Background: Koala Retrovirus-A is a gamma-retrovirus that is spreading across wild koala populations through horizontal and vertical transmission, contributing significantly to genomic diversity across and even within koala populations. Previous studies have estimated that KoRV-A initially integrated into the koala genome less than 50,000 years ago, but the precise origins and the patterns of spread after its endogenization remain unclear.

Results: In this study, we analyzed germline insertions of KoRV-A using whole-genome sequencing data from 405 wild koalas, representing nearly the species' entire geographic range. Our findings reveal an evolutionary trajectory for KoRV-A, suggesting that the initial endogenization might occur near Coffs Harbour on the Mid-north coast of NSW around the middle of the koala's range. As KoRV-A spread, certain subtypes emerged and became prevalent, two of which recombined with an ancient endogenous retrovirus, PhER, resulting in distinct recombination variants in northern and southern koala populations. Additionally, we identified a geographic barrier north of Sydney, which may have slowed the southward spread of KoRV-A into Sydney and beyond.

Conclusions: Our study proposes a comprehensive evolutionary pathway for KoRV-A, beginning with its initial endogenization near Coffs Harbour and highlighting barriers and diversification events that have shaped its distribution and impact on koala populations.