Insect DNA barcoding

Some results from DNA barcoding
of insects from Pasvik
Torbjørn Ekrem et al.
Elisabeth Stur
NTNU University Museum, Trondheim, Norway
Publications:
Insects inhabiting freshwater and humid habitats in Finnmark
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Ekrem, T., Roth, S., Andersen, T., Stur, E., Søli, G. & Halvorsen, G.A. 2012. Insects inhabiting freshwater and humid habitats in
Finnmark, northern Norway. NJE 59, 91–107.
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Kvifte, G.M. & Andersen, T. 2012. Moth flies (Diptera, Psychodidae) from Finnmark, northern Norway. NJE 59, 108–119.
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Kvifte, G.M. 2012. First record of Anomalochaeta guttipennis (Zetterstedt, 1838) from Norway (Diptera, Opomyzidae). NJE 59,
120–121.
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Greve, L. & Andersen, T. 2012. Lacewings (Neuroptera) and Alderflies (Megaloptera) from Finnmark, northern Norway. NJE
59, 122–132.
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Andersen, T. & Hagenlund, L.K. 2012. Caddisflies (Trichoptera) from Finnmark, northern Norway. NJE 59, 133–154.
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Andersen, T. & Kvifte, G. M. 2012. Phantom midges (Diptera, Chaoboridae) from Finnmark, northern Norway. NJE 59, 155–
157.
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Boumans, L. & Brittain, J.E. 2012. Faunistics of stoneflies (Plecoptera) in Finnmark, northern Norway, including DNA barcoding
of Nemouridae. NJE 59, 196–215.
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Søli, G. & Rindal, E. 2012. Fungus gnats (Diptera, Mycetophilidae) from Finnmark, northern Norway. NJE 59,158–181.
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Kjærstad, G., Webb, J.M. & Ekrem, T. 2012. A review of the Ephemeroptera of Finnmark – DNA barcodes identify Holarctic
relations. NJE 59, 182–195.
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Andersen, T., Hagenlund, L.K., Håland, Ø., Kvifte, G.M. & Skartveit, J. New records of lower Diptera (“Nematocera”) from
Finnmark. NJE 60, 182-189.
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Jonassen, T., Andersen, T. & Kvifte, G.M. Empidoidea (Diptera) from Finnmark, northern Norway. NJE 60, 201-245.
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Roth, S. & Coulianos, C-C. 2014. A survey of aquatic and terrestrial Heteroptera in northern Europe with special regard to
Finnmark, Norway (and adjacent regions). NJE 61, 99–116.
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Stur, E. & Borkent A. (in review) When DNA barcoding and morphology mesh: Ceratopogonidae (Diptera) diversity in
Finnmark, Norway. Zookeys
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Stur, E., Halvorsen, G. A. & Ekrem, T. (in preparation) Non-biting midges (Diptera: Chironomidae) from Finnmark, northern
Norway.
Finnmark Ceratopogonidae
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From 0 to 54 species
– 14 likely new to science
– 16 new to Norway
– 1 new to Europe
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DNA barcodes indicate 58 species
– Cryptic diversity
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To be published in ZooKeys
Insecta of Pasvik: DNA barcodes
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Ca. 100 COI sequences Insecta without Diptera
– Ephemeroptera, Heteroptera, Plecoptera, Megaloptera
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Ca. 530 COI sequences Diptera
– Chironomidae (115 sequences, 81 species)
– Ceratopogonidae (179 sequences, 45 species)
– Mycetophilidae (161 sequences, 93 species)
– Psychodidae (20 sequences, 9 species)
– Rest families (95 specimens)
Organisation of DNA barcode data
Specimen
Collection Data
Tissue Sample
Photograph
Extract
DNA
PCR Amplify
Sequence
Web-Accessible Data and
DNA Barcode
Importance of vouchers
The voucher specimen is the key to the library
Identifying of most species is a lot of work
(including slide preparation)
Storing and curating of voucher specimens
in scientific collections is crucial for future revisions
scientific collections increase their value
Analytical success
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Average sequencing success: 91,3%
– well preserved samples: 96-100%
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Sequence quality: 99,6% with < 1%
ambiguity
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17 of 5120 records possible
contaminations
Sequence length summary
Specimen age summary
Challenges
Multiple morphotypes, same cluster
-Hybridisation
-incomplete lineage sorting
-recent radiation/speciation
Multiple clusters, same morphotype
-cryptic species
-Wolbachia infections
Advantages & future prospects
Recognizing of cryptic species
Highlighting regions of interest in chironomid systematics
(speciation, diversification, biogeography, evolution)
Reference library for further studies
(eg. biosurveillance and biomonitoring, environmental barcoding)
Torbjørn Ekrem, Elisabeth Stur, Gaute Kjærstad (NTNU University Museum)
Trond Andersen, Steffen Roth, Gunnar Kvifte (University Museum of Bergen)
Geir Søli (NHM)
Godtfred Anker Halvorsen (UniMiljø, Bergen)
Acknowledgements
Thanks to Linn Hagenlund for sorting the Malaise traps
Paul Hebert and his staff at the Biodiversity Institute, University of Guelph,
Canada for sequencing of partial COI-genes, and the opportunities to join field
work in Churchill
The Norwegian Biodiversity Information Centre for funding