Cookies Policy

This site uses cookies. By continuing to browse the site you are agreeing to our use of cookies.

I accept this policy

Find out more here

Open Access DNA barcodes and morphology reveal unrecognized species in Chironomidae (Diptera)

No metrics data to plot.
The attempt to load metrics for this article has failed.
The attempt to plot a graph for these metrics has failed.
The full text of this article is not currently available.

Brill’s MyBook program is exclusively available on BrillOnline Books and Journals. Students and scholars affiliated with an institution that has purchased a Brill E-Book on the BrillOnline platform automatically have access to the MyBook option for the title(s) acquired by the Library. Brill MyBook is a print-on-demand paperback copy which is sold at a favorably uniform low price.

DNA barcodes and morphology reveal unrecognized species in Chironomidae (Diptera)

  • HTML
  • PDF
Add to Favorites
You must be logged in to use this functionality
image of Insect Systematics & Evolution

Affiliations: 1: Department of Natural History, NTNU University Museum, Norwegian University of Science and Technology, NO-7491, Trondheim, Norway

For over a decade, DNA barcoding has proven an effective modern tool in taxonomy, evolutionary biology and biodiversity research. Many new species have been discovered and described with DNA barcodes as part of their diagnostic features. Using DNA barcodes, we uncovered a number of potential species within the Tanytarsus curticornis and Tanytarsus heusdensis species complexes (Diptera: Chironomidae) and detected morphological differences a posteriori that support the description of new species. Unusually large intraspecific divergence in COI p-distance (up to 10%) was observed for two species complexes. In total, eight species new to science are described and figured: T. adustus sp. n., T. heberti sp. n., T. madeiraensis sp. n., T. pseudoheusdensis sp. n., T. songi sp. n., T. thomasi sp. n., T. tongmuensis sp. n. and T. wangi sp. n.. Tanytarsus reei and T. tamaoctavus are redescribed, and T. tusimatneous is listed as a new junior synonym of T. tamaduodecimus. The diagnostic characters of the remaining species of the complexes are discussed. Keys to males and pupae are given.


Full text loading...


Data & Media loading...

Article metrics loading...

1. Albu P. (1980) "Diptera Fam. Chironomidae – Subfam. Chironominae". Fauna Republicii Socialista România, Insecta 11: 1320.
2. Anderson A.M.,, Stur E., & Ekrem T. (2013) "Molecular and morphological methods reveal cryptic diversity and three new species of Nearctic Micropsectra (Diptera: Chironomidae)". Freshwater Science 32: 892921.
3. Ashfaq M., & Hebert P.D.N. (2016) "DNA barcodes for bio-surveillance: regulated and economically important arthropod plant pests". Genome 59: 933945.
4. Baker C.C.,, Bittleston L.S.,, Sanders J.G., & Pierce N.E. (2016) "Dissecting host-associated communities with DNA barcodes". Philosophical Transactions of the Royal Society B: Biological Sciences 371: 20150328.
5. Ballard J.W.O., & Whitlock M.C. (2004) "The incomplete natural history of mitochondria". Molecular Ecology 13: 729744.
6. Brodin Y.,, Ejdung G.,, Strandberg J., & Lyrholm T. (2013) "Improving environmental and biodiversity monitoring in the Baltic Sea using DNA barcoding of Chironomidae (Diptera)". Molecular Ecology Resources 13: 9961004.
7. Carew M.E., & Hoffmann A.A. (2015) "Delineating closely related species with DNA barcodes for routine biological monitoring". Freshwater Biology 60: 15451560.
8. Carew M.E.,, Marshall S.E., & Hoffmann A.A. (2011) "A combination of molecular and morphological approaches resolves species in the taxonomically difficult genus Procladius Skuse (Diptera: Chironomidae) despite high intra-specific morphological variation". Bulletin of Entomological Research 101: 505519.
9. Carew M.E.,, Pettigrove V.J.,, Metzeling L., & Hoffmann A.A. (2013) "Environmental monitoring using next generation sequencing: rapid identification of macroinvertebrate bioindicator species". Frontiers in Zoology 10: 45.
10. Cranston P.S. (2000) "Monsoonal tropical Tanytarsus van der Wulp (Diptera: Chironomidae) reviewed: New species, life histories and significance as aquatic environmental indicators". Australian Journal of Entomology 39: 138159.
11. Edgar R.C. (2004) "MUSCLE: multiple sequence alignment with high accuracy and high throughput". Nucleic Acids Research 32: 17921797.
12. Edwards F.W. (1929) "British non-biting midges (Diptera, Chironomidae)". Transactions of the Royal Entomological Society of London 77: 279430.
13. Ekrem T. (1999) "Six new Tanytarsus species from Ghana, West Africa (Insecta, Diptera, Chironomidae)". Spixiana 22: 5368.
14. Ekrem T. (2001) "A Review of Afrotropical Tanytarsus van der Wulp (Diptera: Chironomidae)". Tijdschrift voor Entomologie 144: 540.
15. Ekrem T. (2002) "A review of selected South- and East Asian Tanytarsus v.d. Wulp (Diptera: Chironomidae)". Hydrobiologia 474: 139.
16. Ekrem T. (2004) "Immature stages of European Tanytarsus species I. The eminulus-, gregarius-, lugens- and mendax species groups (Diptera, Chironomidae)". Deutsche Entomologische Zeitschrift 51: 97146.
17. Ekrem T.,, Stur E., & Hebert P.D.N. (2010) "Females do count: Documenting Chironomidae (Diptera) species diversity using DNA barcoding". Organisms Diversity & Evolution 10: 397408.
18. Ekrem T.,, Sublette M.F., & Sublette J.E. (2003) "North American Tanytarsus I. Descriptions and Keys to Species in the eminulus, gregarius, lugens and mendax Species Groups (Diptera : Chironomidae)". Annals of the Entomological Society of America 96: 265328.
19. Ekrem T.,, Willassen E., & Stur E. (2007) "A comprehensive DNA sequence library is essential for identification with DNA barcodes". Molecular Phylogenetics and Evolution 43: 530542.
20. Epler J.H.,, Ekrem T., & Cranston P.S., (2013) "The larvae of Chironominae (Diptera: Chironomidae) of the Holarctic region—keys and diagnoses". In: Cederholm L. ed. Chironomidae of the Holarctic Region: Keys and Diagnoses, Part 1: Larvae. Lund, Sweden: Insect Systematics and Evolution, Supplement 66: 387556.
21. Ferrington L.C.,, Blackwood M.A.,, Wright C.A.,, Crisp N.H.,, Kavanaugh J.L., & Schmidt F.J. (1991) "A protocol for using surface-floating pupal exuviae of Chironomidae for rapid bioassessment of changing water-quality". Sediment and Stream Water Quality in a Changing Environment : Trends and Explanation 203: 181190.
22. Folmer O.,, Black M.,, Hoeh W.,, Lutz R., & Vrijenhoek R. (1994) "DNA primers for amplification of mitochondrial cytochrome c oxidase subunit I from diverse metazoan invertebrates". Molecular Marine Biology and Biotechnology 3: 294299.
23. Giłka W., & Paasivirta L. (2009) "Evaluation of diagnostic characters of the Tanytarsus chinyensis group (Diptera: Chironomidae), with description of a new species from Lapland". Zootaxa 2197: 3142.
24. Glover B. (1973) "The Tanytarsini (Diptera: Chironomidae) of Australia". Australian Journal of Zoology Supplementary Series 21: 403478.
25. Goetghebuer M. (1923) "Nouveaux matériaux pour l'étude de la faune des Chironomides de Belgique. 2e note". Annales du Biologie Lacustre 12: 103120.
26. Goetghebuer M. (1928) "Diptères (Nématocères). Chironomidae. III. Chironomariae". Faune de France 18: 1174.
27. Hajibabaei M.,, Baird D.J.,, Fahner N.A.,, Beiko R., & Golding G.B. (2016) "A new way to contemplate Darwin's tangled bank: how DNA barcodes are reconnecting biodiversity science and biomonitoring". Philosophical Transactions of the Royal Society B: Biological Sciences 371: 20150330.
28. Hausmann A.,, Miller S.E.,, Holloway J.D.,, deWaard J.R.,, Pollock D.,, Prosser S.W.J., & Hebert P.D.N. (2016) "Calibrating the taxonomy of a megadiverse insect family: 3000 DNA barcodes from geometrid type specimens (Lepidoptera, Geometridae)". Genome 59: 671684.
29. Hebert P.D.N.,, Cywinska A., & Ball S.L. (2003a) "Biological identifications through DNA barcodes". Proceedings of the Royal Society of London B: Biological Sciences 270: 313321.
30. Hebert P.D.N.,, Penton E.H.,, Burns J.M.,, Janzen D.H., & Hallwachs W. (2004) "Ten species in one: DNA barcoding reveals cryptic species in the neotropical skipper butterfly Astraptes fulgerator". Proceedings of the National Academy of Sciences of the United States of America 101: 1481214817.
31. Hebert P.D.N.,, Ratnasingham S., & de Waard J.R. (2003b) "Barcoding animal life: cytochrome c oxidase subunit 1 divergences among closely related species". Proceedings of the Royal Society of London B: Biological Sciences 270: S96S99.
32. Heckman K.L.,, Mariani C.L.,, Rasoloarison R., & Yoder A.D. (2007) "Multiple nuclear loci reveal patterns of incomplete lineage sorting and complex species history within western mouse lemurs (Microcebus)". Molecular Phylogenetics and Evolution 43: 353367.
33. Hodgetts J.,, Ostojá-Starzewski J.C.,, Prior T.,, Lawson R.,, Hall J., & Boonham N. (2016) "DNA barcoding for biosecurity: case studies from the UK plant protection program 1". Genome 59: 10331048.
34. Kieffer J.J. (1911) "Nouvelles descriptions de chironomides obtenus d’éclosion". Bulletin de la Société d'Histoire naturelle de Metz 27: 160.
35. Knebelsberger T.,, Landi M.,, Neumann H.,, Kloppmann M.,, Sell A.F.,, Campbell P.D.,, Laakmann S.,, Raupach M.J.,, Carvalho G.R., & Costa F.O. (2014) "A reliable DNA barcode reference library for the identification of the North European shelf fish fauna". Molecular Ecology Resources 14: 10601071.
36. Kranzfelder P.,, Anderson A.M.,, Egan A.T.,, Mazack J.E.,, Bouchard R.W.,, Rufer M.M., & Ferrington L.C. (2015) "Use of Chironomidae (Diptera) surface-floating pupal exuviae as a rapid bioassessment protocol for water bodies". JoVE (Journal of Visualized Experiments) : e52558.
37. Langton P. (1994) "If not “filaments” then what?" CHIRONOMUS Newsletter on Chironomidae Research 6: 9.
38. Lee P.S.,, Gan H.M.,, Clements G.R., & Wilson J.J. (2016) "Field calibration of blowfly-derived DNA against traditional methods for assessing mammal diversity in tropical forests". Genome 59: 10081022.
39. Lehmann J. (1981) "Chironomidae (Diptera) aus Fließgewässern Zentralafrikas. Teil II: Die Region um Kisangani, Zentralzaire". Spixiana. Supplement 5: 185.
40. Lin X.L.,, Stur E., & Ekrem T. (2015) "Exploring genetic divergence in a species-rich insect genus using 2790 DNA Barcodes". PloS One 10: e0138993.
41. Lindeberg B. (1960) "Taxonomy, ecology and voltinism of Chironomus neglectus n. sp. (Diptera) and some related species". Annales Entomologici Fennici 26: 6974.
42. Lindeberg B. (1963) "Taxonomy, biology and biometry of Tanytarsus curticornis Kieff. and T. brundini n. sp. (Dipt., Chironomidae)". Annales Entomologici Fennici 29: 118130.
43. Lindeberg B. (1967) "Sibling species delimitation in the Tanytarsus lestagei aggregate Diptera, Chironomidae". Annales Zoologici Fennici 4: 4586.
44. Lindeberg B. (1970) "Tanytarsini (Diptera, Chironomidae) from northern Fennoscandia". Annales Zoologici Fennici 7: 303312.
45. Littlefair J.E., & Clare E.L. (2016) "Barcoding the food chain: from Sanger to high-throughput sequencing". Genome 59: 946958.
46. Macher J.N.,, Salis R.K.,, Blakemore K.S.,, Tollrian R.,, Matthaei C.D., & Leese F. (2016) "Multiple-stressor effects on stream invertebrates: DNA barcoding reveals contrasting responses of cryptic mayfly species". Ecological Indicators 61: 159169.
47. Meier R.,, Wong W.,, Srivathsan A., & Foo M. (2015) "$1 DNA barcodes for reconstructing complex phenomes and finding rare species in specimen-rich samples". Cladistics 32: 111.
48. Miller S.E.,, Hausmann A.,, Hallwachs W., & Janzen D.H. (2016) "Advancing taxonomy and bioinventories with DNA barcodes". Philosophical Transactions of the Royal Society B: Biological Sciences 371: 20150339.
49. Monaghan M.T.,, Balke M.,, Gregory T.R., & Vogler A.P. (2005) "DNA-based species delineation in tropical beetles using mitochondrial and nuclear markers". Philosophical Transactions of the Royal Society of London B: Biological Sciences 360: 19251933.
50. Na K.B., & Bae Y.J. (2010) "New Species of Stictochironomus, Tanytarsus and Conchapelopia (Diptera: Chironomidae) from Korea". Entomological Research Bulletin 26: 3339.
51. Nzelu C.O.,, Caceres A.G.,, Arrunategui-Jimenez M.J.,, Lanas-Rosas M.F.,, Yanez-Trujillano H.H.,, Luna-Caipo D.V.,, Holguin-Mauricci C.E.,, Katakura K.,, Hashiguchi Y., & Kato H. (2015) "DNA barcoding for identification of sand fly species (Diptera: Psychodidae) from leishmaniasis-endemic areas of Peru". Acta Tropica 145: 4551.
52. Ratnasingham S., & Hebert P.D.N. (2007) "BOLD: The Barcode of Life Data System" ( Molecular Ecology Notes 7: 355364.
53. Ratnasingham S., & Hebert P.D.N. (2013) "A DNA-based registry for all animal species: the barcode index number (BIN) system". PloS One 8: e66213.
54. Raunio J.,, Paavola R., & Muotka T. (2007) "Effects of emergence phenology, taxa tolerances and taxonomic resolution on the use of the chironomid pupal exuvial technique in river biomonitoring". Freshwater Biology 52: 165176.
55. Ree H.I.,, Jeong K.Y., & Nam S.H. (2011) "Six New and Four Unrecorded Species of Tanytarsini (Diptera, Chironomidae, Chironominae) Found in Korea". Animal Systematics, Evolution and Diversity 27: 246261.
56. Reiss F., & Fittkau E.J. (1971) "Taxonomie und Ökologie europäisch verbreiteter Tanytarsus-Arten (Chironomidae, Diptera)". Archiv für Hydrobiologie, Supplement 40: 75200.
57. Renaud A.K.,, Savage J., & Adamowicz S.J. (2012) "DNA barcoding of Northern Nearctic Muscidae (Diptera) reveals high correspondence between morphological and molecular species limits". BMC Ecology 12: 24.
58. Roskov, Y., Abucay, L., Orrell, T., Nicolson, D., Bailly, N., Kirk, P.M., Bourgoin, T., DeWalt, R.E., Decock, W., De Wever, A., Nieukerken, E. van, Zarucchi, J. & Penev, L. (2017) "Species 2000 & ITIS Catalogue of Life", 29th May 2017. Digital resource at Species 2000: Naturalis, Leiden, the Netherlands.
59. Roslin T., & Majaneva S. (2016) "The use of DNA barcodes in food web construction—terrestrial and aquatic ecologists unite! 1". Genome 59: 603628.
60. Sæther O.A. (1969) "Some Nearctic Pondonominae, Diamesinae, and Orthocladiinae (Diptera: Chironomidae)". Bulletin of the Fisheries Research Board of Canada 170: 1154.
61. Sæther O.A. (1980) "Glossary of chironomid morphology terminology (Diptera: Chironomidae)". Entomologica scandinavica, Supplement 14: 151.
62. Sanseverino A.M. (2006) "A review of the genus Tanytarsus van der Wulp, 1874 (Insecta, Diptera, Chironomidae) from the Neotropical region". Dissertation zur Erlangung des Doktorgrades der Fakultät für Biologie der Ludwig-Maximilians-Universität, München : pp. 306.
63. Sasa M. (1980) "Studies on chironomid midges of the Tama River. Part 2. Description of 20 species of Chironominae recovered from a tributary". Research Report from the National Institute for Environmental Studies, Japan 13: 9107.
64. Sasa M. (1983) "Studies on chironomid midges of the Tama River. Part 5. An observation on the distribution of Chironominae along the main stream in June, with description of 15 new species". Research Report from the National Institute for Environmental Studies, Japan 43: 167.
65. Sasa M., & Ichimori K. (1983) "Studies on chironomid midges of the Tama River. Part 7. Additional species collected in winter from the main stream". Research Report from the National Institute for Environmental Studies, Japan 43: 101122.
66. Sasa M., & Suzuki H. (1999a) "Studies on the Chironomid Midges of Tsushima and Iki Islands, Western Japan: Part 1. Species of Chironominae Collected on Tsushima". Tropical Medicine 41: 153.
67. Sasa M., & Suzuki H. (1999b) "Studies on the Chironomid Midges of Tsushima and Iki Islands, Western Japan: Part 3. The Chironomid Species Collected on Iki Island". Tropical Medicine 41: 143179.
68. Schmidt S.,, Schmid-Egger C.,, Morinière J.,, Haszprunar G., & Hebert P.D. (2015) "DNA barcoding largely supports 250 years of classical taxonomy: identifications for Central European bees (Hymenoptera, Apoidea partim)". Molecular Ecology Resources 15: 9851000.
69. Silva F.L., & Wiedenbrug S. (2014) "Integrating DNA barcodes and morphology for species delimitation in the Corynoneura group (Diptera: Chironomidae: Orthocladiinae)". Bulletin of Entomological Research 104: 6578.
70. Sinclair C.S., & Gresens S.E. (2008) "Discrimination of Cricotopus species (Diptera: Chironomidae) by DNA barcoding". Bulletin of Entomological Research 98: 555563.
71. Smith M.A.,, Bertrand C.,, Crosby K.,, Eveleigh E.S.,, Fernandez-Triana J.,, Fisher B.L.,, Gibbs J.,, Hajibabaei M.,, Hallwachs W., & Hind K. (2012) "Wolbachia and DNA barcoding insects: patterns, potential, and problems". PloS One 7: e36514.
72. Song C.,, Wang Q.,, Zhang R.L.,, Sun B.J., & Wang X.H. (2016) "Exploring the utility of DNA barcoding in species delimitation of Polypedilum (Tripodura) non-biting midges (Diptera: Chironomidae)". Zootaxa 4079: 534550.
73. Storå R. (1939) "Mitteilungen über die Nematoceren Finnlands II". Notulae entomologicae 19: 1630.
74. Stur E., & Ekrem T. (2015) "A review of Norwegian Gymnometriocnemus (Diptera, Chironomidae) including the description of two new species and a new name for Gymnometriocnemus volitans (Goetghebuer) sensu Brundin". ZooKeys 508: 127142.
75. Tamura K.,, Stecher G.,, Peterson D.,, Filipski A., & Kumar S. (2013) "MEGA6: molecular evolutionary genetics analysis version 6.0." Molecular Biology and Evolution 30: 27252729.
76. Webb J.M.,, Jacobus L.M.,, Funk D.H.,, Zhou X.,, Kondratieff B.,, Geraci C.J.,, DeWalt R.E.,, Baird D.J.,, Richard B.,, Phillips I., & Hebert P.D. (2012) "A DNA barcode library for North American Ephemeroptera: progress and prospects". PloS One 7: e38063.
77. Whitworth T.,, Dawson R.,, Magalon H., & Baudry E. (2007) "DNA barcoding cannot reliably identify species of the blowfly genus Protocalliphora (Diptera: Calliphoridae)". Proceedings of the Royal Society of London B: Biological Sciences 274: 17311739.
78. Willyard A.,, Cronn R., & Liston A. (2009) "Reticulate evolution and incomplete lineage sorting among the ponderosa pines". Molecular Phylogenetics and Evolution 52: 498511.
79. Witt J.D.,, Threloff D.L., & Hebert P.D. (2006) "DNA barcoding reveals extraordinary cryptic diversity in an amphipod genus: implications for desert spring conservation". Molecular Ecology 15: 30733082.
80. Wulp F.M. (1874) "Dipterologische aanteekeningen". Tijdschrift voor Entomologie 17: 109148.
81. Yang Z.F.,, Landry J.F.,, Handfield L.,, Zhang Y.L.,, Alma Solis M.,, Handfield D.,, Scholtens B.G.,, Mutanen M.,, Nuss M., & Hebert P.D.N. (2012) "DNA barcoding and morphology reveal three cryptic species of Anania (Lepidoptera: Crambidae: Pyraustinae) in North America, all distinct from their European counterpart". Systematic Entomology 37: 686705.
82. Zahiri R.,, Lafontaine J.D.,, Schmidt B.C.,, Dewaard J.R.,, Zakharov E.V., & Hebert P.D. (2014) "A transcontinental challenge–a test of DNA barcode performance for 1,541 species of Canadian Noctuoidea (Lepidoptera)". PloS One 9: e92797.
83. Zhou X.,, Frandsen P.B.,, Holzenthal R.W.,, Beet C.R.,, Bennett K.R.,, Blahnik R.J.,, Bonada N.,, Cartwright D.,, Chuluunbat S., & Cocks G.V. (2016) "The Trichoptera barcode initiative: a strategy for generating a species-level Tree of Life". Philosophical Transactions of the Royal Society B: Biological Sciences 371: 20160025.
84. Zhou X.,, Jacobus L.M.,, DeWalt R.E.,, Adamowicz S.J., & Hebert P.D.N. (2010) "Ephemeroptera, Plecoptera, and Trichoptera fauna of Churchill (Manitoba, Canada): insights into biodiversity patterns from DNA barcoding". Journal of the North American Benthological Society 29: 814837.
Submit comment
Comment moderation successfully completed


Can't access your account?
  • Key

  • Full access
  • Open Access
  • Partial/No accessInformation