Fig. 1: The workflow in this study is summarized in this schematic.

Each different coloured background outlines the steps taken to address a given hypothesis. (H1, purple) Identification and confirmation of neo-sex Chr1A behaviour in the coastal Eastern Yellow Robin (EYR) would mirror that seen in the published inland EYR genome and indicate that neo-sex Chr1A arose before the two lineages split. (H2, blue) Identification and classification of Chr1A-derived neo-sex genes into bins with or without mitochondrial functions (N-mt versus non-N-mt genes), followed by testing for differences in protein evolution rates (dNdS) and differentiation levels between lineages (Dxy) would show whether neo-sex N-mt genes are undergoing accelerated evolution relative to non-N-mt genes. (H3, orange) A similar comparison of neo-W versus neo-Z Chr1A-derived genes allows us to draw conclusions about the relative rates of evolution on each neo-sex chromosome, supplemented by an additional search for neo-W candidates under intensified positive selection possibly resulting from accelerated co-evolution with mitochondrial DNA.