116urn:lsid:arphahub.com:pub:619a5b3a-5ec8-5ff7-b0b1-5070a7c17694urn:lsid:zoobank.org:pub:70C65CC0-001D-487B-A05D-B86A205B9582Contributions to EntomologyCTE0005-805X2511-6428Senckenberg Gesellschaft für Naturforschung10.3897/contrib.entomol.74.e127094127094Research ArticleDipteraDNA barcodingTaxonomyThe Lonchaeidae (Diptera) of the GBOL project, with the description of a new Priscoearomyia speciesReimannAndréandre.reimann@senckenberg.dehttps://orcid.org/0009-0006-3087-49311RulikBjörnhttps://orcid.org/0000-0002-2808-93052Senckenberg Naturhistorische Sammlungen Dresden, Museum für Tierkunde, Königsbrücker Landstrasse 159, 01109 Dresden, GermanySenckenberg Naturhistorische Sammlungen Dresden, Museum für TierkundeDresdenGermanyLeibniz-Institut zur Analyse des Biodiversitätswandels, Zoologisches Forschungsmuseum Alexander Koenig, Adenauerallee 127, 53113 Bonn, GermanyLeibniz-Institut zur Analyse des Biodiversitätswandels, Zoologisches Forschungsmuseum Alexander KoenigBonnGermany
Corresponding author: André Reimann (andre.reimann@senckenberg.de)
Academic editor: Frank Menzel
202416082024742165179F143671C-A88E-59D3-8DF5-5C589BF9B6B77DC8F5E0-FD90-4730-ACAB-6A0105AEB1290805202424072024André Reimann, Björn RulikThis 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.http://zoobank.org/7DC8F5E0-FD90-4730-ACAB-6A0105AEB129
The investigation of 331 specimens of the acalypterate dipteran family Lonchaeidae within the GBOL-project resulted in a list of 29 species from which one is new to science, Priscoearomyiabausenbergensissp. nov., and four species represent new records for Germany. For all voucher specimens detailed metadata are provided including validated DNA barcodes. These barcodes build a sound reference basis for future molecular identification of lonchaeid flies and will also allow the inclusion of female specimens in biodiversity studies, when morphological characters for separating these females are not available.
COI barcodesDNA analysisEuropefaunisticsFranceGermanyLonchaeidaenew recordsnew speciestaxonomyIntroduction
Lonchaeidae is a small family of acalypterate flies, which is distributed worldwide. In Europe, approximately 100 species are known to occur with about 60 in Germany (see Maca 1999, Werner 1995, MacGowan 2004, MacGowan 2015, MacGowan et al. 2007, MacGowan and Rotheray 2008, Reimann and Rulik 2014, Reimann and Stuke 2016, MacGowan and Reimann 2021). The adults are small usually shiny black, sometimes somewhat dulled or slightly green/blue metallic flies. The females always have a strongly sclerotized retractable aculeus. The male genitalia vary in shape and structure in and between the genera. There is a wide range of larval habitats, e.g., under the bark of dead wood, in conifer cones, in rotting plant material, in fruits, in bulbs or in galls in the stems of plants. For more details on the biology see Morge (1963) and MacGowan and Rotheray (2008). In many cases, especially in small dipterans, it has been difficult to proper identify female specimens. Often there is a group of closely related species, where females look very similar and the morphological assignment to the respective males is very difficult or even impossible. Barcoding provides a tool for solving this problem. In Lonchaeidae the barcode sequences of the different species analyzed so far are very distinct and females can be identified in comparing their barcode with those of the males. Afterwards it is possible to search for additional morphological characters, which can aid in identification keys. Furthermore, barcoding can be used to identify torn or incomplete specimens which are always present in Malaise trap samples, especially when the emptying cycles are longer than one week.
Since 2011 the German Barcode of Life project (GBOL) acts as a national barcoding campaign and builds up a voucher-based barcode reference library (Geiger et al. 2016). So far roughly half of the German animal diversity is documented, while the megadiverse groups of Diptera or Hymenoptera are highly underrepresented. Both are present with more than 10.000 species in Germany. Therefore, the recent phase – GBOL III: Dark Taxa – is concentrating especially on micro flies and wasps. But even other small dipteran taxa need some serious attention and will reveal more diversity as known before.
In the present study we combine results with reverse identification approaches out of the Global Malaise trap Program (GMP).
Material and methods
The majority of the material examined was caught with three Malaise traps in Rhineland-Palatine in the administrative district Ahrweiler. The remaining samples are from different localities in other federal states of Germany and France (Fig. 1). For a detailed list of localities see Suppl. material 2.
Map with the sample locations for the data presented here. Map created at GPSVisualizer.com. Map data from OpenStreetMap.org under Open Data Commons Open Database License (opendatacommons.org).
https://binary.pensoft.net/fig/1113370
All specimens except the non-GBOL material are stored in purified ethanol at -20 °C. For morphological identification genitalia were dissected, if needed and treated with KOH-solution (5% in aq.) at 70 °C for 30–60 min., then neutralized with acetic acid (5% in aq.) and rinsed in distilled water. The genitalia are stored in small silicon tubes with the respective specimen. From the non GBOL material one specimen is pinned and the genitalia are mounted on a glass slide; the others are stored in denaturated ethanol (70% in aq).
The lonchaeids were morphologically identified using the keys of Collin (1953), Hackman (1956), MacGowan (2014a, 2014b, 2020), MacGowan and Rotheray (2000, 2008), McAlpine (1982), and Morge (1959a, 1959b, 1959c, 1962, 1963). The nomenclature follows MacGowan (2023). All specimens examined in this study were deposited in the Zoologisches Forschungsmuseum Alexander Koenig, Leibniz-Institut zur Analyse des Biodiversitätswandels (LIB) in Bonn or the Senckenberg Naturhistorische Sammlungen Dresden, Museum für Tierkunde in Dresden.
After morphological identification most of the specimens were barcoded following the established GBOL procedure:
Tissue was subsampled from each specimen and transferred into 96 well plates for subsequent DNA extraction. For specimens larger than 2 mm 1–3 legs were used for lysis. For very small specimens (≤ 2 mm) the whole body was used non-destructively for lysis (i.e., subsequent voucher recovery). Genomic DNA was extracted using the BioSprint96 magnetic bead extractor and respective kits by Qiagen (Hilden, Germany). Polymerase chain reaction (PCR) was carried out in a total reaction volume 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 by Qiagen (Hilden, Germany).
Thermal cycling was performed on GeneAmp PCR System 2700 machines (Life Technologies, Carlsbad, 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. As established within GBOL at ZFMK the following standard degenerate barcoding primers LCO1490-JJ: 5´-CHACWAAYCATAAAGATATYGG- 3´ and HCO2198-JJ: 5´-AWACTTCVGGRTGVCCAAARAATCA- 3´ (Astrin and Stüben 2008) LCO1490-JJ2: 5´-CHACWAAYCAYAARGAYATYGG- 3´ and HCO2198-JJ2: 5´-ANACTTCNGGRTGNCCAAARAATCA- 3´ (Astrin et al. 2016) were used. Purification and sequencing were conducted at BGI (Hong Kong, China) by using the amplification primers.
A minor fraction of specimens was processed within the Global Malaise trap Program (GMP) and the CCDB standard barcoding procedures were applied (https://ccdb.ca/resources/).
Sequences were semi-automatically edited, assembled using the MUSCLE alignment approach (Edgar 2004) and checked for the occurrence of stop-codons or hints of nuclear mitochondrial DNA segments (NUMTs) in GENEIOUS version 7.1.9 (http://www.geneious.com; Kearse et al. 2012). All metadata of specimens treated here were deposited in GBIF (https://biocase.zfmk.de/ipt/resource?r=gbolonch); further details like voucher information such as locality data, habitat, collector, identifier, taxonomic classifications, DNA barcode sequences, primer pairs, the sequence data and trace files were deposited in BOLD (https://doi.org/10.5883/DS-GBOLONCH) and subsequently also transferred to GenBank (accession numbers: OP831582–OP831877 and KT781862–KT781863).
A neighbour-joining tree was obtained using the Tamura-Nei model without outgroup (Tamura and Nei 1993). Morphological identifications were checked on taxonomic consistency via the TaxCI script (Rulik et al. 2017). Inconsistent specimens were either morphologically re-identified (males) or assigned by topology within the genetic distance tree (NJ) and BLASTing against reliable barcode compliant entries of BOLD (females). Additionally, the following species delineation methods: Assemble Species by Automatic Partitioning (ASAP; Puillandre et al. 2021) and the BOLD built-in RESL algorithm for Barcode Index Numbers (BINs; Ratnasingham and Hebert 2013) were applied.
Distance statistics were calculated using DiStats (Astrin et al. 2016). For the alignment only barcode sequences with full length of 658 base pairs (n = 274) were used.
The following abbreviations and terms are used in the text or in the Suppl. materials 1, 2: Collection codes:
MTD – "Senckenberg Naturhistorische Sammlungen Dresden, Museum für Tierkunde, Dresden, Germany;
ZFMK – Zoologisches Forschungsmuseum Alexander Koenig, Bonn, Germany.
General abbreviations:
a.s.l. – (elevation) above sea level;
GBOL – German Barcode of Life;
LG1–
LG4 – Barcode custer 1 to 4 of the L.limatula species group;
MG1–
MG3 – Barcode custer 1 to 3 of the L.mallochi species group;
N/A – not available;
Entomol. Soc. Krefeld – Entomological Society Krefeld (= Entomologischer Verein Krefeld);
ZFMK leg. – collected by ZFMK staff;
ZFMK et al. leg. – collected by ZFMK staff and project co-workers. Morphological abbreviations:
bp – basiphallus;
ce – cercus,
dp – distiphallus;
ea – ejaculatory apodeme,
ep – epandrium,
ha – hypandrial apodeme,
hy – hypoproct;
pa – paramere;
ph – phallus;
su – surstylus;
ve – ventral lobe of epandrium. Nomenclatural abbreviations: L. – Lonchaea; P. – Priscoearomyia.
GERMANY • Rhineland-Palatinate; Ahrweiler, 50.55193°N, 7.16998°E; 194 m a.s.l.; Malaise trap; 1 female (GBOL-223965312), 11 March 2014; 1 male (GBOL-223967397), 01 July 2014; both Rulik leg.; both in ZFMK.
GERMANY • Mecklenburg-Western Pomerania; Putbus; 54.32506°N, 13.53878°E; 27 m a.s.l.; Malaise trap; 6 females (ZFMK-TIS-2578147–ZFMK-TIS-25781472578149, ZFMK-TIS-2578151), 05 June 2016; all Rulik leg.; all in ZFMK.
GERMANY • North Rhine-Westphalia; Schladern near Windeck; 50.8°N, 7.585°E; 124 m a.s.l.; Malaise trap; 2 females (ZFMK-TIS-2008888–ZFMK-TIS-2008889), 06 June 2017; both ZFMK et al. leg.; both in ZFMK • North Rhine-Westphalia; Hennef; 50.8°N, 7.585°E; 124 m a.s.l.; Malaise trap; 1 female (ZFMK-TIS-2607649), 30 May 2017; ZFMK leg.; in ZFMK.
GERMANY • Rhineland-Palatinate; Ahrweiler; 50.55175°N, 7.17226°E; 271 m a.s.l.; Malaise trap; 1 male (ZFMK-TIS-2578121), 02 June 2014; 1 male (ZFMK-TIS-2575253), 27 June 2013; both Rulik leg.; both in ZFMK.
GERMANY • Rhineland-Palatinate; Ahrweiler; 50.46541°N, 7.2259°E; 292 m a.s.l.; Malaise trap; 1 male (ZFMK-TIS-2575284), 09 April 2014; Rulik leg.; in ZFMK • Rhineland-Palatinate; Winningen; 50.32°N, 7.493°E; 243 m a.s.l.; Malaise trap; 1 male (GBOL-224009772), 04 April 2013; 2 females (GBOL-224009996, GBOL-224010052), 18 April 2013; all Rulik leg.; all in ZFMK. • Saxony; Dresden; 51.13369°N, 13.81147°E; 210 m a.s.l.; Malaise trap; 1 male (ZFMK-TIS-2516625), 26 March 2014; Reimann leg.; in ZFMK.
Holotype: GERMANY • male (ZFMK-TIS-2575291, ZFMK-DIP-00100412); Rhineland-Palatinate; Ahrweiler; Niederzissen; Bausenberg; 50.4647°N, 7.22215°E; 321 m a.s.l.; Malaise trap; 17 June 2014; Rulik leg.; in ZFMK.
Paratypes: GERMANY • 1 female (ZFMK-TIS-2578106, ZFMK-DIP-00100413); Hesse; Lorch am Rhein; castle Nollig; 50.04912°N, 7.79777°E; 246 m a.s.l.; Malaise trap; 17 June 2015; Niehuis leg.; in ZFMK • 2 males (MTD-Dip-8223, MTD-Dip-8660); Bavaria; Berching; Rappersdorf; Aspen; 49.12318°N, 11.4916°E; 526 m a.s.l.; Malaise trap; 22 May 2020; Hable leg.; in MTD • 2 males (MTD-Dip-8222, MTD-Dip-8659); Bavaria; Limbach; Stephaniter Weiher; Aspen; 49.74018°N, 10.82520°E; 297 m a.s.l.; Malaise trap; 21 May 2020; Grasse and Schmidl leg.; in MTD • 2 males (MTD-Dip-8224, MTD-Dip-8225); Bavaria; Freystadt; mixed forest S of Burggriesbach; Aspen; 49.12375°N, 11.36218°E; 445 m a.s.l.; Malaise trap; 06 June 2021; Hable leg.; in MTD • 1 male (MTD-Dip-8226); Hesse; Oberzell (Sinntal); swampy area with beech and ash trees; 50.31837°N, 9.70374°E; 538 m a.s.l.; hand collecting; 31 May 2021; Kappert leg.; in MTD.
FRANCE • 1 male (MTD-Dip-8021); Centre-Val de Loire; Huisseau-sur-Cosson; 47.57927°N, 1.47151°E; 86 m a.s.l.; Lindgren funnel trap; 19 May 2021; Sallé and Paupe leg.; in MTD • 1 male (MTD-Dip-8052); Centre-Val de Loire; Loury; 48.03878°N, 2.18281°E; 136 m a.s.l.; Lindgren funnel trap; 19 May 2021; Sallé and Paupe leg.; in MTD.
Description.
Male: Head. Eye with sparse very short hairs. Frons and face completely greyish brown dusted. 9 to 10 frontal setulae in a row along the eye margins, 16 scattered interfrontal setulae. Orbital seta strong and long (0.35 mm), as long as outer vertical seta. Inner vertical seta slightly longer than outer vertical seta. Orbital plate dusted, with a shining central streak, bare apart from orbital seta. Ocellar seta strong, as long as orbital seta. Few scattered ocellar setulae, 0.25 times as long as ocellar seta. Lunula bare. Four anterior genal setulae in a single row. Postpedicel short, oval, not reaching mouth edge, 1.5 times as long as wide, distinct orange spot on the basal medial surface, which occupies approximately one third of the inner surface. Arista two times as long as postpedicel, short pubescent.
Thorax. Thoracic dorsum black, dusted greyish brown, covered in setulae one third length of orbital seta. Scutellum completely dusted greyish brown, no obvious contrast to thoracic dorsum, bare apart from the 4 marginal setae. Lateral sclerites completely dusted. Anepisternum with anterodorsal seta absent, three strong posterior setae in a dorsoventral row, approximately 12–16 scattered setulae on disc. Katepisternum with two strong posterior seta and one to two weaker anterior setae in an irregular row close to the dorsal margin, central part bare, approximately 4–7 setulae ventrally. One seta on proepimeron and one on proepisternum. Calypter yellow-white with light brown fringe. Wings yellowish in anterior half, turning slightly brownish towards wing tip, posterior half whitish yellow. Veins yellowish basally turning brownish towards wing tip. All legs entirely black.
Abdomen. Tergite 5 2.2 times the length of tergite 4, funnel-shaped in dorsal view, strongly narrowing in posterior half, with a closely spaced group of 6 stiff setulae on each side before the apex. A small triangular membraneous area on central anterior margin (Fig. 3C). In lateral view anteriorly bulged, abruptly narrowing in posterior half, beak-shaped (Fig. 3D).
Male terminalia. Epandrium (ep) short, belt like, three times as high as long (Figs 2A, 3A), with a terminal fringe of setulae on each side, gradually increasing in length from ventral to dorsal. Ventral lobes (ve) not sclerotized, only present as a membranous strip not exceeding the width of the epandrium, covered in small setulae. Fused cerci (ce) not sclerotized, hardly visible, with a row of setae. Surstyli (su) extending ventromedially from the shell of the epandrium as two lightly sclerotized processes covered in very short thick-based setulae (Fig. 3B). No prensisetae visible. Hypoproct (hy) a cap-shaped plate, ventral margin with two broad triangular apical lobes, almost touching in midline, leaving a narrow oval incision (Figs 2B, 3B), both lobes with a fringe of hairs increasing in length towards the middle. Phallus Z-shaped with a massive basiphallus (bp) attached to a cross shaped plate with a heavy sclerotized rod extending from this plate anteriorly. Distiphallus (dp) long and straight, less sclerotized (Fig. 2C). Parameres (pa) belt like, not well sclerotized, strongly curved before ending close to base of phallus, not fused in midline (Fig. 2C, D). Hypandrial apodeme (ha) strongly sclerotized.
Drawings of male (A–D) and female (E) genitalia of Priscoearomyiabausenbergensis sp. nov.: A. Lateral view of epandrium; B. Posterior view of epandrium and associated structures; C. Lateral view of phallus and parameres; D. Ventral view of phallus and parameres; E. Lateral (above) and dorsal (below) view of female ovipositor.
Abdomen and genital capsule of male holotype of Priscoearomyiabausenbergensis sp. nov.: A. Lateral view of male genital; B. Apicoventral view of male genital; C. Dorsal view of tip of abdomen; D. Lateral view of tip of abdomen.
https://binary.pensoft.net/fig/1113372
Measurements: Body length 3.74 mm. Wing length 3.60 mm.
Female: Head. As in male. Postpedicel oval, reaching mouth edge, 1.6 times as long as wide. Distinct orange spot at the basal inner surface. Arista 1.75 times as long as postpedicel, short pubescent.
Thorax. Thoracic dorsum sub-shining black, completely dusted greyish brown, anteriorly covered in setulae one third length of orbital seta, posterior to the suture some setulae reaching almost half the length of orbital seta. Scutellum completely dusted greyish brown, dusting stronger than on thoracic dorsum, with 4 marginal setae (only bases visible in paratype). Lateral sclerites completely dusted. Anepisternum with no anterodorsal seta and three strong posterior setae in a dorsoventral row. With a few scattered setulae on disc. Katepisternum with two strong posterior setae and two weaker (half the length of the posterior) in an irregular row close to the dorsal margin, central part bare, 7 setulae ventrally. One seta on proepimeron and one on proepisternum. Calypter yellow-white with light brown fringe. Wings yellowish in anterior half, turning slightly brownish towards wing tip, posterior half yellowish white. Veins yellowish basally turning brownish towards wing tip. Only hind legs present in female paratype, entirely black.
Abdomen. Tergites completely brownish dusted. Sternites completely dusted. Aculeus black with yellow tip. Apical segment as in Fig. 2E. In dorsal view slightly constricted in the middle. In side view a long ventro-apical seta with three very small setulae anterior to these on each side and a tiny setula close to the tip. Dorsal surface with a pair of long setae at postion 0.6 of the total length measured from the base of the segment. Apical dorsal setae missing in paratype female specimen.
Measurements: Body length 3.26 mm. Wing length 3.43 mm.
Etymology.
This species is named after the collecting site of the male holotype.
Biology.
Unknown.
Distribution.
Only known from Europe (Germany, France).
Differential diagnosis.
With the plate-like hypoproct and epandrium without well sclerotized ventral lobes, this species belongs to a group with P.greciana (McAlpine, 1983) and P.hermoensis MacGowan & Freidberg, 2008. It differs from both mentioned species by the shape of the phallus, which is more or less U-shaped in P.greciana and P.hermoensis (see MacGowan 2014a, figs 22–31), but Z-shaped in P.bausenbergensis. Furthermore, the shape of the hypoproct is quite different. In P.greciana the hypoproct is a dome-shaped plate with two very short apical lobes. In P.hermoensis these lobes are broader and longer and have numerous finger-like processes on the outer margin (MacGowan 2014a). In P.bausenbergensis the apical lobes are very broad with smooth outer margins and enclosing a narrow oval incision. The extremely narrowed and elongated tip of the abdomen separates the new species from all other known species of Priscoearomyia, except P.greciana, which shows this character to a lesser extent. The small triangular membraneous area on the anterior margin of tergite 5 is not present in P.greciana and clearly seperates P.bausenbergensis from the latter species. The large distinct orange spot at the base of postpedicel separates the described species from all others, except P.withersi MacGowan, 2014, which can have a small spot present. The new species can be included in the key to the Priscoearomyia species by MacGowan (2014a) as follows:
1
Epandrium without an obvious ventral lobe, hypoproct wider than long, in form of a disc, prensisetae absent (greciana group)
2
–
Epandrium with a square or rectangular ventral lobe, hypoproct not disc shaped, prensisetae present (other Priscoearomyia species)
3
2
Abdominal tergite 5 elongated, strongly narrowing before tip
2a
–
Abdominal tergite 5 not elongated. Apical lobes of hypoproct with finger-like projections laterally, outer margin of surstyli obviously serrated. Phallus U-shaped, thickened with basal processes
P.hermonensis MacGowan & Freidberg
2a
Abdominal tergite 5 without median triangular membaneous area. Apical lobes of hypoproct rounded, without processes, surstyli with multiserial rows of denticles along outer margin and extending over posterior surface. Phallus narrow and sinuous
P.greciana McAlpine
–
Abdominal tergite 5 with median triangular membaneous area. Apical lobes of hypoproct very broad, without processes, medially enclosing oval incision. Surstyli extending ventromedially from the shell of the epandrium as two lightly sclerotized processes covered in very short thick-based setulae. Phallus Z-shaped
GERMANY • Hesse; Lorch; 50.04912°N, 7.79777°E; 246 m a.s.l.; Malaise trap; 1 female (ZFMK-TIS-2578107), 17 June 2015; Niehuis leg.; in ZFMK • North Rhine-Westphalia; Wesel; 51.64820°N, 6.70026°E; 27 m a.s.l.; Malaise trap; 1 male (ZFMK-TIS-2603019); 3 females (ZFMK-TIS-2603006, ZFMK-TIS-2603008, ZFMK-TIS-2603013), 09 June 2016; all Entomol. Soc. Krefeld leg.; all in ZFMK • Rhineland-Palatinate; Ahrweiler; 50.55175°N, 7.17226°E; 271 m a.s.l.; Malaise trap; 1 male (ZFMK-TIS-2575246), 27 June 2013; Rulik leg.; in ZFMK • Saxony-Anhalt; Harzgerode; 51.66887°N, 11.16425°E; 281 m a.s.l.; Malaise trap; 1 male (ZFMK-TIS-2603070), 28 July 2017; Geiger et al. leg.; in ZFMK.
GERMANY • Rhineland-Palatinate; Ahrweiler; 50.55175°N, 7.17226°E; 271 m a.s.l.; Malaise trap; 1 male (ZFMK-TIS-2575245), 1 female (ZFMK-TIS-2575250), 27 June 2013; both Rulik leg.; both in ZFMK • Saxony-Anhalt; Harzgerode; 51.66887°N, 11.16425°E; 281 m a.s.l.; Malaise trap; 1 male (ZFMK-TIS-2603071), 2 females (ZFMK-TIS-2603072, ZFMK-TIS-2603073), 28 July 2017; all Geiger et al. leg.; all in ZFMK.
GERMANY • Mecklenburg-Western Pomerania; Zartwitz; car-net; 1 male (ZFMK-TIS-2584167), 12 June 2015; Kleeberg leg.; in ZFMK • North Rhine-Westphalia; Hennef; 50.8°N, 7.585°E; 124 m a.s.l.; Malaise trap; 1 male (ZFMK-TIS-2607650), 30 May 2017; 1 male (ZFMK-TIS-2607636), 9 females (ZFMK-TIS-2607637–ZFMK-TIS-2607641, ZFMK-TIS-2607693–ZFMK-TIS-2607696), 13 June 2017; all ZFMK leg.; all in ZFMK • North Rhine-Westphalia; Schladern near Windeck; 50.8°N, 7.585°E; 124 m a.s.l.; Malaise trap; 1 male (ZFMK-TIS-2627290), 1 female (ZFMK-TIS-2627291), 23 May 2017; 2 females (ZFMK-TIS-2606587, ZFMK-TIS-2606588), 06 June 2017; 1 male (ZFMK-TIS-2627293), 20 June 2017; 3 females (ZFMK-TIS-2606571, ZFMK-TIS-2606573, ZFMK-TIS-2606574), 18 July 2017; all ZFMK et al. leg.; all in ZFMK • North Rhine-Westphalia; Wesel; 51.64820°N, 6.70026°E; 27 m a.s.l.; Malaise trap; 1 male (ZFMK-TIS-2602997), 19 May 2016; Entomol. Soc. Krefeld leg.; in ZFMK • Saxony; Dresden; 51.12879°N, 13.78985°E; 203 m a.s.l.; hand collecting; 1 female (ZFMK-TIS-2584169), 01 July 2008; Reimann leg.; in ZFMK • Saxony; Großenhain; 51.2929°N, 13.4721°E; 117 m a.s.l.; Malaise trap; 1 female (ZFMK-TIS-2584168), 23 June 2011; Reimann leg.; in ZFMK.
GERMANY • Rhineland-Palatinate; Ahrweiler; 50.55175°N, 7.17226°E; 271 m a.s.l.; Malaise trap; 3 females (ZFMK-TIS-2574266, ZFMK-TIS-2575265, ZFMK-TIS-2575266), 24 April 2014; Rulik leg.; in ZFMK • North Rhine-Westphalia; Hennef; 50.8°N, 7.585°E; 124 m a.s.l.; Malaise trap; 1 male (ZFMK-TIS-2607672), 09 May 2017; 1 male (ZFMK-TIS-2607652), 30 May 2017; ZFMK leg.; in ZFMK.
GERMANY • Lower Saxony; Lüchow-Dannenberg; 53.04133°N, 11.47986°E; 19 m a.s.l.; Malaise trap; 1 female (ZFMK-TIS-2602984), 20 August 2013; Schütte et al. leg.; in ZFMK • North Rhine-Westphalia; Hennef; 50.8°N, 7.585°E; 124 m a.s.l.; Malaise trap; 2 females (ZFMK-TIS-2607642, ZFMK-TIS-2607643), 30 May 2017; 1 female (ZFMK-TIS-2607682), 13 June 2017; 1 female (ZFMK-TIS-2607656), 11 July 2017; all ZFMK leg.; all in ZFMK • North Rhine-Westphalia; Schladern near Windeck; 50.8°N, 7.585°E; 124 m a.s.l.; Malaise trap; 1 female (ZFMK-TIS-2606575), 18 July 2017; ZFMK et al. leg.; in ZFMK • Rhineland-Palatinate; Ahrweiler; 50.55175°N, 7.17226°E; 271 m a.s.l.; Malaise trap; 1 male (ZFMK-TIS-2574288), 1 female (ZFMK-TIS-2575255), 27 June 2013; 2 females (ZFMK-TIS-2574245, ZFMK-TIS-2574246), 25 July 2013; 2 females (ZFMK-TIS-2574230, ZFMK-TIS-2574236), 08 August 2013; all Rulik leg.; all in ZFMK • Rhineland-Palatinate; Ahrweiler; 50.55175°N, 7.17226°E; 271 m a.s.l.; Malaise trap; 1 female (ZFMK-TIS-2578124), 02 June 2014; Rulik leg.; in ZFMK.
GERMANY • Rhineland-Palatinate; Ahrweiler; 50.55175°N, 7.17226°E; 271 m a.s.l.; Malaise trap; 3 males (ZFMK-TIS-2574267–ZFMK-TIS-2575269), 24 April 2014; 1 male (ZFMK-TIS-2575316), 16 May 2014; all Rulik leg.; all in ZFMK.
GERMANY • North Rhine-Westphalia; Hennef; 50.8°N, 7.585°E; 124 m a.s.l.; Malaise trap; 1 male (ZFMK-TIS-2607654), 11 July 2017; ZFMK leg.; in ZFMK • North Rhine-Westphalia; Wesel; 51.64820°N, 6.70026°E; 27 m a.s.l.; Malaise trap; 1 female (ZFMK-TIS-2602999), 09 June 2016; Entomol. Soc. Krefeld leg.; in ZFMK.
GERMANY • Rhineland-Palatinate; Ahrweiler; 50.55175°N, 7.17226°E; 271 m a.s.l.; Malaise trap; 1 male (ZFMK-TIS-2574289), 27 June 2013; 1 female (ZFMK-TIS-2578155), 15 July 2014; 1 female (ZFMK-TIS-2574237), 08 August 2013; 1 female (ZFMK-TIS-2578100), 16 September 2013; all Rulik leg.; all in ZFMK • Rhineland-Palatinate; Winningen; 50.32°N, 7.493°E; 243 m a.s.l.; Malaise trap; 1 male (GBOL-224008221), 20 September 2012; Rulik leg.; in ZFMK.
GERMANY • North Rhine-Westphalia; Schladern near Windeck; 50.8°N, 7.585°E; 124 m a.s.l.; Malaise trap; 1 female (ZFMK-TIS-2606578), 06 June 2017; ZFMK et al. leg.; in ZFMK • Rhineland-Palatinate; Ahrweiler; 50.55175°N, 7.17226°E; 271 m a.s.l.; Malaise trap; 3 females (ZFMK-TIS-2574268, ZFMK-TIS-2574269, ZFMK-TIS-2575274); 1 female (ZFMK-TIS-2575317), 24 April 2014; all Rulik leg.; all in ZFMK.
GERMANY • North Rhine-Westphalia; Hennef; 50.8°N, 7.585°E; 124 m a.s.l.; Malaise trap; 1 female (ZFMK-TIS-2607635), 13 June 2017; ZFMK leg.; in ZFMK • North Rhine-Westphalia; Wesel; 51.64820°N, 6.70026°E; 27 m a.s.l.; Malaise trap; 1 female (ZFMK-TIS-2602995), 19 May 2016; 1 female (ZFMK-TIS-2603001), 09 June 2016; both Entomol. Soc. Krefeld leg.; both in ZFMK.
GERMANY • North Rhine-Westphalia; Hennef; 50.8°N, 7.585°E; 124 m a.s.l.; Malaise trap; 1 female (ZFMK-TIS-2607653), 11 July 2017; ZFMK leg.; in ZFMK • North Rhine-Westphalia; Schladern near Windeck; 50.8°N, 7.585°E; 124 m a.s.l.; Malaise trap; 1 female (ZFMK-TIS-2606576), 18 July 2017; ZFMK et al. leg.; in ZFMK • Saxony-Anhalt; Harzgerode; 51.66887°N, 11.16425°E; 281 m a.s.l., Malaise trap; 1 male (ZFMK-TIS-2603063), 28 July 2017; Geiger et al. leg.; in ZFMK.
GERMANY • Rhineland-Palatinate; Ahrweiler; 50.55175°N, 7.17226°E; 271 m a.s.l.; Malaise trap; 1 female (ZFMK-TIS-2575328), 16 May 2013; 1 female (ZFMK-TIS-2574286), 27 June 2013; both Rulik leg.; both in ZFMK • Rhineland-Palatinate; Ahrweiler; 50.55193°N, 7.16998°E; 194 m a.s.l.; Malaise trap; 1 female (GBOL-223960563), 08 August 2013; Rulik leg.; in ZFMK • North Rhine-Westphalia; Schladern near Windeck; 50.8°N, 7.585°E; 124 m a.s.l.; Malaise trap; 1 female (ZFMK-TIS-2627287), 30 May 2017; 1 female (ZFMK-TIS-2606580), 06 June 2017; both ZFMK et al. leg.; both in ZFMK.
GERMANY • North Rhine-Westphalia; Hennef; 50.8°N, 7.585°E; 124 m a.s.l.; Malaise trap; 1 female (ZFMK-TIS-2607644), 30 May 2017; ZFMK leg.; in ZFMK • Rhineland-Palatinate; Ahrweiler; 50.55175°N, 7.17226°E; 271 m a.s.l.; Malaise trap; 1 male (ZFMK-TIS-2574271), 24 April 2014; Rulik leg.; in ZFMK.
Unclarified species and unnamed clusters in <italic><tp:taxon-name><tp:taxon-name-part taxon-name-part-type="genus" reg="Lonchaea">Lonchaea</tp:taxon-name-part></tp:taxon-name></italic>AnimaliaDipteraLonchaeidaeE862E1AA-6D44-5A87-ACB4-7F56CC615EE0Lonchaeacf.spicataMacGowan, 2008Material.
GERMANY • Hesse; Sinntal; 50.29290°N, 9.55060°E; 374 m a.s.l.; net sweeping; 1 male (ZFMK-TIS-2616851), 27 April 2018; Kappert leg.; in ZFMK.
Remarks.
This specimen could not be assigned to L.spicata with confidence and holotype comparison was not possible due to unavailability. Therefore, we could only assign the specimen to L.cf.spicata.
<italic><tp:taxon-name><tp:taxon-name-part taxon-name-part-type="genus" reg="Lonchaea">L.</tp:taxon-name-part> <tp:taxon-name-part taxon-name-part-type="species" reg="limatula">limatula</tp:taxon-name-part></tp:taxon-name></italic> Collin, 1953 - group
Note. There are four barcode clusters (LG1–LG4) with specimens that show morphological similarities to L.limatula.
Cluster LG1
Material. GERMANY • Rhineland-Palatinate; Ahrweiler; 50.55175°N, 7.17226°E; 271 m a.s.l.; Malaise trap; 1 female (ZFMK-TIS-2574229), 08 August 2013; Rulik leg.; in ZFMK.
Remarks. There was no respective male in our sampling. Therefore, this female could not be assigned to species level by barcoding.
Cluster LG2
Material. GERMANY • Saxony-Anhalt; Harzgerode; 51.66887°N, 11.16425°E; 281 m a.s.l.; Malaise trap; 1 female (ZFMK-TIS-2603065), 28 July 2017; Geiger et al. leg.; in ZFMK.
Remarks. There was no respective male in our sampling. Therefore, this female could not be assigned to species level by barcoding.
Cluster LG3
Material. GERMANY • Rhineland-Palatinate; Ahrweiler; 50.55175°N, 7.17226°E; 271 m a.s.l.; Malaise trap; 1 female (ZFMK-TIS-2578146), 15 July 2014; Rulik leg.; in ZFMK.
Remarks. There was no respective male in our sampling. Therefore, this female could not be assigned to species level by barcoding.
Cluster LG4
Material. GERMANY • Mecklenburg-Western Pomerania; Putbus; 54.32506°N, 13.53878°E; 27 m a.s.l.; Malaise trap; 1 female (ZFMK-TIS-2578150), 05 June 2016; Rulik leg.; in ZFMK • Lower Saxony; Lüchow-Dannenberg; 53.04133°N, 11.47986°E; 19 m a.s.l.; Malaise trap; 1 female (ZFMK-TIS-2602985), 20 August 2013; Schütte et al. leg.; in ZFMK • North Rhine-Westphalia; Hennef; 50.8°N, 7.585°E; 124 m a.s.l.; Malaise trap; 1 female (ZFMK-TIS-2607655), 11 July 2017; ZFMK leg.; in ZFMK • North Rhine-Westphalia; Schladern near Windeck; 50.8°N, 7.585°E; 124 m a.s.l.; Malaise trap; 1 female (ZFMK-TIS-2606577), 18 July 2017; ZFMK et al. leg.; in ZFMK • Rhineland-Palatinate; Ahrweiler; 50.55175°N, 7.17226°E; 271 m a.s.l.; Malaise trap; 1 female (ZFMK-TIS-2574239), 08 August 2013; Rulik leg.; in ZFMK.
Remarks. There was no respective male in our sampling. Therefore, these females could not be assigned to species level by barcoding. Comparison to data on BOLD indicates, that this is most probably Lonchaeacontigua Collin, 1953.
GERMANY • Baden-Württemberg; Oberbergen; 48.09677°N, 7.67622°E; 387 m a.s.l.; pitfall trap; 1 female (ZFMK-TIS-2593340), October 2011; Gack leg.; in ZFMK • Bavaria; Euerbach; 50.05242°N, 10.11571°E; 243 m a.s.l.; Malaise trap; 2 females (ZFMK-TIS-2549003, ZFMK-TIS-2549005), 30 August 2011; Wagner leg.; in ZFMK • Bavaria; Landau; 48.60401°N, 12.73976°E; 421 m a.s.l.; Malaise trap; 2 females (ZFMK-TIS-2548999, ZFMK-TIS-2549001), 20 June 2011; Wagner leg.; in ZFMK • Lower Saxony; Schwanewede; 53.23498°N, 8.59133°E; 7 m a.s.l.; yellow pan trap; 2 females (ZFMK-TIS-2549011, ZFMK-TIS-2549015), 27 June 2001; Rudzinski leg.; in ZFMK • North Rhine-Westphalia; Hennef; 50.8°N, 7.585°E; 124 m a.s.l.; Malaise trap; 1 female (ZFMK-TIS-2607681), 08 May 2017; 1 female (ZFMK-TIS-2607648), 30 May 2017; both ZFMK leg.; both in ZFMK • North Rhine-Westphalia; Schladern near Windeck; 50.8°N, 7.585°E; 124 m a.s.l.; Malaise trap; 2 females (ZFMK-TIS-2627280, ZFMK-TIS-2627281), 30 May 2017; 1 female (ZFMK-TIS-2606586); all ZFMK et al. leg.; all in ZFMK • North Rhine-Westphalia; Wesel; 51.64820°N, 6.70026°E; 27 m a.s.l.; Malaise trap; 1 male (ZFMK-TIS-2602996), 19 May 2016; 1 female (ZFMK-TIS-2602998), 09 June 2016; both Entomol. Soc. Krefeld leg.; both in ZFMK • Rhineland-Palatinate; Ahrweiler; 50.55193°N, 7.16998°E; 194 m a.s.l.; Malaise trap; 1 female (ZFMK-TIS-1800146), 25 July 2013; 1 male (ZFMK-TIS-1800756), 08 August 2013; 1 male (GBOL-223959994), 26 August 2013; 1 female (ZFMK-TIS-1801372), 11 February 2014; 1 female (GBOL-223967436), 01 July 2014; 1 male (GBOL-223970864), 15 July 2014; 1 male (GBOL-223971662), 29 July 2014; 1 female (ZFMK-TIS-1803982), 29 July 2014; 1 female (GBOL-223971994), 29 July 2014; all Rulik leg.; all in ZFMK • Rhineland-Palatinate; Ahrweiler; 50.55175°N, 7.17226°E; 271 m a.s.l.; Malaise trap; 1 female (ZFMK-TIS-2574287), 27 June 2013; 1 male (ZFMK-TIS-2578093), 2 females (ZFMK-TIS-2574228, ZFMK-TIS-2578094), 08 August 2013; 1 female (ZFMK-TIS-2578095), 16 September 2013; 1 female (ZFMK-TIS-2578133), 16 May 2014; 2 females (ZFMK-TIS-2578122, ZFMK-TIS-2578123), 02 June 2014; 2 females (ZFMK-TIS-2578143, ZFMK-TIS-2578154), 15 July 2014; all Rulik leg.; all in ZFMK • Rhineland-Palatinate; Ahrweiler; 50.4647°N, 7.22215°E; 321 m a.s.l.; Malaise trap; 2 females (ZFMK-TIS-2574232, ZFMK-TIS-2575290), 29 July 2014; Rulik leg.; in ZFMK • Rhineland-Palatinate; Ahrweiler; 50.46541°N, 7.2259°E; 292 m a.s.l.; Malaise trap; 3 females (ZFMK-TIS-2575331−ZFMK-TIS-2575333), 16 September 2013; Rulik leg.; in ZFMK • Rhineland-Palatinate; Winningen; 50.32°N, 7.493°E; 243 m a.s.l.; Malaise trap; 1 male (GBOL-224008644), 25 July 2013; Rulik leg.; in ZFMK.
Genetic analysis
Our sampling includes 331 specimens of which 68 are males and 263 are females. Barcoding resulted in COI-sequences of 298 specimens, which allowed us to identify most of the female specimens which could not be identified by morphology. All specimens and the according BOLD and GenBank accession numbers for successfully sequenced material are listed in Suppl. material 2.
Average COI sequence length for the 298 sequences was 653 bp of 658 bp full barcode length, including twenty-four shorter sequences composed from 561 to 648 residues, respectively.
Among nucleotides, there was a compositional bias towards AT: 66.6% especially at third codon positions (average 90.1%) which is close to levels previously reported for other Diptera groups (e.g., Bernasconi et al. 2000; Cywinska et al. 2010; Rivera and Currie 2009). In detail, overall base composition was: A 28.5, C 16.7, G 16.7, T 38.1%.
Altogether, 37,401 pairwise distances were computed for our dataset: of these, 6,560 were intraspecific distances.
Maximum intraspecific distances averaged 0.52% (range 0–2.13%) while the nearest neighbor distance averaged 6.93% (range 2.28–13.07% – 14.3%), roughly 13-fold higher than the maximum intraspecific distance (Fig. 4) indicating an existing barcode gap for our dataset.
Barcode gab analysis. Relationship between maximum intraspecific and nearest neighbor p-distances. Points above the diagonal line indicate species with a barcode gap. Molecular species delimitation by ASAP and BINs suggests 38 putative species, which fits well with our morphological findings (Fig. 5 and Suppl. material 1).
Neighbor joining tree of Priscoearomyia species. Data taken from the German Barcode of Life project (GBOL) and identification verified by morphology; cluster of P.withersi collapsed, with the number of sequences indicated in brackets thereafter.
https://binary.pensoft.net/fig/1113374
Our combined analysis resulted in a list of 29 species from which one, Priscoearomyiabausenbergensis is new to science. Furthermore, it includes eight well separated female only molecular clusters (see Suppl. material 1). Four species are new to the list of German Lonchaeidae. Included in the list are also those clusters of female specimens which could not be identified to species level. If possible, a closer relation to known species is given. One specimen could only be assigned with cf. due to taxonomic issues.
Discussion
In our sampling there were many more females than males. For most genera in the Lonchaeidae aerial swarming behavior of the males is known (McAlpine and Munroe 1968). This might be a more general habit in this taxon as already mentioned in MacGowan and Rothery (2008) and would partly explain why fewer males are obtained by Malaise trap sampling at ground level. The females are actively searching for substrates suitable for egg laying and may therefore be more often encountered by this method. As already mentioned, it was possible to identify most of the female specimens by barcode comparison or group females based on clustering. This is especially important in those species groups where the females are extremely similar and a morphological identification is impossible at the moment, but as far as barcodes of the corresponding males become available this may change immediately. Otherwise, we would not have been able to identify all the female specimens of the genus Priscoearomyia and the female specimens of e.g. the fraxina group of species within the genus Lonchaea (L.fraxina and L.iona in our sampling). Additionally, the assignment of the female paratype of P.bausenbergensis with certainty was only possible based on the barcoding results. The finding of a new species in our sampling is not that surprising. As already mentioned by MacGowan and Reimann (2021), there were several new species descriptions in the genus Priscoearomyia in the last decades. At the moment the species can only be separated by means of the male genitalia. This indicates that there may be many more distinct species in collections, which are not yet recognized. Another reason, of course, is the lack of taxonomic expertise in Lonchaeidae and in a broader sense in many other acalypterate dipteran taxa. Here barcoding of trap samples or other material can aid in finding new species. If there are clusters of specimens which cannot be assigned to known species, these clusters are worthy of deeper morphological investigation. But, at first, there is the need of barcode references for as many species as possible. Our study provides a sound reference basis for future molecular identification of lonchaeid specimens. This can significantly influence the data accessible for biogeographic studies, if all data of female records can be included.
Acknowledgments
We would like to thank all collectors and providers of lonchaeid material to the GBOL project and the collectors of additional material: Claudia Gack, Nicola Grasse, Joachim Hable, Olaf Jäger, Jürgen Kappert, Andreas Kleeberg, Oliver Niehuis, Frauke Nielsen, Anastasia Paupe, Hans-Georg Rudzinski, Aurélian Salle, Jürgen Schmidl, Kai Schütte, Matthias Schwabe, Christian Wagner and the Entomological Society of Krefeld. We are also grateful to Jana Thormann and Laura von der Mark (both ZFMK) for doing the lab work. Genetic 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 / #01LI1501A). We are indebted to the Struktur- und Genehmigungsdirektion Nord, especially Axel Schmidt, granting the collection permit for Rhineland-Palatinate. Map data copyrighted OpenStreetMap contributors and available from https://www.openstreetmap.org. We thank the reviewers and editors for their valuable comments.
ReferencesAstrinJJHöferHSpeldaJHolsteinJBayerSHendrichLHuberBAKielhornKHKrammerHJLemkeMMonjeJCMorinièreJRulikBPetersenMJanssenHMusterC (2016) Towards a DNA Barcode Reference Database for Spiders and Harvestmen of Germany. PLOS ONE 11 e0162624. https://doi.org/10.1371/journal.pone.0162624AstrinJJStübenPE (2008) Phylogeny in cryptic weevils: molecules, morphology and new genera of western Palaearctic Cryptorhynchinae (Coleoptera: Curculionidae).Invertebrate Systematics22: 503–522. https://doi.org/10.1071/IS07057BernasconiMVValsangiacomoCPiffarettiJ-CWardPI (2000) Phylogenetic relationships among Muscoidea (Diptera: Calyptratae) based on mitochondrial DNA sequences.Insect Molecular Biology9: 67–74. https://doi.org/10.1046/j.1365-2583.2000.00158.xCywinskaAHannanMAKevanPGRoughleyREIranpourMHunterFF (2010) Evaluation of DNA barcoding and identification of new haplomorphs in Canadian deerflies and horseflies.Medical and Veterinary Entomology24: 382–410. https://doi.org/10.1111/j.1365-2915.2010.00896.xHackmanW (1956) The Lonchaeidae (Dipt.) of Eastern Fennoscandia.Notulae Entomologicae36: 89–115.MacGowanI (2004) New species of Palearctic Lonchaeidae (Diptera).Dipterists Digest, second series11: 25–32.MacGowanI (2014a) Three new species of Protearomyia McAlpine, 1962 (Diptera: Lonchaeidae) with a key to males of the Palearctic species.Zootaxa3796: 337–348. https://doi.org/10.11646/zootaxa.3796.2.6MacGowanI (2014b) A review of the Lonchaeafraxina group of species (Diptera: Lonchaeidae) with description of a new species.Entomologisk Tidskrift135: 179–186.MacGowanI (2015) Protearomyiawithersi MacGowan (Diptera, Lonchaeidae) new to the British Isles.Dipterists Digest, second series22: 17–20.MacGowanI (2020) European Lonchaea test key. https://lonchaeidae.myspecies.info/node/2373 [Downloaded 2020]MacGowanI (2023) World Catalogue of the family Lonchaeidae (Diptera, Cyclorapha, Acalyptratae).Zootaxa5307: 1–96. https://doi.org/10.11646/zootaxa.5307.1.1MacGowanIMerzBWermelingerB (2007) Six species of Lonchaea Fallén (Diptera, Lonchaeidae new to Switzerland.Mitteilungen der Schweizerischen Entomologischen Gesellschaft80: 31–35.MacGowanIReimannA (2021) A new species of Protearomyia (Diptera, Lonchaeidae) with a review of the genus in the Palearctic.Zootaxa4966: 487–493. https://doi.org/10.11646/zootaxa.4966.4.8MacGowanIRotherayG (2000) New species, additions and possible deletions to British Lonchaea Fallén (Diptera, Lonchaeidae).Dipterists Digest, second series7: 37–49.MacGowanIRotherayG (2008) British Lonchaeidae. Diptera. Cyclorrhapha, Acalyptratae.Handbooks for the identification of British insects 10(15), Royal Entomological Society, St Albans, 142 pp.MácaJ (1999) Lonchaeidae. In: SchumannHBährmannRStarkA (Eds) Entomofauna Germanica 2.Checkliste der Dipteren Deutschlands. Studia dipterologica Supplement 2, Halle an der Saale, Ampyx-Verlag, 158–159.McAlpineJF (1982) Review of the genus Chaetolonchaea Czerny, with the description of a new species from North America (Diptera: Lonchaeidae).Memoirs of the Entomological Society of Washington10: 118–124.McAlpineJFMunroeDD (1968) Swarming of lonchaeid flies and other insects, with description of four new species of Lonchaeidae (Diptera).The Canadian Entomologist100: 1154–1178. https://doi.org/10.4039/Ent1001154-11MorgeG (1959a) Monographie der paläarktischen Lonchaeidae (Diptera).Beiträge zur Entomologie9: 1–92. https://doi.org/10.21248/contrib.entomol.9.1-2.1-92MorgeG (1959b) Monographie der paläarktischen Lonchaeidae (Diptera).Beiträge zur Entomologie9: 323–371. https://doi.org/10.21248/contrib.entomol.9.3-4.323-371MorgeG (1959c) Monographie der paläarktischen Lonchaeidae (Diptera).Beiträge zur Entomologie9: 909–945. https://doi.org/10.21248/contrib.entomol.9.7-8.909-945MorgeG (1962) Monographie der paläarktischen Lonchaeidae (Diptera).Beiträge zur Entomologie12: 381–434. https://doi.org/10.21248/contrib.entomol.12.3-4.381-434MorgeG (1963) Die Lonchaeidae und Pallopteridae Österreichs und der angrenzenden Gebiete. Eine Revision auf Grundlage der Linzer Sammlung und anderer österreichischer Kollektionen. 1. Teil: Die Lonchaeidae.Naturkundliches Jahrbuch der Stadt Linz9: 123–312.ReimannARulikB (2014) Dasiopscalvus Morge (Diptera: Lonchaeidae), a lance fly new to the German fauna, revealed by the GBOL-project.Studia Dipterologica21: 283–287.ReimannAStukeJ-H (2016) Die Lanzenfliegen (Diptera: Lonchaeidae) Niedersachsens und Bremens (Nordwest-Deutschland).Studia dipterologica23: 3–11.RiveraJCurrieDC (2009) Identification of Nearctic black flies using DNA barcodes (Diptera: Simuliidae).Molecular Ecology Resources9: 224–236. https://doi.org/10.1111/j.1755-0998.2009.02648.xPuillandreNBrouilletSAchazG (2021) ASAP: assemble species by automatic partitioning.Molecular Ecology Resources21: 609–620. https://doi.org/10.1111/1755-0998.13281RatnasinghamSHebertPD (2013) A DNA-based registry for all animal species: the Barcode Index Number (BIN) system. PLOS ONE 8: e66213. https://doi.org/10.1371/journal.pone.0066213RulikBEberleJvon der MarkLThormannJJungMKöhlerFApfelWWeigelAKopetzAKöhlerJFritzlarFHartmannMHadullaKSchmidtJHörrenTKrebsDThevesFEulitzUSkaleARohwedderDKleebergAAstrinJJGeigerMFWägeleWJGrobePAhrensD (2017) Using taxonomic consistency with semi-automated data pre-processing for high quality DNA barcodes.Methods in Ecology and Evolution Oxford8: 1878–1887. https://doi.org/10.1111/2041-210X.12824TamuraKNeiM (1993) Estimation of the number of nucleotide substitutions in the control region of mitochondrial-DNA in humans and chimpanzees.Molecular Biology and Evolution10: 512–526. https://doi.org/10.1093/oxfordjournals.molbev.a040023WernerD (1995) Lamprolonchaeasmaragdei (Walker, 1849) – Erstnachweis für Deutschland (Dipt., Lonchaeidae).Entomologische Nachrichten und Berichte39: 154–155.Supplementary materials10.3897/contrib.entomol.74.e127094.suppl175B895E0-061C-5372-B922-AC96034C1298
Results of species delimitation analysis
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fig. S1. Results of species delimitation analysis (ASAP, BINs and morphology) plotted on the COI genetic distance tree.
https://binary.pensoft.net/file/1113375This dataset is made available under the Open Database License (http://opendatacommons.org/licenses/odbl/1.0). The Open Database License (ODbL) is a license agreement intended to allow users to freely share, modify, and use this Dataset while maintaining this same freedom for others, provided that the original source and author(s) are credited.André Reimann, Björn Rulik10.3897/contrib.entomol.74.e127094.suppl2E5F29C3A-4158-572E-B583-AFD2248E9AE4
Metadata of all specimens used in this study including locality details and BOLD and GenBank references
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The table includes all metadata of the specimens used in the study. The locality data are given in detail with all information including locality description, date, method and collector. For those specimens with barcode the BOLD-ID and GenBank accession numbers ar given..
https://binary.pensoft.net/file/1113376This dataset is made available under the Open Database License (http://opendatacommons.org/licenses/odbl/1.0). The Open Database License (ODbL) is a license agreement intended to allow users to freely share, modify, and use this Dataset while maintaining this same freedom for others, provided that the original source and author(s) are credited.André Reimann, Björn Rulik