DNA Barcoding and integrative taxonomy of bees Packer L.; Gibbs J. York University Agapostemon virescens
DNA Barcoding and integrative
taxonomy of bees Packer L.;
Gibbs J.
York
University
Agapostemon
virescens
OUTLINE
Importance of bees
The taxonomic impediment
Some detailed examples of the use of DNA barcoding in bees
Progress towards barcoding the bees of the world (problems, prognosis)
Importância das abelhas
O ImpedimentoTaxonômico
Exemplos da aplicaçãodo DNA barcoding aoestudo de abelhas
Avanços no projeto de barcoding das abelhas do mundo
SUMÁRIO
A Importância das Abelhas
Polinazação de frutas e vegetais
Polinazação de plantas nativas
Biologia dapolinazação
Monitoramentoambiental
Bees are responsible for25%, 33% (?) of our food.Abelhas são reponsaveis por 25%, 33% (?) dos alimentos produzidos.
“If the bee disappeared off the surface
of the globe then man would only
have four years of life left.”Einstein (?)
& without
flies!
The Taxonomic Impediment
O Impedimento Taxonômico
There are over 19,500 described bee species
There are several genera with more than 1,000 species
There are very few people capable of identifying more than a small proportion of the total
Há mais de 19.500 espécies de abelhasdescritas
Há vários gêneros com mais de 1.000 espécies
Há muito poucas pessoascapazes de identificarmais que uma pequenaparte deste total
Bee Experts
% correct identification as a bee by experts!% de identificação correta como abelhas, por especialistas
% correct identification as a bee by experts!% de identificação correta como abelhas, por especialistas
Hyleoides – 57%
Euryglossina
leyburnensis – 15%
Experts got 75% right, beginners 64%Sucesso de 75% para especialistas e 64% para iniciantes
Photo Steve
Buchman
Um exemplo de um erro
Em uma coleção, eu encontrei uma únicaespécie identificada como pertencendo a trêsgêneros diferentes, de duas famílias distintas, por três taxônomos diferentes (um especialistada fauna local, um especialista da fauna do paíse um especialista da fauna mundial de abelhas)
Em uma coleção, eu encontrei uma únicaespécie identificada como pertencendo a trêsgêneros diferentes, de duas famílias distintas, por três taxônomos diferentes (um especialistada fauna local, um especialista da fauna do paíse um especialista da fauna mundial de abelhas)
Todas as identificações genéricas estavamincorretas.
Um exemplo de um erro
Sometimes there are no keys for identificationÀs vezes não há chaves de identificação
Example: the bee family Colletidae
146 higher level taxa
30% have no revision or key
45% of colletid species are not identifiable with available literature
32/33 keys for one subfamily were written by one researcher
Only four groups have been revised more than once
Exemplo: família Colletidae
146 táxons supraespecíficos
30% nunca foram revisados; não há chaves disponíveis paraidentificação
45% das espécies de Colletidaenão são identificáveis apenascom a literatura taxonômica
32/33 chaves para uma das subfamílias foram propostaspor um único pesquisador
Apenas quatro táxons foramrevisados mais de uma vez
Some examples of
the success of DNA
barcoding
An early example: Halictus ligatus
In North America is two species
As duas espécies são
indistinguíveis
morfologicamente,
mas possuem diferenças
genéticas reconhecíveis:
possuem 8 diferenças
fixadas,
dentre 32 locos de alozimas
Morphologically theyare exactly the same
but genetically,they have 8 fixed differences out of32 allozyme loci
Halictus ligatus/poeyi
The morphological gap is 0.0%
The allozyme gap is 25%
The barcode gap is 4%
There are probably >3 species
A diferençamorfológica é 0,0%
As diferençasdetectáveis com alozimas são 25%
As diferençasdetectáveis com obarcode são de 4%
Provavelmente há ≥3 espécies
DNA BARCODING A NIGHTMARE TAXONDNA Barcoding de um pesadelotaxonômico
Subfamily Halictinae
“sweat bees”
3rd most speciose subfamily (3423 spp.)
Abundant, cosmopolitan
Often 40 – 80 % of bees in surveys
Funding provided by:
The Halictinae
(Sweat bees):
Apresentam enormevariação de comportamentosocial
Muitas espéciessolitárias
Algumas espécieseusociais, com ninhos que abrigammais de 1000 operárias.
DNA Barcoding de um pesadelo taxonômico
Morphological characters are often subtle
Revision of the Canadian Dialictus 80 species in total
16 new species
48 new synonymies
3 new combinations
10 new Canadian records
776 ms pages
235 colour plates
Zootaxa in review
Integrative taxonomy
DNA barcodes facilitate taxonomy
Funding provided by:
75 %
L. (D.) tegulare
L. tegulare (Robertson): one of the most easily identifiable Dialictus species
Widespread in eastern USA
L. (D.) tegulare species group But...its actually FIVE species!
Funding provided by:
Character based identification using DNA barcodes
L. tegulare (Robertson) easily identifiable Dialictus species
Widespread in eastern USA
UNIQUE FIXED NUCLEOTIDE SUBSTITUTIONS
tegulare = 2
ellisiae = 7
lepidii = 4
puteulanum = 2
carlinvillense = 3
surianae = 13
L. (D.) petrellum species group
Similar result for L. petrellum species group
Funding provided by:
New synonymies
L. (D.) disparile (Cresson)
Described from Texas
= Dialictus brassicae Mitchell
Described from North Carolina
= Halictus albitarsus Cresson
Described from male
17% of species have identical barcodes
Funding provided by:
More cryptic diversity?
8 %
Dialictus: barcoding aids their taxonomy even without a clear “barcodegap”
Morphological identification accuracy?
0-40%
Barcodingaccuracy?
≥ 75%
Acurácia daidentificaçãomorfológica?
0-40%
Acurácia do barcoding?
≥ 75%
It’s not always such a struggle
Megachile pugnata: image
courtesy Theresa Pitts-Singer
DNA barcoding é excelente para:
Associating sexes
Associating larvae with adults
Associating castes
Finding clusters in problematic taxa to aid in search for morphological differences
Associação de sexos
Associação de larvas eadultos
Associação de castas
Reconhecimento de agrupamentosgeneticamenteidentificáveis, paraauxiliar a busca pordiferenças mofológicas
Resurrecting incorrect synonymies
Providing an independent test of taxonomic hypotheses
Reavaliação de sinonímias incorretas
Fornece um testeindepende das hipóteses taxonômicas
DNA barcoding é excelente para:
May 12th
– 14th
2008 12 – 14 Mayo, 2008
12 – 14 Maio, 2008 5
Inaugural Meeting Reunión Inagural
Reunião Inaugural
Progress We have a
steering committee
We have a website (currently being improved upon)
And we have a lot of data
Melissodes sp.
Progress: Steering committee – in
approximate geographic location
The website: www.bee-bol.org
We have data from:
>27 collaborators
> 76 countries
> 60% of bee
genera
Distribution of specimens barcoded
Stenotritidae:
2 gêneros 21 espécies
2 genera and
3 species barcoded
Ctenocolletes smaragdinus
Colletidae: 55 genera, 2506 species
34 genera and 403 species barcodedXeromelissa rozeni
Colletes inaequalis
Andrenidae: 40 genera, 2895 species
30 genera and 353 species barcodedAndrena erythrogaster
Calliopsis anomoptera
Halictidae: 76 genera, 4238 species
56 genera and 835 species barcodedHalictus ligatus? Agapostemon splendens
Melittidae: 15 genera, 183 species
7 genera and 17 species barcoded
Meganomia binghamiRediviva sp.
Rediviva sp.
Megachilidae: 75 genera 3973 species
37 genera and 631 species barcoded
Anthidium manicatum Megachile sp.
Apidae: 176 genera, 5687 species
141 genera and 1226 species barcodedTriepeolus sp. Apis mellifera
% de espécies barcoded
0
5
10
15
20
25
%
Preliminary
analyses on the
data for all
records of
Colletidae in
the database:
740 sequences
The barcode
gap for the
prelminary
data set for
all Colletidae
“Intraspecific”
“Interspecific”1 2 3 4 >5
5 10 15 20
10%
50%
25%
5%
74%
7%
12%
7%
Acurácia do barcoding/taxônomo: Colletidae, n = 740 sequências
Barcoding/Taxonomist accuracy: Colletidae, n = 740 sequences
consistent
identified by barcoding
unidentified
inconsistent
10%
72%
6% 12%
Razões para inconsistência, n = 50 sequênciasCauses of inconsistency, n = 50 sequences
nonsense
misidentification
uncertain assignment
wrong sequence
75%
3%
22% consistent
inconsistent
unidentified
Acurácia do barcoding/taxônomo Accurácia, n = 250 "clusters“
Barcoding/Taxonomist Accuracy, n = 250 clusters
12%
25%
63%
Razões para inconsistência: n = 8 “espécies”Causes of inconsistency: n = 8 “species”
new species
synonymies
diff spp. = sequences
Problemas Curating all of the incoming
data is impossible on a part-time basis.
Trabalho de curadoria de todo o material recebido é impossível quando esta é uma dentre várias funçõesdo pesquisador.
Obtaining accurate identifications.
Identificações confiáveis.
Problems
Specimens are difficult to obtain from some countries.
Dificuldade de obtenção de material de alguns países
Some preferred collection methods are suboptimal for barcoding.
Alguns dos métodos maispopulares para coleta de espécimes são subóptimospara barcoding.
$$
Soluções para os Problemas Full-time assistance is now available for 6 months
for database curation. Material transfer agreements are being developed. Museum specimens, new collections. A team of experts is being amalgamated.
Auxílio técnico “full-time” disponibilizado por 6 meses/ano para curadoria dos bancos de dados.
Acordos para envio de material estão sendofirmados.
Espécimes de museus, expedições de coleta. Um extenso grupo de especialistas está sendo
formado.
Prognosis Caupolicana fulvicollis
With funds…
- Excellent!Nolanomelissa toroi
CONCLUSÕES Identificação de abelhas é uma tarefa difícil
O impedimento taxonômico é enorme
Barcoding funciona para a maior parte da melitofauna do Canadá 100% sucesso para Megachile
100% sucesso para Dufourea
100% sucesso para abelhas de Nova Scotia
75% sucesso para o “grupo pesadelo”–Dialictus
Associação de sexos, castas, estágios do desenvolvimento, correção de sinonímiaserrôneas, testes de hipóteses taxonômicas
Barcoding is a useful tool for taxonomy even in the absence of a “barcode gap”
Barcoding é uma ferramenta útilpara taxonomia mesmo quando umaseparação pelo barcode (“barcode gap”) não é evidente.
Barcoding works most of the time for the Canadian fauna 100% success in Megachile
100% success in Dufourea
100% for the bees of Nova Scotia
75% success for the nightmare group –Dialictus
Associates sexes, castes, life stages, correcting erroneous synonymies, tests taxonomic hypotheses
It certainly does not threaten the future of traditional taxonomy, indeed, in Canada, it has saved it from extinction.
Barcoding não representa um perigopara o futuro da taxonomiatradicional. Na verdade, no Canadá, o perigo de extinção da taxonomiaestá sendo eliminado graças ao
barcoding.
There are problems to overcomeHá problemas a seremsuperados
They are all related to time and money.
Todos eles relacionam-se a tempo e dinheiro
Acknowledgements Sam Droege (United States Geological Survey, Patuxent Wildlife Research Center), Terry Griswold (United States Department of
Agriculture, Agricultural Research Service), Ralph Grundel (United States Geological Survey, Great Lakes Science Center), Andrea Patenaude (University of Manitoba), Julianna Tuell (Michigan State University), John Ascher (American Museum of Natural History), Rob Jean (Indiana State University), Joan Milan (University of Massachusetts Amherst), Elizabeth Elle (Simon Fraser University), Miriam Richards (Brock University), Amy Wolf (University of Wisconsin Green Bay), Peter Hallett (University of Toronto), Bryan Danforth (Cornell University), Mike Arduser (Missouri Department of Conservation), Jennifer Hopwood (University of Kansas), Charles Michener (Kansas University Natural History Museum), Bob Minckley (University of Rochester), Paul Catling (Agriculture and Agri-Food Canada), Victoria MacPhail (University of Guelph), Rebecca Andres (North Dakota State University), Stephen Hendrix (University of Iowa), Matthias Buck (Royal Alberta Museum), Jack Neff (Central Texas Melittological Institute), Doug Yanega (University of California at Riverside), Jen Frye (Maryland Department of Natural Resources)
ANSP: Academy of Natural Sciences, Philadelphia, Pennsylvania (J. Weintraub); AMNH: American Museum of Natural History, New York, New York (J.G Rozen, Jr., and J.S. Ascher); ARC: Albert J. Cook Arthropod Research Collection, Michigan State University, Lansing, Michigan (G.L. Parsons); BCLU: Utah State University Bee Biology and Systematics Laboratory, Logan, Utah (T.L. Griswold); BDUC: University of Calgary–Entomology, Calgary, Alberta (J. Swann and R. Longair); BLNP: Badlands National Park, Interior, South Dakota (M. Cherry); BMNH: Natural History Museum, London, England (D. Notton); CAS: California Academy of Science, San Francisco, California (W.J. Pulawski and V. Lee); CNC: Canadian National Collection of Insects, Arachnids and Nematodes, Ottawa, Ontario (A. Bennett); CTMI: Central Texas Melittological Institute, Austin, Texas (J.L. Neff); CUIC: Cornell University Insect Collection, Ithaca, New York (E.R. Hoebeke and B.N. Danforth); DEBU: University of Guelph Insect Collections, Guelph, Ontario (S.A. Marshall); FSCA: Florida State Collection of Arthropods, Gainesville, Florida (L. Stange and J. Wiley); GSNP: Great Smoky Mountains National Park, Gatlinburg, Tennessee (A. Mayor); IDNL: Indiana Dunes National Lakeshore, Porter, Indiana (R. Grundel); INHS: Illinois Natural History Survey, Champaign, Illinois (P.P. Tinarella); JHRC: James Hanula research collection, USDA Forest Service, Southern Research Station, Athens, Georgia (J. Hanula). (Eventually to be deposited in Georgia Museum of Natural History); MCZ: Harvard University Museum of Comparative Zoology, Cambridge, Massachusetts (P.D. Perkins); NCSU: North Carolina State University, Raleigh, North Carolina (B.Blinn); NMNH: National Museum of Natural History, Washington, D.C. (D. Furth, B. Harris, and S.G. Brady); PCYU: Packer Collection at York University, Toronto, Ontario (L. Packer). (Subsamples of this material will eventually be deposited in institutional collections managed by C. Sheffield, L. Best, and the author when appropriate); PHPC: Peter Hallett private collection, Toronto, Ontario (P.E. Hallett); PMAE: Royal Alberta Museum, Edmonton, Alberta (M. Buck); RBCM: Royal British Columbia Museum, Victoria, British Columbia (R. Cannings); ROM: Royal Ontario Museum, Toronto, Ontario (B. Hubley); RWRU: Rachael Winfree research collection, Rutgers University, New Brunswick, New Jersey (R. Winfree); SEMC: Snow Entomological Museum, (Kansas University Natural History Museum), Lawrence, Kansas (C.D. Michener and J.C. Thomas); UCMC: University of Colorado Museum of Natural History, Boulder, Colorado (V.L. Scott); UCR: University of California Riverside Entomology Research Museum, Riverside, California (D. Yanega); UNSM: University of Nebraska State Museum, Lincoln, Nebraska (B. Ratcliffe and M.J. Paulsen); UWGB: University of Wisconsin (Richter Museum of Natural History), Green Bay, Wisconsin (T. Erdman)
CCDB and BIO at the University of Guelph for sequencing. Funding from Genome Canada, NSERC and other sponsors listed at www.BOLNET.ca is greatly appreciate.
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.
• CBOL – funded the May workshop
• York University
EDUARDO (EDDY) ALMEIDA for translation
• Sujeevan Ratnasingham (BOLD)
PARTNERS IN FUNDING AND IN-KIND SUPPORT
¡Gracias!
Photo
courtesy Dr.
S.F. Sakagami