20While the influence of Pleistocene climatic changes on divergence and speciation 21 has been well-documented across the globe, complex spatial interactions between 22 hydrology and eustatics over longer timeframes may also determine species 23 evolutionary trajectories. Within the Australian continent, glacial cycles were not 24 associated with changes in ice cover and instead largely resulted in fluctuations from 25 moist to arid conditions across the landscape. Here, we investigate the role of 26 hydrological and coastal topographic changes brought about by Plio-Pleistocene 27 climatic changes on the biogeographic history of a small Australian freshwater fish, 28 the southern pygmy perch Nannoperca australis. Using 7,958 ddRAD-seq (double 29 digest restriction-site associated DNA) loci and 45,104 filtered SNPs, we combined 30 phylogenetic, coalescent and species distribution analyses to investigate the relative 31 roles of aridification, sea level and tectonics and their associated biogeographic 32 changes across southeast Australia. Sea-level changes since the Pliocene and 33 reduction or disappearance of large waterbodies throughout the Pleistocene were 34 determining factors in strong divergence across the clade, including the initial 35 formation and maintenance of a cryptic species, N. 'flindersi'. Isolated climatic 36 refugia and fragmentation due to lack of connected waterways maintained the 37 identity and divergence of inter-and intraspecific lineages. Our historical findings 38 suggest that predicted increases in aridification and sea level due to anthropogenic 39 climate change might result in markedly different demographic impacts, both 40 spatially and across different landscape types. 41 42