Avian Tree Research Articles

Why Do Phylogenomic Data Sets Yield Conflicting Trees? Data Type Influences the Avian Tree of Life more than Taxon Sampling

Phylogenomics, the use of large-scale data matrices in phylogenetic analyses, has been viewed as the ultimate solution to the problem of resolving difficult nodes in the tree of life. However, it has become clear that analyses of these large genomic data sets can also result in conflicting estimates of phylogeny. Here, we use the early divergences in Neoaves, the largest clade of extant birds, as a "model system" to understand the basis for incongruence among phylogenomic trees. We were motivated by the observation that trees from two recent avian phylogenomic studies exhibit conflicts. Those studies used different strategies: 1) collecting many characters [$\sim$ 42 mega base pairs (Mbp) of sequence data] from 48 birds, sometimes including only one taxon for each major clade; and 2) collecting fewer characters ($\sim$ 0.4 Mbp) from 198 birds, selected to subdivide long branches. However, the studies also used different data types: the taxon-poor data matrix comprised 68% non-coding sequences whereas coding exons dominated the taxon-rich data matrix. This difference raises the question of whether the primary reason for incongruence is the number of sites, the number of taxa, or the data type. To test among these alternative hypotheses we assembled a novel, large-scale data matrix comprising 90% non-coding sequences from 235 bird species. Although increased taxon sampling appeared to have a positive impact on phylogenetic analyses the most important variable was data type. Indeed, by analyzing different subsets of the taxa in our data matrix we found that increased taxon sampling actually resulted in increased congruence with the tree from the previous taxon-poor study (which had a majority of non-coding data) instead of the taxon-rich study (which largely used coding data). We suggest that the observed differences in the estimates of topology for these studies reflect data-type effects due to violations of the models used in phylogenetic analyses, some of which may be difficult to detect. If incongruence among trees estimated using phylogenomic methods largely reflects problems with model fit developing more "biologically-realistic" models is likely to be critical for efforts to reconstruct the tree of life. [Birds; coding exons; GTR model; model fit; Neoaves; non-coding DNA; phylogenomics; taxon sampling.].

Birds in a bush: Toward an avian phylogenetic network

ABSTRACT Reconstructing the avian tree of life has become one of the major goals in ornithology. The use of genomic tools seemed a promising approach to reach this goal, but, instead, phylogenetic analyses of large numbers of genes uncovered high levels of incongruence between the resulting gene trees. This incongruence can be caused by several biological processes, such as recombination, hybridization, and rapid speciation (which can lead to incomplete lineage sorting). These processes directly or indirectly amount to deviations from tree-like patterns, thereby thwarting the use of phylogenetic trees. Phylogenetic networks provide an ideal tool to deal with these difficulties. We illustrate the usefulness of phylogenetic networks to capture the complexity and subtleties of diversification processes by discussing several recent genomic analyses of birds in general and the well-known radiation of Darwin's finches. With the increasing amount of genomic data in avian phylogenetic studies, capturing the evolu...

The phylogenomic forest of bird trees contains a hard polytomy at the root of Neoaves

Birds have arguably been the most intensely studied animal group for their phylogenetic relationships. However, the recent advent of genome‐scale phylogenomics has made the forest of bird phylogenies even more complex and confusing. Here, in this perspective piece, I show that most parts of the avian Tree of Life are now firmly established as reproducible phylogenetic hypotheses. This is to the exception of the deepest relationships among Neoaves. Using phylogenetic networks and simulations, I argue that the very onset of the super‐rapid neoavian radiation is irresolvable because of eight near‐simultaneous speciation events. Such a hard polytomy of nine taxa translates into 2 027 025 possible rooted bifurcating trees. Accordingly, recent genome‐scale phylogenies show extremely complex conflicts in this (and only this) part of the avian Tree of Life. I predict that the upcoming years of avian phylogenomics will witness many more, highly conflicting tree topologies regarding the early neoavian polytomy. I further caution against bootstrapping in the era of genomics and suggest to instead use reproducibility (e.g. independent methods or data types) as support for phylogenetic hypotheses. The early neoavian polytomy coincides with the Cretaceous–Paleogene (K‐Pg) mass extinction and is, to my knowledge, the first empirical example of a hard polytomy.

Re‐evaluating the distribution of cooperative breeding in birds: is it tightly linked with altriciality?

The evolution of cooperative breeding (CB) in birds has aroused intensive interest for decades, largely due to the paradox that some adults forgo independent breeding to help others. While much effort has been directed at understanding the adaptive significance of CB behavior, much less effort has been spent on understanding its origin. Ligon and Burt argued that the evolution of altriciality played a key role in the origin of CB since CB occurs more frequently in altricial lineages than expected if developmental mode and CB evolved independently and that both traits arose early in the avian tree of life. We mapped presence or absence of CB, and precocial or altricial development on a recent phylogeny of all birds to re‐evaluate their conclusions. Our results suggest altriciality may be more recently derived than previously thought, and that CB species clustered in a derived land bird clade (especially within Passeriformes) where we reconstructed many gains and losses. We did find a link between cooperative breeding and altriciality. However, since CB also occurs in precocial species, has not evolved in many altricial clades, and may have evolved prior to altriciality (based on some classifications of which species have CB), it is not clear whether altriciality is linked to other factors, such as benefits to group living, that are necessary for the acquisition of CB behavior, or whether altriciality may have been a driving force in the evolution of CB itself. The relative importance of these other factors versus altriciality for the origin of CB needs to be considered.

Comment on "Statistical binning enables an accurate coalescent-based estimation of the avian tree".

Mirarab et al. (Research Article, 12 December 2014, p. 1250463) introduced statistical binning to improve the signal in phylogenetic methods using the multispecies coalescent model. We show that all forms of binning-naïve, statistical, and weighted statistical-display poor performance and are statistically inconsistent in large regions of parameter space, unlike unbinned sequence data used with species tree methods.

Response to Comment on "Statistical binning enables an accurate coalescent-based estimation of the avian tree".

Liu and Edwards argue against the use of weighted statistical binning within a species tree estimation pipeline. However, we show that their mathematical argument does not apply to weighted statistical binning. Furthermore, their simulation study does not follow the recommended statistical binning protocol and has data of unknown origin that bias the results against weighted statistical binning.

Avian Diversification Patterns across the K-Pg Boundary: Influence of Calibrations, Datasets, and Model Misspecification1

Birds represent the most diverse extant tetrapod clade, with ca. 10,000 extant species, and the timing of the crown avian radiation remains hotly debated. The fossil record supports a primarily Cenozoic radiation of crown birds, whereas molecular divergence dating analyses generally imply that this radiation was well underway during the Cretaceous. Furthermore, substantial differences have been noted between published divergence estimates. These have been variously attributed to clock model, calibration regime, and gene type. One underappreciated phenomenon is that disparity between fossil ages and molecular dates tends to be proportionally greater for shallower nodes in the avian Tree of Life. Here, we explore potential drivers of disparity in avian divergence dates through a set of analyses applying various calibration strategies and coding methods to a mitochondrial genome dataset and an 18-gene nuclear dataset, both sampled across 72 taxa. Our analyses support the occurrence of two deep divergences (i.e., the Palaeognathae/Neognathae split and the Galloanserae/Neoaves split) well within the Cretaceous, followed by a rapid radiation of Neoaves near the K-Pg boundary. However, 95% highest posterior density intervals for most basal divergences in Neoaves cross the boundary, and we emphasize that, barring unreasonably strict prior distributions, distinguishing between a rapid Early Paleocene radiation and a Late Cretaceous radiation may be beyond the resolving power of currently favored divergence dating methods. In contrast to recent observations for placental mammals, constraining all divergences within Neoaves to occur in the Cenozoic does not result in unreasonably high inferred substitution rates. Comparisons of nuclear DNA (nDNA) versus mitochondrial DNA (mtDNA) datasets and NT- versus RY-coded mitochondrial data reveal patterns of disparity that are consistent with substitution model misspecifications that result in tree compression/tree extension artifacts, which may explain some discordance between previous divergence estimates based on different sequence types. Comparisons of fully calibrated and nominally calibrated trees support a correlation between body mass and apparent dating error. Overall, our results are consistent with (but do not require) a Paleogene radiation for most major clades of crown birds.

Building the avian tree of life using a large-scale, sparse supermatrix

Birds are the most diverse tetrapod class, with about 10,000 extant species that represent a remarkable evolutionary radiation in which most taxa arose during a short period of time. There has been a tremendous increase in the amount of molecular data available from birds, and more than two-thirds of these species have some sequence data available. Here we assembled these available sequence data from birds to estimate a large-scale avian phylogeny. We performed an unconstrained maximum likelihood analysis of a sparse supermatrix comprising 22 nuclear loci and seven mitochondrial regions from 6714 species. We inferred a phylogeny with a backbone remarkably similar to that obtained by detailed analyses of multigene datasets, yet with the addition of thousands of more taxa. All orders were monophyletic with generally high support. While most families and genera were well supported, a number of them, especially within the oscine passerines, had little or no support. This likely reflects problems with the circumscription of these genera and families. Our results indicate that the amount of sequence data currently available is sufficient to produce a robust estimate of the avian tree of life using current methods of inference. The availability of a tree that is unconstrained by prior information, with branch lengths that have a direct connection to the underlying data, should be useful for comparative methods, taxonomic revisions, and prioritizing taxa that should be targeted for additional data collection.

Statistical binning enables an accurate coalescent-based estimation of the avian tree.

Gene tree incongruence arising from incomplete lineage sorting (ILS) can reduce the accuracy of concatenation-based estimations of species trees. Although coalescent-based species tree estimation methods can have good accuracy in the presence of ILS, they are sensitive to gene tree estimation error. We propose a pipeline that uses bootstrapping to evaluate whether two genes are likely to have the same tree, then it groups genes into sets using a graph-theoretic optimization and estimates a tree on each subset using concatenation, and finally produces an estimated species tree from these trees using the preferred coalescent-based method. Statistical binning improves the accuracy of MP-EST, a popular coalescent-based method, and we use it to produce the first genome-scale coalescent-based avian tree of life.

One hundred new universal exonic markers for birds developed from a genomic pipeline

The functional constraint imposed on exonic sequences increases marker universality and improves sequence alignment accuracy, yet introns have been favoured recently in Avian Tree of Life studies because of the increased sequence coverage that would be required to yield similar resolving power with exons. In this study, we address this shortcoming by developing a pipeline to identify a large number of exonic markers from five avian genomes. Markers were targeted to maximize phylogenetic informativeness and to exclude multi-copy genes. We demonstrate the universality of these markers by testing a portion of them on a taxonomically diverse assemblage of avian specimens.

Reweaving the tapestry: a supertree of birds.

Our knowledge of the avian tree of life remains uncertain, particularly at deeper levels due to the rapid diversification early in their evolutionary history. They are the most abundant land vertebrate on the planet and have been of great historical interest to systematists. Birds are also economically and ecologically important and as a result are intensively studied, yet despite their importance and interest to humans around 13% of taxa currently on the endangered species list perhaps as a result of human activity. Despite all this no comprehensive phylogeny that includes both extinct and extant species currently exists. Here we present a species-level supertree, constructed using the Matrix Representation with Parsimony method, of Aves containing approximately two thirds of all species from nearly 1000 source phylogenies with a broad taxonomic coverage. The source data for the tree were collected and processed according to a strict protocol to ensure robust and accurate data handling. The resulting tree topology is largely consistent with molecular hypotheses of avian phylogeny. We identify areas that are in broad agreement with current views on avian systematics and also those that require further work. We also highlight the need for leaf-based support measures to enable the identification of rogue taxa in supertrees. This is a first attempt at a supertree of both extinct and extant birds, it is not intended to be utilised in an overhaul of avian systematics or as a basis for taxonomic re-classification but provides a strong basis on which to base further studies on macroevolution, conservation, biodiversity, comparative biology and character evolution, in particular the inclusion of fossils will allow the study of bird evolution and diversification throughout deep time.

Identifying localized biases in large datasets: A case study using the avian tree of life

Large-scale multi-locus studies have become common in molecular phylogenetics, with new studies continually adding to previous datasets in an effort to fully resolve the tree of life. Total evidence analyses that combine existing data with newly collected data are expected to increase the power of phylogenetic analyses to resolve difficult relationships. However, they might be subject to localized biases, with one or a few loci having a strong and potentially misleading influence upon the results. To examine this possibility we combined a newly collected 31-locus dataset that includes representatives of all major avian lineages with a published dataset of 19 loci that has a comparable number of sites (Hackett et al., 2008. Science 320, 1763–1768). This allowed us to explore the advantages of conducting total evidence analyses, and to determine whether it was also important to analyze new datasets independent of published ones. The total evidence analysis yielded results very similar to the published results, with only slightly increased support at a few nodes. However, analyzing the 31- and 19-locus datasets separately highlighted several differences. Two clades received strong support in the published dataset and total evidence analysis, but the support appeared to reflect bias at a single locus (β-fibrinogen [FGB]). The signal in FGB that supported these relationships was sufficient to result in their recovery with bootstrap support, even when combined with 49 loci lacking that signal. FGB did not appear to have a substantial impact upon the results of species tree methods, but another locus (brain-derived neurotrophic factor [BDNF]) did have an impact upon those analyses. These results demonstrated that localized biases can influence large-scale phylogenetic analyses but they also indicated that considering independent evidence and exploring multiple analytical approaches could reveal them.

The global diversity of birds in space and time

Current global patterns of biodiversity result from processes that operate over both space and time and thus require an integrated macroecological and macroevolutionary perspective. Molecular time trees have advanced our understanding of the tempo and mode of diversification and have identified remarkable adaptive radiations across the tree of life. However, incomplete joint phylogenetic and geographic sampling has limited broad-scale inference. Thus, the relative prevalence of rapid radiations and the importance of their geographic settings in shaping global biodiversity patterns remain unclear. Here we present, analyse and map the first complete dated phylogeny of all 9,993 extant species of birds, a widely studied group showing many unique adaptations. We find that birds have undergone a strong increase in diversification rate from about 50 million years ago to the near present. This acceleration is due to a number of significant rate increases, both within songbirds and within other young and mostly temperate radiations including the waterfowl, gulls and woodpeckers. Importantly, species characterized with very high past diversification rates are interspersed throughout the avian tree and across geographic space. Geographically, the major differences in diversification rates are hemispheric rather than latitudinal, with bird assemblages in Asia, North America and southern South America containing a disproportionate number of species from recent rapid radiations. The contribution of rapidly radiating lineages to both temporal diversification dynamics and spatial distributions of species diversity illustrates the benefits of an inclusive geographical and taxonomical perspective. Overall, whereas constituent clades may exhibit slowdowns, the adaptive zone into which modern birds have diversified since the Cretaceous may still offer opportunities for diversification.

A locally funded Puerto Rican parrot (Amazona vittata) genome sequencing project increases avian data and advances young researcher education.

BackgroundAmazona vittata is a critically endangered Puerto Rican endemic bird, the only surviving native parrot species in the United States territory, and the first parrot in the large Neotropical genus Amazona, to be studied on a genomic scale.FindingsIn a unique community-based funded project, DNA from an A. vittata female was sequenced using a HiSeq Illumina platform, resulting in a total of ~42.5 billion nucleotide bases. This provided approximately 26.89x average coverage depth at the completion of this funding phase. Filtering followed by assembly resulted in 259,423 contigs (N50 = 6,983 bp, longest = 75,003 bp), which was further scaffolded into 148,255 fragments (N50 = 19,470, longest = 206,462 bp). This provided ~76% coverage of the genome based on an estimated size of 1.58 Gb. The assembled scaffolds allowed basic genomic annotation and comparative analyses with other available avian whole-genome sequences.ConclusionsThe current data represents the first genomic information from and work carried out with a unique source of funding. This analysis further provides a means for directed training of young researchers in genetic and bioinformatics analyses and will facilitate progress towards a full assembly and annotation of the Puerto Rican parrot genome. It also adds extensive genomic data to a new branch of the avian tree, making it useful for comparative analyses with other avian species. Ultimately, the knowledge acquired from these data will contribute to an improved understanding of the overall population health of this species and aid in ongoing and future conservation efforts.

Homoplastic microinversions and the avian tree of life.

BackgroundMicroinversions are cytologically undetectable inversions of DNA sequences that accumulate slowly in genomes. Like many other rare genomic changes (RGCs), microinversions are thought to be virtually homoplasy-free evolutionary characters, suggesting that they may be very useful for difficult phylogenetic problems such as the avian tree of life. However, few detailed surveys of these genomic rearrangements have been conducted, making it difficult to assess this hypothesis or understand the impact of microinversions upon genome evolution.ResultsWe surveyed non-coding sequence data from a recent avian phylogenetic study and found substantially more microinversions than expected based upon prior information about vertebrate inversion rates, although this is likely due to underestimation of these rates in previous studies. Most microinversions were lineage-specific or united well-accepted groups. However, some homoplastic microinversions were evident among the informative characters. Hemiplasy, which reflects differences between gene trees and the species tree, did not explain the observed homoplasy. Two specific loci were microinversion hotspots, with high numbers of inversions that included both the homoplastic as well as some overlapping microinversions. Neither stem-loop structures nor detectable sequence motifs were associated with microinversions in the hotspots.ConclusionsMicroinversions can provide valuable phylogenetic information, although power analysis indicates that large amounts of sequence data will be necessary to identify enough inversions (and similar RGCs) to resolve short branches in the tree of life. Moreover, microinversions are not perfect characters and should be interpreted with caution, just as with any other character type. Independent of their use for phylogenetic analyses, microinversions are important because they have the potential to complicate alignment of non-coding sequences. Despite their low rate of accumulation, they have clearly contributed to genome evolution, suggesting that active identification of microinversions will prove useful in future phylogenomic studies.

Climate: Baselines for the Biological Effects of Environmental Change

Establishing biological baselines requires access to organisms which lived earlier in, or before, the present episode of anthropogenic change. Specimens of a bryozoan collected on Scott's Antarctic expeditions, and subsequently, provide clear evidence of recent increases in growth rate after 80 years of constancy.

Are Transposable Element Insertions Homoplasy Free?: An Examination Using the Avian Tree of Life

Are Transposable Element Insertions Homoplasy Free?: An Examination Using the Avian Tree of Life

Clarifying the systematics of an enigmatic avian lineage: What is a bombycillid?

Avian Tree of Life (TOL) projects using multilocus sequence data have begun to reweave Sibley and Ahlquist’s (1990) “tapestry” of the systematic relationships among birds, which was based on DNA-DNA hybridization. Many of these studies have focused on relationships among members of the most species-rich avian order, Passeriformes, or the perching birds (e.g., Barker et al., 2002, 2004; Cibois and Cracraft, 2004; Voelker and Spellman, 2004; Alstrom et al., 2006). Bringing direct sequence-based character evidence to bear on the systematic relationships among passerine birds has demonstrated that monophyly cannot be assumed for many lineages (Barker et al., 2004), from suborder to tribe, proposed by Sibley and Ahlquist (1990) and Sibley and Monroe (1990). However, many of the paraphyletic passerine lineages identified by modern sequence-based studies represent taxa identified as difficult to place or otherwise problematic by Sibley and Ahlquist (1990). One of the consistently problematic groups has been the Bombycillidae (waxwings and allies). Sibley and Ahlquist (1990, p. 630) ultimately placed the Bombycillidae within the superfamily Muscicapoidea, but they were not confident of this placement. Recent studies have only confirmed the ambiguous placement of Bombycillidae, removing the family from the “core Muscicapoidea” because of poorly supported (low Bayesian credibility and bootstrap values) relationships with the core Muscicapoidea (Barker et al., 2002, 2004; Cibois and Cracraft, 2004; Voelker and Spellman, 2004), but keeping it in the superfamily. A potentially confounding factor for placing the Bombycillidae within the passerine phylogeny is that membership in and relationships among members of the family have long been controversial; therefore, a more thorough sampling of the group is desirable. Many early taxonomies recognized the morphological similarities among waxwings (Bombycilla, 3 spp.), silky-flycatchers (Phainopepla, Phainoptila, and Ptilogonys, 4 spp.), and the palmchat (Dulus, 1 sp.) and proposed them as members of a single family Bombycillidae (Arvey, 1951; Beecher, 1953). However, studies of egg white proteins in the waxwings, silky-flycatchers, and palmchat indicated a close relationship between the former two, but not the latter (Sibley, 1970). The DNA-DNA hybridization analyses of Sibley and Ahlquist (1990) supported the close association among these three groups as indicated by morphology, leading these authors and Sibley and Monroe (1990) to maintain these species as members of the Bombycillidae, while also recognizing their differences by labeling them as distinct tribes (Bombycillini, Ptilogonatini, and Dulini; Table 1). Other modern taxonomies of the bombycillids (Voous, 1977; Cramp, 1988; AOU, 1998) have followed Wetmore (1930) in emphasizing skeletal differences among the three groups and elevating each to the family level (Bombyicillidae, Ptilogonatidae, and Dulidae). If the waxwings, silky-flycatchers, and palmchat are considered the “core” bombycillids (but relationships among the core groups are still unknown), then the remaining ambiguity lies in placement of the only other species ever to be considered among the bombycillids, the grey hypocolius (Hypocolius ampelinus). The grey hypocolius is generally considered to belong to a monotypic family, Hypocoliidae; however, at various points in its taxonomic history it has been considered closely allied with shrikes of the families Prionopidae and Laniidae (morphological characters; Mayr and Amadon, 1951), cuckoo-shrikes (jaw musculature; Campephagidae, Beecher, 1953), bulbuls (morphology; Pycnonotidae, Sibley and Ahlquist, 1990; Sibley and Monroe, 1990; Clements, 1991), and the Bombycillidae (plumage and morphological characters; Lowe, 1947; Delacour and Amadon, 1949). The most recent taxonomic treatment of waxwings, silky-flycatchers, palmchat, and grey hypocolius listed each as a separate but closely related family (Bombycillidae, Ptilogonatidae, Dulidae, and Hypocoliidae), reflecting the continued ambiguity of relationships among the groups (del Hoyo, 2005). Table 1 Specimen information.

Bird evolution: testing the metaves clade with six new mitochondrial genomes

BackgroundEvolutionary biologists are often misled by convergence of morphology and this has been common in the study of bird evolution. However, the use of molecular data sets have their own problems and phylogenies based on short DNA sequences have the potential to mislead us too. The relationships among clades and timing of the evolution of modern birds (Neoaves) has not yet been well resolved. Evidence of convergence of morphology remain controversial. With six new bird mitochondrial genomes (hummingbird, swift, kagu, rail, flamingo and grebe) we test the proposed Metaves/Coronaves division within Neoaves and the parallel radiations in this primary avian clade.ResultsOur mitochondrial trees did not return the Metaves clade that had been proposed based on one nuclear intron sequence. We suggest that the high number of indels within the seventh intron of the β-fibrinogen gene at this phylogenetic level, which left a dataset with not a single site across the alignment shared by all taxa, resulted in artifacts during analysis. With respect to the overall avian tree, we find the flamingo and grebe are sister taxa and basal to the shorebirds (Charadriiformes). Using a novel site-stripping technique for noise-reduction we found this relationship to be stable. The hummingbird/swift clade is outside the large and very diverse group of raptors, shore and sea birds. Unexpectedly the kagu is not closely related to the rail in our analysis, but because neither the kagu nor the rail have close affinity to any taxa within this dataset of 41 birds, their placement is not yet resolved.ConclusionOur phylogenetic hypothesis based on 41 avian mitochondrial genomes (13,229 bp) rejects monophyly of seven Metaves species and we therefore conclude that the members of Metaves do not share a common evolutionary history within the Neoaves.

Mitochondrial Genomes and Avian Phylogeny: Complex Characters and Resolvability without Explosive Radiations

We improve the taxon sampling for avian phylogeny by analyzing 7 new mitochondrial genomes (a toucan, woodpecker, osprey, forest falcon, American kestrel, heron, and a pelican). This improves inference of the avian tree, and it supports 3 major conclusions. The first is that some birds (including a parrot, a toucan, and an osprey) exhibit a complete duplication of the control region (CR) meaning that there are at least 4 distinct gene orders within birds. However, it appears that there are regions of continued gene conversion between the duplicate CRs, resulting in duplications that can be stable for long evolutionary periods. Because of this stable duplicated state, gene order can eventually either revert to the original order or change to the new gene order. The existence of this stable duplicate state explains how an apparently unlikely event (finding the same novel gene order) can arise multiple times. Although rare genomic changes have theoretical advantages for tree reconstruction, they can be compromised if these apparently rare events have a stable intermediate state. Secondly, the toucan and woodpecker improve the resolution of the 6-way split within Neoaves that has been called an "explosive radiation." An explosive radiation implies that normal microevolutionary events are insufficient to explain the observed macroevolution. By showing the avian tree is, in principle, resolvable, we demonstrate that the radiation of birds is amenable to standard evolutionary analysis. Thirdly, and as expected from theory, additional taxa breaking up long branches stabilize the position of some problematic taxa (like the falcon). In addition, we report that within the birds of prey and allies, we did not find evidence pairing New World vultures with storks or accipitrids (hawks, eagles, and osprey) with Falconids.