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A phylogeny of the species in the genera Chondrostoma and Squalius was constructed based on the complete mitochondrial cytochrome b gene (1140pb). The molecular phylogeny was used to test the effect of the Mediterranean Lago Mare dispersal theory on the processes of divergence and speciation of European freshwater fishes. Phylogenetic relationships among Squalius samples and the molecular clock revealed that the ancestor of the current Iberian Squalius species inhabited a wide geographic area in the central and southwestern part of the former Iberian Peninsula during the Miocene before the Lago Mare phase. Similarly, the four main Iberian lineages of the genus Chondrostoma originated in the Middle-Upper Miocene. Hence, the Lago Mare phase of the Mediterranean Sea seems to have been a too recent paleogeographic event to have had any major impact on the dispersion of Squalius and Chondrostoma species. However, the reduction of the water-bodies during the Tortonian and Messinian may have intensified the isolation of populations. The Operational Biogeographic Units recovered from the Squalius and Chondrostoma phylogenies also reject the Lago Mare dispersal theory and support the idea that the differentiation processes were due to both the formation of the current hydrographical basin during the Plio-Pleistocene as well as to an earlier endorrheism event that occurred prior to hydrographical configuration.

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Testing freshwater Lago Mare dispersal theory on the phylogeny relationships of iberian cyprinid genera &lt;i&gt;Chondrostoma&lt;/i&gt; and &lt;i&gt;Squalius&lt;/i&gt; (Cypriniformes, Cyprinidae)