A New Species of Deogyu Fat-minnow, Rhynchocypris deogyuensis (Cypriniformes: Cyprinidae) from Korea
Seung-Rok Lee
1* and Jae-Hwan Sim
21Chiaksan National Park Office, National Park Service, Wonju 26304, Korea
2Seoyeong University, Physical Therapy, Gwangju 61268, Korea
한국산 잉어과 버들치속 어류 1 신종, Rhynchocypris deogyuensis
이승록
1*·심재환21국립공원관리공단 치악산국립공원사무소
2서영대학교 물리치료과
Abstract :A new species of deogyu fat-minnow, Rhynchocypris deogyuensis, is described from the Geum River in Korea. This species is distinguished from Rhynchocypris kumgangensis by the following characteristics: shorter interorbital width; shorter eye diameter; longer caudal peduncle depth; number of scales above lateral line; and black transverse bar on the first unbranched ray in dorsal fin. The lower band of yellowish longitudinal two bands on lateral body is interrupted and unsharpened. Numerous silverly white spots are on blackish dorso-lateral body. Analyses of 1,141 bp length of mitochondrial Cyt b gene proved that, R. deogyuensis is clearly distinguished from R. kumgangensis in that the former showed a high level (94 bootstrap) of reliability.
Key words : New species, Rhynchocypris deogyuensis, Fat-minnow, Cyprinidae, Geum River
요 약 :한국의 금강수계에 서식하는 본 종을 Rhynchocypris deogyuensis (국명신칭: 덕유모치)로 신종 기재하였다. R.
deogyuensis는 짧은 양안간격, 짧은 안경, 높은 미병고, 측선상부린수 및 등지느러미 제1 불분지연조의 흑색반문 등의 분류형질에 의해서 R. kumgangensis와 구분된다. 몸의 측면을 따라 있는 2줄의 황갈색 세로무늬에서 아래 줄은 비연속 적이고 불분명하다. 옅은 검은색의 체측상단부에는 다수의 은백색 반점이 있다. 미토콘드리아 Cyt b 유전자 1,141 bp 서 열 분석에서도 R. deogyuensis는 높은 수치의 신뢰도(94 bootstrap)를 나타내면서 R. kumgangensis와 명확하게 구분된다.
주요어 :신종, Rhynchocypris deogyuensis(국명신칭: 덕유모치), Fat-minnow, 잉어과, 금강
Introduction
The subfamily Leuciscinae, including about 68 genera, are small fish distributed in the North America, Eurasia except India and Southeast Asia (Nelson 2006). In the Korean Peninsula, 6 genera and 11 species have been recorded (Kim 1997; Youn 2002; Kim et al. 2005). Among them, Rhyncho- cypris kumgangensis is endemic species, which inhabits middle and upper stream of the Han River and the Geum River in Korea (Kim 1997; Youn 2002). For the first time, Uchida (1939) recorded information on morphometric characters, ecology, life cycle and distribution of the species
under the scientific name “Moroco sp.” based on specimens from the Yalu River, the Jeokbyeok River and the Han River.
Kim (1980) reported a new species as Phoxinus kumgangensis on the basis of specimens collected from the Daedong River in North Korea. The scientific name of R. kumgangensis has been named differently by many ichthyologists (Chyung 1961; Chyung 1977; Kim 1980; Kim et al. 1985; Kim and Kang 1986; Kim 1997; Youn 2002; Kim et al. 2005). Suh and Yang (1989) provided genetic and morphological variation of Moroco sp. by isozyme, general protein and morphometric characters.
In this study, thirty-eight specimens of Rhynchocypris were
*Corresponding author E-mail: LSL7007@knps.or.kr
collected from the upper stream of the Geum River and compared with original description of congeneric species and the Han River populations. The findings include differentia- tion in measurements and meristic count characters, body coloration and black transverse bar in dorsal-fin. In this paper, we define Rhynchocypris as a distinct new species by comparing it with R. kumgangensis.
Materials and Methods
Specimens were collected from the middle and upper streams of the Han River and the Geum River in Korea from April to June 2013. The specimens were fixed in 10%
formalin after taking body pictures. The counts and measure- ments of the specimens were followed by Hubber and Lager (1964) and Kim (1997). The measurement characters were made with dial calipers (nearest 0.1 mm). The number of fin rays and band patterns of dorsal-fin were counted from microscope (Olympus, SZH10, Japan). Specimens examined were deposited at National Park Research Institute (NPRI).
Genomic DNA was extracted from dorsal muscle tissues of specimens using the DNeasy Blood & Tissue Kit (QIAGEN Co., USA).
The fragments of mitochondrial Cyt b gene sequences were amplified by polymerase chain reaction(PCR) using the primer set (npsf3: forward 5'-ACC GAA ACC TGT GAT TTG AAG A-3', reverse 5'-ACC ACA AAT CCG AGC AAA TCT-3', npsf5: forward 5'-ACA CGA AAC CGG ATC AAA CA-3', reverse 5'-AAA TCT CCT TCT TTT GGG CG- 3'). The reactions included Primix 10 µl, Genomic DNA 1 µl, Primer 10 pmol forward 1 µl, Primer 10 pmol reverse 1 µl, D/W 7 µl (G-Tag PCRPrimix Kit, COSMO geneteck, Korea). The temperature profile for amplifying Cyt b gene
sequences was as follows: denaturation at 95oC for 10 min, 35 cycles of 95oC for 30 sec, 53oC for 30 sec, 73oC for 1 min;
final extension of 72oC for 10 min. PCR products were electrophoresed on a 1% agarose gel and purified with a PCR purification Kit (COSMO geneteck, Korea).
The mitochondrial Cyt b gene sequenced were aligned using Clustal × 1.8 (Thompson et al. 1997). Neighbor Joining (NJ) analysis was performed as implemented in MEGA 5.0 (Tamura et al. 2011). NJ tree was inferred using Kimura's 2- parameter model (Kimura 1980), with bootstrapping searches of 1,000 replicates.
Rhynchocypris deogyuensis sp. nov. (Figure 1) (English name: Deogyu fat-minnow)
(New Korean name: Deog-yu-mo-chi, 국명신칭: 덕유모치) Holotype. NPRI 148, male, 68.9 mm SL (Standard Length);
Deogyusan National Park, Geum River, Samgong-ri, Solcheon- myon, Muju-gun, Jeollabuk-do, Korea, April 25, 2013, Dr.
S.R. Lee and Dr. J.H. Sim.
Paratypes. NPRI 111-147(37), 58.1-74.9 mm SL; Same data as holotype.
Diagnosis. Rhynchocypris deogyuensis sp. nov. is distin- guished from R. kumgangensis by the following characteristics:
shorter interorbital width (22.7-27.6% HL); shorter eye diameter (21.3-26.2% HL); longer caudal peduncle depth (10.1-13.6% SL); number of scales above lateral line (12-15);
black transverse bar on the first unbranched ray in dorsal fin (Figure 3A). The lower band of yellowish longitudinal two bands on lateral body is interrupted and unsharpened.
Numerous silverly white spots are on blackish dorso-lateral body.
Description. Meristic and morphometric data for holotype and paratypes summarized in Table 1. Dorsal fin rays iii, 7;
Figure 1. Rhynchocypris deogyuensis sp. nov. Holotype from the Geum River, Muju, Korea. NPRI 148, male, 68.9 mm SL. Scale bar=10 mm.
anal fin rays iii, 8; lateral line scales 53-61; scales above lateral line 12-15; gill raker on first arch usually 6-8;
proportion of % SL: head length 25.1-28.0 (mean 26.7);
body depth 19.6-26.5 (mean 21.9); body width 9.9-14.4 (mean 11.8), length of predorsal 51.1-56.5 (mean 53.6), length of preanal 59.8-67.6 (mean 63.3); length of prepelvic 41.0-48.0 (mean 44.5); length of prepectoral 22.5-26.5 (mean 24.1); caudal peduncle length 22.6-27.5 (mean 25.3), caudal peduncle depth 10.1-13.6 (mean 12.1); proportion of % HL:
snout length 29.1-36.8 (mean 31.5); eye diameter 21.3-26.2 (mean 23.5), interorbital width 22.7-27.6 (mean 26.2). Body is small, elongated and compressed. Head is moderate depressed, snout is produced, mouth is inferior, upper jaw is protruded than lower jaw without barbel. Posterior margin of operculum is slightly roundish. Eye moderate size. Eye has
diameter is smaller than interorbital width. Dorsal fin is posteriorly located, its origin posterior to pelvic fin origin.
Posterior margin of caudal fin is forked. Lateral line is complete, well developed, running along the middle of the body. Scales are cycloid, covering entire body, absent from head. Scales of breast and belly are embedded in flesh, difficult to detect. Dorsal fin has a broad black transverse bar which is interrupted posteriorly. Black transverse bar on the first unbranched ray in dorsal fin (Figure 2A-a, Figure 3A).
Dorsal half blackish and ventral half grayish, with yellowish longitudinal two bands on lateral body. Upper band is extended from the posterior margin of operculum to the base of caudal fin. Lower band is extended from the origin of pectoral fin to the base of anal fin. Lower band is interrupted and unsharpened.
Table 1. Morphometric and meristic data of Rhynchocypris deogyuensis and R. kumgangensis.
Characters R. deogyuensis sp. nov. R. kumgangensis R. kumgangensis P. kumgangensis Moroco sp.
Present study Present study Kim(1997) Kim(1980) Uchida(1939)
Type status Holotype Paratype − − Holotype −
Number 1 37 110 − − 15
Standard length(mm) 68.9 58.1-74.9 42.1-80.2 − − 32.0-76.0
In % of standard length
Head length 26.0 26.7±0.7
(25.1-28.0)
26.1±1.0
(23.1-29.2) 24.2-27.4 − −
Body depth 22.3 21.9±1.4
(19.6-26.5)
21.7±1.9
(17.5-25.8) 22.1-25.8 − −
Body width 12.6 11.8±1.2
(9.9-14.4)
13.3±1.48
(9.8-16.8) − − −
Length of Predorsal 52.4 53.6±1.4 (51.1-56.5)
54.2±1.4
(50.5-58.0) 49.0-53.8 − −
Length of Preanal 60.3 63.3±1.8 (59.8-67.6)
63.7±1.5
(58.9-68.3) − − −
Length of Prepelvic 43.5 44.5±1.8 (41.0-48.0)
46.5±1.2
(43.6-51.8) − − −
Length of Prepectoral 23.4 24.1±0.9 (22.5-26.5)
24.3±0.9
(21.7-26.9) − − −
Caudal peduncle length 26.0 25.3±1.2 (22.6-27.5)
25.1±1.2
(21.5-28.5) 24.6-28.5 − −
Caudal peduncle depth 12.2 12.1±0.6 (10.1-13.6)
11.2±0.5
(8.9-13.0) 10.6-12.6 − −
In % of head length
Snout length 33.0 31.5±1.3
(29.1-36.8)
31.1±1.2
(28.3-34.5) 31.0-35.4 − −
Eye diameter 23.4 23.5±1.2
(21.3-26.2)
26.9±2.0
(23.1-35.6) 23.0-27.2 − −
Interorbital width 27.3 26.2±0.9 (22.7-27.6)
31.3±1.8
(28.2-40.5) 29.7-32.4 − −
No. of dorsal fin ray ⅲ, 7 ⅲ, 7 ⅲ, 7 ⅲ, 7 ⅲ, 6-7 ⅲ, 6-7
No. of anal fin ray ⅲ, 8 ⅲ, 8 ⅲ, 8 ⅲ, 7-8 ⅲ, 7-8 ⅲ, 7-8
No. of lateral line scale 56 56.4±1.9 (53-61)
56.7±2.3
(50-63) 59-66 58-65 58-65
No. of scales above lateral
line 14 13.5±0.9
(12-15)
12.2±0.7
(10-15) 12-14 − −
No. of Gill raker 7 6.8±0.5
(6-8)
6.7±0.6
(5-8) 7-9 6-8 6-10
Distribution. Rhynchocypris deogyuensis sp. nov. is restricted to the middle and upper stream of the Geum River, Korea:
Samgong-ri, Solcheon-myon, Muju-gun, Jeollabuk-do (Figure 4).
Sexual dimorphism. During the spawning season, upper and lower yellowish bands on lateral body of male are brighter than those of female. Urogenital papilla of male extend beyond anus. Breeding tubercles in both sexes are absent.
Coloration. In life, the side body above the lateral line is blackish, and below the lateral line and ventral grayish.
Blackish dorsal region has numerous silverly white spots (Figure 2A-b). Suborbital and operculum region is grayish white (little more bright and yellowish in breeding season) (Figure 2A-d). Longitudinal two bands of lateral body are slightly yellowish. The color of lateral body of between longitudinal two bands is slightly grayish. Yellowish upper band is extended from the posterior margin of operculum to the base of caudal fin, but the lower band is unsharpened and is extended from the origin of pectoral fin to the base of anal fin (Figure 2A-c). In breeding season, the longitudinal two bands are turned brighter. The male has brighter yellow pigmentation on the lateral and ventral portion of the body
than female. The color of the origin of all fins is yellowish (Figure 2).
Etymology. The name deogyuensis, distributed in Deogyusan National Park, refers to the geographical occurrence of the species in Korea.
Remarks
The genus Rhynchocypris is represented by five species in Korea: R. kumgangensis (Kim), R. oxycephalus (Sauvage and Dabry), R. steindachneri (Dybowski), R. percnurus (Pallas) and R. semotilus (Jordan and Starks). Among them, R.
kumgangensis has the closest resemblance with R. deogyuensis sp. nov. in the external morphology. But it is distinguished significantly from meristics, morphometrics and pigmentation.
The species are further distinguished by relatively shorter interorbital width (22.7-27.6% HL in R. deogyuensis vs.
28.2-40.5% HL in R. kumgangensis); shorter eye diameter (21.3-26.2 % HL vs. 23.1-35.6); longer caudal peduncle depth (10.1-13.6% SL vs. 8.9-13.0% SL); number of scales above lateral line (12-15 vs. 10-15); black transverse bar on the first unbranched ray of dorsal fin in R. deogyuensis vs. no Figure 2. Nuptial coloration, A; Rhynchocypris deogyuensis, 68.9 mm, SL, male, collected April 25, 2013, from the Muju. B; R.
kumgangensis, 68.2 mm, SL, male, collected April 16, 2013, from the Pyeongchang. Scale bar=10 mm.
black bar on the 1st to 3rd unbranched ray of dorsal fin in R.
kumgangensis (Figure 3); The lower band of yellowish longitudinal two bands on lateral body is interrupted and unsharpened (vs. complete and bright in R. kumgangensis).
Numerous silverly white spots are on blackish dorso-lateral body (absent in R. kumgangensis).
R. kumgangensis was distributed in upper stream which drains into Yellow Sea of the Korean Peninsula, while other species of the congener were inhabitated some other rivers:
R. oxycephalus in the Han River, the Nakdong River, the Mangyeong River, the Geum River and the Semjin River. R.
percnurus in the Bukdae Stream and the Seongcheon River, R. steindachneri in the Namdae Stream and drained into the East Sea of Korean Peninsula, R. semotilus in the Songhyeon Stream, the Nam River and the Jeokbyeok River (Uchida 1939; Jeon 1980; Kim 1997; Youn 2002). But R. deogyuensis is found yet only in the Geum River, showing the most limited distribution among the fishes of the Rhynchocypris.
A 1,141bp length of the the Cyt b gene were analyzed. As a result, twenty-seven haplotypes were defined. According to the NJ (Neighbor Joining) analysis, Rhynchocypris populations revealed clearly two separated lineages. R. deogyuensis was clearly distinguished from R. kumgangensis (Figure 5). The genetic difference between R. kumgangensis (the populations of the Han River) and R. deogyuensis (the Geum River) identified preceding research analysis (Lee and Kim 2016), DNA polymorphism between the populations of Han River and the population of Geum River was 27 to 73bp (2.4-6.4%) and the identified pairwise genetic distance was 0.029 to
0.036.
Key to the species of the genus Rhynchocypris in Korea
1a. No black band on Dorsal fin···2 b. Black band on Dorsal fin ···4 2a. Eye diameter 25.6-27.8(proportion of % HL), body depth 27.0-31.3(proportion of % SL)··· R. percnurus b. Eye diameter 15.4-21.1(proportion of % HL), body depth 19.6-23.6(proportion of % SL) ···3 3a. Lateral line scales 80-83, scales above lateral line 23-28
··· R. steindachneri b. Lateral line scales 72-78, scales above lateral line 18-22
··· R. oxycephalus 4a. No longitudinal band on lateral body··· R. semotilus b. Longitudinal two bands on lateral body···5 5a. Interorbital width 28.2-40.5(proportion of % HL), Figure 3. Two types black transverse bars of dorsal fin between
Rhynchocypris deogyuensis and R. kumgangensis; A type (Geum River type), B type (Han River type).
Figure 4. Map showing distribution of the Rhynchocypris deogyuensis and R. kumgengensis in Korea Peninsula.
caudal peduncle depth 8.9-13.0(proportion of % SL), no black transverse bar on the 1st and 3rd unbranched ray in dorsal-fin; the lower longitudinal band of lateral body is uninterrupted ···R. kumgangensis b. Interorbital width 22.7-27.6(proportion of % HL);
Caudal peduncle depth 10.1-13.6(proportion of % SL);
Black transverse bar on the first unbranched ray in dorsal-fin; the lower longitudinal band of lateral body is interrupted ···R. deogyuensis sp. nov.
Comparative Materials
Rhynchocypris kumgangensis: NPRI 001-010(10), 53.6- 80.2 mm SL; Bukhan River, Jeokmok-ri, Buk-myon, Gapyeong-gun, Gyeonggi-do, Korea, June 3, 2013, Dr. S.R.
Lee and Dr. J.H. Sim; NPRI 011-020(10), 42.1-57.6mm SL;
Bukhan River, Chiljeon-ri, Bangsan-myon, Yanggu-gun, Gangwon-do, Korea, April 15, 2013, Dr. S.R. Lee; NPRI
021-030(10), 50.0-64.5 mm SL; Bukhan River, Bangdong-ri, Kirin-myon, Inje-gun, Gangwon-do, Korea, June 4, 2013, Dr.
S.K. Park and Dr. J.H. Sim; NPRI 031-040(10), 43.4-74.5 mm SL; Bukhan River, Hangye-ri, Buk-myon, Inje-gun, Gangwon-do, Korea, June 3, 2013, Dr. S.R. Lee and Dr. J.H.
Sim; NPRI 041-050(10), 49.2-65.1 mm SL; Bukhan River, Saenggok-ri, Seoseok-myon, Hongcheon-gun, Gangwon-do, Korea, April 16, 2013, Dr. S.K. Park; NPRI 051-060(10), 57.0-77.3 mm SL; Namhan River, Dongsan-ri, Jinbu-myon, Pyeongchang-gun, Gangwon-do, Korea, April 16, 2013, Dr.
S.R. Lee; NPRI 061-070(10), 53.7-73.8 mm SL; Namhan River, Bongsan-ri, Imgyeo-myon, Jeongseon-gun, Gangwon- do, Korea, June 5, 2013. Dr. S.K. Park; NPRI 071-080(10), 51.4-68.1 mm SL; Namhan River, Molun-ri, Hwaam-myon, Jeongseon-gun, Gangwon-do, Korea, June 5, 2013. Dr. S.R.
Lee; NPRI 081-090(10), 44.6-76.6 mm SL; Namhan River, Seongnam-ri, Sinrim-myon, Wonju-si, Gangwon-do, Korea, June 5, 2013. Dr. S.K. Park; NPRI 091-100(10), 45.5-60.2 Figure 5. Phylogenetic tree of relationship between Rhynchocypris deogyuensis and R. kumgangensis by the NJ analysis. Number above node represent bootstrap value.
mm SL; Namhan River, Marak-ri, Dansan-myon, Yeongju-si, Gyeongbuk-do, Korea, June 5, 2013, Dr. S.R. Lee. and Dr.
J.H. Sim; NPRI 101-110(10), 62.1-70.5 mm SL; Namhan River, Uguchi-ri, Chunyang-myon, Bonghwa-gun, Gyeongbuk- do, Korea, June 5, 2013. Dr. S.K. Park.
Acknowledgements
We are very thanks to Dr. S.K. Park for assistance in sampling. and we also thanks Mr. J. J. Kim for drawing of the holotype. This study was supported by the nature resource research in the Deogyusan National Park of National Park Research Institute (NPRI).
References
Chyung MK. 1961. Illustrated Encyclopedia of the Fauna of Korea. 2. Fishes. Ministry of Education. Seoul (in Korea).
Chyung MK. 1977. The Fishes of Korea. Iljisa. Seoul (in Korea).
Hubbs CL, KF Lager. 1964. Fish of Great lakes region. Ann Arbor. University of Michigan Press.
Jeon SR. 1980. Studies on the distribution of fresh-water fishes from Korea. Chung-Ang University Press.
Kim IS, EJ Kang. 1986. The Osteology of Moroco keumgang Uchida (Pisces, Cyprinidae) in Korea. Jeonbuk National University Press. 28: 319-337.
Kim IS, GY Lee, SY Yang. 1985. Systematic Study of the Subfamily Leuciscinae (Cyprinidae) from Korea. Korea Fish Science. 18: 381-400.
Kim IS, Y Choi, CL Lee, YJ Lee, BJ Kim, JH Kim. 2005.
Illustrated Book of Korean Fishes. Kyohak Publishing Company, Ltd. Seoul (in Korea).
Kim IS. 1997. Illustrated Encyclopedia of Fauna and Flora of
Korea Vol. 37 Freshwater Fishes. Ministry of Education.
Seoul (in Korea).
Kim LT. 1980. A New species of Genus Phoxinus From D. P. R.
of Korea. Biology. 2: 27-29.
Kimura M. 1980. A simple method for estimating evolutionary rates of base substitutions through comparative studies of nucleotide sequence. Journal of Molecular Evolution. 16: 111- 120.
Lee SR, JJ Kim. 2016. Molecular Phylogeny Study of Rhyncho- cypris kumgangensis in Korea Based on the Mitochondrial DNA Cytochrom b Gene. Journal of National Park Research.
7: 147-153.
Nelson JS. 2006. Fishes of the world (4th ed), John Wiley & Sons, New York.
Suh JH, SY Yang. 1989. Studies on the Geographic Variation and Speciation of Moroco sp. Department of Biology, College of Science Inha University. 10: 183-192.
Tamura K, D Peterson, N Peterson, G Stecher, M Nei, S Kumar.
2011. MEGA5; molecular evolution genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Molecular Biology and Evolution. 28:
2731-2739.
Thompson JD, TJ Gibson, F Plewniak, F Jeanmougin, Dg Higgins. 1997. The Clustal X windows interface: flecible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic acids Research. 24: 4876-4882.
Uchida K. 1939. The fishes of Tyosen, Part 1, Nematognathi, Eventognathi, Bulletin of the Fisheries Experiment Station of the Government General of Tyosen (in Japanese).
Youn CH. 2002. Fish of Korea with pictorial key and systematic list. Academybook Publishing Company. Seoul (in Korea).
(2016년 11월 29일 접수; 2016년 12월 30일 수정;
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