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

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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