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This is the first report of Fowlerichthys scriptissimus (Lophiiformes, Antennariidae) from Korea. A single specimen (291.0 mm SL) was collected off the coast of Jejudo Island by gill net on 28 March 2012 and identified with morphological and molecular approaches. The specimen is characterized by having all five pelvic fin rays bifurcate and possessing 20 vertebrae, 13 pectoral-fin rays, and a basidorsal ocellus on the side of the body. This species is distinguishable from other Korean taxa by the number of pectoral fin rays, the bifurcate form of the pelvic rays, and the vertebral count. We add this species to the Korean fish fauna and suggest new Korean names, “Byeol-ssin-beng-i-sok” and “Byeol-ssin-beng-i” for the genus and species, respectively. Keywords: Fowlerichthys scriptissimus, Antennariidae, New record, Jejudo Island, Korea Background A single specimen of Fowlerichthys scriptissimus,which The frogfishes (Antennariidae), which belong to order is otherwise unknown from Korea, was collected in the Lophiiformes, occur in all tropical and subtropical seas ex- coastal waters of Jejudo Island by gill net on 28 March cept the Mediterranean Sea (Nelson 2006). Worldwide, the 2012. Here, we describe the morphological characters of family includes 46 species in 13 genera (Arnold and Pietsch F. scriptissimus and report the results of a molecular 2012) and four species in two genera in Korea (Kim et al. barcode determination of the specimen’s identification 2005; Kim et al. 2011). All members of Antenariidae have using the COI gene. the first dorsal fin spine modified into a fishing pole (illicium) and gill openings below or behind the base of the Methods pectoral fin (Nelson 2006; Arnold and Pietsch 2012). The Counts and measurements followed the method of genus Fowlerichthys was originally suggested by Barbour Hubbs and Lagler (1964). When conducting this study, (1941) when he described the new species, Fowlerichthys we adhered to the ethical guideline of the International floridanus. However, as some of his morphological Council for Laboratory Animal Science (ICLAS) for descriptions were not clear, most ichthyologists regarded researchers. The specimen was fixed in 10% buffered the genus Fowlerichthys as a synonym of Antennarius formalin and then transferred to 70% ethanol. Vertebrae (Pietsch 1984; Senou 2002; Manilo and Bogorodsky 2003). were counted from radiographs (REX-525R, listem). The Later, Arnold and Pietsch (2012) reconstructed evolution- examined specimen was deposited at the Jeju National ary relationships within Antennariidae with molecular University (JNU), Korea, and is available from the phylogenetics and recognized that Fowlerichthys as a valid corresponding author by reasonable request. genus. They demonstrated that it can be separated from Total DNA was extracted from 25 mg of the muscle Antennarius by having all five pelvic fin rays bifurcate tissue with an AccuPrep Genomic DNA Extraction Kit instead of one in the genus Antennarius. (Bioneer Inc.) according to manufacturer’s protocol. The mitochondrial cytochrome c oxidase subunit I (COI) gene was PCR-amplified with the primers, Asn-F1 (AAA HWC * Correspondence: firstname.lastname@example.org College of Ocean Sciences, Jeju National University, Jeju 63243, Korea Full list of author information is available at the end of the article © The Author(s). 2017 Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. Han et al. Fisheries and Aquatic Sciences (2017) 20:2 Page 2 of 5 TTA GTT AAC AGC TAA) and Ser-R1 (GGG GTT CDA 128 (Rarotanga, Cook Islands); Stewart 2015, 886 YTC CYC CCT TTC T). The polymerase chain reaction (New Zealand). (PCR) was performed on a thermal cycler (TP600, Takara The genus Fowlerichthys includes five species diagnosed Bio Inc.) with a final volume of 40 μL in a 0.2 mL PCR by the following combination of characters: one or three tube containing 10 mM Tris-HCl (pH 8.3), 50 mM KCl, darkly pigmented ocelli on side of body; illicium about as 2 mM MgCl2, 0.2 mM dNTP mix, 0.5 mM of each primer, long as second dorsal spine; esca in form of tuft of slender 50 ng template DNA solution, and 1 U Ex Taq DNA filaments, a simple, oval-shaped appendage, escal pigment polymerase (Takara Bio Inc.) The PCR cycles consisted of spots absent; dorsal rays 12–14 (usually 13); pectoral rays an initial denaturing step of 94 °C for 2 min, followed by 11–14 (usually 12 or 13); all five pelvic rays bifurcate; anal 30 cycles of 30 s at 94 °C for denaturation, 1 min at 54 °C rays 7–10 (usually 8), all bifurcate; vertebrae 20 for primer annealing, and 1 min and 50 s at 72 °C for (Pietsch and Grobecker 1987; Arnold and Pietsch 2012). extension, and an additional 7 min interval at 72 °C for a final extension. The PCR products were sequenced using F. scriptissimus (Jordan 1902) the BigDye Terminator v3.1 Cycle Sequencing Kit (New Korean name, Byeol-ssin-beng-i) (Table 1; Fig. 1) (Applied Biosystems Inc.) and ABI PrismTM 3730XL Antennarius scriptissimus Jordan, 1902, 373 (type DNA Analyzer. Sequencing reactions were conducted locality, Bōsō Peninsula, Chiba Prefecture, Japan); with the amplification primers in two directions. The Pietsch 2000, 597 (South China Sea); Senou 2002, 456 DNA sequence obtained was deposited in the National (Japan); Manilo and Bogorodsky 2003, S99 (Oman). Center for Biotechnology Information (NCBI) as GenBank Antennarius sarasa Pietsch 1984, 36 (Japan); Araga accession number, KY195977. For molecular identifica- 1984, 103 (Japan); Pietsch and Grobecker 1987, 123 tion, we compared the specimen’s COI sequence with the (Réunion Island, New Zealand, and Philippines). GenBank DNA sequences of the seven anglerfish species: F. scriptissimus Arnold and Pietsch 2012, 128 (Rarotanga, Antennarius hispidus (FJ582855), Antennarius pictus Cook Islands) (FJ582858), Antennarius striatus (AB282828), Histrio histrio (AB282829), Fowlerichthys avalonis (DQ0279840), Material examined F. scriptissimus (GU188480), and Lophius litulon JNU-637, one specimen, 291.0 mm in standard length (KJ020931). The DNA sequences were aligned and (SL), gill net, Hanlim-eup, Jeju-si, Jejudo Island, Korea, edited using Clustal W (Thompson et al. 1994) and 28 March 2012. BioEdit version 7 (Hall 1999). Genetic distances were calculated using MEGA 6 (Tamura et al. 2013) based on the Kimura two-parameter (K2P) model (Kimura Description 1980). A neighbor-joining (NJ) tree was constructed Meristic counts appear in Table 1. Measurements as a using the K2P model and 10,000 bootstrap replications percentage of SL are as follows: body depth 67.6; body in MEGA 6. width 21.3; snout length 24.0; eye diameter 5.1; interor- bital length 14.7; illicium length 6.9; first predorsal fin Results length 18.3; second predorsal fin length 20.1; third Genus Fowlerichthys Barbour 1941 predorsal fin length 35.5; prepectoral fin length 36.7; (New Korean genus name, Byeol-ssin-beng-i-sok) preanal fin length 67.3; prepelvic fin length 9.5; length of Fowlerichthys Barbour 1941; 12 (type species, Fowlerichthys second dorsal spine 8.4; length of third dorsal spine 10.8; floridanus Barbour 1941). Arnold and Pietsch 2012, length of pectoral fin ray 13.0; length of anal fin ray Table 1 Morphological characters compared between the present specimen and previous studies on F. scriptissimus Morphological characters Present study Jordon (1902) Araga (1984) Pietsch and Grobecker (1987) Senou (2002) Total length (mm) 350.0 (n =1) – 350.0 (n =1) - (n =5) – Standard length (mm) 291.0 –– 99.0–280.0 280.0 Count Dorsal fin rays (bifurcate rays) 13 (7) III-12 III-13 13 (7–9) 13 Pectoral fin rays 13 – 13 13 13 Pelvic fins rays I, 5 – 5 5 I, 5 Anal fin rays 8 8 8 8 8 Caudal fin rays 9 –– 99 Han et al. Fisheries and Aquatic Sciences (2017) 20:2 Page 3 of 5 Fig. 1 F. scriptissimus (Jordan 1902), JNU-637, 291.0 mm SL, gill net, Fig. 2 Radiograph of JNU-637, F. scriptissimus. Scale bar =50 mm Hanlim-eup, Jeju-si, Jejudo Island, Korea. Scale bar =50 mm 12.5; caudal peduncle depth 12.1; and caudal peduncle Molecular identification length 9.2. To confirm and verify the accuracy of the morphological Body round and compressed; skin of body very species identification, we also analyzed 634 base pairs of rough and covered with close-set dermal spinules; the mitochondrial COI gene. The DNA sequence of the head and eyes small; eyes lateral; mouth large and ex- specimen was almost identical to a previously published tremely oblique; first dorsal spine (illicium) shorter sequence (GU188480) of F. scriptissimus (genetic than second spine; esca simple and oval-shaped with distance, d = 0.003). The NJ tree also clustered the speci- a tuft; second and third dorsal spine rough and men with the GenBank sample of F. scriptissimus, with a curved posteriorly; gill opening located below base of strong 100% bootstrap value (Fig. 4). pectoral fin; depression between second and third dorsal spines; membrane behind second dorsal spine Discussion extending posteriorly, dividing area between second The specimen in question has all the previously reported and third dorsal spines and nearly reaching to base of diagnostic morphological characters of F. scriptissimus, third (Fig. 3). including five bifurcate pelvic fin rays, 20 vertebrae, 13 pectoral fin rays, and a basidorsal ocellus on the side of the body (Table 1 and Fig. 2). The form of the esca as a Live coloration simple, oval-shaped appendage with numerous, more or Whole body uniformly greenish brown; entire head, less parallel, vertically aligned folds also matches the body, and fins with mottled dark brown reticulations morphology of F. scriptissimus. Additionally, the form of except for inner surface of paired fins; a single, darkly the membrane behind the second dorsal spine matches pigmented, basidorsal ocellus on each side of body; F. scriptissimus, by lacking division into naked dorsal scattered beige spots on body; esca with dark pigment at base. Color in preservative Body uniformly pale greenish brown; mottled dark brown spots and ocellus; beige spots no longer apparent. Distribution Widely known from Indo-West Pacific, including Réunion Island, New Zealand, the Philippines (Pietsch and Grobecker 1987), the Cook Islands (Arnold and Fig. 3 Morphology of the dorsal fin spines of JNU-637, F. scriptissimus, showing the pattern of dermal spinules on the membrane behind Pietsch 2012), the South China Sea (Pietsch 2000), Japan the second spine (A) and membrane dividing the area between the (Jordan 1902; Pietsch 1984; Araga 1984), and Korea second and third dorsal spines (B) (Jejudo Island, present study). Han et al. Fisheries and Aquatic Sciences (2017) 20:2 Page 4 of 5 Fig. 4 Neighbor-joining tree showing the relationship of the specimen with the seven species of anglerfishes. Numbers above nodes indicate bootstrap probabilities based on 10,000 replications. Bar indicates a K2P genetic distance of 0.02 and ventral portions, extending posteriorly to the area four simple and one bifurcate rays). It can be further between the second and third dorsal spines, and nearly separated from all members of Antennarius except reaching to the base of the third spine (Fig. 3). Thus, the Antennarius commerson by possessing 20 vertebrae morphological characteristics of the specimen clearly fit (Table 2). Currently, the genus Fowlerichthys is the species descriptions given by previous studies recognized as valid, with five known species worldwide (Table 1). In the molecular analysis of the mitochondrial Fowlerichthys radiosus, F. avalonis, Fowlerichthys COI gene, the small genetic distance (d = 0.003) between senegalensis, Fowlerichthys ocellatus,and F. scriptissimus the new specimen and a previously sequenced specimen (Froese and Pauly 2016). of F. scriptissimus, and the close clustering of those two F. scriptissimus is easily distinguished from the other samples in a neighbor-joining tree (Fig. 4) including the Korean frogfishes (A. pictus, A. commerson, A. macula- seven anglerfish species confirmed the morphological tus, and H. histrio) by having 13 pectoral fin rays (vs. 10 identification. Thus, both morphological and molecular in A. commerson,11in A. maculatus,10in A. pictus, approaches indicated the specimen in the present study and 10 in H. histrio), five bifurcate pelvic rays (vs. one in to be F. scriptissimus. the genus Antennarius and none in H. histrio), and 20 The genus Fowlerichthys was firstly suggested when vertebrae (vs. 19 in H. histrio and all members of Barbour (1941) described Fowlerichthys floridanus, but Antennarius except A. commerson) (Table 2). his description and diagnosis of the genus were unclear. We propose new Korean names, “Byeol-ssin-beng-i-sok” He mentioned that the strong and sharp dorsal spines of and “Byeol-ssin-beng-i” for the genus and species, the species were unlike the dorsal fin rays in the species respectively. The Korean name “Byeol-ssin-beng-i” of Antennarius, but whether these spines were bare in was given for F. scriptissimus because of its scattered life is difficult now to determine certainly. Thus, most beigespots on thebodythatlookalikestars.The ichthyologists have regarded the genus Fowlerichthys as Korean words “byeol” and “ssin-beng-i” mean star a synonym of Antennarius. Recently, Arnold and Pietsch and frogfish, respectively. (2012) demonstrated that Fowlerichthys is a valid genus that can be separated from Antennarius by having all Conclusions five pelvic fin rays bifurcate (all other antennariids have Not applicable. Table 2 Comparison in the number of pectoral fin rays, bifurcate pelvic fin rays, and vertebrae among five frogfish species inhabiting Korea Species Pectoral fin rays Bifurcate pelvic rays Vertebrae References F. scriptissimus 13 5 20 Pietsch and Grobecker (1987) A. commerson 10–11 (usually 11) 1 19–20 (usually 19) A. maculatus 10–11 (usually 10) 1 19 A. pictus 9–11 (usually 10) 1 19 H. histrio 9–11 (usually 10) 0 18–19 (usually 19) Han et al. Fisheries and Aquatic Sciences (2017) 20:2 Page 5 of 5 Abbreviations Senou H. Antennariidae. In: Nakabo T, editor. Fishes of Japan with pictorial keys COI: Cytochrome c oxidase subunit 1; d: Genetic distance to the species. Tokyo: Tokai Univ Press; 2002. p. 454–8. Stewart AL. Family Antennariidae. In: Roberts CD, Stewart AL, Struthers CD, editors. The Fishes of New Zealand, vol. 3. 2015. p. 577–1152. Acknowledgements Tamura K, Stecher G, Peterson D, Filipski A, Kumar S. MEGA6: molecular This work was supported by a grant from the National Institute of Fisheries evolutionary genetics analysis version 6.0. Mol Bio Evol. 2013;30:2725–9. Science (R2016034). We thank Prof. B. Sidlauskas, Oregon State University, for Thompson JD, Higgins DG, Gibson TJ. CLUSTAL W: improving the sensitivity of the valuable discussion. progressive multiple sequence alignment through sequence weighting, position specific gap penalties and weight matrix choice. Nucl Acids Res. Funding 1994;22:4673–80. This study was funded by a grant from the National Institute of Fisheries Science (R2016034). Availability of data and materials All datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request. Authors’ contributions SHH performed experiment and wrote the manuscript. JSK collected the sample. CBS conceived of the study and helped to write the manuscript. All authors read and approved the final manuscript. Competing interests The authors declare that they have no competing interests. Consent for publication Not applicable. Ethics approval and consent to participate Not applicable. Author details Jeju Fisheries Research Institute, National Institute of Fisheries Science, Jeju 63068, Korea. Korea Fisheries Resources Agency, Jeju Branch, Jeju 63005, Korea. College of Ocean Sciences, Jeju National University, Jeju 63243, Korea. Received: 10 August 2016 Accepted: 2 February 2017 References Araga C. Family Antennarioidae. In: Masuda H, Amaoka K, Araga C, Uyeno U, Yoshino T, editors. 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