a regional barcode database for bees cory s. sheffield department of biology, york university...

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