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DNA barcoding and evolutionary relationships in Accipiter Brisson,1760 (Aves, Falconiformes: Accipitridae) with a focus on African and Eurasian representatives. Nomenclature. A. gentilis : Nl; havik, Eng; gosshawk, Fr; Autour des palombes
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DNA barcoding and evolutionary relationships in Accipiter Brisson,1760 (Aves,Falconiformes: Accipitridae) with a focus on African and Eurasian representatives
Nomenclature • A. gentilis: Nl; havik, Eng; gosshawk, Fr; Autour des palombes • A. nisus: Nl; sperwer, Eng; sparrowhawk, Fr; epervier d’Europe • Buteo buteo: Nl; buizerd, Eng; Common buzzard Fr; Buse variable • Buteo albicaudatus; Nl; witstaart buizerd, Eng; white tailed hawk, Fr; Buse à queue blanche
DNA barcoding a short introduction General DNA barcoding Mini barcodes 10X threshold, best compromise threshold BCM and BM criteria Character based identification Coverage Sample origins The dataset Methods Character based method (CB) The 10 X intraspecific distance threshold (10 X) The Best compromise threshold (BCTh) Best match and best close match explained Results for DNA barcoding Character based identification Different thresholds Phylogeny reconstruction 291 bp dataset 647 bp dartaset Discussion Conclusion Content
DNA barcoding • Identification of species with an agreed upon (part of a) gene (COI) • 5’-3’ part of Cytochrome C Oxidase subunit I • ~650bp in length • Polymerase chain reactions and Sanger sequencing
Mini barcodes • Genetic material from museums is often degraded • Shorter fragments are needed • E.g. sequences of 250-300bp or, in some cases even shorter, also work for identification purposes Sonet et al. in press
Sample coverage and origins • African and European taxa are well represented • The new world partially represented • Australia and S-E Asia poorly represented • Most samples from RMCA/RBINS (n=83/9) • Supplemented with available genbank sequences (n=28)
The resulting dataset • 25 European and African Accipiter species with a total of 140 specimens • sequenced for a 291bp (mini) and 647bp (standard) BC fragment of the COI gene • three datasets analysed: • dataset A: 25 species, 291bp • dataset B: 19 species, 647bp • dataset C: 19 species, 291bp
Character based indentification • Easy it is about detecting unique differences • May be unreliable with insufficient sampling • No threshold whatsoever is used
Average 10 X intraspecific distances and the barcoding gap • Distances • Arbitrary • Does not always hold (Tephritidae) • However, works well in birds
The Best compromise threshold (BCTh) • Based on the intercept between cumulative intra and interspecific distances • Based on the dataset under study rather than a predetermined threshold • Results from one dataset not applicable to another Lefébure et al. 2006
BM and Best Close match • Best Match (BM), assignes the species name of its best-matching sequence regardless of how similar the query and reference sequences were. Identification is considered correct when both sequences were from the same species, incorrect if the query species differed from the closest reference species or ambiguous if multiple species were the BM of the query species • Best close match (BCM) also considers the threshold
BM vs. BCM example • 4 sequences • 3 species: A. francesiae (n=2) A. badius (n=1) & A. toussenelii (n=1) • BM would assign A. badius and A. toussenelii to the “ambiguous” or “wrong” (dependent on the threshold) categories • BCM would classify them as impossible when the difference is larger than the threshold
Results BCTh vs. 10 X distance threshold • Different thresholds • Partial overlap • Caused by • A. nisus/rufiventris • A. virgatus/gularis • A. cooperii/gundlachi • At or just below the BC treshold are • A. badius subsp • A. gentilis subsp
A. nisus A. rufiventris Results BCTh vs. 10 X distance threshold: Accipiter nisus example
Results BCTh vs. 10 X distance threshold: A. Badius and A. brevipes example
Phylogeny reconstruction • Hypothesis of evolution • Clades are units of supported taxa (related by direct descent) • This dataset is only suited for low taxonomic level inferences • One genegene tree rather than species tree • Comparison with the current taxonomy based on morphology and morphometrics
Phylogeny based on 291bp • 11 supported clades • [tachiro] • A. tachiro subclades • A. minullus • A. francesiae/ soloensis • [cooperii] • A. badius/ brevipes • A. nisus/ striatus/ erythronemius • A. ovampensis/ madagascariensis • [virgatus] including A. fasciatus
Phylogeny based on 291bp a closer look: Accipiter tachiro example
Phylogeny based on 291bp a closer look: Accipiter badius and A. brevipes example
Phylogeny based on 647bp • 19 supported clades • [tachiro] • A. tachiro • A. toussenelii & subsp • A. soloensis &A. francesiae subsp • [gentilis] • [cooperii] • [nisus] (without A. ovampensis/ madagascariensis • [minullus] (without A. erythropus) • [virgatus] including A. fasciatus
Phylogeny based on 647bp a closer look: The example from Accipiter soloensis and A. francesiae
Phylogeny based on 647bp a closer look: an example with Accipiter gentilis and A. melanoleucus
Discussion • Thresholds • The 10 X threshold would have considered A. brevipes and A. badius, as well as A. gentilis and A. melanoleucus as belonging to the same species and • Species concepts • Morphological • Distances based • Phylogenetic • Support values • Models and interpretation remain issues of discussion • Sampling • Incomplete or lacking in some cases
Conclusions • Molecular techniques provide a powerfull method for reviewing taxonomy • Museum material works! • The content of the dataset matters • COI can identify 19 out of the 25 species included • COI carries a phylogenetic signal suitable for molecular taxonomy on a low taxonomic level • Parts of genus Accipiter may need revision