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DNA Barcodes reveal incredible dipteran diversity at a site in Canada's Arctic.
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DNA Barcodes Reveal Incredible Dipteran Diversity at a Site in Canada’s Arctic
Jinjing Wang, Valerie Levesque-Beaudin, Paul Hebert, Ding Yang, Sarah Adamowicz
Jinjing Wang in Churchill
Place of Diptera in Insect Biodiversity
161 families
152244 species
Characters Used in Diptera Taxonomy
• Pollination• Agricultural Pests & Natural Enemies• Infectious Disease Vectors………………
Importance of Diptera
Churchill Northern Studies CentreChurchill Northern Studies Centre
Churchill, Manitoba, Canada
Sub Arctic
Low Arctic
High Arctic
Barcoding All Multicellular Eukaryotes at Churchill
Barcoding Biotas - iBOL >150 researchers and students >50K specimens, >6K species
Previous Knowledge of Diptera of Churchill
32 Families; 280 Species
• How many species of Diptera are there in Churchill?
• How does Diptera species composition differ among tundra, fen, and boreal forest sites?
• Build a DNA barcode library for Diptera from Churchill
Research Questions
Field Collections 2004-2010
Year Period Malaisetrap
Pantrap
Hand-sweep Pitfall Light
trapLiver trap
2004 July-August Yes Yes Yes
2005 July-August Yes Yes Yes Yes Yes
2006 July-August Yes Yes
2007 July-August Yes Yes Yes
2008 July-August Yes Yes Yes
2009 July-August Yes Yes Yes Yes
2010 June-August Yes Yes Yes Yes Yes
Collection Methods
June-August 2010 Malaise Traps
June-August 2010 Malaise Traps
• For the ecological analysis (5 localities), 3 specimens of each provisional morphospecies per Malaise trap sample were barcoded.
• From other localities, 3 specimens of each provisional morphospecies per site were barcoded.
• All the samples were extracted and sequenced in the CCDB according to standard DNA barcoding protocols.
Specimen Sorting and DNA Barcoding
• BINs (Barcode Index Number = barcode cluster) used as provisional species
• BIN accumulation curves were drawn for the order and for families having >100 DNA barcodes. The rate of accumulation of new BINs in the last 10 specimens was calculated. A threshold of 0.01 was used to assess whether a family was well sampled.
Biodiversity Assessment
1600
1800
0
1400
1200
1000
800
600
400
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2000
0 2000 4000 6000 8000 10000 12000 14000 16000
Number of Specimens
56 Families; 1817 BINs
Taxonomic Group Specimens Barcoded Species/ Barcode clusters
Diptera 15,700 1,800
Hymenoptera 10,700 1,300
Lepidoptera 3,300 275
Coleoptera 2,300 250
Trichoptera 2,700 85
Ephemeroptera 650 40
Other insects 2,800 350
Collembola 1,050 100
Arachnids 5,600 775
Crustaceans 1,500 170
Molluscs 580 50
Annelids 1,200 130
Other marine invertebrates 340 60
Vertebrates 220 24
Plants 900 310
Seaweeds 270 65
Total 50,000 5,800
Family BINs Specimens Singletons Doubletons SlopeChironomidae 379 1977 165 52 0.084Sciaridae 140 448 65 28 0.147Mycetophilidae 120 409 67 12 0.164Cecidomyiidae 110 157 85 13 0.456Phoridae 106 418 52 13 0.125Muscidae 97 1573 21 14 0.014Anthomyiidae 92 839 28 14 0.034Empididae 75 958 20 8 0.021Syrphidae 69 923 16 10 0.018Ceratopogonidae 68 243 26 14 0.098Dolichopodidae 63 902 12 10 0.016Chloropidae 48 808 10 4 0.013Tipulidae 41 457 8 4 0.018Sciomyzidae 41 425 8 4 0.019Scathophagidae 32 568 8 6 0.012Ephydridae 31 408 13 2 0.032Tachinidae 31 118 10 3 0.087Simuliidae 20 577 3 4 0.006Limoniidae 20 176 7 3 0.041Culicidae 18 772 5 3 0.007Tabanidae 14 314 4 0 0.012Pipunculidae 12 127 3 2 0.026Calliphoridae 10 153 1 0 0.007
BIN composition of Diptera families from Churchill
Site Families BINs Specimens SingletonsDoubleton
sChao2 Mean
Landing Lake Tundra 35 253 631 163 45 548.21
Launch Road Fen 25 183 439 124 29 448.1
Twin Lake Fen 29 214 484 139 43 438.66
Ramsey Creek Boreal Forest 31 300 741 190 55 628.18
Twin Lake Boreal Forest 33 192 451 129 35 402.22
Total 45 677 2746 344 117 1182.71
Observed and estimated diversity at 5 locations sampled during 2010 via Malaise trapping
Observed and estimated diversity at 5 locations sampled during 2010 via Malaise trapping
Site Families BINs Specimens SingletonsDoubleton
sChao2 Mean
Landing Lake Tundra 35 253 631 163 45 548.21
Launch Road Fen 25 183 439 124 29 448.1
Twin Lake Fen 29 214 484 139 43 438.66
Ramsey Creek Boreal Forest 31 300 741 190 55 628.18
Twin Lake Boreal Forest 33 192 451 129 35 402.22
Total 45 676 2746 344 117 1182.71
Temporal distribution of Diptera BIN richness collected in 2010 using all methods
Venn diagram showing BIN composition overlap among 5 locations sampled in a standardize way in summer 2010
Similarity in species composition of Diptera among five locations measured by Sørensen’s Similarity Index
The proportion of BINs found in 1, 2 and 3 periods at each location sampled in 2010 by Malaise trapping
1817
Increase in species/BIN richness after this
study
1817 Churchill 1.8K BINs
Earth 152K species
Increase in species/BIN richness after this
study
Churchill is Home to >1% of Diptera Biodiversity?
Place of Diptera in Insect Biodiversity?
161 families
152244 species
Experience plays an important role in sampling
Amateur Expert
Example of Muscidae
Number of BINs in each location in each period for 5 locations from 2010