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Baeopterogyna mihalyii Matile (Diptera, Mycetophilidae): association of sexes using morphological and molecular approaches with the first description of females.
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PMID:  21976994     Owner:  NLM     Status:  PubMed-not-MEDLINE    
Abstract/OtherAbstract:
Both males and females of Baeopterogyna mihalyiiMatile, 1975 are recorded from northern Greece. Females are described for the first time providing photographs of the general facies and terminalia. In contrast to the single congener with stenopterous females - Baeopterogyna nudipes Vockeroth, 1972 - Baeopterogyna mihalyii is shown to have normally developed wings in both sexes. Association of sexes is based on both morphological characters and sequence data from cytochrome oxidase subunit one (COI). DNA sequences are used for the first time for the association of sexes in Mycetophilidae.
Authors:
Olavi Kurina; Erki Ounap; Gordon Ramel
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Type:  Journal Article     Date:  2011-06-30
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Title:  ZooKeys     Volume:  -     ISSN:  1313-2970     ISO Abbreviation:  Zookeys     Publication Date:  2011  
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Created Date:  2011-10-06     Completed Date:  2011-11-10     Revised Date:  2013-05-29    
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Nlm Unique ID:  101497933     Medline TA:  Zookeys     Country:  Bulgaria    
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Languages:  eng     Pagination:  15-27     Citation Subset:  -    
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Institute of Agricultural and Environmental Sciences, Estonian University of Life Sciences, Riia st 181, 51014 Tartu, Estonia.
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Journal ID (nlm-ta): Zookeys
Journal ID (publisher-id): ZooKeys
ISSN: 1313-2989
ISSN: 1313-2970
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Olavi Kurina, Erki Õunap, Gordon Ramel
creative-commons-attribution:
Received Day: 7 Month: 4 Year: 2011
Accepted Day: 27 Month: 5 Year: 2011
collection publication date: Year: 2011
Electronic publication date: Day: 30 Month: 6 Year: 2011
Issue: 114
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ID: 3130343
PubMed Id: 21976994
DOI: 10.3897/zookeys.114.1364

Baeopterogyna mihalyii Matile (Diptera, Mycetophilidae): association of sexes using morphological and molecular approaches with the first description of females
Olavi Kurina1
Erki Õunap123
Gordon Ramel4
1Institute of Agricultural and Environmental Sciences, Estonian University of Life Sciences, Riia st 181, 51014 Tartu, Estonia
2Institute of Ecology and Earth Sciences, University of Tartu, Vanemuise 46, EE-51014 Tartu, Estonia
3Museum of Zoology, University of Tartu, Vanemuise 46, EE-51014 Tartu, Estonia
4Ploy Plu Residency, 165/5 Moo 9, Pho Khum Rd., Rob Wiang Sub-District, Muang District, Chiang Rai. 57000, Thailand
Correspondence: Corresponding author: Olavi Kurina (olavi.kurina@emu.ee).
[edited-by] Academic editor: Vladimir Blagoderov

Introduction

[taxon-name: Baeopterogyna] Vockeroth, 1972 is a small genus of [taxon-name: Mycetophilidae] including only two species: [taxon-name: Baeopterogyna nudipes] Vockeroth, 1972 from the Nearctic and [taxon-name: Baeopterogyna mihalyii]Matile, 1975 from the Palaearctic region, respectively. The genus belongs to the subfamily [taxon-name: Sciophilinae] and is most closely related to the [taxon-name: Neuratelia] Rondani (Vockeroth 1972, Matile 1975a) distinguished from it by absence of distinct tibial bristles, setosity of the thorax and wing (Søli et al. 2000), and structure of the male terminalia. Both known species of [taxon-name: Baeopterogyna] have simple slender gonostyli compared to the complex convoluted gonostyli of [taxon-name: Neuratelia]. The type species – [taxon-name: Baeopterogyna nudipes] – was described by Vockeroth (1972) from North America (Yukon Territory and Alaska) but since then it has not been recorded. The European species – [taxon-name: Baeopterogyna mihalyii] – was described from Hungarian material (Matile 1975a) and subsequently it has been recorded in the Czech and Slovak Republics (Ševčík 1999, Chandler 2005). Females of [taxon-name: Baeopterogyna nudipes] are described as stenopterous (Vockeroth 1972) while females of [taxon-name: Baeopterogyna mihalyii] were so far unknown but, analogically to the congener, were also supposed to have reduced wings (e.g. Søli et al. 2000).

Since the introduction of the ‘DNA barcoding’ approach (Hebert et al. 2003), it has repeatedly been shown for different groups of organisms that the intraspecific genetic distances of the mitochondrial COI gene are in most cases at least a magnitude smaller than the interspecific genetic distances. Moreover, it has been demonstrated that sharing the same COI haplotype between different species is rare (Hebert et al. 2010, Raupach et al. 2010, Hausmann et al. 2011). Exceptions to these rules are not frequent and it has been shown that ambiguities have often been derived from taxonomically poorly known groups (Handfield and Handfield 2006; Ståhls and Savolainen 2008, Alexander et al. 2009). These findings have allowed the use of COI sequences in different approaches. In addition to being useful in identifying and delimiting species that are otherwise hard to distinguish (Janzen et al. 2005, Burns et al. 2008, Pauls et al. 2010), these so-called ‘DNA barcodes’ have been shown to be powerful tools in e.g. clarifying the status of morphologically different sexes or races of polymorphic species (Foottit et al. 2009, Janzen et al. 2009) and associating different life stages of insects (Zhou et al. 2007, Pauls et al. 2010). These two practices have repeatedly been implemented in earlier studies of [taxon-name: Diptera] (e.g. Carew et al. 2005, Ekrem et al. 2010, Stur and Ekrem 2011), but to the best of our knowledge, no attempts have been made to utilize them in research of [taxon-name: Mycetophilidae]. In this article, we implemented section from the 3’ end of the COI gene, commonly used in the phylogenetic studies of the [taxon-name: Mycetophilidae] (Rindal et al. 2007, 2009). Despite this part of the COI does not overlap with the so-called ‚barcoding fragment’ from near the 5’ end of the gene, it still is influenced by the identical evolutionary processes as part of the same gene. Therefore, all predictions and conclusions regarding to the systematic utility of the ‚barcoding fragment’ can be attributed to the 3’ end of COI.

The current study was initiated by finding both sexes of [taxon-name: Baeopterogyna mihalyii] in Malaise trap samples from northern Greece. The aims of this article are to describe the so far unknown female of [taxon-name: Baeopterogyna mihalyii] and introduce a possibility of using COI sequence data for association of females and males of fungus gnat species.


Material and methods
Collection, illustration and morphological study

All [taxon-name: Baeopterogyna mihalyii] material was collected by GR from the Kerkini Lake area in Northern Greece south of the Bulgarian border. Despite an extensive [taxon-name: Mycetophilidae] material collected from the area during a survey of invertebrates from 2003 to 2009 (for details see Ramel et al. 2008 and http://www.ramel.org/lake-kerkini/project.html), only samples from one home-made Malaise trap yielded 11 specimens of [taxon-name: Baeopterogyna mihalyii]. The trap was situated over the bog from where the Sultanitsa stream springs and faced down hill into a beech ([taxon-name: Fagus sylvatica]) forest. All material was initially collected and preserved in 70% ethanol while the final preservation method of the studied specimens is indicated in Table 1. For detailed study of terminalia they were detached and cleared in solution of KOH, followed by neutralization in acetic acid and washing in distilled water (see also Kurina 2003). The remaining chitinous parts were either (1) inserted into glycerine for study and photography and thereafter preserved as glycerine preparations in polyethylene micro vials or (2) slide-mounted in Euparal following the method described by Kurina (2008). After detaching the terminalia, the remaining part of the abdomen was used for molecular study while the rest of body was slide-mounted. Some male specimens are preserved in alcohol or dry-mounted in accordance with the method dercribed by Vockeroth (1966). The habitus photos were taken in alcohol using a Canon EOS7D camera fitted with a Canon MP-E65 (F2.8 1–5 x) lens. Illustrations of male and female terminalia are combined using Helicon Focus 4.7 software, from several partly focused images taken with a Leica DFC295 camera attached to an Olympus CX31 compound microscope. Morphological terminology follows Søli (1997).

The material has been deposited in IZBE (Institute of Agricultural and Environmental Science, Estonian University of Life Sciences, former Institute of Zoology and Botany) and all specimen data have been inserted into the database of Estonian animal collections (Abarenkov et al. 2010, see also http://elurikkus.ut.ee/collections.php?lang=eng). All specimen information including photographs is available also on the Fungus Gnats Online website (www.sciaroidea.info/taxonomy/45717).

Molecular techniques

The genomic DNA was extracted using a High Pure PCR Template Preparation Kit (Roche Diagnostics GmbH, Mannheim, Germany). Anterior segments of the abdomen that had been stored after genitalia dissection were crushed and used for the extraction. This process was carried out following the manufacturer’s instructions for extraction of genetic material from mammalian tissue.

A 762-bp fragment of cytochrome C oxidase subunit 1 (COI), corresponding to positions 2228–2989 of the mitochondrial genome of [taxon-name: Drosophila melanogaster] Meigen, 1830 (RefSeq NC_001709) was amplified and sequenced using primers C1-J-2195 (5’-TTGATTTTTTGGTCACCCTGAAGT-3’) and TL2-N-3014 (5’-TCCAATGCACTAATCTGCCATATTA-3’) (Simon et al. 1994). PCR was performed in a total volume of 20 µl, with the reaction mixture containing 1X BD Advantage 2 PCR buffer, 1U BD Advantage 2 Polymerase mix (BD Biosciences, San Jose, USA), 0.2 mM dNTP (Fermentas, Vilnius, Lithuania), 4 pmol of primers and 20–80 ng of purified genomic DNA. PCR was carried out in a Biometra T1 Thermocycler (Biometra, Göttingen, Germany), its conditions were an initial denaturation at 94°C for 2 min, 35 cycles of 30 s at 94°C, 30 s at 50°C and 1 min at 68°C, followed by a final extension at 68°C for 7 min. PCR products were visualised on a 1.6% agarose gel, and 10 μl of the PCR solution was treated with fast alkaline phosphatase and exonuclease I (Fermentas). DNA cycle sequencing was performed in a total volume of 10 μl using the Big Dye Terminator v.3.1 Cycle Sequencing Kit (Applied Biosystems, Foster City, USA). Cycling conditions were: initial denaturation for 1 min at 96°C followed by 25 cycles of 10 s at 95°C, 15 s at 47°C and 4 min at 60°C. Both DNA strands were sequenced using 1.6 pmol of primers. The sequences were resolved on a 3730xl DNA Analyzer (Applied Biosystems).

Phylogenetic analysis

In total, 9 specimens including three species of fungus gnats from the subfamily [taxon-name: Sciophilinae] were analysed. In addition to 3 males and 2 females of [taxon-name: Baeopterogyna mihalyii], both sexes of [taxon-name: Allocotocera pulchella] (Curtis 1837) and both sexes of [taxon-name: Sciophila nigronitida] Landrock, 1925, the latter as an outgroup, were included. For detailed information about specimens see Table 1.

Consensus sequences were created with the program Consed (Gordon et al. 1998) using sequence data from both DNA strands. Sequences were double-checked by eye and aligned with ClustalW (Thompson et al. 1994), using BioEdit (Hall 1999) as a sequence editor. Modeltest 3.06 (Posada and Crandall 1998) was used to search for the optimal model of DNA substitution. Bayesian phylogenetic inference, maximum likelihood (ML), maximum parsimony (MP) and neighbour-joining (NJ) approaches were all used to evaluate the robustness of the phylogenetic analysis. The GTR+I model, selected by Modeltest using Akaike Information Criterion, was implemented for NJ and ML analysis in PAUP*4.0b10 (Swofford 1998). Branch supports were assessed using 1000 bootstrap replicates. MP analysis with simple addition of taxa was also performed in PAUP and resulted in a single most parsimonious tree. Branch supports for this tree were assessed using 1000 bootstrap replicates, with 10 heuristic searches and simple addition of taxa used for each replicate. ML, NJ and MP trees were visualised in TreeView 1.6.6 (Page 1996).

Bayesian phylogenetic analysis implementing the GTR+I model was performed using MrBayes 3.1 (Ronquist and Huelsenbeck 2003). Four simultaneous Markov chains (one cold and three heated) were run for 4 million generations, with trees sampled every 1000 generations. Likelihood values were inspected, and the first 1000 sampled trees were discarded as ‘burn-in’. To estimate posterior probabilities of recovered branches, a 50% majority rule was applied. Phylograms were created as average-branch-length consensus trees and visualised in TreeView 1.6.6.


Results and discussion

For determination of male material of [taxon-name: Baeopterogyna mihalyii], the key to mycetophilid genera by Søli et al. (2000) was used successfully. In addition, male specimens were compared with type material of [taxon-name: Baeopterogyna nudipes] (paratypes, 2♂♂, in MNHN; see also Vockeroth 1972) and they were found to be congeneric. Primary association of sexes of [taxon-name: Baeopterogyna mihalyii] was based on simultaneous finding of females and males that were morphologically similar, except characters in terminalia, in the same sample. However, these females have normally developed wings while those of [taxon-name: Baeopterogyna nudipes] are stenoperous (Matile 1975a, Søli et al. 2000). Moreover, they lack setae on the upper part of the anepisternum which are present in males, a character that is considered diagnostic of the genus by Søli et al. (2000). Therefore, additional support by COI sequence data was needed to associate the sexes of [taxon-name: Baeopterogyna mihaylii] unambiguously.

All specimens identified preliminarily as [taxon-name: Baeopterogyna mihalyii] according to their morphological characteristics carried identical COI haplotypes, and the same applied for both [taxon-name: Allocotocera pulchella] individuals, thus proving that morphology-based identification was correct. The [taxon-name: Sciophila nigronitida] specimens, however, carried different COI haplotypes at one locus corresponding to position 2508 of the full mitochondrial genome of [taxon-name: Drosophila melanogaster] (RefSeq NC_001709); the male had an adenine nucleotide, whereas the female had a guanine nucleotide. Since the genetic distance between these two specimens is only 0,13%, i. e. significantly below the average pairwise distance between individuals belonging to different species (Hebert et al. 2010, Raupach et al. 2010, Hausmann et al. 2011), we conclude that their conspecificity is not under question. Due to the non-existing or minimal genetic distance between the conspecific individuals, all three species formed clearly monophyletic well-supported lineages in the phylogenetic trees (Figure 1). Concerning the main question of the current article, we conclude that it has been proven that the hitherto unknown females of [taxon-name: Baeopterogyna mihalyii] have fully developed wings in contrast to the stenopterous females of its only known congener, [taxon-name: Baeopterogyna nudipes].

Among the species used for phylogenetic analysis also [taxon-name: Sciophila nigronitida] is representing the first record from Greece (for collecting details see Table 1).


Taxonomy
[taxon-name[genus: Baeopterogyna][species: mihalyii]]

[taxon-authority: Matile, 1975]

http://species-id.net/wiki/Baeopterogyna_mihalyii

Figs 23

Material studied:

9♂♂ 2♀♀, for collecting data see Table 1: voucher numbers from IZBE0200004 to IZBE0200014.

Female (Figs 2a, 2c – 2f, 3a – 3c).

Description.

Length of body 4.65 – 4.94 mm (n=2).

Head brown with dark setae. Three equally sized ocelli in a triangular arrangement. Clypeus subrounded. Palpus with 5 light brown setose segments with ratios of 1: 1.17:1.58: 2.25:4.33. Mouthparts brownish. Antenna with 2+14 segments. Scape, pedicel and base of first flagellar segment light brown, rest of flagellomeres brown. Scape with sparse setae including dorsoapicals extending to middle of pedicel. Pedicel with sparse and all flagellomeres with dense setae. First flagellomere 3 times as long as wide, succeeding segments gradually shorter. Apical flagellomere cylindrical, about three times as long as wide.

Thorax brown. Scutum covered with pale setae including long lateral hairs. Lateral parts of thorax slightly paler than scutum. Antepronotum with numerous long pale hairs. Proepisternum with numerous shorter setae. Laterotergite and mediotergite with upward directed hairs. Anepimeron and metepisternum with short setae, anepisternum bare. Scutellum with setae not in distinct pairs.

Legs. Fore coxa light brown with hind margin and apical fourth yellow. Mid and hind coxae brown, apically slightly paler. All trochanters brown. All femora and tibiae yellow with apical brown markings. All tarsi dark brown. Tibiae with irregularly arranged setae but without distinct bristles. Fore tibia with a spur 0.18 of basitarsus length. Mid and hind tibiae both with two equal spurs, 0.19 and 0.21 of basitarsus length, respectively. Ratio of femur to tibia for fore, mid and hind legs: 1.00; 0.82; 0.81. Ratio of tibia to basitarsus for fore, mid and hind legs: 1.15; 1.57; 1.86.

Wing hyaline. Length of wing 4.00 – 4.23 mm (n=2). Ratio of length to width 2.83. Veins light brown, setose on both surfaces. Wing membrane with dense irregularly arranged microtrichia and with few macrotrichia in anal area and close to wing tip below of R1 and R5. C not produced beyond apex of R5, which is strongly sinuate. Sc ends in C at the level of beginning of medial fork. Sc2 situated at the level of middle of bM-Cu. M1basally obsolete. Cubital fork begins slightly before the base of r-m. Haltere pale with brownish knob. Both, stem and knob with short setae.

Abdomen brown with segments I–IV lighter. Terminalia (Figs 3a, 3b, 3c) light brown. Cercus distinctly two-segmented, segments with subequal length, proximal segment more than twice as wide as distal. Tergite VIII subquadrate, as large as tergite IX and tergite X together. Sternite VIII with deep ventral cleft. Gonapophysis IX well sclerotized and visible in lateral view. Hypoproct with apical incision, exposed in ventral view.

Male (Figs 2b, 3d).

Length of body 5.36 – 6.32, 5.65 mm (n=5). Length of wing 4.37 – 5.30, 4.76 mm (n=5), ratio of length to width 2.55 – 2.82, 2.64 (n=5). Coloration and other non-terminal characters similar to female except anepisternum, which has short setae on upper part. Terminalia brown. Gonostylus simple, without any additional lobes or spines, slender, tapering, curved medially and covered with short setae.


Conclusive remarks

Vockeroth (1972) described [taxon-name: Baeopterogyna nudipes] in both sexes. In contrast to the normally developed wings in male specimens, the wings in females were greatly reduced: narrow and extending only to the apex of second abdominal tergite. Female halteres were also reduced, with a weak knob (cf. Vockeroth 1972: Figs 2, 5, 6). As the description of females based on two specimens from separate samples (although from the same locality: Herschel Island in Yukon Territory, Canada), this peculiarity cannot be argued as a possible aberration. Hackman (1964) summarized the knowledge of wing reduction in [taxon-name: Diptera] and discussed, among others, low temperature and wind-exposed habitats like oceanic islands as the causes. In addition to [taxon-name: Baeopterogyna nudipes] from an arctic habitat, only a few fungus gnats ([taxon-name: Sciaroidea] excl. [taxon-name: Sciaridae]) are described with reduced wings. [taxon-name: Macrocera crozetensis] Colless, 1970 with stenopterous females (cf. Matile 1975b: Figs 2, 3) has been described from the subantarctic Crozet Islands and [taxon-name: Mycetophila brachyptera] Duret, 1989 with two forms, one of them brachypterous (cf. Duret 1989: Figs 9 – 15), has been described from cold and wet forest of the Argentine-Chilean Patagonia. [taxon-name: Moriniola grilloti] Matile, 1976 with reduction of wing width in females (cf. Matile 1976: Fig. 1) has been described from the Afrotropics (Mayombe, Congo). In most cases, the reduction is obviously caused by the extreme habitat in which the species lives and is a characteristic only of females. This is an adaptive response to environmental pressure and can provide fitness advantages as shown in female flightlessness of some geometrid moths (e.g. Wahlberg et al. 2010). This type of reduction cannot be expected in congeners that inhabit more favourable habitats and finding [taxon-name: Baeopterogyna mihalyii] with normally developed wings of both sexes in central Europe is therefore not surprising.


Acknowledgments

The study was funded by Estonian Science Foundation grants 7558 and 8583, targeted financing projects SF0170160s08 and SF0180122s08, and by the European Union through the European Regional Development Fund (Center of Excellence FIBIR). OK is grateful to the European Commission’s Research Infrastructure for funding the study visit to MNHN via the SYNTHESYS programme (FR-TAF-5005) and is much obliged to the curator Dr. Christophe Daugeron (MNHN) for the opportunity to work with the collections in Paris. We thank Tuuli Reisberg (Estonian Biocentre, Tartu, Estonia) for technical assistance with sequencing. Peter Chandler (Melkshamn, United Kingdom) and Peter Kerr (Sacramento, USA) are acknowledged for their comments and suggestions to the manuscript.


References
Abarenkov K,Tedersoo L,Nilsson RH,Vellak K,Saar I,Veldre V,Parmasto E,Prous M,Aan A,Ots M,Kurina O,Ostonen I,Jõgeva J,Halapuu S,Põldmaa K,Toots M,Truu J,Larsson, K-H,Kõljalg U. (Year: 2010) PlutoF – a web based workbench for ecological and taxonomic research, with an online implementation for fungal ITS sequences.Evolutionary Bioinformatics6:189-196
Alexander LC,Delion M,Hawthorne DJ,Lamp WO,Funk DH. (Year: 2009) Mitochondrial lineages and DNA barcoding of closely related species in the mayfly genus Ephemerella (Ephemeroptera: Ephemerellidae).Journal of the North American Benthological Society28:584-595doi:10.1899/08-150.1
Burns JM,Janzen DH,Hajibabaei M,Hallwachs W,Hebert PDN. (Year: 2008) DNA barcodes and cryptic species of skipper butterflies in the genus Perichares in Area de Conservacio´ n Guanacaste, Costa Rica.Proceedings of the National Academy of Sciences of the United States of America105:6350-6355doi:10.1073/pnas.071218110518436645
Carew ME,Pettigrove V,Hoffmann AA. (Year: 2005) The utility of DNA markers in classical taxonomy: using cytochrome oxidase I markers to differentiate Australian Cladopelma (Diptera: Chironomidae) midges.Annals of the Entomological Society of America98:587-594doi:10.1603/0013-8746(2005)098[0587:TUODMI]2.0.CO;2
Chandler PJ.. (Year: 2005) Fauna Europaea: Mycetophilidae. – In: de Jong H (Ed) Fauna Europaea: Diptera, Nematocera. Fauna Europaea version 1. 2. Available from: http://www.faunaeur.org [accessed 15.02.2011]
Colless D. (Year: 1970) A new species of Macrocera from the Crozet Islands (Diptera: Mycetophilidae).Proceedings of the Royal Entomological Society of London (B)39:27-28doi:10.1111/j.1365-3113.1970.tb00249.x
Duret JP. (Year: 1989) El genero Mycetophila Meigen, 1803, en la Patagonia (Diptera, Mycetophilidae). Parte 13. Descripcion de Mycetophila brachyptera sp. n.Boletin de la Sociedad de Biologia de Concepción60:111-122
Ekrem T,Stur E & Hebert PDN. (Year: 2010) Females do count: Documenting Chironomidae (Diptera) species diversity using DNA barcoding.Organisms Diversity & Evolution10:397-408doi:10.1007/s13127-010-0034-y
Foottit RG,Maw HEL,Pike KS. (Year: 2009) DNA barcodes to explore diversity in aphids (Hemiptera Aphididae and Adelgidae).RediaXCII: 87–91
Gordon D,Abajian C,Green P.. (Year: 1998) Consed: a graphical tool for sequence finishing.Genome Research8:195-2029521923
Hackman W. (Year: 1964) On reduction and loss of wings in Diptera.Notulae Entomologicae44:73-93
Hall TA. (Year: 1999) BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT.Nucleic Acids Symposium Series41:95-98
Handfield D,Handfield L. (Year: 2006) A new species of Plusia (Lepidoptera: Noctuidae) from North America.The Canadian Entomologist138:853-859doi:10.4039/n06-041
Hausmann A,Haszprunar G,Hebert PDN. (Year: 2011) DNA Barcoding the Geometrid Fauna of Bavaria (Lepidoptera): Successes, Surprises, and Questions.PLoS ONE6(2): e17134doi:10.1371/journal.pone.0017134
Hebert PDN,Cywinska A,Ball SL,deWaard JR. (Year: 2003) Biological identification through DNA barcodes.Proceedings of the Royal Society B: Biological Sciences270: 313–321doi:10.1098/rspb.2002.2218
Hebert PDN,deWaard JR,Landry J. (Year: 2010) DNA barcodes for 1/1000 of the animal kingdom.Biology Letters6:359-362doi:10.1098/rsbl.2009.084820015856
Janzen DH,Hajibabaei M,Burns JM,Hallwachs W,Remigio E,Hebert PDN. (Year: 2005) Wedding biodiversity inventory of a large and complex Lepidoptera fauna with DNA barcoding.Philosophical Transactions of the Royal Society B360:1835-1845doi:10.1098/rstb.2005.1715
Janzen DH and 44 authors. (Year: 2009) Integration of DNA barcoding into an ongoing inventory of complex tropical biodiversity.Molecular Ecology Resources9(s1): 1–26doi:10.1111/j.1755-0998.2009.02628.x21564558
Kurina O. (Year: 2003) Notes on the Palaearctic species of the genus Polylepta Winnertz (Diptera : Mycetophilidae) with a new synonymization.Entomologica Fennica,14 (2):91-97
Kurina O. (Year: 2008)Cluzobra matilei sp. n. from French Guyana, with notes on congeners (Diptera: Mycetophilidae).Zootaxa1874:63-68
Matile L. (Year: 1975a) Découverte du genre Baeopterogyna en Région paléarctique; description d’une espèce nouvelle de Hongrie (Diptera, Mycetophilidae).Cahiers des naturalistes, Bull. N. P., n. s.29(1973): 87–88
Matile L. (Year: 1975b) Un nouveau cas de brachyptérisme chez les Diptères Mycetophilidae: la femelle de Macrocera crozetensis Colless.Bulletin de la Société entomologique de France80:2-6
Matile L. (Year: 1976) Un genre nouveau de Mycomyini à nervation alaire reduite; diagnose préliminaire (Dipt., Mycetophilidae Sciophilinae).Bulletin de la Société entomologique de France81:139-140
Page RD. (Year: 1996) TreeView: an application to display phylogenetic trees on personal computers.Computer Applications in the Biosciences12:357-3588902363
Pauls SU,Blahnik RJ,Zhou X,Wardwell CT,Holzenthal RW. (Year: 2010) DNA barcode data confirm new species and reveal cryptic diversity in Chilean Smicridea (Smicridea) (Trichoptera: Hydropsychidae).Journal of the North American Benthological Society29:1058-1074doi:10.1899/09-108.1
Posada D,Crandall KA. (Year: 1998) MODELTEST: testing the model of DNA substitution.Bioinformatics14:817-818doi:10.1093/bioinformatics/14.9.8179918953
Ramel G,Baquero E,Jordana R. (Year: 2008)Biodiversity of the Collembola Fauna of Wetland Kerkini (N. Greece), with description of the sexual dimorphism of Entomobrya atrocincta Schött 1896 (Collembola: Entomobryomorpha).Annales de la Société Entomologique de France (N.S.),44(1):113-128
Raupach MJ,Astrin JJ,Hannig K,Peters MK,Stoeckle MY,Wägele J. (Year: 2010) Molecular species identification of Central European ground beetles (Coleoptera: Carabidae) using nuclear rDNA expansion segments and DNA barcodes.Frontiers in Zoology7: 26doi:10.1186/1742-9994-7-26
Rindal E,Søli GEE,Kjærandsen J,Bachmann L. (Year: 2007) Molecular phylogeny of the fungus gnat tribe Exechiini (Mycetophilidae, Diptera).Zoologica Scripta36:327-335doi:10.1111/j.1463-6409.2007.00285.x
Rindal E,Søli GEE,Bachmann L. (Year: 2009) On the systematics of the fungus gnat subfamily Mycetophilinae (Diptera): a combined morphological and molecular approach.Journal of Zoological Systematics and Evolutionary Research47:227-233doi:10.1111/j.1439-0469.2008.00498.x
Ronquist F,Huelsenbeck JP. (Year: 2003) MrBayes 3: Bayesian phylogenetic inference under mixed models.Bioinformatics19:1572-1574doi:10.1093/bioinformatics/btg18012912839
Simon C,Frati F,Beckenbach A,Crespi B,Liu H,Flook P. (Year: 1994) Evolution, weighing, and phylogenetic utility of mitochondrial gene sequences and a compilation of conserved polymerase chain reaction primers.Annals of the Entomological Society of America87:651-707
Søli GEE. (Year: 1997) On the morphology and phylogeny of Mycetophilidae, with a revision of Coelosia Winnertz (Diptera, Sciaroidea).Entomologica Scandinavica Supplement50:1-139
Søli GEE,Vockeroth RJ,Matile L. (Year: 2000) A. 4. Families of Sciaroidea. In: Papp ,Darvas B. (Eds) Contribution to a Manual of Palaearctic Diptera. Appendix.Science Herald, Budapest, 49–92
Ståhls G,Savolainen E. (Year: 2008) MtDNA COI barcodes reveal cryptic diversity in the Baetis vernus group (Ephemeroptera, Baetidae).Molecular Phylogenetics and Evolution46:82-87doi:10.1016/j.ympev.2007.09.00918023596
Stur E,Ekrem T. (Year: 2011) Exploring unknown life stages of Arctic Tanytarsini (Diptera: Chironomidae) with DNA barcoding.Zootaxa2743:27-39
Swofford DL. (Year: 1998) PAUP*: Phylogenetic Analysis Using Parsimony (*And Other Methods), Version 4.0b10. Sinauer Associates, Massachusetts.
Ševčík J. (Year: 1999) Fifty species of fungus gnats (Diptera: Mycetophilidae) new for the Czech Republic and/or Slovakia, including a new species of Allodia Winnertz.Časopis Slezského zemského muzea, Opava (A)48:97-105
Thompson JD,Higgins DG,Gibson TJ. (Year: 1994) ClustalW: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position specific gap penalties and weight matrix choice.Nucleic Acids Research22:4673-4680doi:10.1093/nar/22.22.46737984417
Vockeroth JR. (Year: 1966) A method of mounting insects from alcohol.The Canadian Entomologist98:69-70doi:10.4039/Ent9869-1
Vockeroth JR. (Year: 1972) A new Nearctic genus of Mycetophilidae (Diptera) with a stenopterous female.The Canadian Entomologist104:1529-1533doi:10.4039/Ent1041529-10
Wahlberg N,Snäll N,Viidalepp J,Ruohomäki K,Tammaru T. (Year: 2010) The evolution of female flightlessness among Ennominae of the Holarctic forest zone (Lepidoptera, Geometridae).Molecular Phylogenetics and Evolution55 (3):929-938doi:10.1016/j.ympev.2010.01.02520123129
Zhou X,Kjer KM,Morse JC. (Year: 2007). Associating larvae and adults of Chinese Hydropsychidae caddisflies (Insecta:Trichoptera) using DNA sequences.Journal of the North American Benthological Society26:719-742doi:10.1899/06-089.1

Figures

[Figure ID: F1]
Figure 1. 

Bayesian phylogenetic tree (GTR+I model) of selected [taxon-name: Mycetophilidae] taxa, based on a 762 bp fragment of a COI gene. Bayesian posterior probabilities are given above the branches; bootstrap support for the ML/NJ/MP trees, which exhibited identical topology, are presented below the branches.



[Figure ID: F2]
Figure 2. 

[taxon-name: Baeopterogyna mihalyii]. a female b male c female wing d female antenna e female thorax f female head (last palpal segments absent). Scale = 1 mm (a, b, c), 0.2 mm (d, e, f).<br/> anepm = anepimeron; anepst = anepisternum; aprnt = antepronotum; htl = halter; ltg = laterotergite; mtepst = metepisternum; mtg = mediotergite; proepst = proepisternum; preepst = preepisternum; sc = scutum.



[Figure ID: F3]
Figure 3. 

[taxon-name: Baeopterogyna mihalyii]. a female terminalia, lateral view b female terminalia, dorsal view c female terminalia, ventral view d male terminalia, ventral view. Scale = 0.2 mm.<br/> cerc = cercus; gc = gonocoxite; gp = gonapophysis; gst = gonostylus; hyp = hypoproct; st = sternite; tg = tergite



Tables
[TableWrap ID: T1] Table 1. 

Details of specimens used for taxonomic study and molecular analysis


Voucher No Species Sex Collecting site, collecting method and collector Date Method of preservation GenBank acc. code for COI
IZBE0200002 [taxon-name: Allocotocera pulchella] (Curtis, 1837) Estonia, Palupõhja [dwc:verbatimCoordinates: 58°25'54.68"N 26°14'28.90"E], Malaise trap, Soon, V. leg. 25.vii – 4. viii 2009 Abdomen used for DNA sequencing; terminalia in glycerin; rest of body dry mounted from ethanol JN007851
IZBE0200003 Abdomen used for DNA sequencing; terminalia in glycerin; rest of body dry mounted from ethanol JN007851
IZBE0200004 [taxon-name: Baeopterogyna mihalyii] Matile, 1975 Greece, Central Macedonia, Kerkini lakes area, village Neo Petritsi, Sultanitsa site, [dwc:verbatimCoordinates: 41°19'02.1"N 023°12'05.0"E], 1485 m a.s.l., Malaise trap, Ramel G. leg. 12 – 18.v 2008 In ethanol
IZBE0200005 In ethanol
IZBE0200006 Abdomen used for DNA sequencing; terminalia in glycerin; rest of body in ethanol JN007850
IZBE0200007 19 – 25. v 2008 Abdomen used for DNA sequencing; rest of body and terminalia slide mounted JN007850
IZBE0200008 Abdomen used for DNA sequencing; terminalia in glycerin; rest of body in ethanol JN007850
IZBE0200009 Slide mounted under 5 different coverslips
IZBE0200010 In ethanol
IZBE0200011 In ethanol
IZBE0200012 25.v – 1. vi 2008 Abdomen used for DNA sequencing; rest of body and terminalia slide mounted JN007850
IZBE0200013 In ethanol
IZBE0200014 Abdomen used for DNA sequencing; terminalia in glycerin; rest of body slide mounted JN007850
IZBE0200015 [taxon-name: Sciophila nigronitida] Landrock, 1925 Greece, Central Macedonia, Kerkini lakes area, village Neo Petritsi, Farfara site, [dwc:verbatimCoordinates: 41°19'30.5"N 023°15'00.1"E], 750 m a.s.l., Malaise trap, Ramel G. leg. 16 – 22. vi 2008 Abdomen used for DNA sequencing; terminalia in glycerin; rest of body dry mounted from ethanol JN007853
IZBE0200016 Abdomen used for DNA sequencing; terminalia in glycerin; rest of body dry mounted from ethanol JN007852


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Keywords: Keywords  Diptera, Mycetophilidae, Baeopterogyna, systematics, Europe, COI.

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