Research Article |
Corresponding author: Seong Myeong Yoon ( smyun@chosun.ac.kr ) Academic editor: Greg Rouse
© 2017 Hyun Ki Choi, Jong Guk Kim, Dong Won Kang, Seong Myeong Yoon.
This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Citation:
Choi HK, Kim JG, Kang DW, Yoon SM (2017) A new species of Leodice from Korean waters (Annelida, Polychaeta, Eunicidae). ZooKeys 715: 53-67. https://doi.org/10.3897/zookeys.715.20448
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A new eunicid species, Leodice duplexa sp. n., from intertidal and subtidal habitats in the eastern coast of South Korea is described. The new species is assigned to the C-2 group, and is similar to Leodice antennata, the type species of the genus, in having the following combination of characteristics: moniliform antennae and palps, bidentate compound falcigers, articulated peristomial and notopodial cirri, pectinate branchiae showing bimodal distribution of branchial filaments, and yellow aciculae. However, L. duplexa sp. n. is readily distinguished from L. antennata by the following features: the aciculae are 2–4 in number, with blunt or pointed tips and hammer-headed or bifid tips, and the subacicular hooks are paired in some chaetigers. A detailed description and illustrations are provided for the new species. The validity of the new species is also supported by a genetic comparison using sequences of the mitochondrial cytochrome c oxidase subunit I (COI). A revised key to known Leodice species is provided with a comparison of their morphological characteristics.
COI, eunicid, Korea, polychaete, taxonomy
The genus Leodice Savigny in Lamarck, 1818, a member of genera belonging to family Eunicidae Berthold, 1827, had been previously considered an invalid taxon (
While studying the polychaetes from Korean waters as a part of the ‘Securement, Analysis and Evaluation of Marine Invertebrate Bioresources’, a new species of the genus Leodice was found. In the present paper, detailed description and illustrations of the new species are provided with a genetic comparison between the new species and other Leodice species in the barcode region of the mitochondrial cytochrome c oxidase subunit I (COI). A taxonomic comparison of known Leodice species is also presented, with a revised taxonomic key based on the previous literature (
Samples were collected from rocky bottoms of the intertidal and subtidal zone. Specimens were sorted using sieves with pore size of 0.5 mm, fixed initially with 5% formaldehyde-seawater solution, and transferred to 85% ethyl alcohol. The characteristics of the whole body were observed with appendages dissected in a petri dish using dissection forceps or surgical knives and needles under stereomicroscope (SMZ1500; Olympus, Tokyo, Japan). Dissected specimens were mounted onto temporary slides using glycerol or permanent slides using polyvinyl lactophenol solution. Drawings were made under the stereomicroscope and light microscope (LABOPHOT-2; Nikon, Tokyo, Japan) with the aid of drawing tubes. Photographs were taken of the appendages in a permanent slide. Images of appendages were captured using an image system (i-SOLUTION/LITE, iMTechnology®, Vancouver, Canada). Specimens for scanning electron microscopy (SEM) were dehydrated by t-BuOH freeze dryer (VFD-21S; Vacuum Device, Ibaraki, Japan). They were mounted on stubs and coated with gold-palladium. SEM observations were carried out using a scanning electron microscope (SU3500; Hitachi, Tokyo, Japan). Type material and additional material examined were deposited in the National Marine Biodiversity Institute of Korea (MABIK).
Genomic DNA was extracted from the posterior segments of three specimens selected among additional materials using a DNeasy Blood and Tissue Kit (Qiagen, Hilden, Germany) according to the manufacture’s protocol. Amplifications of partial sequences of the mitochondrial cytochrome c oxidase subunit I (COI) from gDNA were carried out by polymerase chain reaction (PCR) method using a set of primers: ACOIAF 5’- CWAATCAYA AAGATATTGGAAC-3’ and ACOIAR 5’- AATATAWACTTCWGGGTGACC -3’ (Zonal et al. 2010). PCR amplification was conducted in a total volume of 20 µL: 10 µL of 2x DyeMIX-Tenuto (Enzynomics), 0.5 µL of each primer, 1 µL of gDNA, and 8 µL of sterile water. PCR condition was determined based on the work of Zonal et al. (2010) as follows: 5 min at 94°C, followed by 35 cycles of 1 min at 94°C, 1 min at 45°C, and 2 min at 72°C, with a final extension of 7 min at 72°C. PCR products were purified with a QIAquick® PCR Purification Kit (Qiagen, Chatsworth, CA, USA). Sequences for the new species were obtained by an Applied Biosystems 3730 DNA sequencer, and deposited in GenBank under accession number MF669544–MF669546. These sequences were aligned with those of other Leodice species and outgroup taxa using Geneios Pro v.9.1.8 (Biomatters, Auckland, New Zealand). The genetic distances of the new species from other species and the phylogenetic tree of them were produced by MEGA v.6.06 (
South Korea, Gyeongsangbuk-do Province: Gyeongju-si County, Gampo-eup, Oryu 1-ri, 35°48'13"N, 129°32'21"E, intertidal rocky bottom.
Holotype. complete specimen (53.0 mm long, 4.8 mm width), cat no. MABIKNA00146045. Paratypes. one complete specimen (24.0 mm long, 4.9 mm width), cat no. MABIKNA00146046; one incomplete specimen (15.0 mm long, 3.3 mm width), cat no. MABIKNA00146047. All type material was collected from intertidal rocky bottom at the type locality on 9 April 2014.
South Korea, Gyeongsangbuk-do Province: 5 specimens, Ulleung-gun County, Ulleung-eup, Dokdo-ri, 37°14’31”N, 131°52’06”E, 05 Sep. 2016, subtidal rocky bottom at 10–15 m depth; 5 specimens, Pohang-si County, Homigot-myeon, Guman-ri, 36°04'35"N, 129°34'31"E, 19 May 2015; 2 specimens, Yeongdeok-gun County, Chuksan-myeon, Gyeongjeong-ri, 36°27'40"N, 129°32'34"E, 17 Sep. 2014., intertidal rocky bottom.
Prostomium with three antennae and one pair of lateral palps arranged in crescent pattern; palpostyles and ceratostyles regularly articulated and moniliform, and with ring-shaped palpo- and ceratophores. Peristomial cirri with four weak articulations, not extending middle part of first peristomial ring. Pectinate branchiae from chaetiger VI to near posterior end, with maximum of 7–8 branchial filaments. Limbate chaetae slender, with narrow wings. Heterodont pectinate chaetae with 5–10 teeth. Compound falcigers bidentate, with hoods marginally serrated. Aciculae yellow, with both blunt and hammer-headed or bifid tip, and 2–4 per parapodium. Subacicular hooks yellow, tridentate, present from chaetiger XXIII to last chaetiger, and 1–2 per parapodium. Pygidium with two pairs of anal cirri with four articulations.
Holotype: complete specimen with cylindrical body, slightly flattened dorsoventrally in posterior segments, and with approximately 94 segments.
Prostomium bilobed, distinctly shorter and narrower than peristomium, and about as deep as 1/2 of peristomium; prostomial lobes anteriorly rounded, dorsally flattened, separated by shallow and narrow notch. Prostomial appendages consisted of three antennae and two lateral palps, arranged in shallow semicircle, evenly spaced, similar in thickness; palpophores and ceratopores ring-shaped without articulation; palpostyles and ceratostyles tapering, regularly articulated, with up to 12 moniliform articulations in A-I extending to anterior edge of chaetiger I, but incomplete distally with 8–11 moniliform articulations in others except A-I (Fig.
Eyes black, spherical, and located between bases of palps and lateral antennae (Fig.
Peristomium cylindrical, divided into first and second ring; first ring 3–4 times longer than second one; peristomial cirri with four weak articulations, not extending middle part of first peristomial ring (Fig.
Maxillary formula (in paratype): Mx I 1+1; Mx II 6+6; Mx III 7+0; Mx IV 6+7; Mx V 1+1; Mx III located at frontal end of distal arc with left Mx IV. Mandibles flat (Fig.
Branchiae pectinate, slightly longer than dorsal cirri, and present on more than 65% of total number of chaetigers from chaetiger VI to near posterior end. Branchial filaments bimodal distribution, single at first branchial chaetiger, reaching maximum of eight in number between chaetigers IX to XXIII, reduced to 3–4 in number on chaetigers XXXI to XLVIII, increasing to 5–6 in number on chaetigers XLIX to LXIV, and thereafter decreasing to 2–4 in number on posterior chaetigers. Last six chaetigers without branchiae (Fig.
Dorsal cirri tapering and digitiform, with 2–3 weak articulations (Fig.
Leodice duplexa sp. n., A–C, F–N holotype (MABIKNA00146045) D, F paratype (MABIKNA00146046) A anterior end, dorsal view B anterior end, lateral view C posterior end, dorsal view D maxillae E mandible F left parapodium VI, anterior view G left parapodium XX, anterior view H right parapodium XXXVIII, anterior view I right parapodium LX, anterior view J limbate chaeta on parapodium XLVII K pectinate chaeta on parapodium XLVII L compound falciger on parapodium XLVII M subacicular hook on parapodium XXXVIII N aciculae with blunt and hammer-headed tips on parapodium LXXXV. Scale bars: 1.0 mm (A, B), 0.5 mm (C–E), 0.4 mm (F–I), 0.05 mm (J–N).
Anterior neuropodial lobes truncate and distally rounded with aciculae emerging near midline; pre- and postchaetal lobes low, transverse folds. Ventral cirri on anterior chaetigers thick and ovoid-shaped, with slightly inflated base; median ones with inflated base more than anterior ones; posterior ones slightly elongated with smaller base than median ones (Fig.
Limbate chaetae slender and elongate, longer than other chaetae, and with narrow wings (Fig.
Pygidium with two pairs of anal cirri; dorsal pair as long as last five chaetigers with up to four cylindrical articulations, ventral pair reduced to small bump (Fig.
Leodice duplexa sp. n., A–D paratype (MABIKNA00146047) A yellow aciculae with pointed tips B yellow and paired subacicular hooks C parapodium with four aciculae D aciculae with hammer-headed or bifid tips E compound falcigers with bidentate appendages F pectinate chaeta. Scale bars: 0.05 mm (A–C), 0.025 mm (D–F).
The epithet of the specific name, duplexa, is derived from the Latin duplex, meaning ‘double’. This name refers to the presence of paired subacicular hooks.
The East Sea (or the Sea of Japan) of South Korea.
Leodice species were redefined by
The distal shape of aciculae has been considered as a useful character for distinguishing eunicid species (
The aciculae with both blunt or pointed and hammer-headed or bifid tips, which appear in the new species, have been often described in the eunicid species. In the species of the C-2 group of the eunicids (
We obtained three partial COI sequences of a total 664 bp size from three individuals of Leodice duplexa sp. n., respectively. All COI sequences obtained are identical. For the genetic comparison on the new species, we sort out the sequences of 14 eunicid species including 12 Leodice species, which were originally registered as Eunice species, and two non-Leodice species as outgroup taxa, Eunice norvegica (Linnaeus, 1767) and Marphysa sanguinea (Montagu, 1813), from GenBank (
Comparison of morphological characteristics among known Leodice species.
Species | Eyes | Articulations of antennae and palpal styles | Peristomial cirri | Branchiae | Notopodial cirri | Subacicular hooks | Compound falcigers | Aciculae | Data source | ||||||
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Shape | Starting chaetiger | Distribution | Number of filaments | Shape | Starting chaetiger | Number per parapodium | shape | Number per parapodium | |||||||
L. americana ( |
present | regularly cylinderical | smooth | pectinate | 3 | less than 55% of body | 2–20 | articulated | yellow, tridentate | 25 | single | bidentate, with mucros | yellow, bently blunt tips | paired |
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L. antarctica (Baird, 1869) | absent | irregularly cylinderical | smooth | pectinate | 3 | less than 55% of body | 1–5 | articulated | yellow, bidentate | 31–44 | single | bidentate | yellow, blunt tips | paired |
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L. antennata Lamarck, 1818 | present | moniliform | with articulations | pectinate | 5–7 | more than 65% of body | 2–7 | articulated | yellow, tridentate | 24–26 | single | bidentate | yellow, blunt or pointed tips | paired |
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L. antillensis (Ehlers, 1887) | present | irregularly cylinderical | with articulations | pectinate | 4 | less than 55% of body | 1–6 | articulated | yellow, bidentate | 33 | 1–2 | bidentate | yellow, flattened and expanded tips | paired |
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L. duplexa sp. n. | present | moniliform | with articulations | pectinate | 5–6 | more than 65% of body | 1–8 | articulated | yellow, tridentate | 24–26 | 1–2 | bidentate | yellow, both blunt and hammer-headed or bifid tips | 2–4 | Present study |
L. gracilicirrata Treadwell, 1922 | unknown | irregularly cylinderical | with articulations | pectinate | 3 | less than 55% of body | 1–7 | articulated | yellow, bidentate | 63 | single | bidentate | yellow, both pointed with mucros and bluntly rounded tips | paired |
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L. harassii (Audouin & Milne Edwards, 1933) | faintly present | regularly cylinderical | smooth | pectinate | 4 | more than 65% of body | 1–10 | smooth | light brown, bidentate | 28 | single | bidentate | yellow, turned dark brown, pointed tips | paired | Fauchald, 1992, |
L. laurillardi (Quatrefages, 1866) | present | regularly cylinderical | with articulations | pectinate | 2–3 | more than 65% of body | 1–10 | articulated | dark brown, bidentate | 29–32 | single | bidentate | dark brown, blunt tips | paired | Fauchald, 1992, |
L. limosa (Ehlers 1868) | unknown | regularly cylinderical | smooth | pectinate | 3 | less than 55% of body | 1–12 | smooth | yellow, tridentate | before 30 | unknown | bidentate, with mucros | yellow, blunt tips | unknown |
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L. lucei (Grube, 1856) | present | moniliform | with articulations | pectinate | 5 | more than 65% of body | 1–8 | articulated | yellow, tridentate | 24–34 | single | bidentate, with mucros | yellow, distinctly hammer-headed or bifid tips | paired |
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L. marcusi (Zanol et al., 2000) | present | moniliform | with articulations | palmate | 4 | more than 65% of body | 1–4 | articulated | black, bidentate | 19–26 | single | bidentate | black, pointed tips | paired | Zanol et al. 2000 |
L. miurai (Carrera-Parra & Salazar-Vallejo, 1998) | present | moniliform | with articulations | pectinate | 5–6 | less than 55% of body | 1–3 | articulated | yellow, tridentate | 18–23 | single | bidentate to tridentate | black, bifid tips | unknown | Carrera-Parra & Salazar-Vallejo 1998, |
L. rubra (Grube, 1856) | present | moniliform | with articulations | pectinate | 4–6 | more than 65% of body | 1–21 | articulated | yellow, tridentate | 27 | 1–2 | bidentate | yellow, both pointed or blunt and bifid tips | paired |
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L. thomasiana (Augener, 1922) | present | basally cylindrical and distally ovate | with articulations | palmate | 4 | more than 65% of body | 1–3 | articulated | brown, bidentate | 22–24 | 1–2 | bidentate | brown, blunt tips | paired |
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L. torquata (Quatrefages, 1866) | present | moniliform | with articulations | pectinate | 3 | more than 65% of body | 2–7 | articulated | brown, bidentate | 22 | 1–2 | bidentate | brown, blunt tips | paired |
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L. valens Chamberlin, 1919 | present | regularly cylinderical | with articulations | pectinate | 3 | less than 55% of body | 1–11 | articulated | yellow, bidentate | 43 | 1–2 | bidentate | yellow, blunt tips | 2–4 |
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Genetic distance (K2P) based on 664 bp size of COI sequence among 13 Leodice species with two outgroup taxa (Eunice norvegica and Marphysa sanguinea).
No. | Species | Accession No. | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 | 13 | 14 | 15 | 16 | Data source |
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1 | Leodice duplexa sp. n. | MF669544 | Present study | ||||||||||||||||
2 | Leodice duplexa sp. n. | MF669545 | 0.000 | " | |||||||||||||||
3 | Leodice duplexa sp. n. | MF669546 | 0.000 | 0.000 | " | ||||||||||||||
4 | L. americana | GQ497561 | 0.095 | 0.095 | 0.095 |
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5 | L. antarctica | GQ497532 | 0.088 | 0.088 | 0.088 | 0.043 | " | ||||||||||||
6 | L. antennata | DQ317858 | 0.097 | 0.097 | 0.097 | 0.103 | 0.124 |
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7 | L. cf. antillensis | GQ497533 | 0.144 | 0.144 | 0.144 | 0.102 | 0.088 | 0.159 |
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8 | L. harassii | GQ497535 | 0.108 | 0.108 | 0.108 | 0.062 | 0.043 | 0.115 | 0.095 | " | |||||||||
9 | L. cf. limosa | GQ497531 | 0.124 | 0.124 | 0.124 | 0.104 | 0.110 | 0.102 | 0.153 | 0.116 | " | ||||||||
10 | L. lucei | GQ497529 | 0.082 | 0.082 | 0.082 | 0.088 | 0.095 | 0.062 | 0.115 | 0.088 | 0.102 | " | |||||||
11 | L. miurai | GQ497530 | 0.104 | 0.104 | 0.104 | 0.109 | 0.116 | 0.096 | 0.145 | 0.123 | 0.153 | 0.083 | " | ||||||
12 | L. rubra | GQ497528 | 0.083 | 0.083 | 0.083 | 0.089 | 0.109 | 0.012 | 0.144 | 0.101 | 0.102 | 0.062 | 0.111 | " | |||||
13 | L. thomasiana | GQ497563 | 0.143 | 0.143 | 0.143 | 0.094 | 0.081 | 0.136 | 0.108 | 0.068 | 0.129 | 0.128 | 0.150 | 0.121 | " | ||||
14 | L. torquata | GQ497539 | 0.114 | 0.114 | 0.114 | 0.094 | 0.068 | 0.121 | 0.094 | 0.055 | 0.144 | 0.128 | 0.135 | 0.107 | 0.024 | " | |||
15 | L. valens | GQ497534 | 0.116 | 0.116 | 0.116 | 0.089 | 0.069 | 0.139 | 0.075 | 0.075 | 0.103 | 0.095 | 0.132 | 0.124 | 0.129 | 0.115 | " | ||
16 | Eunice norvegica (outgroup) | GQ497541 | 0.188 | 0.188 | 0.188 | 0.143 | 0.121 | 0.165 | 0.150 | 0.129 | 0.157 | 0.180 | 0.180 | 0.165 | 0.144 | 0.129 | 0.143 | " | |
17 | Marphysa sanguinea (outgroup) | GQ497547 | 0.172 | 0.172 | 0.172 | 0.135 | 0.101 | 0.157 | 0.115 | 0.101 | 0.150 | 0.121 | 0.157 | 0.142 | 0.136 | 0.121 | 0.114 | 0.101 | " |
1 | Antennae and palps regularly articulated | 2 |
– | Antennae and palps irregularly articulated | 13 |
2 | Subacicular hooks bidentate | 3 |
– | Subacicular hooks tridentate | 7 |
3 | Peristomial cirri and notopodial cirri articulated | 4 |
– | Peristomial cirri and notopodial cirri smooth | L. harassii (Audouin & Milne Edwards, 1933) |
4 | Branchiae present on more than 65% of body | 5 |
– | Branchiae present on less than 55% of body | L. valens Chamberlin, 1919 |
5 | Parapodia with pectinate branchiae | 6 |
– | Parapodia with palmate branchiae | L. marcusi (Zanol et al., 2000) |
6 | Subacicular hooks usually paired | L. torquata (Quatrefages, 1866) |
– | Subacicular hooks always single | L. laurillardi (Quatrefages, 1866) |
7 | Branchiae with up to 20–21 filaments | 8 |
– | Branchiae with less than 12 filaments | 9 |
8 | Aciculae with bifid or hammer-headed tips absent | L. americana (Hartman, 1944) |
– | Aciculae with bifid or hammer-headed tips present | L. rubra (Grube, 1856) |
9 | Guards of compound falcigers with mucros | 10 |
– | Guards of compound falcigers without mucros | 11 |
10 | Palpostyles and ceratostyles cylindrical; peristomial cirri smooth | L. limosa (Ehlers 1868 |
– | Palpostyles and ceratostyles moniliform; peristomial cirri articulated | L. lucei (Grube, 1856) |
11 | Compound falcigers tridentate in posterior chaetigers | L. miurai (Carrera-Parra & Salazar-Vallejo, 1998) |
Compound falcigers only bidentate | 12 | |
12 | Subacicular hooks always single; aciculae paired | L. antennata Lamarck, 1818 |
– | Subacicular hooks paired in some chaetigers; aciculae 2–4 in number | L. duplexa sp. n. |
13 | Subacicular hooks and acicular light; branchiae pectinate | 14 |
– | Subacicular hooks and acicular dark; branchiae palmate | L. thomasiana (Augener, 1922) |
14 | Subacicular hooks paired in some chaetigers | L. antillensis (Ehlers, 1887) |
– | Subacicular hooks always single | 15 |
15 | With finely hooded aciculae | L. gracilicirrata Treadwell, 1922 |
– | Without finely hooded aciculae | L. antarctica (Baird, 1869) |
This work was supported by the National Marine Biodiversity Institute of Korea as a part of the ‘Securement, Analysis and Evaluation of Marine Invertebrate Bioresources (2017M00600)’.