OCBIL theory provides a basis for understanding of the evolution and ecology of biota on old, climatically buffered, infertile landscapes (OCBILs) worldwide. Here, we test a genetic hypothesis presented in OCBIL theory that predicts highly differentiated intraspecific population systems on OCBILs vs. more limited differentiation on young, often disturbed, fertile landscapes (YODFELs). We examined patterns of genomic and morphological divergence in Banksia seminuda across OCBILs and YODFELs in south-western Australia. We also used these data to determine whether these OCBIL and YODFEL populations represent distinct subspecific lineages, a point of previous contention among taxonomists. As hypothesized, genomic analyses based on 3466 SNP loci revealed strong structuring within B. seminuda, with high differentiation across narrow geographic scales among OCBIL populations vs. lower differentiation across much larger geographic scales among YODFEL populations. In addition, genomic and morphological divergence was found between OCBIL and YODFEL populations, providing comprehensive quantitative evidence for two subspecies. These findings have taxonomic implications for the species and provide support for OCBIL theory and its insights into the evolution, ecology and conservation of biota on ancient landscapes.