Mediterranean climate river systems are among the most threatened ecosystems worldwide, due to a long history of anthropogenic impacts and alien invasive species introductions. Many of such rivers naturally exhibit a non-perennial flow regime, with distinct seasonal, inter-annual and spatial heterogeneity. The present study seeks to detect diatom community patterns and to understand the processes that cause these structures in an Austral Mediterranean river system among different months and river sections. In general, most environmental variables showed an increasing trend downstream for both months, with the exception of pH, dissolved oxygen, PO₄3− and substrate embeddedness, which decreased downstream. A total of 110 diatom species between the two study months (October – 106 taxa; January – 78 taxa) were identified, dominated by 30 species with at least >2% abundance. Diatom community structure differed significantly across river zones, while no significant differences were observed between the study months. A boosted regression trees model showed that B (43.3%), Cu (20.8%), Fe (3.4%) and water depth (3.2%) were the most significant variables structuring diatoms. Diatom species communities reflected environmental variables (i.e., sediment and water chemistry) in this Mediterranean climate river system, as sediment metals such as B, Cu and Fe were found to be important in structuring diatom communities. Biotic influences from fish communities had little effect on diversity, but shifted diatom community structure. Therefore, the current study highlights how river systems have complex interactions that play an important role in determining diatom species composition. Display omitted •Sediment and water chemistry variables increased downstream in both events.•110 diatom species across the two study events identified.•Significant differences observed in diatom community across river zones and events.•Boron and copper most important variables in structuring diatoms.