Leonardo Campagna

Distinguishing Noise from Signal in Patterns of Genomic Divergence in a Highly Polymorphic Avian Radiation

Recently diverged taxa provide the opportunity to search for the genetic basis of the phenotypes that distinguish them. Genomic scans aim to identify loci that are diverged with respect to an otherwise weakly differentiated genetic background. These loci are candidates for being past targets of selection because they behave differently from the rest of the genome that has either not yet differentiated or that may cross species barriers through introgressive hybridization. Here we use a reduced‐representation genomic approach to explore divergence among six species of southern capuchino seedeaters, a group of recently radiated sympatric passerine birds in the genus Sporophila. For the first time in these taxa, we discovered a small proportion of markers that appeared differentiated among species. However, when assessing the significance of these signatures of divergence, we found that similar patterns can also be recovered from random grouping of individuals representing different species. A detailed demographic inference indicates that genetic differences among Sporophila species could be the consequence of neutral processes, which include a very large ancestral effective population size that accentuates the effects of incomplete lineage sorting. As these neutral phenomena can generate genomic scan patterns that mimic those of markers involved in speciation and phenotypic differentiation, they highlight the need for caution when ascertaining and interpreting differentiated markers between species, especially when large numbers of markers are surveyed. Our study provides new insights into the demography of the southern capuchino radiation and proposes controls to distinguish signal from noise in similar genomic scans.

Genomic approaches to understanding population divergence and speciation in birds

ABSTRACT The widespread application of high-throughput sequencing in studying evolutionary processes and patterns of diversification has led to many important discoveries. However, the barriers to utilizing these technologies and interpreting the resulting data can be daunting for first-time users. We provide an overview and a brief primer of relevant methods (e.g., whole-genome sequencing, reduced-representation sequencing, sequence-capture methods, and RNA sequencing), as well as important steps in the analysis pipelines (e.g., loci clustering, variant calling, whole-genome and transcriptome assembly). We also review a number of applications in which researchers have used these technologies to address questions related to avian systems. We highlight how genomic tools are advancing research by discussing their contributions to 3 important facets of avian evolutionary history. We focus on (1) general inferences about biogeography and biogeographic history, (2) patterns of gene flow and isolation upon secondary contact and hybridization, and (3) quantifying levels of genomic divergence between closely related taxa. We find that in many cases, high-throughput sequencing data confirms previous work from traditional molecular markers, although there are examples in which genome-wide genetic markers provide a different biological interpretation. We also discuss how these new data allow researchers to address entirely novel questions, and conclude by outlining a number of intellectual and methodological challenges as the genomics era moves forward.

Repeated divergent selection on pigmentation genes in a rapid finch radiation

Selection acted repeatedly on regions that may regulate the expression of genes underlying coloration differences in seedeaters. Instances of recent and rapid speciation are suitable for associating phenotypes with their causal genotypes, especially if gene flow homogenizes areas of the genome that are not under divergent selection. We study a rapid radiation of nine sympatric bird species known as capuchino seedeaters, which are differentiated in sexually selected characters of male plumage and song. We sequenced the genomes of a phenotypically diverse set of species to search for differentiated genomic regions. Capuchinos show differences in a small proportion of their genomes, yet selection has acted independently on the same targets in different members of this radiation. Many divergent regions contain genes involved in the melanogenesis pathway, with the strongest signal originating from putative regulatory regions. Selection has acted on these same genomic regions in different lineages, likely shaping the evolution of cis-regulatory elements, which control how more conserved genes are expressed and thereby generate diversity in classically sexually selected traits.

Correlated patterns of genetic diversity and differentiation across an avian family

Comparative studies of closely related taxa can provide insights into the evolutionary forces that shape genome evolution and the prevalence of convergent molecular evolution. We investigated patterns of genetic diversity and differentiation in stonechats (genus Saxicola), a widely distributed avian species complex with phenotypic variation in plumage, morphology and migratory behaviour, to ask whether similar genomic regions have become differentiated in independent, but closely related, taxa. We used whole‐genome pooled sequencing of 262 individuals from five taxa and found that levels of genetic diversity and divergence are strongly correlated among different stonechat taxa. We then asked whether these patterns remain correlated at deeper evolutionary scales and found that homologous genomic regions have become differentiated in stonechats and the closely related Ficedula flycatchers. Such correlation across a range of evolutionary divergence and among phylogenetically independent comparisons suggests that similar processes may be driving the differentiation of these independently evolving lineages, which in turn may be the result of intrinsic properties of particular genomic regions (e.g. areas of low recombination). Consequently, studies employing genome scans to search for areas important for reproductive isolation or adaptation should account for corresponding regions of differentiation, as these regions may not necessarily represent speciation islands or evidence of local adaptation.

Genomic evidence of demographic fluctuations and lack of genetic structure across flyways in a long distance migrant, the European turtle dove

BackgroundUnderstanding how past climatic oscillations have affected organismic evolution will help predict the impact that current climate change has on living organisms. The European turtle dove, Streptopelia turtur, is a warm-temperature adapted species and a long distance migrant that uses multiple flyways to move between Europe and Africa. Despite being abundant, it is categorized as vulnerable because of a long-term demographic decline. We studied the demographic history and population genetic structure of the European turtle dove using genomic data and mitochondrial DNA sequences from individuals sampled across Europe, and performing paleoclimatic niche modelling simulations.ResultsOverall our data suggest that this species is panmictic across Europe, and is not genetically structured across flyways. We found the genetic signatures of demographic fluctuations, inferring an effective population size (Ne) expansion that occurred between the late Pleistocene and early Holocene, followed by a decrease in the Ne that started between the mid Holocene and the present. Our niche modelling analyses suggest that the variations in the Ne are coincident with recent changes in the availability of suitable habitat.ConclusionsWe argue that the European turtle dove is prone to undergo demographic fluctuations, a trait that makes it sensitive to anthropogenic impacts, especially when its numbers are decreasing. Also, considering the lack of genetic structure, we suggest all populations across Europe are equally relevant for conservation.

Supergenes: The Genomic Architecture of a Bird with Four Sexes

Supergenes are clusters of physically linked, co-evolving genes that often control complex traits. A new study clarifies the origin and possible fate of a fascinating supergene that determines the coloration and mating behavior of a widespread North American bird.

A flicker of hope: Genomic data distinguish Northern Flicker taxa despite low levels of divergence

ABSTRACT Next-generation sequencing technologies are increasingly being employed to explore patterns of genomic variation in avian taxa previously characterized using morphology and/or traditional genetic markers. The hybridization dynamics of the Northern Flicker complex have received considerable attention, primarily due to the conspicuous plumage differences among these birds and the geographically extensive hybrid zone between the Red-shafted (Colaptes auratus cafer) and Yellow-shafted (Colaptes auratus auratus) flickers in the Great Plains region of North America. However, no traditional molecular techniques have been able to differentiate these 2 morphologically well-defined taxa from one another, or conclusively from the closely related Gilded Flicker (Colaptes chrysoides). Here, we use a next-generation sequencing approach to assess the genetic diversity and evolutionary history of these 3 taxa. We confirm the overall low levels of differentiation found using traditional molecular markers, but are able to distinguish between the 3 taxa for the first time, using a dataset of thousands of SNP loci distributed across the genome. Through demographic modeling and phylogenetic reconstructions, we find that Red-shafted and Yellow-shafted flickers are likely sister taxa, and that their divergence from the Gilded Flicker was comparatively older. The low level of divergence and lack of fixed differences in our dataset between Red-shafted and Yellow-shafted flickers, in particular, suggests whole-genome re-sequencing may be necessary to assess the dynamics of their hybridization and identify the genetic basis of their striking differences in plumage.

Transgressive phenotypes and evidence of weak postzygotic isolation in F1 hybrids between closely related capuchino seedeaters

Postzygotic reproductive isolation may become strong only once the process of speciation is in its advanced stages. For taxa in the early stages of speciation, prezygotic reproductive isolation barriers may play a predominant role in maintaining species boundaries. Here, we study the recent capuchino seedeater biological radiation, a group of highly sympatric species from the genus Sporophila that have diversified during the Pleistocene in Neotropical grasslands. Capuchinos can be diagnosed by adult male coloration patterns and song, two sets of characters known to contribute to pre-mating reproductive isolation. However, it remains unknown whether potzygotic incompatibilities contribute to maintaining species limits in this group. Here we use existing breeding records from captive individuals to test for patterns consistent with F1 inviability. We compare hatching success, fledging success, and the sex ratio at adulthood between conspecific and hybrid capuchino pairs. We observed a trend towards lower numbers of the heterogametic sex among adult hybrids, consistent Haldanes rule, but this was supported by only one of our statistical tests. Our study is the first to document hybrid male capuchino phenotypes based on known crosses. We observed phenotypes that were similar or intermediate to those of the parental species, as well as novel plumage patterns that have not been described in the wild. One cross produced a plumage pattern that has been observed at low frequencies in natural populations. We discuss the implications of our results for understanding the relative importance of the mechanisms of reproductive isolation in capuchino seedeaters.

Population Genomics of Birds: Evolutionary History and Conservation

The use of genome-scale data to understand the evolutionary history of birds has provided important progress in the field of evolutionary biology and conservation. Here we review the conceptual advances of avian genomics, along with key examples from the literature. In each section, we contrast studies that utilized only a small number of genetic markers to studies that incorporated many independent loci across the genome. We discuss the important characteristics of avian genome architecture, and we explore the connections between DNA sequence variation and ecologically relevant phenotypes, such as color and morphology. We ask how environmental factors have left their mark on the genomes of birds and how genomic data can be used to reconstruct histories across multiple species. We outline how admixture and reticulate evolutionary histories have been an important source of variation and review cases in which hybridization has possibly led to the formation of new species. Finally, we discuss how genomic data have helped delineate population structure and inform conservation actions in declining avian species. Like in other taxonomic groups, the ever-expanding molecular toolbox for avian biologists is at once becoming more accessible in cost and more powerful in its applications. Therefore, the study of avian genomes will continue to provide important insights into many aspects of ecology, evolutionary history, and conservation biology.

Double-digest RAD sequencing outperforms microsatellite loci at assigning paternity and estimating relatedness: A proof of concept in a highly promiscuous bird

Information on genetic relationships among individuals is essential to many studies of the behaviour and ecology of wild organisms. Parentage and relatedness assays based on large numbers of single nucleotide polymorphism (SNP) loci hold substantial advantages over the microsatellite markers traditionally used for these purposes. We present a double‐digest restriction site‐associated DNA sequencing (ddRAD‐seq) analysis pipeline that, as such, simultaneously achieves the SNP discovery and genotyping steps and which is optimized to return a statistically powerful set of SNP markers (typically 150–600 after stringent filtering) from large numbers of individuals (up to 240 per run). We explore the trade‐offs inherent in this approach through a set of experiments in a species with a complex social system, the variegated fairy‐wren (Malurus lamberti) and further validate it in a phylogenetically broad set of other bird species. Through direct comparisons with a parallel data set from a robust panel of highly variable microsatellite markers, we show that this ddRAD‐seq approach results in substantially improved power to discriminate among potential relatives and considerably more precise estimates of relatedness coefficients. The pipeline is designed to be universally applicable to all bird species (and with minor modifications to many other taxa), to be cost‐ and time‐efficient, and to be replicable across independent runs such that genotype data from different study periods can be combined and analysed as field samples are accumulated.