AbstractTheoretical and empirical studies revealed that fast evolving microsatellite markers might be successfully employed in phylogenetic reconstruction. In this study we used seven highly polymorphic microsatellite loci and six different genetic distances to infer the performance of microsatellites in estimation of phylogenetic relationships between closely related crested newt species (Triturus cristatus superspecies). Many intraspecific distances exceeded interspecific values likely due to loss of genetic distance linearity in time, revealing that none of them was able to estimate divergence between the species. The Neighbor-joining trees constructed on the basis of genetic distances showed trichotomies (three-way polychotomies) and low bootstrap support at the species level. This study thus revealed limits of microsatellites to estimate phylogenetic relationships between the crested newt taxa.