Gammarus leopoliensis (Crustacea: Amphipoda) is considered a north-eastern Carpathian endemic species and therefore can be regarded as an appropriate model for testing the hypothesis of Quaternary glacial survival in northern microrefugia. However, 250km south, the south-western Carpathians harbour populations that resemble phenotypically both G.leopoliensis and Gammarus kischineffensis, a similar species distributed east of the Carpathians.
We used maximum-likelihood and Bayesian methods to evaluate the phylogenetic relationships of these three taxa based on mitochondrial and nuclear markers, and quantitatively compared diversity patterns, phylogeography and divergence times among north-eastern and south-western Carpathian taxa. Results indicate that G.leopoliensis and the south-western populations form together a strongly supported group (G.leopoliensis s.l.) which, along with G.kischineffensis, belongs to the Gammarus balcanicus clade.
This group contains 12 lineages mainly of Pliocene age. G.leopoliensis consists of two widely distributed and recently expanded allopatric sister lineages that diverged from the southern ones ca. 4Ma, indicating long-term survival in northern microrefugia.
The southern lineages are micro-endemic and display a scattered distribution, suggesting a more ancient, relict pattern. We conclude that the contrasting diversity patterns between the disjunct distributional areas of G.leopoliensis s.l. reflect differential survival of lineages across the latitudinal gradient, offering a promising system for comparing the evolutionary ecology of lineages persisting in latitudinally disconnected microrefugia.
These results fill an important gap in the knowledge of European gammarid biogeography and reveal that all Carpathian Gammarus taxa are ancient and diverse species complexes.