Bee Barcoding (Bee-BOL) Campaign

J. Gibbs, L. Packer, C. Sheffield York University

Bee-BOL: the campaign to barcode the bees of the world.◦ Its beginning◦ Its current status

Example studies:◦A whole bee community◦A whole (nightmare) taxon

iBOL and pollinators.◦How to barcode the bees of the world

◦How you can help

May 12th – 14th 2008 12 – 14 Mayo, 200812 – 14 Maio, 2008 ２００８年　 5 月１２－１４日

Inaugural Meeting Reunión Inagural Reunião Inaugural 創立

総会

Methods York6 boxes – 516 specimens from 28 nations

Ninety-six well DNA isolation protocols

Hierarchy of primer decisions; including degenerate primer

cocktails

% success amplifying barcode

sequence

Ano ch etus sp .|A mb a toAno ch etus sp .|Am batoAno ch etus sp .|A mb atoAno ch etus sp .|Am bato

Ano ch etus sp .|A mba toAn o ch etus sp. |Ant sah abeAn o ch etus sp. |An tsah ab eAn o ch etus sp. |Ants ah abeAno ch etus sp .|Bina raAno ch etus sp .|Bina raAno ch etus sp .|Bi naraAn o ch etus sp. |An ts ah abeAno ch etus sp .|B in a ra

An o ch etus sp. |Ants ah abeAn o ch etus sp. |Ants ah abe

An o ch etus sp. |Ants ah abeAn o ch etus sp. |An tsah abeAn o ch etus sp. |An tsah ab eAno ch etus sp .|B inaraAn o ch etus sp .|M ar oje jyAn o ch etus sp .|M ar oje jy

Od o nt omachus sp .| Maroj ej yOd o nt omachus sp.| Maroj ej y

Chilicola|1406-H06Chilicola|1399-H06Chilicola|1406-H03Chilicola|1399-H03Hylaeus sp.|1408-H09

Hylaeus sp.|1407-H09Hylaeus calvus|1407-F07Hylaeus elatus|1407-E12Nomiocolletes|1406-G06

Nomiocolletes|1399-G06Andrena|1408-H01Lasioglossum|1408-G08Lasioglossum|1408-G06Andrena|1407-H01Lasioglossum|1407-G08Lasioglossum|1407-G06

Andrena clarkella|1406-A08Andrena clarkella|1399-A08Andrena|1406-E10

Andrena|1399-E10Andrena ilicis|1408-F10Andrena ilicis|1407-F10

Andrena cubiceps|1406-E08Andrena cubiceps|1399-E08Calliopsis fracta|1408-F09

Calliopsis fracta|1407-F09Andrenidae|1408-01Andrenidae|1407-01Dufourea tinsleyi|1408-E05

Dufourea tinsleyi|1407-E05Sphecodes|1399-A10Nomia pavonura|1408-C11

Nomia pavonura|1407-C11Pseudapis|1406-A06Pseudapis|1399-A06Nomiine|1399-B01

Pseudaugochlora sp.|1408-H12Pseudaugochlora sp.|1407-H12Augochloropsis|1408-07Augochloropsis|1407-07

Augochlora|1408-B05Augochlora|1407-B05Neucorynura|1408-08Neucorynura|1407-08

Caenohalictus|1406-H10Caenohalictus|1399-H10Corynura chloris|1406-D06Corynura chloris|1399-D06Corynura prothysteres|1406-C11Corynura prothysteres|1399-C11

Corynura chilensis|1406-D05Corynura chilensis|1406-C12Corynura chilensis|1399-D05

Corynura chilensis|1399-C12Augochlorella|1408-B04Augochlorella|1407-B04

Chlerogelloides femoratus|1406-H12Chlerogelloides femoratus|1399-H12Halictus|1408-G09

Halictus|1407-G09Halictus maculatus|1406-C09Halictus maculatus|1399-C09Halictus tumulorum|1406-B10Halictus tumulorum|1399-B10

Halictus africanus|1408-G02Halictus africanus|1407-G02Halictus seladonia sp.|1406-F02

Seladonia|1406-A05Seladonia|1399-A05Halictus hesperus|1406-H08Halictus hesperus|1399-H08

Halictus quadricinctus|1406-F01Halictus quadricinctus|1399-F01Habralictus sp.|1408-H11Habralictus sp.|1407-H11Caenohalictus|1406-H11

Caenohalictus|1399-H11Rhinetula denticrus|1406-H09Rhinetula denticrus|1399-H09

Agapostemon semimelleus|1408-H10Agapostemon semimelleus|1407-H10

Lasioglossum kumejimense|1407-C12Patellapis zonalioctus sp.|1408-G01Patellapis zonalioctus sp.|1407-G01

Lasioglossum sp.|1408-E01Lasioglossum sp.|1407-E01Lasioglossum|1408-F12

Lasioglossum|1408-04Lasioglossum|1407-04Lasioglossum discum|1408-E09

Lasioglossum discum|1407-E09Lasioglossum zonulum|1406-B06Lasioglossum zonulum|1399-B06

Lasioglossum|1408-G12Lasioglossum|1408-G11Lasioglossum|1407-G12Lasioglossum|1407-G11Lasioglossum|1408-G07Lasioglossum|1407-G07Lasioglossum evylaeus|1408-E06

Lasioglossum evylaeus|1407-E06Lasioglossum|1408-G10Lasioglossum evylaeus|1408-E07Lasioglossum|1407-G10

Lasioglossum|1407-05Evylaeus dybowski|1406-C06Evylaeus dybowski|1399-C06

Lasioglossum morio|1406-C02Lasioglossum morio|1399-C02

Lasioglossum nitidulum|1406-C01Lasioglossum nitidulum|1399-C01Lasioglossum evylaeus|1408-E11

Lasioglossum evylaeus|1407-E11Lasioglossum politum|1406-C03Lasioglossum politum|1399-C03

Lasioglossum apristum|1408-C08Lasioglossum apristum|1407-C08Lasioglossum hoffmanni|1408-C07

Lasioglossum hoffmanni|1407-C07Lasioglossum sibiracum|1407-C06Lasioglossum affine|1408-C05

Lasioglossum affine|1407-C05Lasioglossum calceatum|1406-C04Lasioglossum calceatum|1399-C04

Lasioglossum malachurum|1406-E12Lasioglossum malachurum|1399-E12

Lasioglossum sakagami|1407-D01Lasioglossum sp. 5|1408-C03Lasioglossum sp. 5|1407-C03

Lasioglossum sp. 6|1408-C02Lasioglossum sp. 6|1407-C02Dialictus|1406-B02Dialictus|1399-B02Lasioglossum sp. 4|1408-C01

Lasioglossum sp. 4|1407-C01Lasioglossum|1406-G11Lasioglossum|1399-G11Lasioglossum|1406-G10Lasioglossum|1399-G10

Lasioglossum|1406-G12Lasioglossum|1399-G12Lasioglossum jultscinicum|1408-F04Lasioglossum jultscinicum|1407-F04

Lasioglossum equestre|1408-E10Lasioglossum equestre|1407-E10Lasioglossum xanthopus|1408-F03Lasioglossum xanthopus|1407-F03Lasioglossum xanthopus|1406-B11Lasioglossum xanthopus|1399-B11

Lasioglossum caspicum|1408-E08Lasioglossum caspicum|1407-E08Lasioglossum titusi|1407-F05

Xeromelissa rozeni|1408-H08Xeromelissa rozeni|1407-H08Xeromelissa laureli|1408-H06

Xeromelissa laureli|1407-H06Xeromelissa laureli|1408-H05Xeromelissa laureli|1407-H05

Xeromelissa new sp.|1408-H03Xeromelissa new sp.|1408-H02Xeromelissa new sp.|1407-H02

Xeromelissa new sp.|1407-H03Xeromelissa chusmiza|1408-H04Xeromelissa chusmiza|1407-H04Colletes seminitudus|1406-D04

Colletes seminitudus|1406-C10Colletes seminitudus|1399-C10Colletes seminitudus|1406-D03

Colletes seminitudus|1399-D04Colletes seminitudus|1399-D03Ptiloglossa|1408-06

Ptiloglossa|1407-06Camptopoeum|1406-E09Camptopoeum|1399-E09Caupolicana|1406-G04

Caupolicana|1399-G04Caupolicana chalybaea|1406-G05Caupolicana chalybaea|1399-G05

Chilicola|1406-H07Chilicola|1399-H07Chilicola|1399-H05Chilicola|1406-H05

Chilicola|1406-H04Chilicola|1399-H04Melitta americana|1408-F06

Melitta americana|1407-F06Centris brethesi|1408-D10 MACN09Centris|1408-B01

Centris|1407-B01Diadasia pereyrae|1408-D02 MACN01Diadasia baraderensis|1407-D03 MACN02Tetrapedia|1406-H01

Tetrapedia|1399-H01Caenonomada|1406-G01Caenonomada|1399-G01

Caenonomada|1406-F12Caenonomada|1399-F12

Svastrides bauni|1408-D08 MACN07Svastrides bauni|1407-D08 MACN07Melissoptila bonariensis|1408-D07 MACN06Melissoptila bonariensis|1407-D07 MACN06Exomalopsis|1408-11

Exomalopsis|1407-11Megachile frigida|1408-F01

Megachile frigida|1407-F01Epeolus amabilis|1406-F05Thyreus|1406-F07

Thyreus|1399-F07Epiclopus|1406-G08Epiclopus|1399-G08

Ericrocis pintada|1406-F11Ericrocis pintada|1399-F11Ericrocis lata|1406-F10

Ericrocis lata|1399-F10Xylocopa|1408-G03Xylocopa|1407-G03

Xylocopa caffra|1406-F04Xylocopa caffra|1399-F03Xylocopa caffra|1406-F03 Apidae|1408-E02

Apidae|1408-D12Apidae|1407-E02Apidae|1407-D12

Apidae|1408-B02Apidae|1407-B02Apidae|1406-H02

Apidae|1399-H02Bombus|1408-B03Bombus|1407-B03Bombus brasiliensis|1406-G03Bombus atratus|1406-G02Bombus brasiliensis|1399-G03Bombus atratus|1399-G02

Bombus atratus|1399-E07Bombus atratus|1406-E07Bombus brasiliensis|1406-E06Bombus brasiliensis|1399-E06

Lasioglossum sakagami|1408-D01Apis florea|1408-03Apis florea|1407-03

Apis florea|1406-D01Apis florea|1399-D01Apis sp.|1406-E05

Apis|1406-B08Apis dorsata|1406-E04

Apis dorsata|1399-E04Apis nigricinctus|1406-D02Apis nigricinctus|1399-D02

Apis|1406-B09Apis dorsata|1399-E03Apis sp.|1399-E02

Macrogalea candida|1408-G04Macrogalea candida|1407-G04Ceratina sp. 2|1408-B09Ceratina sp. 2|1407-B09

Ceratina sp. 8|1408-B08Ceratina sp. 8|1407-B08Ceratina sp. 6|1408-B07Ceratina sp. 6|1407-B07Ceratina sp. 3|1408-B06Ceratina sp. 3|1407-B06

Ceratina hieroglyphica|1406-D12Ceratina hieroglyphica|1399-D12Ceratina hieroglyphica|1399-F09

Tetrapedia|1408-10Tetrapedia|1407-10 Amegilla acraensis|1406-F08

Lithurgus rufiventris|1408-D05 MACN04Lithurgus rufiventris|1407-D05 MACN04Microthurge|1406-G07

Microthurge|1399-G07Anthidiellum|1406-B03Anthidiellum|1399-B03Megachilidae|1408-G05

Megachilidae|1407-G05Anthodioctes|1408-09Anthodioctes|1407-09

Megachilidae|1408-F11Megachilidae|1407-F11Osmia glauca|1408-E04

Osmia glauca|1407-E04Megachilidae|1408-02

Megachilidae|1407-02Anthidiine|1406-A11Anthidiine|1399-A11

Stelis|1406-A07Stelis costalis|1399-A07Megachile integrella|1408-F08Megachile integrella|1407-F08

Megachile|1408-B10Megachile|1407-B10Megachile|1408-B11Megachile|1407-B11

2 %

BeeBol: 6 submitted plates: 423 sequences

• 423/561 with some sequence (75%)• ~1200 sequencing reactions• 235 with >600 bp• 85 with < 400 bp• Taxonomic patterns

• 6 families, 76 genera• Age patterns• 6 boxes

• 297 P1 (LepF1/LepR1)• 215 P2 (LepF1/C_ANTMR1D)• 130 P3 (RonMWASPDeg_T1/LepR1)• 85 P3 (MLepF1/LepR1)• Folmer primers (not tried on these

specimens)• 10 Wolbachia CO1 sequences that

amplified with P1• All produced insect CO1 with P2 and P3• 3 mouse sequences• Taq contamination

BeeBol – – within taxa comparisions

Within species (as originally defined)

Between sister species

BeeBol– length / age

The Bee-BOL website

The Bee-BOL website•You will be able to find

•All power point presentations or read texts from Toronto and Durban meetings•Meeting documents, including guidelines on how you can help•The world bee species list, with each species directly linked to its page on the discover life website•Images and Key to Andreninae•List of contributors, contact information, lists of projects etc.

•www.bee-bol.org

Bee Imaging Project

Ctenocolletes smaragdinus (Smith)Caupolicana fulvicollis Spinola

BEE STATS

6,752 Barcodes

>1,542 Species Barcoded

452 Andrenidae1,996 Apidae 638 Colletidae

2,838 Halictidae 912 Megachilidae 11 Melittidae 5 Stenotritidae

A community-wide survey:the bees of Nova Scotia, Canada

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%

Colletidae Sheffield et al. submitted to Molecular Ecology Resources

Whole taxon studies

Dialictus: DNA barcoding a nightmare taxon

Revision of the Canadian and Eastern USA Dialictus fauna

Describe all species in these regions (~100)◦New synonymies

20 so far◦Resurrections

4 so far◦New species

20 so far

Typical DNA barcoding results Clear agreement between

morphology and molecules Little variation within

species across large geographical areas

Hidden diversity

Deep divergences within a ‘single’ species◦ Erroneous synonymies◦ Cryptic species

L. tegulare species group

Gibbs (submitted to Zootaxa)

Unexpected results

Different morphologies Same sequence! L. rohweri and L. callidum

Bee-BOL Goals

• Barcode all bee species (19231 + undescribed spp.)•Combination of regional studies and whole taxon studies•Gain specimens from collections•Focused collecting for rare species and under sampled localities•www.bee-bol.org

www.dnabarcoding.org

10 specimens each from 500,000 species, within 5 years of activationWe hope to barcode all bees for ~$5M.

iBOL and pollinators BEE-BOL a major component Flower flies Other economically important groups◦Eg. Cacao midges

High impact groups Meliponini

Xylocopa

How you can help

SPECIMENS!

Coloured bowls for easy collecting

Requirements and Benefits

Requirements Specimens to be barcoded must:◦ Have full collection data◦ Have an associated image (we can take these)◦ At least one exemplar of each species must be kept in an

institution that will be able to loan it if further morphological taxonomic study is needed

All materials can be returned to their country of origin Benefits◦ Data◦ Identifications◦ Publications associated with release of full data sets◦ Training, workshop/conference attendance for students

BEE-BOL www.bee-bol.org

FUNDING• The Gordon and Betty Moore Foundation, Natural Sciences and

Engineering Research Council (Canada), Canada Research Chairs program, Ontario Foundation for Innovation and Genome Canada through the Ontario Genomics Institute to PDNH

• Fonds québécois de la recherche sur la nature et les technologies B3 postdoctoral fellowship to MAS.

• CBOL – funded the May workshop• York University • Sujeevan Ratnasingham (BOLD)• Kate Crosby and Anibal Castillo (diligent, careful and enthusiastic lab

work)COLLABORATIONS• John Ascher and John Pickering and others that contributed to the bee

species list on Discover Life• Sam Droege• All workshop attendees who submitted samples• Numerous collaborators throughout the world who have sent in samples

PARTNERS IN FUNDING AND IN-KIND SUPPORT

MANY THANKS TO THE ORGANIZERS

AND THANKS FOR LISTENING