169.5 mm) (IOCAS 135FB00144-3), Sanya, Hainan 3.5 Coloration Island, China; 1 ex. (142.8 mm) (135FB00144-4), Sanya, Hainan Island, China. Upper part of body pinkish, lower flanks silvery; belly Other materials examined – 1 ex. (171.0 mm SL) silvery from lower jaw to base of pelvic fin, rest yellow; (NTM S. 11003-001), Sunda Strait, Indonesia; 2 ex. top of head behind eye with a golden sheen; 11 to 12 pale (89.5–90.8 mm SL) (BMNH 1880.4.21.42-43), Batavia golden-yellow stripes along body from behind head to (=Jakarta), Java, Indonesia; 2 ex. (121.8–127.0 mm SL) base of caudal fin; an obvious red blotch below origin of (CAS SU 61463), Surubaya, Indonesia; 1 ex. (161.0 mm lateral line; margin of dorsal fin yellow, edged with red; a SL) (BMNH 1984.8.20.10), SE-Java, Indonesia; 1 ex. yellow stripe near base of dorsal fin, this stripe narrow (197.0 mm SL) (NTM S. 10735-005), Bali Strait, anteriorly and widening on posterior part of fin; anal fin Indonesia; 1 ex. (150.7 mm SL) (NTM S. 10678-006), whitish with yellow broken lines or scribbles over most of Kuching, Sarawak, Malaysia; 2 ex. (153.0–181.7 mm SL) fin; pectoral fin translucent pinkish; pelvic fins whitish (NTM S. 10677-014), Kota Kinabulu, Sabah, Malaysia; 1 with yellow axillary scale; caudal fin pink, upper tip and ex. (195.0 mm SL) (SMF 19975), Bangkok, Thailand; 1 filament yellow. ex. (109.7 mm SL) (USNM 112811), Iloilo, Philippines; 1 ex. (142.6 mm SL) (USNM 122158), Corregidor I., 3.6 Distribution Philippines; 1 ex. (170.0 mm SL) (USNM 122148), San Widespread throughout the Indian Ocean and West Pa- Fernando, Philippines; 2 ex. (102.0–132.0 mm SL) (CAS cific ranging from East Africa, including the Persian SU 6100), Tolo Channel, Hong Kong, China; 3 ex. (Arabian) Gulf and Red Sea, to the Indo-Malay Archi- (109.0–174.0 mm) (NTM S. 10676-002), Aberdeen market, pelago. In China, occurs in Beibu Gulf (Gulf of Tokin), Hong Kong, China; 1 ex. (107.0 mm SL, holotype of South China Sea (northern part) and Taiwan Strait.

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more frequent than transversions (sv=19) with an average 3.7 Sequence Characters and Gene Tree for Nemip- ratio (si/sv) of 2.79 (ingroup). terus The gene trees constructed using the two methods showed a similar topology. The species of Nemipterus Clustal W was used to align the three new 655 bp se- were grouped into three major clades by both NJ (Fig.2) quences for N. japonicus. Once aligned with the data and ML (Fig.3) with a bootstrap value higher than 50%: from GenBank, the records produced a 652bp alignment. (1) N. peronii and N. furcosus; (2) N. japonicus and, (3) Of these sites 476 were conserved, 176 were variable, 149 the other three species. Although this analysis strongly were parsimony informative and 27 were singletons. Nu- supported the monophyly of all specimens of N. japoni- cleotide frequencies were A=23.4%, T=31.8%, G= 18.1% cus, two geographical isolates were apparent. and C =26.7%. As expected, transitions (si=54) were

Fig.2 NJ tree based on the analysis of COI sequences from six species of Nemipterus. Bootstrap values are shown near the nodes.

Fig.3 ML tree based on the analysis of COI sequences from six species of Nemipterus. Bootstrap values are shown near the nodes. genetic distance in marine fish species (Ward et al., 3.8 Genetic Distances Between Different Nemipterus 2005). Species Table 1 summarizes the genetic distances between spe- 3.9 Comparison cies of Nemipterus. Interspecific distances ranged from The specimens of N. japonicus from the South China 8.6%–20.0%, while intraspecific distances ranged from Sea and the Indian Ocean are morphologically indistin- 0.0–0.3%, with the exception of N. japonicus where the guishable based on counts and proportional measure- Indian seas specimens showed 2.7% divergence from ments (Table 2). However, there appears be some small those collected in the South China Sea. This divergence but consistent differences in colour pattern, with speci- value is considerably higher than the normal intraspecific mens in the South China Sea having a silver belly, while

NING et al. / J. Ocean Univ. China (Oceanic and Coastal Sea Research) 2015 14 5

Table 1 Genetic distances (K2P model) among six taxa belonging to the genus Nemipterus.

Species group 1 2 3 4 5 6 7 1. N. japonicus, Indian sea 2. N. japonicus, South China sea 2.7% 3. N. mesoprion 15.6% 15.3% 4. N. virgatus 15.6% 15.7% 11.6% 5. N. hexodon 16.6% 17.0% 9.7% 13.1% 6. N. peronii 16.3% 15.4% 18.2% 17.8% 19.8% 7. N. furcosus 17.5% 17.8% 18.2% 16.9% 20.0% 8.6% 8. 17.6% 18.4% 20.9% 22.1% 23.8% 19.8% 19.0% Ingroup distance 0.3% 0.1% 0.1% 0.0% — — — Table 2 Comparison of morphological characters of N. japonicus from the Pacific and Indian Ocean. N. japonicus N. japonicus N. japonicus Holotypea (South China Sea) (western Pacific Ocean) (Indian Ocean) b Measurment SL/ Body depth 2.5 2.7–3.4 2.7–3.5 2.8–3.5 SL/ Head length 2.8 3.1–3.5 3.1–3.5 2.9–3.4 SL/ Dorsal fin length 2.2 1.8–2.0 1.8–2.1 1.8–2.1 SL/ Anal fin length 5.1 4.9–5.8 4.9–5.8 4.8–5.7 HL/ Snout length 4.0 3.1–3.9 2.8–3.9 2.8–3.8 HL/ Suborbital depth 4.9 3.7–5.7 3.7–6.7 4.2–6.7 HL/ Pectoral fin length - 0.9–1.1 0.9–1.3 1.0–1.3 HL/ Pelvic fin length - 1.0–1.3 1.0–1.6 1.2–1.6 snout length/ Diameter of eye 0.9 0.8–1.3 0.8–1.5 0.9–1.5 Diameter of eye/ Suborbital depth 1.3 0.9–1.9 0.9–1.9 1.0–1.9 Peduncle length/ caudal peduncle depth 1.1 1.3–1.5 1.0–1.5 1.0–1.5 Counts Dorsal fin rays X, 9 X, 9 X, 9 X, 9 Anal fin rays III, 7 III, 7 III, 7 III, 7 Pectoral fin rays 18 17 17–18 17–18 Pelvic fin rays i, 5 i, 5 i, 5 - Caudal fin rays 18 17 17 - Lateral-line scales - 43–47 43–48 45–48 Transverse scale rows - 3/10 3/10 3/10 Gill rakers - 14–17 14–17 14–17 a Some measurements can not be made because of the uncomplete fish body. The count data are from the origin description by Bloch (1791). b Data from Russell (1986). those from the Indian Ocean isolate specimen have a yel- mens have been reported from Japan. Russell (1986, 1993) low coloration (Fig.4). noted the distribution of the species in Japan based on five specimens from the Ryukyu Islands (USNM 6368), 4 Discussion but the presence of N. japonicus in Japan is questionable, and it is likely that the Ryukyu specimens might have Sparus japonicus was described by Bloch (1791) based been caught in Taiwan waters for sale in Japan (H. Mo- on a single dried and mounted specimen (Fig.5) deposited tomura pers. com. 2013) . in the Berlin Museum (ZMB 8147). Bloch’s original de- As is the case with many of Bloch’s , it seems scription is brief and his illustration (Bloch, 1791: plate likely that Nemipterus japonicus came from East Indies 277, fig.1. shown here as Fig.6) is inaccurate in some (Java). As pointed out by Paepke (1999: 27), many of respects (eg the lateral line scale count is incorrect, it Bloch’s species represent tropical species unknown in lacks a filamentous extension to the upper caudal lobe, Japanese waters. Indeed, the vernacular ‘Japanese’ names and the greenish body color is not seen in life). However, of several fishes mentioned by Bloch are based on the the length of the pectoral and pelvic fins and the many Malay name ‘ikan’ for fish (Pethiyagoda and Gill, 2012). yellow stripes along body from behind the head to the Cuvier (1828: 100) observed that though Bloch believed base of caudal fin are characteristic of N. japonicus and his specimens to be of Japanese origin, he might have together distinguish it from all other species of Nemip- been misled by the Dutch traders who sold him the terus. specimens. Bloch (1791) failed to mention where the type speci- The 5’ region of the cytochrome c oxidase subunit I men was collected, although the name japonicus suggests (COI) has been adopted as the standard DNA barcode its origin from Japanese waters. However, in the more region for members of the animal kingdom. It has a than two centuries since its description, very few speci- strong track record in revealing cryptic species (Hebert et

6 NING et al. / J. Ocean Univ. China (Oceanic and Coastal Sea Research) 2015 14 al., 2004; Ward et al., 2005). We used this gene region to terns in other Indo-Pacific marine fishes clearly indicate examine genetic diversity in N. japonicus. The NJ and the role of shifting sea levels linked to climatic oscilla- ML trees both strongly support the conclusion that indi- tions during the Pleistocene as an important cause of viduals of this species form two geographical isolates population isolation and differentiation (Gaither et al., (South China Sea, Indian Ocean) with 2.7% divergence. 2011). In this study, the level of divergence between re- gional populations of N. japonicus suggests that gene flow between populations of N. japonicus on the conti- nental shelves of the Pacific Ocean and the Indian Ocean was interrupted by the Indo-Pacific Barrier (Fleminger, 1986) during Pleistocene sea level fluctuations. Further effort is needed to carry out an extensive collaborative study using a multi-genic analysis to examine the phy- logeny of a wide range of Nemipterus species and geo- graphical isolates. Work also is needed to ascertain the respective geographical distributions of the West Pacific and Indian Ocean populations of Nemipterus japonicus.

Acknowledgements Fig.4 Nemipterus japonicus: A, NPPF1159, 90.4mm SL, This investigation was supported by IOCAS funding Nayband National Park Coast, Bushehr, Iran, one of specimens from the Indian Ocean examined by Asgharian (2012IO060102). We are very grateful to Dr. Peter et al. 2011. Scale bar=2.0 cm; B, NPPF 1160, 88.5mm SL, Bartsch, Curator of Fishes at the Museum fűr Naturkunde, Nayband National Park Coast, Bushehr, Iran, one of Berlin, and to Professor Michael Tuerkay, Senckenberg specimens from the Indian Ocean examined by Asgharian Forschungsinstitut, Frankfurt, for providing a photograph et al. 2011. Scale bar=2.0 cm; C, IOCAS 135FB00144-4, of the holotype ZMB 8147, a copy of Bloch’s original 142.8mm SL, Sanya City, Hainan Island, China. Scale description and other help with this paper. Dr Hiromitsu bar=1.0 cm. The belly’s color from lower jaw to base of Motomura, Kagoshima University Museum, Japan, pro- pelvic fin (1, 2) is silver in the South China Sea speci- vided information and photographs of specimens from the mens, while yellow in the Indian Ocean specimens; The Philippines. We also thank Lian-Mei Shuai of the Marine belly’s color from base of pelvic fin to caudal peduncle (3) is yellow in both two form of specimens. The pictures of Biological Museum, Chinese Academy of Sciences for A and B are deposited in BOLD. her help in sorting the fish specimens. Prof. Jin-Xian Liu provided helpful suggestions on this paper, while Dr. Wei Jiang provided the photographs of the specimens from Hainan Island. Finally, we thank Drs. Chuan-Yan Yang, and Da-Hai Gao for their aid on various aspects of this study.

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