Research Article |
Corresponding author: Jan Ševčík ( sevcikjan@hotmail.com ) Academic editor: Jukka Salmela
© 2016 Jan Ševčík, David Kaspřák, Björn Rulik.
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:
Ševčík J, Kaspřák D, Rulik B (2016) A new species of Docosia Winnertz from Central Europe, with DNA barcoding based on four gene markers (Diptera, Mycetophilidae). ZooKeys 549: 127-143. https://doi.org/10.3897/zookeys.549.6925
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A new species of Docosia Winnertz, D. dentata sp. n., is described and illustrated, based on a single male specimen collected in Muránska planina National Park in Central Slovakia. DNA sequences (COI, COII, CytB, and ITS2) are included and compared for 13 species of Docosia. There was found only little congruence between the molecular results and previous scarce data about interspecific relationships based on morphology. The COI and CytB gene markers showed the highest interspecific gene distances while ITS2 showed the lowest ones. An updated key to the 23 Central European species of Docosia is also presented.
Bibionomorpha , Sciaroidea , fungus gnats, Palaearctic Region, taxonomy, DNA sequences, identification key
The species in the genus Docosia Winnertz, 1863 are rather uniform, medium sized fungus gnats (Diptera: Mycetophilidae) with dark body and unmarked hyaline wings. Their identification is mainly based on the characters on the male and female terminalia (cf.
Data on the biology of Docosia are scarce, with the exception of the common mycophagous species Docosia gilvipes (Walker, 1856), see the recent reviews by
Although Docosia is a principally Holarctic genus, several species have also been reported from the Neotropical, Afrotropical and Oriental regions (
A further new species of Docosia from Slovakia, tentatively announced by
The holotype of the new species was collected by the senior author by sweeping in Muránska planina National Park in central Slovakia. This national park belongs to the most valuable protected areas in Slovakia with regard to biodiversity, with many rare and thermophilous species, often reaching there the northernmost limit of their distribution (cf.
The material used for DNA extraction was collected with Malaise traps or sweep net at different localities of the Czech and Slovak Republics in the years 2012–2015. The voucher specimens are deposited in the collection of Jan Ševčík (JSEVC) or in the
All the specimens were collected in 70% ethanol. The holotype is stored in the glycerine medium in a plastic pinned microvial. Before placing in glycerol the specimen was incubated in proteinase K to extract DNA.
Genomic DNA was extracted using NucleoSpin Tissue Kit (Macherey-Nagel, Düren, Germany) following the included protocol. The specimens were cleaned with PBS (Phosphate Buffered Saline) and left in lysis buffer with proteinase K overnight at 56 °C. The primers used for the PCR amplifications were as follows: LCO1490 (5′-GGTCAACAAATCATAAAGATATTGG-3′) and HCO2198 (5′-TAAACTTCAGGGTGACCAAAAAATCA-3′) (
German material discussed here was obtained within the German Barcode of Life Project (GBOL). Specimens used for DNA extraction were collected with Malaise traps at two different localities in Germany in the years 2013 and 2015. All the specimens were collected in pure 96% ethanol.
Genomic DNA was extracted from legs of the specimens using the BioSprint96 magnetic bead extractor by Qiagen (Hilden, Germany). Polymerase chain reaction (PCR) was carried out in total reaction mixes of 20 μl, including 2 μl of undiluted DNA template, 0,8 μl of each primer (10 pmol/μl), 2 μl of ‘Q-Solution’ and 10 μl of ‘Multiplex PCR Master Mix’, containing hot start Taq DNA polymerase and buffers. The latter components are available in the Multiplex PCR kit from Qiagen (Hilden, Germany). PCR reactions were run individually and not multiplexed.
Thermal cycling was performed on GeneAmp PCR System 2700 (Applied Biosystems, Foster City, CA, USA) as follows: hot start Taq activation: 15 min at 95 °C; first cycle set (15 repeats): 35-s denaturation at 94 °C, 90-s annealing at 55 °C (−1 °C/cycle) and 90-s extension at 72 °C. Second cycle set (25 repeats): 35-s denaturation at 94 °C, 90-s annealing at 40 °C and 90-s extension at 72 °C; final elongation 10 min at 72 °C using the primers LCO1490-JJ: 5´-CHACWAAYCATAAAGATATYGG- 3´ with HCO2198-JJ: 5´-AWACTTCVGGRTGVCCAAARAATCA- 3´ respectively (
Sequencing of the unpurified PCR products in both directions was conducted at Beijing Genomics Institute (Hongkong, CN) by using the amplification primers. Barcode sequence analysis was done using the Geneious® software version 7.1.7 (http://www.geneious.com).
Vouchers were deposited in the collection of the Zoologisches Forschungsmuseum Alexander Koenig, Bonn, Germany.
The sequences were aligned using MAFFT version 7 (
Genetic distances were calculated in MEGA6 (
Species | Voucher code | Sampling locality and year | GenBank accession numbers | BOLD Process ID |
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---|---|---|---|---|---|---|---|
COI | COII | CYTB | ITS2 | ||||
Boletina nasuta (Haliday, 1839) | JSGS18 | Slovakia, 2013 | KT923571 | KT923585 | KT923598 | KT923614 | JSDO011-15 |
Gnoriste bilineata Zetterstedt, 1852 | JSGS4 | Czech Republic, 2009 | KT316839 | KT923584 | KT923597 | KT923613 | JSDO014-15 |
Docosia dentata sp. n. | JSDO1 | Slovakia, 2012 | KT923562 | KT923575 | KT923600 | KT923604 | JSDO001-15 |
D. diutina Plassmann, 1996 |
|
Germany, 2013 | KU146854 | KU146860 | KU146858 | KU146856 | SRDOC001-15 |
D. flavicoxa Strobl, 1900 | JSDO9a | Slovakia, 2012 | KT923570 | KT923583 | KT923596 | KT923612 | JSDO009-15 |
D. fumosa Edwards, 1925 |
|
Germany, 2015 | KU146855 | KU146861 | KU146859 | KU146857 | SRDOC002-15 |
D. fuscipes (Roser, 1840) | JSDO2 | Slovakia, 2015 | KT923563 | KT923576 | KT923590 | KT923605 | JSDO002-15 |
D. gilvipes (Walker, 1856) | JSGS29 | Slovakia, 2013 | KT923572 | KT923586 | KT923599 | KT923615 | JSDO012-15 |
D. landrocki Laštovka & Ševčík, 2006 | JSDO7 | Slovakia, 2014 | KT923568 | KT923581 | KT923594 | KT923610 | JSDO007-15 |
D. lastovkai Chandler, 1994 | JSDO4 | Slovakia, 2013 | KT923565 | KT923578 | KT923591 | KT923607 | JSDO004-15 |
D. montana Laštovka & Ševčík, 2006 | JSDO5 | Slovakia, 2013 | KT923566 | KT923579 | KT923592 | KT923608 | JSDO005-15 |
D. moravica Landrock, 1916 | JSDO6 | Slovakia, 2013 | KT923567 | KT923580 | KT923593 | KT923609 | JSDO006-15 |
D. muranica Kurina & Ševčík, 2011 | JSM10 | Slovakia, 2013 | KC435639 | KT923587 | KC435683 | KC435708 | JSDO013-15 |
D. sciarina (Meigen, 1830) | JSDO8 | Czech Republic, 2014 | KT923569 | KT923582 | KT923595 | KT923611 | JSDO008-15 |
D. setosa Landrock, 1916 | JSDO3 | Slovakia, 2015 | KT923564 | KT923577 | KT923601 | KT923606 | JSDO003-15 |
Holotype male, in a pinned microvial with glycerol. SLOVAKIA, Muránska planina National Park, Muráň env., Šiance National Nature Reserve, sweeping along forest edge, N48°46'12", E20°04'20", 1005 m a.s.l., 25. May 2012 (J. Ševčík leg.) [
Male (n = 1). Length of wing 4.2 mm.
Head blackish brown with numerous pale setae. Three ocelli, with lateral ones almost touching compound eyes, separated from the eye margins by their own diameter. Clypeus blackish, with setae pale. Mouthparts light brownish. Palpus brownish yellow, basally and apically darker. Scape, pedicel and all flagellomeres dark brown. Flagellomeres cylindrical, flagellomeres 1 to 7 about twice as long as broad, apical flagellomeres (8 to 14) slightly conical, three times as long as broad.
All parts of thorax blackish brown. All bristles and setae yellowish white. Scutellum with several marginal and submarginal pale bristles and with numerous setae. Antepronotum and proepisternum with pale bristles and short dark setae. Upper part of antepronotum with a strong pale bristle reaching to the ocellus. Laterotergite and other pleural parts bare. Haltere pale yellow.
Legs. All coxae entirely yellow. Femora yellow with hind femur brownish only around its tip. All trochanters blackish brown. Tibiae and tarsi yellow, tarsal segments seemingly brownish because of dense setulae. Fore tibia apicomedially with a semicircular tibial organ (anteroapical depressed area), without strong setae, only densely covered with fine setulae. Mid tibia with 5 anterior, 4 dorsal, 3 anteroventral and 5–6 posterior setae. Hind tibia with 16 anterior, 12 dorsal, 5 anteroventral and 6 posterior setae.
Wings hyaline, unmarked. Radial veins and r-m brown, other veins paler while m-stem and the base of M1 are faint, almost not traceable. Sc, Rs and basal third of cu-stem asetose, the other veins setose. Costa reaches to 0.45 of the distance between R5 and M1. Sc ends in R at the level of beginning of m-stem. Posterior fork begins before anterior fork, approximately at the level of the middle of r-m.
Abdomen all dark brown. Terminalia (Figs
Female. Unknown.
Unknown.
The species name refers to the structure of gonostylus.
The genetic distances among the 13 species of Docosia and two outgroup species for particular gene markers are presented in Tab.
Kimura 2-parametr genetic distances for COI, COII, CytB and ITS2. The lowest and highest values among Docosia species are highlighted in bold.
COI | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 | 13 | 14 | |
1 | D. dentata sp. n. | ||||||||||||||
2 | D. diutina | 8,6% | |||||||||||||
3 | D. flavicoxa | 9,8% | 12,4% | ||||||||||||
4 | D. fumosa | 13,3% | 13,1% | 13,5% | |||||||||||
5 | D. fuscipes | 8,2% | 9,4% | 11,1% | 12,8% | ||||||||||
6 | D. gilvipes | 13,9% | 14,6% | 14,1% | 12,8% | 13,5% | |||||||||
7 | D. landrocki | 9,3% | 9,9% | 12,3% | 13,3% | 10,6% | 15,9% | ||||||||
8 | D. lastovkai | 12,6% | 13,5% | 15,2% | 13,9% | 11,9% | 14,8% | 14,0% | |||||||
9 | D. montana | 7,5% | 10,6% | 11,4% | 15,3% | 10,4% | 14,8% | 11,7% | 14,4% | ||||||
10 | D. moravica | 13,2% | 15,1% | 15,4% | 19,1% | 14,7% | 19,6% | 15,1% | 17,0% | 16,4% | |||||
11 | D. muranica | 10,2% | 13,0% | 14,6% | 16,6% | 11,3% | 16,2% | 13,3% | 15,5% | 12,0% | 17,8% | ||||
12 | D. sciarina | 9,4% | 11,5% | 11,8% | 14,6% | 11,2% | 14,3% | 12,0% | 13,1% | 9,0% | 15,7% | 14,4% | |||
13 | D. setosa | 6,0% | 10,0% | 10,3% | 14,1% | 10,3% | 14,7% | 10,6% | 13,9% | 7,1% | 14,7% | 12,3% | 9,0% | ||
14 | Boletina nasuta | 15,9% | 17,8% | 16,2% | 18,6% | 16,3% | 16,1% | 17,4% | 16,4% | 18,0% | 20,1% | 16,2% | 15,7% | 16,1% | |
15 | Gnoriste bilineata | 16,1% | 17,9% | 18,4% | 15,4% | 17,0% | 15,6% | 18,2% | 17,5% | 18,6% | 19,8% | 20,3% | 17,5% | 17,2% | 14,9% |
COII | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 | 13 | 14 | |
1 | D. dentata sp. n. | ||||||||||||||
2 | D. diutina | 6,6% | |||||||||||||
3 | D. flavicoxa | 7,5% | 8,2% | ||||||||||||
4 | D. fumosa | 14,4% | 17,2% | 14,9% | |||||||||||
5 | D. fuscipes | 7,5% | 8,9% | 11,0% | 17,4% | ||||||||||
6 | D. gilvipes | 7,2% | 10,2% | 11,9% | 16,8% | 8,6% | |||||||||
7 | D. landrocki | 6,8% | 5,6% | 7,8% | 15,2% | 8,4% | 10,7% | ||||||||
8 | D. lastovkai | 11,7% | 12,9% | 14,2% | 19,3% | 11,7% | 9,7% | 14,8% | |||||||
9 | D. montana | 6,2% | 7,3% | 9,4% | 15,8% | 9,3% | 10,3% | 8,4% | 13,5% | ||||||
10 | D. moravica | 6,2% | 8,8% | 7,3% | 16,6% | 9,1% | 9,3% | 9,1% | 12,2% | 7,3% | |||||
11 | D. muranica | 8,6% | 11,0% | 12,9% | 16,0% | 9,1% | 4,3% | 11,5% | 12,2% | 11,1% | 11,0% | ||||
12 | D. sciarina | 6,7% | 8,7% | 8,4% | 17,4% | 10,1% | 8,9% | 8,9% | 13,0% | 6,9% | 7,3% | 11,6% | |||
13 | D. setosa | 5,1% | 6,4% | 7,7% | 15,2% | 8,2% | 10,0% | 6,4% | 13,5% | 5,6% | 6,6% | 11,0% | 7,4% | ||
14 | Boletina nasuta | 23,1% | 22,8% | 20,9% | 21,9% | 21,2% | 22,4% | 22,4% | 23,1% | 21,2% | 22,2% | 20,0% | 22,6% | 21,0% | |
15 | Gnoriste bilineata | 17,7% | 18,5% | 18,2% | 18,2% | 17,7% | 19,7% | 18,8% | 25,4% | 18,8% | 18,0% | 17,4% | 18,6% | 17,9% | 19,1% |
cytB | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 | 13 | 14 | |
1 | D. dentata sp. n. | ||||||||||||||
2 | D. diutina | 8,3% | |||||||||||||
3 | D. flavicoxa | 11,0% | 14,5% | ||||||||||||
4 | D. fumosa | 19,3% | 22,0% | 21,0% | |||||||||||
5 | D. fuscipes | 10,9% | 12,5% | 14,9% | 24,1% | ||||||||||
6 | D. gilvipes | 19,6% | 20,6% | 19,9% | 19,7% | 21,9% | |||||||||
7 | D. landrocki | 10,1% | 9,7% | 14,3% | 23,6% | 12,2% | 19,9% | ||||||||
8 | D. lastovkai | 15,5% | 16,4% | 15,8% | 26,0% | 18,7% | 21,0% | 15,4% | |||||||
9 | D. montana | 9,4% | 12,8% | 17,1% | 22,9% | 13,1% | 22,3% | 13,5% | 21,9% | ||||||
10 | D. moravica | 10,0% | 11,6% | 12,0% | 17,8% | 14,0% | 19,5% | 12,0% | 16,1% | 14,5% | |||||
11 | D. muranica | 7,8% | 12,6% | 13,6% | 21,6% | 10,4% | 19,4% | 12,9% | 16,2% | 13,1% | 13,9% | ||||
12 | D. sciarina | 7,8% | 11,0% | 10,8% | 21,7% | 11,4% | 19,9% | 10,1% | 16,1% | 11,6% | 10,5% | 10,7% | |||
13 | D. setosa | 6,6% | 10,3% | 10,0% | 21,5% | 11,0% | 16,7% | 9,4% | 15,7% | 10,8% | 10,3% | 10,2% | 7,4% | ||
14 | Boletina nasuta | 17,4% | 18,8% | 19,0% | 21,9% | 22,2% | 18,9% | 22,3% | 20,8% | 25,0% | 17,9% | 23,9% | 18,5% | 19,9% | |
15 | Gnoriste bilineata | 16,0% | 18,8% | 19,0% | 18,9% | 20,6% | 20,4% | 20,3% | 21,1% | 21,2% | 16,0% | 20,2% | 18,9% | 18,4% | 17,1% |
ITS2 | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 | 13 | 14 | |
1 | D. dentata sp. n. | ||||||||||||||
2 | D. diutina | 1,8% | |||||||||||||
3 | D. flavicoxa | 2,6% | 3,4% | ||||||||||||
4 | D. fumosa | 9,6% | 9,2% | 10,4% | |||||||||||
5 | D. fuscipes | 0,8% | 2,3% | 3,5% | 8,7% | ||||||||||
6 | D. gilvipes | 7,2% | 6,9% | 9,1% | 11,3% | 7,2% | |||||||||
7 | D. landrocki | 2,0% | 1,3% | 3,7% | 9,2% | 2,9% | 6,0% | ||||||||
8 | D. lastovkai | 4,3% | 5,2% | 5,8% | 12,7% | 4,7% | 7,9% | 5,2% | |||||||
9 | D. montana | 2,0% | 3,4% | 3,2% | 9,6% | 2,1% | 8,7% | 3,7% | 6,4% | ||||||
10 | D. moravica | 1,3% | 3,2% | 1,8% | 9,2% | 1,9% | 8,3% | 3,5% | 4,4% | 2,6% | |||||
11 | D. muranica | 0,3% | 2,1% | 2,9% | 9,6% | 0,5% | 6,9% | 2,3% | 4,1% | 2,3% | 1,3% | ||||
12 | D. sciarina | 1,3% | 2,1% | 3,4% | 9,4% | 2,1% | 7,9% | 2,3% | 5,8% | 2,1% | 2,6% | 1,5% | |||
13 | D. setosa | 1,3% | 2,1% | 2,3% | 9,0% | 2,1% | 8,2% | 2,3% | 5,8% | 1,8% | 2,4% | 1,5% | 1,0% | ||
14 | Boletina nasuta | 23,7% | 23,3% | 23,6% | 22,0% | 23,4% | 21,2% | 23,3% | 25,1% | 23,2% | 23,1% | 23,3% | 21,7% | 21,7% | |
15 | Gnoriste bilineata | 20,5% | 20,5% | 20,3% | 19,3% | 20,2% | 20,1% | 20,5% | 21,1% | 19,6% | 19,7% | 20,1% | 19,0% | 18,5% | 12,8% |
The phylogenetic tree for the concatenated dataset is presented in Fig.
According to the key by
Considering the genetic distance, the closest relative of Docosia dentata is D. setosa, based on COI (6.0 %), COII (5.1%) and CytB (6.6%) gene markers. In the phylogenetic tree based on all the four gene markers (Fig.
The comparison of the four variable gene regions revealed that the genetic distances between Docosia species are quite high for CytB and COI, with the average values being 14.9% and 12.8%, respectively. From this point of view, CytB performs as the best barcoding marker for Docosia species, followed by the traditional animal barcoding region (COI), COII and ITS2. The ITS2 sequences show remarkably high similarity in the genus Docosia and do not appear as a suitable barcode marker in this case. The high uniformity of ITS2 has been reported in several studies (e.g.
There are many studies comparing the utility of various gene markers for DNA barcoding and the identification of species (e.g.
The CytB region has mostly been used in the studies devoted to vertebrates but it was recently used also for barcoding of Calliphoridae (
There has been no previous phylogenetical hypothesis for Docosia species published to date but if we take into account the characters used in most available keys, there is only little congruence between those morphological characters and the molecular results presented here. For example, Docosia fumosa does not appear as particularly separated from the other species by morphological characters. It belongs to the group of species with pubescent lateroterites, together with D. flavicoxa, D. gilvipes, D. moravica, D. sciarina, and several other species, not represented in our molecular dataset. These latter species do not constitute a monophyletic group in the molecular tree (Fig.
The second widely used character in the keys is the coloration of legs, mainly the coxae, which also does not help much in defining any group congruent with the clades in the molecular tree presented here.
Further, it is difficult to find clear morphological synapomorphies for the highly supported clades in the tree (Fig.
Interestingly, in two of the four trees based on one gene region only (in COI and ITS2, data not shown), D. gilvipes (not D. fumosa) branched basally as a sister group to the rest of Docosia, a result which would be expected on the basis of morphological and ecological data (D. gilvipes is the only species in the dataset with Sc setose and ending free, different structure of male terminalia, endomycophagous larvae etc.).
Nevertheless, the number of species included in this analysis is rather limited to reach any final conclusion. A more comprehensive phylogeny of the genus, as well as of other genera of fungus gnats, is thus needed.
1 | Laterotergite pubescent | 2 |
– | Laterotergite bare | 9 |
2 (1) | Sc setose and ending free | 3 |
– | Sc bare and ending in R | 4 |
3 (2) | Tergite 9 subrectangular, gonostylus with a patch of fine subapical setae | D. gilvipes (Walker, 1856) |
– | Tergite 9 distinctly broadened posteriorly, gonostylus without a patch of subapical setae ( |
D. pseudogilvipes Kurina, 2008 |
4 (2) | Hind femur completely black or dark brown | 5 |
– | Hind femur at least partly yellow | 6 |
5 (4) | All coxae and palpi black | D. carbonaria Edwards, 1941 |
– | Coxae yellowish brown, wings slightly smoked | D. fumosa Edwards, 1925 |
6 (4) | Coxae largely dark (at least basal ½), laterotergal setae short | D. sciarina (Meigen, 1830) |
– | Coxae at most parts yellow, laterotergal setae long as usua | 7 |
7 (6) | Mid tibia dorsally with longitudinal black band | D. tibialis Laštovka & Ševčík, 2006 |
– | Mid tibia dorsally yellowish | 8 |
8 (7) | Hind coxa basally brown, apical 1/5 and proximoventral part of hind femur darkened | D. moravica Landrock, 1916 |
– | Hind coxa at most with basal margin darkened, hind femur dark only at tip, its ventral surface yellow | D. flavicoxa Strobl, 1910 |
9 (1) | All coxae dark brown, hind femur largely dark | 10 |
– | Coxae with at least apical half pale | 11 |
10 (9) | Haltere yellowish, male gonocoxites broadly rounded posteriorly ( |
D. fuscipes (von Roser, 1840) |
– | Haltere with a dark knob, male gonocoxites with a deep median incision ( |
D. morionella Mik, 1884 |
11 (9) | All coxae yellow, hind coxa dark at most on basal 1/6 | 12 |
– | Hind coxa blackish brown at least on the basal third | 17 |
12 (11) | Antenna with long basal segments, flagellomere 1 to 4 about twice as long as wide, lateral ocelli distant from eyes | 13 |
– | Antennal segments shorter, flagellomere 1 to 4 about 1.5 as long as wide, ocelli touching eyes | 16 |
13 (12) | Pedicel yellow | D. chandleri Ševčík & Laštovka, 2008 |
– | Pedicel dark brown | 14 |
14 (13) | Posteroventral margin of gonocoxites almost straight, with posterolateral projections, gonostylus short, subtriangular (Figs |
D. dentata sp. n. |
– | Posteroventral margin of gonocoxites medially with a distinct rounded projection, gonostylus elongated | 15 |
15 (14) | Gonostylus with several unusually thick black megasetae along its ventral margin ( |
D. matilei Ševčík & Laštovka, 2008 |
– | Gonostylus with black megasetae only apically ( |
D. montana Laštovka & Ševčík, 2006 |
16 (12) | Costa extends to ½ distance from R5 to M1 or slightly more, apical 1/5 of hind femur dark | D. lastovkai Chandler, 1994 |
– | Costa extends only to 2/5 distance from R5 to M1, only tip of hind femur dark | D. landrocki Laštovka & Ševčík, 2006 |
17 (11) | Hind coxa dark only on basal third, haltere entirely yellowish | 18 |
– | Basal half or slightly more of hind coxa blackish brown | 19 |
18 (17) | Lateral ocelli distant from eyes (about a diameter of ocellus), mid coxa dark on basal third, pedicel dark, male tergite 9 subtrapezoidal ( |
D. setosa Landrock, 1916 |
– | Lateral ocelli touching eyes, mid coxa dark on basal fourth or less, pedicel usually pale, male tergite 9 subcircular ( |
D. expectata Laštovka & Ševčík, 2006 |
19 (17) | Costa extends about 2/5 from R5 to M1, palpi yellow, haltere darkened | D. nigra Landrock, 1928 |
– | Costa extends only about 2/7 from R5 to M1, palpi darkened, haltere yellow | 20 |
20 (19) | Male terminalia with tergite 9 short, as long as broad, trapezoid, posterior margin of gonocoxites without distinct medioventral process ( |
D. muranica Kurina & Ševčík, 2011 |
– | Male terminalia with tergite 9 long, about twice as long as broad, posterior margin of gonocoxites with distinct medioventral process, hind tibia with a dark patch of modified setae | 20 |
21 (20) | Male terminalia with lateral lobes of gonocoxites in ventral view distinctly longer than medioventral process of gonocoxites ( |
D. diutina Plassmann, 1996 |
– | Male terminalia with lateral lobes of gonocoxites shorter, at most as long as medioventral process of gonocoxites | 21 |
22 (21) | Tergite 9 pear-shaped, distinctly broader in posterior half ( |
D. pannonica Laštovka & Ševčík, 2006 |
– | Tergite 9 subrectangular, not distinctly broader in posterior half ( |
D. rohaceki Ševčík, 2006 |
We are grateful to Dr. Jindřich Roháček (Opava, Czech Republic) for his assistance in the field and to the Administration of the Muránska Planina National Park (Revúca, Slovakia) for providing the accommodation and the permission for collecting material. This study was supported by the Ministry of Culture of the Czech Republic by institutional financing of long-term conceptual development of the research institution (the Silesian Museum, MK000100595), internal grant of the Silesian Museum No. IGS201505/2015, and by the internal grant for specific research of the University of Ostrava (SGS28/PřF/2015). Part of the results presented here were achieved in the frame of the German Barcode of Life, a project of the Humboldt Ring, grant-funded by the German Federal Ministry for Education and Research (GBOL1: BMBF #01LI1101A). Peter J. Chandler (Melksham, U.K.) and Olavi Kurina (Tartu, Estonia) provided useful comments and suggestions to the text, especially to the key. P. J. Chandler also kindly checked the English of this paper.