Apomorphic and plesiomorphic ITS-1 rDNA patterns in morphologically similar snails (Stylommatophora: Vallonia), with estimates of divergence time

Morphological classification of snail shells is sometimes arbitrary and misleading in correct species delimitation. This topic is investigated in four nominal species of the minute gastropods of the genus Vallonia ( V. pulchella , V. excentrica , V. costata and V. enniensis ). First, individuals are classified according to their shell morphology. Second, variation in ribosomal internal transcribed spacer 1 DNA (ITS-1 rDNA) is analysed. Using a cladistic approach, the nucleotide positions of the ITS-1 are elucidated on apomorphic and plesiomorphic character states (outgroup= Acanthinula aculeata ; Valloniidae). The molecular data yield evidence that the morphospecies V. excentrica is a paraphyletic group, comprising just a loose arrangement of individuals with similar shells. Data on fossil shells and putative divergence time are also elucidated. In the monograph on Vallonia it has been suggested that the split between the branch of V. costata and the branch of V. pulchella , V. excentrica and V. enniensis could have been in the upper Cretaceous. Based on the ITS-1 sequence alignment and the number of variable nucleotide positions, it is suggested that the last common ancestor of both branches lived at some time during the Miocene or even the Pliocene. Therefore, the fossil stem lines of Vallonia should be newly interpreted.