Mitogenomic Phylogenetic Analysis Supports Continental-Scale Vicariance in Subterranean Thalassoid Crustaceans

Many continental subterranean water crustaceans ("stygobionts") display extreme disjunct distributions, where different species in the same genus are isolated on continents or islands separated by broad oceanic expanses. Despite their freshwater habitat, most of these taxa appear to be most closely related to typical marine groups ("thalassoid" origin). Among the hadzioids-thalassoid amphipods including the stygobiont families Hadziidae, Pseudoniphargidae, and Metacrangonyctidae-several genera are restricted to inland groundwaters ranging from the Caribbean region to the Mediterranean and Middle East, including interspersed oceanic islands. This distribution might have arisen from Tethyan vicariance triggered by the sequential occlusion of the former Tethys Sea, a vast circumtropical ocean existing from the Middle Jurassic up to 20 million years ago (mya). Previous studies have been based on morphological analyses or limited DNA sequence data, making it difficult to test this hypothesis. We used complete mitochondrial protein-coding gene sequences, mainly obtained by next-generation sequencing methods and a nuclear ribosomal gene to resolve the phylogeny and to establish a time frame for diversification of the family Metacrangonyctidae (Amphipoda). The results were consistent with the plate tectonics vicariance hypothesis, with major diversifications occurring between 96 and 83 mya.