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A Regional Barcode Database for Bees
Cory S. SheffieldDepartment of Biology, York University
Toronto, ON
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?
Insect Diversity
“Biological Identifications through DNA Barcodes” published in 2002
Paul D.N. Hebert
University of Guelph
A Taxonomic ImpedimentMany 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…
Bees
Almost 20,000 described bee species in the world
Bee taxonomy requires intensive work!!!
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
Nova Scotia55, 491 km2
Connected to Canada by 30 kms of land
Separated by the
Bay of Fundy
Number of NS Bee SpeciesPre-2000:
139 bee species recorded in NS based on all literature examined
2003-04:
19 species added to published list
2008:
Over 200 species
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
Colletidae
Hylaeus affinis / modestus
Hylaeus affinis / modestus
Hylaeus basalis
Hylaeus annulatus
Hylaeus mesillae
Hylaeus verticallis
Colletes compactus
Colletes inaequalis
Colletes simulans
Macropis nuda
Perdita octomaculata
2%
AndrenidaeCalliopsis andreniformis
Andrena nivalisAndrena vicina
Andrena kalmiaeAndrena melanochroa
Andrena crataegi
Andrena hirticincta
Andrena dunningiAndrena carlini
Andrena regularisAndrena frigida
Andrena carolinaAndrena mandibularisAndrena milwaukeensis
Andrena thaspiiAndrena barbilabris
Andrena robertsoniiAndrena wheeleri
Andrena alleghaniensisAndrena forbesii
Andrena rugosaAndrena hippotesAndrena sigmundiAndrena wilkella
Andrena erigeniaeAndrena algida
Andrena wscriptaProtandrena aestivalisProtandrena andrenoides
Andrena asteris
5%
Andrena canadensisAndrena canadensisAndrena canadensis
Andrena nubeculaAndrena nubeculaAndrena nubecula
Andrena miserabilisAndrena miserabilisAndrena miserabilisAndrena miserabilis
Lasioglossum leucozoniumLasioglossum zonulum
Lasioglossum cinctipesLasioglossum acuminatum
Lasioglossum coriaceumLasioglossum divergens
Lasioglossum pectoraleLasioglossum cressonii
Lasioglossum foxiiLasioglossum quebecense
Agapostemon virescenAugochlorella aurata
Dufourea novaangliae
Halictidae
Halictus rubicundusHalictus ligatus
Halictus confusus
Sphecodes ranunculiSphecodes confertus
Sphecodes autumnalisSphecodes atlantis
Sphecodes solonis
Sphecodes cressoniiSphecodes clematidis
Sphecodes prosphorusSphecodes minorSphecodes dichrous
2%
Sphecodes carolinusSphecodes coronus
Lasioglossum athabascense
Association of Sexes
Higher taxonomic classification
+ Geographic overlap
+ DNA barcoding
________________________
Together provide data which could dramatically change regional and global
species lists
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
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
MegachilidaeCoelioxys moesta
Coelioxys funerariaCoelioxys rufitarsis
Coelioxys sodalisMegachile pugnata
Megachile latimanusMegachile gemula
Megachile frigidaMegachile melanophaea
Megachile inermisMegachile centuncularis
2%
Megachile montivagaMegachile rotundata
Heriades carinataHoplitis pilosifronsHoplitis producta
Hoplitis spoliataOsmia lignaria
Osmia coerulescensOsmia atriventris
Osmia bucephala
Osmia proximaOsmia inermisOsmia tersula
Osmia simillima
Osmia virgaStelis foederalis
Stelis subemarginata
ApidaeApis mellifera
Bombus ashtoni
Bombus fernaldaeBombus citrinus
Bombus insularisBombus impatiens
Bombus ternariusBombus terricola
Bombus borealisBombus fervidus
Bombus rufocinctusBombus perplexus
Bombus sandersoniBombus vagansEpeoloides pilosula
Holcopasites calliopsidisCeratina dupla
Ceratina calcarataCeratina new sp.?
Anthophora bomboidesAnthophora furcata
Melissodes desponsaMelissodes druriella
Epeolus autumnalisEpeolus scutellaris
Triepeolus brittaini
2%
Cryptic / New Species?
Ceratina complex:
C. strenua
C. calcarata
C. dupla
C. new sp. ?
% Divergence
Within Species Within Genus
% Divergence
Fre
quen
cy (
%)
The Nomada ProblemOver 300 species in North America
Very difficult….many known from one sex and/or few specimens
Host associations of most are unknown
Barcoding on a Global-level
DNA Quality
Sequence Length (bp)
Fre
que
ncy
(%)
Age versus # Base Pairs
Age (years)
# of
Seq
uenc
es
More an artifact of collecting methods and killing agents
Collecting / Killing Methods
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: 21-24Quicke et al. 1999. Zoologica Scripta 28: 261-267
Epeoloides pilosula
Why not all the NS Species?
Rare species….some of the specimens are literature records only
Macropis nuda
Log-normal Plots
Very rare species
Rare species
Why not all the Species?
Rare species….some of the specimens are literary records only
But most of the species
we have from elsewhere
Bees in CanadaProvince/
Territory
No. of
Species
NS 206
NB 147
PE 65
NF 55
QC 243
ON 379
MB 170
SK 161
AB 286
BC 352
NU 8
NT 80
YT 52
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
Barcoding Bees - Conclusions
Taxonomic studies
Great potential for
ecological studies: – diversity
– monitoring
– conservation
– invasive species
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)
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)
Thank You
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