Song evolution in light of ecosystem differences: exploring effects of urbanization and ecology on temporal and frequency traits of Spotted and Eastern towhee songs

The Eastern towhee (Pipilo erythrophthalmus) and Spotted towhee (Pipilo maculatus) are large New World sparrows found across North America. These two species were previously classified as a single species, the Rufous-sided towhee, which was separated in 1995 based on differences in plumage, geographic range, and song. Previous studies have shown that ecological factors, such as urbanization and climate, can affect learned vocalizations, particularly frequency-related song characteristics such as minimum frequency; however, most studies have been conducted on one species in a specific location. The extensive geographic distributions of these towhee species, along with their ample publicly available song-recording data, give us the unique opportunity to assess whether ecological pressures influence song variation. Here, we extract frequency-related song features from 2916 Spotted and Eastern towhee recordings and investigate whether geography and ecology--including recording location (latitude and longitude), tree cover, urbanization (nighttime lights, human population density, and distance to road), climate zones, elevation, and ecoregions--explain patterns in these song frequency variables of this sister-species pair. Our results show that geographic location, particularly longitude, contributes more strongly to the variation of song frequency features than urbanization, environment, and climate, suggesting that culturally transmitted differences in learned song, not ecology or anthropogenic structure, drive this variation. However, there was not a clear pattern of isolation-by-distance despite the geographic patterns that we found in their songs. Further, we did not find strong support for behavioral adaptation to habitat structure, but we did find a weak signal that urbanization was associated with increased frequency in Spotted towhees. Overall, we provide a detailed study on the interactions between ecology and song evolution, and provide new insight into the evolution of birdsong.