This study investigated the causes of high level of intra-and inter-population variation known to occur in the morphology of fish in the genus Hypseleotris Eleotride in southern Australia, particularly within the Murray-Darling river system. The three major objectives of the study were, identify the number and distribution of species,determine the genetic structure of the populations and analyse relationships between species and consider the process of speciation in this species complex. The investigation of morphological variation in Hypseleotris confirmed the presence of two well known species i.e. H. compressa and H. galli, in the coastal rivers and also of the inland species H. klunzingeri. Populations of Hypseleotris klunzigeri sensu lato in inland river were found to be very highly variable and analysis using discriminant functions and principle component analysis showed the widespread presence of three forms (A, B1 and B2). The analysis was confused by the presence of north/south clines and upstream/downstream variation in characteristic in each form. After these factors were removed, there was still a great deal of variation in each population. The presence of hybrids between each pair of inland species, identified by both morphological and genetic data, further confused the analysis and makes identification of all specimens to species in the field difficult. Examination of type material of H. Klunzingeri showed that this belonged to form B2. The other forms can be related to the undescribed species, Midgley's carp gudgeon and Lake's carp gudgeon. Keys to the species in the complex in southeastern Australia are given. The morphological and genetic data show that H. compressa and H. klunzingeri are sister species, primarily separated by the eastern uplands. Similarly, the coastal species, H. galli is related to form B1 and more distantly, to form A. Possible scenarios for the complex are given. / Doctor of Philosophy (PhD)