1. We performed spatial genetic analyses, incorporating landscape genetic methods using microsatellite data and phylogeographic analyses using mtDNA data, to identify the principal factors that determine population heterogeneity of the tropical freshwater fish, Mogurnda mogurnda, in the Daly River, northern Australia. We tested the individual and interactive effects of several environmental variables on spatial genetic patterns, including metrics relating to connectivity (i.e. stream distance, maximum stream gradient and elevation), habitat size (i.e. mean annual discharge) and a categorical variable relating to population history, as determined by mtDNA phylogeographic analyses. The Daly River is geomorphologically and hydrologically complex, and M. mogurnda has life history traits that limit its dispersal potential at river basin scales. Thus, we predicted that variables relating to connectivity would be the most important landscape factors driving population structure of the species.
2. Tree-based phylogeographic analyses indicated four divergent mtDNA lineages within M. mogurnda in the Daly River, although three of the lineages were sympatric in various combinations and did not correspond with microsatellite groups identified by assignment tests. The allopatric mtDNA lineage detected in the uppermost part of the catchment was also identified as being highly differentiated by the microsatellite data, strongly suggesting that it may be a cryptic species. This site was therefore excluded from subsequent landscape genetic analyses.
3. Analyses of Molecular Variance indicated that M. mogurnda has a hierarchical population structure in the Daly River, thus supporting theoretical expectations that hierarchically arranged river habitats in dendritic systems impose hierarchal population structures on lotic species.
4. All landscape genetic analyses rejected stream distance, and supported stream gradient, as the major determinant of spatial genetic variation in M. mogurnda in the Daly River. Support for elevation as a determinant of spatial genetic patterns differed among the landscape genetic methods. Several of the landscape genetic methods also indicate that population history, including secondary contact between divergent and formerly allopatric genetic lineages, has a strong influence on spatial genetic patterns within M. mogurnda in the Daly River.
5. This study demonstrates the need to consider multiple environmental factors, especially factors relating to connectivity, and their interactions in spatial genetic analysis, rather than just geographic distance. Importantly, it demonstrates the need to account for population history and evolutionary divergences in landscape genetic analyses.