Research Article |
Corresponding author: Yeon Jae Bae ( yjbae@korea.ac.kr ) Academic editor: Pavel Starkevic
© 2023 Jisoo Kim, Yeon Jae Bae.
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:
Kim J, Bae YJ (2023) Description of two new species of Dicranomyia (Erostrata) crane fly (Diptera, Limoniidae) from Korea, with remarks on DNA barcoding and updated taxonomic key. ZooKeys 1157: 193-206. https://doi.org/10.3897/zookeys.1157.90792
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Two new crane fly species, Dicranomyia (Erostrata) jejuensis sp. nov. and D. (E.) koreana sp. nov., from Korea are described on the basis of morphology and mitochondrial COI sequences. DNA barcode sequences for other four D. (Erostrata) species from Korea are also provided for the first time. The identification key for all known D. (Erostrata) species is presented.
Dicranomyia (Erostrata) jejuensis, Dicranomyia (Erostrata) koreana, DNA barcode, Limoniinae, taxonomy
Genus Dicranomyia Stephens, 1829, is the largest genus of the Limoniidae and, as such, contains 1,136 species and 24 subgenera, including the subgenus D. (Erostrata) Savchenko, 1976. Twelve species of this subgenus have been reported from the Palearctic, Nearctic, and Oriental regions (
Here, two new D. (Erostrata) crane fly species are described from Korea, providing identification for all Korean members of the subgenus. A DNA barcode (COI) dataset for six Dicranomyia species of the subgenus Erostrata from Korea is also presented for the first time.
Crane fly adults were collected using insect nets or Malaise traps and preserved in 80% ethanol (Table
Collection data of the Korean Dicranomyia (Erostrata) species used in the barcode analyses of this study.
Species* | GenBank accession number | Specimen code | Locality | Date | Collector(s) | Coordinates |
---|---|---|---|---|---|---|
D. (E.) globithorax | OM102980 | CF21-0149 | Gangwon-do, Wonju-si | 3 Sep. 2021 | J. Kim, D. Lee | 37°29'50.88"N, 130°53'23.78"E |
D. (E.) jejuensis sp. nov. | OM102981 | CF21-0150H | Jeju-do, Seoguipo-si | 14 Jun.–4 Aug. 2021 | Y. J. Bae | 33°19'49.07"N, 126°37'28.08"E |
OM102982 | CF21-0150P | Jeju-do, Seoguipo-si | 4 Aug.–8 Sep. 2021 | Y. J. Bae | 33°19'49.07"N, 126°37'28.08"E | |
OM102983 | CF21-0151 | Jeju-do, Seoguipo-si | 4 Aug.–8 Sep. 2021 | Y. J. Bae | 33°19'49.07"N, 126°37'28.08"E | |
D. (E.) koreana sp. nov. | OM102979 | CF21-0148 | Jeju-do, Seoguipo-si | 13 Jun.–4 Aug. 2021 | Y. J. Bae | 33°20'57.10"N, 126°29'43.29"E |
OP081140 | CF21-0152 | Gyeongsangnam-do, Sancheong-gun | 28 Jul. 2021 | J. Kim, C. Lim, D. Lee, W. Lee | 35°18'37.83"N, 127°45'05.47"E | |
D. (E.) submelas | OM102978 | CF21-0133 | Jeju-do, Seoguipo-si | 4 Aug.–8 Sep. 2021 | Y. J. Bae | 33°19'49.07"N, 126°37'28.08"E |
D. (E.) tabashii | OM102975 | CF21-0115 | Gyeongsangnam-do, Sancheong-gun | 28 Jul. 2021 | J. Kim, C. Lim, D. Lee, W. Lee | 35°18'37.83"N, 127°45'05.47"E |
OM102976 | CF21-0115f | Gyeongsangnam-do, Sancheong-gun | 28 Jul. 2021 | J. Kim, C. Lim, D. Lee, W. Lee | 35°18'37.83"N, 127°45'05.47"E | |
D. (E.) yazuensis | OM102977 | CF21-0132 | Gangwon-do, Pyeongchang-gun | 28 Jul.–15 Sep. 2020 | Y. J. Bae | 37°47'05.67"N, 128°34'16.97"E |
D. (D.) kandybinae | OP093621 | CF21-0099 | Gangwon-do, Wonju-si | 23 Jul.–3 Sep. 2021 | Y. J. Bae | 37°17'26.50"N, 128°04'54.77"E |
The terminologies used to describe the morphology generally follow
Specimen depositories are as follows:
Total genomic DNA was extracted from the leg muscle of using the DNeasy Blood & Tissue Kit (Qiagen, Hilden, Germany) according to the manufacturer’s instructions. COI sequences were amplified and sequenced following
DNA barcode analysis was performed using 11 COI sequences (Table
Dicranomyia (Erostrata) canis (Alexander, 1931b)
Dicranomyia (Erostrata) cnephosa (Alexander, 1959)
Dicranomyia (Erostrata) congesta (Alexander, 1967)
Dicranomyia (Erostrata) cynotis (Alexander, 1931a)
Dicranomyia (Erostrata) globithorax Osten Sacken, 1869
Dicranomyia (Erostrata) globulithorax Alexander, 1924
Dicranomyia (Erostrata) jejuensis sp. nov.
Dicranomyia (Erostrata) koreana sp. nov.
Dicranomyia (Erostrata) melas (Alexander, 1934)
Dicranomyia (Erostrata) reniformis Kato, Tachi & Gelhaus, 2018
Dicranomyia (Erostrata) striopleura (Edwards, 1919)
Dicranomyia (Erostrata) submelas Kato, Tachi & Gelhaus, 2018
Dicranomyia (Erostrata) tabashii (Alexander, 1934)
Dicranomyia (Erostrata) yazuensis Kato, Tachi & Gelhaus, 2018
1 | Tarsi with white bands | Dicranomyia (Erostrata) congesta (India) |
– | Tarsi without bands | 2 |
2 | Pleuron with broad, blackish brown lateral stripe | Dicranomyia (Erostrata) striopleura (Indonesia, Malaysia) |
– | Pleuron without lateral stripe | 3 |
3 | Scutellum obscure yellow | 4 |
– | Scutellum yellowish brown to blackish brown | 6 |
4 | Rostrum black. Wing strongly blackened | Dicranomyia (Erostrata) cnephosa (Nepal) |
– | Rostrum pale. Wing tinged with pale brown | 5 |
5 | Palpus 2-segmented. Male seventh sternite with strongly darkened internal sac | Dicranomyia (Erostrata) tabashii (Japan, Korea, Russia) |
– | Palpus 3-segmented. Male seventh sternite with slightly darkened internal sac with rounded entrance | Dicranomyia (Erostrata) koreana sp.nov. (Korea) |
6 | Gonostylus with black spines on mesal face | 7 |
– | Gonostylus without black spines | 12 |
7 | Gonostylus narrowed to a point, triangular | 8 |
– | Gonostylus not as above | 9 |
8 | Mesal face of gonostylus densely covered with black setae | Dicranomyia (Erostrata) cynotis (Philippines) |
– | Mesal face of gonostylus with black setae restricted to distal 1/2 | Dicranomyia (Erostrata) canis (Philippines) |
9 | Paramere distally with rounded tip | 10 |
– | Paramere distally with pointed tip | 11 |
10 | Gonostylus elongate with truncated apex | Dicranomyia (Erostrata) globithorax (Canada, Japan, Korea, USA) |
– | Gonostylus elongate with angled apex, length of angled apex ca 1/5 of gonostylus | Dicranomyia (Erostrata) globulithorax (Japan, Korea, Russia) |
11 | Gonocoxite with apically rounded ventromesal lobe. Gonostylus shallowly concaved at inner apical edge, the area 1/5 as long as gonostylus | Dicranomyia (Erostrata) melas (Taiwan) |
– | Gonocoxite with apically truncated ventromesal lobe. Gonostylus deeply emarginated at inner apical edge, the area 1/3 as long as gonostylus | Dicranomyia (Erostrata) submelas (Japan, Korea) |
12 | Gonostylus stout and reniform, tip of ventral surface covered with dense, fine setae | Dicranomyia (Erostrata) reniformis (Japan) |
– | Gonostylus not reniform, tip of ventral surface without dense, fine setae | 13 |
13 | Palpus 2-segmented. Gonostylus tapered in distal 1/3, without apical spine | Dicranomyia (Erostrata) yazuensis (Japan, Korea) |
– | Palpus 3-segmented. Gonostylus tapered in distal 1/2, with black apical spine | Dicranomyia (Erostrata) jejuensis sp. nov. (Korea) |
Family Limoniidae Speiser, 1909
Subfamily Limoniinae Speiser, 1909
Dicranomyia (Erostrata)
Savchenko in
Dicranomyia globithorax Osten Sacken, 1869 (original designation).
Rostrum is very short or reduced. Number of palpomeres ranges from one to three. Wings have no patterns, even in stigmal region. Third and fourth tarsomere are slightly swollen. Internal sac or notch is located on the male seventh sternite. Gonocoxite has ventromesal lobe. Gonostylus is one paired, with one or two setae arising from small tubercle on outer surface.
Holotype
: Korea • ♂; Jeju-do, Seogwipo-si, Namwon-eup, Sillye-ri, Iseungi-oreum Volcanic Cone; 33°20.24'N, 126°37.25'E; alt. 450 m; 4 Aug.–8 Sep. 2021; Y. J. Bae leg.; Malaise trap; GenBank: OM102981; CF21-0150H;
Paratypes
: Korea • 1 ♀; same data as holotype, 14 Jul.–4 Aug. 2021; GenBank: OM102983; CF21-0151;
Palpus is 3-segmented. Male seventh sternite has shallow V-shaped notch. Outer face of gonostylus has single seta arising from tubercle. Distal lobe of paramere has a hooked tip with a subapical process.
Male (holotype). Body length 4.3 mm, wing length 4.6 mm, antenna length 0.9 mm. General body coloration pale yellow to yellowish brown (Fig.
Dicranomyia (Erostrata) jejuensis sp. nov. A habitus, male (paratype) B head, female (paratype) C wing, male (paratype) D male seventh sternite, ventral view E male terminalia, dorsal view F aedeagal complex, dorsal view G paramere, lateral view H female terminalia, lateral view I tip of hypovalva, lateral view. Abbreviations: ad – aedeagus; cc – cercus; fl – flagellum; gc – gonocoxite; gf – genital fork; gs – gonostylus; hv – hypovalva; pd – pedicel; pl – palpus; pm – paramere; sc – scape; tb – tubercle; t9 – ninth tergite; vlg – ventromesal lobe of gonocoxite. Scale bars: 2 mm (A, C); 0.5 mm (B, D, E, H); 0.1 mm (F, G, I).
Head
(Fig.
Thorax. Prescutum and presutural scutum yellow. Postsutural scutum, scutellum and mediotergite yellowish brown. Pleuron uniformly dull yellow, without lateral stripes. Wing (Fig.
Abdomen. Tergites yellowish brown, sternites 1–4 yellow, remaining yellowish brown. Seventh sternite with shallow V-shaped notch with darkened margin (Fig.
Male terminalia
(Fig.
Female. Body length 4.5 mm, wing length 4.8 mm, antenna length 0.9 mm (N = 1). General body coloration brighter than male. Femur I 2.8 mm; II 3.2 mm; III 3.4 mm; tibia I 3.2 mm; I: 3.1 mm; III 3.4 mm; tarsus I 3.0 mm; II 2.6 mm; III 2.4 mm.
Female terminalia
(Fig.
Specific name “jejuensis” refers to the type locality, Jejudo Island, Korea.
The species is currently only known from Jejudo Island, Korea.
Adults were collected from June through early September.
Dicranomyia (E.) jejuensis sp. nov. is morphologically similar to D. (E.) yazuensis based on the male genital structures, but it can be distinguished by the following characters: pleuron entirely dull yellow (vs dark dorsally); palpus 3-segmented (vs 2-segmented); distal 1/2 of gonostylus tapered to tip (vs distal 2/3 strongly narrowed toward tip); posterior margin of male seventh sternite with shallow V-shaped notch (vs long triangular notch); distal part of paramere with hooked tip (vs straight tip).
Holotype
: Korea • ♂; Jeju-do, Seogwipo-si, Hawon-dong, Mt. Hallasan; 33°20.95'N, 126°29.72'E; alt. 1220 m; 13 Jun.–4 Aug. 2021; Y. J. Bae leg.; Malaise trap; GenBank: OM102979; CF21-0148;
Paratypes
: Korea • 1 ♂; Gyeonggi-do, Gapyeong-si, Buk-myeon, Jeokmok-ri, Garim-gyo (Br.); 37°58.60'N, 127°26.55'E; alt. 300 m; 25 Jul.–1 Aug. 2015; Y. J. Bae leg.; Malaise trap; published as D. (E.) tabashii by
Palpus is 3-segmented. Center of male seventh sternite has a deep conical internal sac that has a wide, round entrance. Outer face of gonostylus has two setae arising from a small tubercle. Paramere is elongated and narrow, distally with a darkened tip.
Male (holotype). Body length 3.5 mm, wing length 4.5 mm, antenna length 0.7 mm. General body coloration yellow (Fig.
Dicranomyia (Erostrata) koreana sp. nov. A habitus, male (holotype) B head, male (paratype) C wing, male (paratype) D male seventh sternite, ventral view E male terminalia, dorsal view F gonostylus, ventral view G aedeagal complex, dorsal view H paramere, lateral view I female terminalia, lateral view J tip of hypovalva, lateral view. Abbreviation: tb – tubercle. Scale bars: 2 mm (A, C); 0.5 mm (B, D, E, I); 0.1 mm (F, G, H, J).
Head
(Fig.
Thorax. Prescutum, scutum and scutellum yellow. Mediotergite yellowish brown. Pleuron entirely pale yellow, without lateral stripes. Wing (Fig.
Abdomen. Tergites yellow except pale eighth tergite; sternites paler. Seventh sternite (Fig.
Male terminalia
(Fig.
Female. Body length 3.8 mm, wing length 4.7 mm, antenna length 0.7 mm (N = 1). General body coloration lighter than male.
Female terminalia
(Fig.
Specific name “koreana” refers to the country of its discovery, Korea.
The species is widely distributed in Korea, including Jejudo Island.
This species is found along intermittent mountain streams in moist mixed forests with grassy vegetation (Fig.
Adults are mainly active from July through August.
In terms of the shape of the male terminalia, D. (E.) koreana sp. nov. is similar to D. (E.) tabashii, but it can be distinguished by the following characters: palpus 3-segmented (vs 2-segmented); male seventh sternite with weakly darkened, conical internal sac with round entrance (vs strongly darkened, U-shaped internal sac); paramere with darkened tip (vs without). This species is also similar to another species, D. (E.) jejuensis sp. nov. based on the male genital structures, but it can be distinguished by the following characters: male seventh sternite with a deep, conical internal sac (vs shallow, V-shaped notch); gonostylus with two setae from tubercle (vs a single seta); paramere without hook at tip (vs with hook).
The male genitalia of D. (E.) koreana sp. nov. from Mount Bangtaesan differs from other materials of the species based on the shape of the seventh sternite internal sack (shallow conical without rounded mouth) and paramere distal lobe (pointed tip). However, additional specimens are needed to determine whether this difference is due to intra- or interspecific variation.
The 676-bp COI sequences contained 190 variable sites, of which 156 were parsimony-informative. The interspecific divergences (p-distances) within subgenus D. (Erostrata) ranged from 11.54% to 16.42%, with a mean distance of 13.17% across the entire dataset (Table
Estimates of genetic divergence (%) between sequences. The number of base differences per site from between sequences are shown. Standard errors (%) are shown above the diagonal and were obtained by a bootstrap procedure (1,000 replicates). All positions containing gaps and missing data were eliminated (complete delete option).
Species | Accession number | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | |
---|---|---|---|---|---|---|---|---|---|---|---|---|---|
1 | D. (E.) globithorax | OM102980 | – | 1.36 | 1.36 | 1.36 | 1.36 | 1.37 | 1.28 | 1.42 | 1.41 | 1.37 | 1.34 |
2 | D. (E.) jejuensis sp. nov. | OM102981 | 13.61 | – | 0 | 0.15 | 1.37 | 1.35 | 1.36 | 1.34 | 1.33 | 1.33 | 1.45 |
3 | OM102982 | 13.61 | 0 | – | 0.15 | 1.37 | 1.35 | 1.36 | 1.34 | 1.33 | 1.33 | 1.45 | |
4 | OM102983 | 13.76 | 0.15 | 0.15 | – | 1.36 | 1.34 | 1.36 | 1.34 | 1.33 | 1.34 | 1.45 | |
5 | D. (E.) koreana sp. nov. | OM102979 | 15.24 | 14.94 | 14.94 | 14.79 | – | 0.30 | 1.38 | 1.21 | 1.20 | 1.31 | 1.45 |
6 | OP081140 | 15.38 | 14.64 | 14.64 | 14.50 | 0.59 | – | 1.40 | 1.22 | 1.21 | 1.31 | 1.45 | |
7 | D. (E.) submelas | OM102978 | 11.54 | 14.64 | 14.64 | 14.79 | 15.09 | 15.68 | – | 1.43 | 1.43 | 1.37 | 1.36 |
8 | D. (E.) tabashii | OM102975 | 16.12 | 14.94 | 14.94 | 14.79 | 11.69 | 11.83 | 16.42 | – | 0.15 | 1.29 | 1.44 |
9 | OM102976 | 15.98 | 14.79 | 14.79 | 14.64 | 11.54 | 11.69 | 16.27 | 0.15 | – | 1.30 | 1.44 | |
10 | D. (E.) yazuensis | OM102977 | 15.24 | 12.43 | 12.43 | 12.57 | 13.46 | 13.46 | 13.76 | 12.87 | 13.02 | – | 1.30 |
11 | D. (D.) kandybinae | OP093621 | 14.50 | 16.27 | 16.27 | 16.42 | 16.12 | 16.12 | 14.50 | 15.68 | 15.53 | 12.57 | – |
This is the first study to use DNA barcoding for the delimitation of the D. (Erostrata) species. The present study identified two new species using both morphological and molecular data. According to the NJ tree (Fig.
Based on our morphological examinations of the materials, we also found that some specimens identified as D. (E.) tabashii by
We are deeply grateful to Dr Sigitas Podenas (Nature Research Centre and Vilnius University, Vilnius, Lithuania), who provided photographs of the D. (E.) tabashii type specimens, and to Dr Pjotr Oosterbroek (University of Amsterdam, Amsterdam, the Netherlands), who provided valuable information through his website, Catalogue of the Crane flies of the World (https://ccw.naturalis.nl/index.php). We are very grateful to members of the Laboratory of Biodiversity and Ecology, and the Korean Entomology Institute (Korea University, Seoul, Korea) for their assistance during field trips. We are also grateful to Dr Pavel Starkevich and Dr Daichi Kato for offering valuable comments and improving the manuscript. This work was supported by a grant from the National Institute of Biological Resources (