Repeated behavioural evolution is associated with convergence of gene expression in cavity-nesting songbirds.

Uncovering the genomic bases of phenotypic adaptation is a major goal in biology, but this has been hard to achieve for complex behavioural traits. Here we leverage the repeated, independent evolution of obligate cavity nesting in birds to test the hypothesis that pressure to compete for a limited breeding resource has facilitated convergent evolution in behaviour, hormones and gene expression. We used an integrative approach, combining aggression assays in the field, testosterone measures and transcriptome-wide analyses of the brain in wild-captured females and males. Our experimental design compared species pairs across five avian families, each including one obligate cavity-nesting species and a related species with a more flexible nest strategy. We find behavioural convergence, with higher levels of territorial aggression in obligate cavity nesters, particularly among females. Across species, levels of testosterone in circulation were not associated with nest strategy nor aggression. Phylogenetic analyses of individual genes and co-regulated gene networks revealed more shared patterns of brain gene expression than expected by drift, although the scope of convergent gene expression evolution was limited to a small percentage of the genome. When comparing our results to other studies that did not use phylogenetic methods, we suggest that accounting for shared evolutionary history may reduce the number of genes inferred as convergently evolving. Altogether, we find that behavioural convergence in response to shared ecological pressures is associated with largely independent evolution of gene expression across different avian families, punctuated by a narrow set of convergently evolving genes.