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Abstract In song-learning birds, vocalizations are species recognition signals and may act as premating reproductive barriers; for allopatric taxa, testing how the signals can influence the speciation processes is quite challenging. This study aims to understand genomic divergence and species recognition via songs in 2 allopatric taxa, eastern and western Nashville warblers (Leiothlypis ruficapilla ruficapilla vs. Leiothlypis ruficapilla ridgwayi). We performed playback experiments to assess their reciprocal behavioral responses, which suggests an asymmetric barrier: the eastern L. r. ruficapilla discriminates between the 2 songs, but the western L. r. ridgwayi does not. Using whole-genome sequencing, we also examined the extent of the taxa’s genomic divergence and estimated their demographic history. We identified dozens of highly differentiated genomic regions, as well as fluctuations in historical effective population sizes that indicate independent demographic trajectories during the Pleistocene. To contextualize the magnitude of divergence between L. ruficapilla subspecies, we applied the same genomic analyses to 2 additional eastern-western pairs of parulid warblers, Setophaga virens vs. Setophaga townsendi and Setophaga coronata coronata vs. Setophaga coronata auduboni, which have existing behavior studies but are not in strict allopatry. Our findings provide insights into the role of vocalizations in defining within-pair relationship and the important legacy of isolation during the Pleistocene. 

Introgression between species has the potential to shape evolutionary trajectories in important ways, but uncovering complex introgression dynamics has only recently been made possible by advances in genomics. Warblers of the avian family Parulidae exemplify rapid diversification and sexual trait divergence, and we endeavored to study historical introgression in the family. We sequenced multiple genomes of nearly every species, constructed a phylogeny for the family, and investigated gene flow across the genome and at genes known for controlling feather color. We found that DNA haplotypes including the geneBCO2, which encodes an enzyme that degrades yellow carotenoids, have spread among genera multiple times—fromVermivoratoGeothlypisand fromLeiothlypisto multipleCardellinaandSetophagaspecies. Patterns of inheritance in the latter case point to introgression that occurred 0.5 to 2 million years ago, and the shared haplotype among recipient species is less than 100 nucleotides long. Separately, we found evidence of introgression from redCardellinaspecies to both of the two redMyioborusspecies atBDH1Land from one redMyioborusspecies to the other atCYP2J19; both are key genes in the pathway that converts yellow carotenoids to red ketocarotenoids. Our results show that introgression is a common mechanism for the evolution of colorful plumage in parulid warblers and hint at complex histories of gene flow behind some of the Western Hemisphere’s most colorful birds. 

The Kirtland's warbler (Setophaga kirtlandi) is a rare migratory passerine species and habitat specialist of the North American Jack Pine Forests. Their near extinction in the 1970s classified them as endangered and protected under the Endangered Species Act of 1973. After decades of intense conservation management, their population size recovered, and they were delisted from federal protection in 2019. We explore the genomic consequences of this harsh bottleneck and recovery by comparing the genomic architecture of two closely related species whose population sizes have remained large and stable, Hooded Warblers (Setophaga citrina) and American Redstarts (Setophaga ruticilla). We used whole‐genome sequencing to characterize the distribution of runs of homozygosity and deleterious genetic variation. We find evidence that Kirtland's warblers exhibit genetic patterns consistent with recent inbreeding. Our results also show that Kirtland's warblers carry an excess proportion of deleterious variation, which could complicate management for this conservation‐reliant species. This analysis provides a genetically informed perspective that should be thoroughly considered when delisting other species from federal protections. Through the increasing accessibility of genome sequencing technology, it will be more feasible to monitor the genetic landscape of recovering populations to ensure their long‐term survival independent of conservation intervention. 

Vertebrates host complex microbiomes that impact their physiology. In many taxa, including colourful wood-warblers, gut microbiome similarity decreases with evolutionary distance. This may suggest that as host populations diverge, so do their microbiomes, because of either tight coevolutionary dynamics, or differential environmental influences, or both. Hybridization is common in wood-warblers, but the effects of evolutionary divergence on the microbiome during secondary contact are unclear. Here, we analyse gut microbiomes in two geographically disjunct hybrid zones between blue-winged warblers (Vermivora cyanoptera) and golden-winged warblers (Vermivora chrysoptera). We performed 16S faecal metabarcoding to identify species-specific bacteria and test the hypothesis that host admixture is associated with gut microbiome disruption. Species identity explained a small amount of variation between microbiomes in only one hybrid zone. Co-occurrence of species-specific bacteria was rare for admixed individuals, yet microbiome richness was similar among admixed and parental individuals. Unexpectedly, we found several bacteria that were more abundant among admixed individuals with a broader deposition of carotenoid-based plumage pigments. These bacteria are predicted to encode carotenoid biosynthesis genes, suggesting birds may take advantage of pigments produced by their gut microbiomes. Thus, host admixture may facilitate beneficial symbiotic interactions which contribute to plumage ornaments that function in sexual selection. 

Bachman’s warbler (Vermivora bachmanii)—last sighted in 1988—is one of the only North American passerines to recently go extinct. Given extensive ongoing hybridization of its two extant congeners—the bluewinged warbler (V. cyanoptera) and golden-winged warbler (V. chrysoptera)—and shared patterns of plumage variation between Bachman’s warbler and hybrids between those extant species, it has been suggested that Bachman’s warbler might have also had a component of hybrid ancestry. Here, we use historic DNA (hDNA) and whole genomes of Bachman’s warblers collected at the turn of the 20th century to address this. We combine these data with the two extant Vermivora species to examine patterns of population differentiation, inbreeding, and gene flow. In contrast to the admixture hypothesis, the genomic evidence is consistent with V. bachmanii having been a highly divergent, reproductively isolated species, with no evidence of introgression. We show that these three species have similar levels of runs of homozygosity (ROH), consistent with effects of a small long-term effective population size or population bottlenecks, with one V. bachmanii outlier showing numerous long ROH and a FROH greater than 5%. We also found—using population branch statistic estimates—previously undocumented evidence of lineage-specific evolution in V. chrysoptera near a pigmentation gene candidate, CORIN, which is a known modifier of ASIP, which is in turn involved in melanic throat and mask coloration in this family of birds. Together, these genomic results also highlight how natural history collections are such invaluable repositories of information about extant and extinct species.