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Disentangling the main drivers of species richness and community composition is a central theme in ecology. Freshwater biodiversity patterns have been poorly explored; yet, it has been shown that different freshwater biota have different, often contrasting responses to environmental gradients. In this study, we investigated the relative contribution of geographical and environmental (habitat-, climate- and water quality-related) factors/gradients in shaping the α- and β-diversity patterns of macrophytes and fish in sixteen natural freshwater lakes of an unexplored Balkan biodiversity hotspot, the Southern Balkan Peninsula. We employed generalized linear modeling to identify drivers of α-diversity, and generalized dissimilarity modeling to explore commonalities and dissimilarities of among-biota β-diversity. Species richness of both biota was significantly associated with lake surface area, whereas macrophytes had an inverse response to altitude, compared to fish. Both species turnover and nestedness significantly contributed to the total β-diversity of macrophytes. In contrast, species turnover was the most significant contributor to the total fish β-diversity. We found that the compositional variation of macrophytes is primarily limited by dispersal and ultimately shaped by environmental drivers, resulting in spatially structured assemblages. Fish communities were primarily shaped by altitude, highlighting the role of species sorting. We conclude that among-biota diversity patterns are shaped by different/contrasting factors, and, thus, effective/sustainable conservation strategies should encompass multiple aquatic biota.

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α- and β-Diversity Patterns of Macrophytes and Freshwater Fishes are Driven by Different Factors and Processes in Lakes of the Unexplored Southern Balkan Biodiversity Hotspot