1. A Regional Barcode Database for Bees Cory S. Sheffield Department of Biology, York University Toronto, ON

2. The Diversity of Life Estimates of up to 10-15 million species (some are much higher) How can we hope to get an accurate picture of the true richness of life? How much are we losing?

4. Insect Diversity

5. “Biological Identifications through DNA Barcodes” published in 2002 Paul D.N. Hebert University of Guelph

7. A Taxonomic Impediment Many groups of bees are morphologically similar and very difficult to distinguish Some of the bee fauna of Western North America is particularly difficult DNA barcoding for bees…

8. Bees Almost 20,000 described bee species in the world Bee taxonomy requires intensive work!!!

9. A Well Known Fauna T.B. Mitchell “Bees of the Eastern United States” Vol. 1 (1960) & Vol. 2 (1962) Species level identification for bees in Northeastern NA

10. Nova Scotia 55, 491 km 2 Connected to Canada by 30 kms of land Separated by the Bay of Fundy

11. Number of NS Bee Species Pre-2000: 139 bee species recorded in NS based on all literature examined 2003-04: 19 species added to published list 2008: Over 200 species

12. Bee Genera in Nova Scotia Colletes (9) Hylaeus (7) Andrena (41) Protandrena (2) Perdita (1) Calliopsis (1) Dufourea (1) Agapostemon (1) Halictus (3) Lasioglossum (36) Augochlora (1) Augochlorella (1) Macropis (1) Anthidium (1) Heriades (1) Hoplitis (4) Osmia (10) Megachile (11) Ceratina (3?) Melissodes (4) Anthophora (2) Bombus (10) Apis (1) Sphecodes (17) Stelis (6) Coelioxys (5) Nomada (17) Epeolus (2) Triepeolus (1) Epeoloides (1) Bombus (Psithyrus) (4) 12 genera represented by one species

13. Colletidae Hylaeus affinis / modestus Hylaeus basalis Hylaeus annulatus Hylaeus mesillae Hylaeus verticallis Colletes compactus Colletes inaequalis Colletes simulans Macropis nuda Perdita octomaculata 2%

14. Andrenidae Calliopsis andreniformis Andrena nivalis Andrena vicina Andrena kalmiae Andrena melanochroa Andrena crataegi Andrena hirticincta Andrena dunningi Andrena carlini Andrena regularis Andrena frigida Andrena carolina Andrena mandibularis Andrena milwaukeensis Andrena thaspii Andrena barbilabris Andrena robertsonii Andrena wheeleri Andrena alleghaniensis Andrena forbesii Andrena rugosa Andrena hippotes Andrena sigmundi Andrena wilkella Andrena erigeniae Andrena algida Andrena wscripta Protandrena aestivalis Protandrena andrenoides Andrena asteris 5% Andrena canadensis Andrena nubecula Andrena miserabilis

15. Lasioglossum leucozonium Lasioglossum zonulum Lasioglossum cinctipes Lasioglossum acuminatum Lasioglossum coriaceum Lasioglossum divergens Lasioglossum pectorale Lasioglossum cressonii Lasioglossum foxii Lasioglossum quebecense Agapostemon virescen Augochlorella aurata Dufourea novaangliae Halictidae Halictus rubicundus Halictus ligatus Halictus confusus Sphecodes ranunculi Sphecodes confertus Sphecodes autumnalis Sphecodes atlantis Sphecodes solonis Sphecodes cressonii Sphecodes clematidis Sphecodes prosphorus Sphecodes minor Sphecodes dichrous 2% Sphecodes carolinus Sphecodes coronus Lasioglossum athabascense

16. Association of Sexes Higher taxonomic classification + Geographic overlap + DNA barcoding ________________________ Together provide data which could dramatically change regional and global species lists

17. Unknown Sexes We recently discovered and described the unknown male of Megachile nivalis ID was confirmed with DNA barcoding Sheffield & Westby 2007. J. Hym. Res. 16: 178-191. Megachile nivalis Megachile relativa

18. Association of Sexes 50 additional species of Megachile in NA (37%) were known from one sex (Sheffield & Westby 2007) Gonzalez & Griswold 2007: - described two unknown males - synonymy of two females with another species Sheffield et al. (in preparation): - describing unknown sexes of three additional Megachile

19. Megachilidae Coelioxys moesta Coelioxys funeraria Coelioxys rufitarsis Coelioxys sodalis Megachile pugnata Megachile latimanus Megachile gemula Megachile frigida Megachile melanophaea Megachile inermis Megachile centuncularis 2% Megachile montivaga Megachile rotundata Heriades carinata Hoplitis pilosifrons Hoplitis producta Hoplitis spoliata Osmia lignaria Osmia coerulescens Osmia atriventris Osmia bucephala Osmia proxima Osmia inermis Osmia tersula Osmia simillima Osmia virga Stelis foederalis Stelis subemarginata

20. Apidae Apis mellifera Bombus ashtoni Bombus fernaldae Bombus citrinus Bombus insularis Bombus impatiens Bombus ternarius Bombus terricola Bombus borealis Bombus fervidus Bombus rufocinctus Bombus perplexus Bombus sandersoni Bombus vagans Epeoloides pilosula Holcopasites calliopsidis Ceratina dupla Ceratina calcarata Ceratina new sp.? Anthophora bomboides Anthophora furcata Melissodes desponsa Melissodes druriella Epeolus autumnalis Epeolus scutellaris Triepeolus brittaini 2%

21. Cryptic / New Species? Ceratina complex: C. strenua C. calcarata C. dupla C. new sp. ?

22. % Divergence Within Species Within Genus % Divergence Frequency (%)

23. The Nomada Problem Over 300 species in North America Very difficult….many known from one sex and/or few specimens Host associations of most are unknown

24. Barcoding on a Global-level

25. DNA Quality Sequence Length (bp) Frequency (%)

26. Age versus # Base Pairs Age (years) # of Sequences More an artifact of collecting methods and killing agents

27. Collecting / Killing Methods

28. Specimen Preparation Ethyl Acetate Ethanol / Room Temp “Hit and Miss” but mostly miss Generally degraded DNA which did not PCR Cyanide Ethanol / Cold Temp. Critical Point Drying Propylene Glycol Salt water / Ethanol DNA from pinned specimens was mostly fine after these treatments Dillon et al. 1996. Insect Molecular Biology 5: 21-24 Quicke et al. 1999. Zoologica Scripta 28: 261-267

29. Epeoloides pilosula Why not all the NS Species? Rare species….some of the specimens are literature records only Macropis nuda

30. Log-normal Plots Very rare species Rare species

31. Why not all the Species? Rare species….some of the specimens are literary records only But most of the species we have from elsewhere

32. Bees in Canada Province/ Territory No. of Species NS206 NB147 PE65 NF55 QC243 ON379 MB170 SK161 AB286 BC352 NU8 NT80 YT52

33. Barcoding Bees of North America Presently about 30% of the North American bee fauna is barcoded (including ca. 60% Canadian species) Many problematic taxa will be resolved using barcoding

34. Barcoding Bees - Conclusions Taxonomic studies Great potential for ecological studies: –diversity –monitoring –conservation –invasive species

35. Acknowledgements Paul Hebert (University of Guelph) Peter Kevan (University of Guelph) Laurence Packer (York University) John Ascher (AMNH) Terry Griswold (Logan Bee Lab) Jason Gibbs (York University) Sam Droege (USGS)

36. Acknowledgements Biodiversity Institute of Ontario Canadian Barcode of Life Network (www. BOLNET.ca) Genome Canada Ontario Genomics Institute Natural Sciences and Engineering Research Council (NSERC)

37. Thank You