Identification of endangered Mediterranean cyprinodontiform fish by means of DNA inter-simple sequence repeats (ISSRs)

We used inter-simple sequence repeat (ISSR) technique to obtain species-specific molecular markers for the cyprinodontiform fish Valencia hispanica, Valencia letourneuxi and Aphanius fasciatus. The aims were (i) to assess the effectiveness of ISSRs in discriminating the three species and (ii) to identify tissues of two unidentified fish suspected to belong to one of the three above species by comparing ISSR genotypes. Nine ISSR primers produced 101 scorable polymorphic loci with a large number of species-specific bands. UPGMA cluster analysis and nonmetric multidimensional scaling of inter-individual genetic dissimilarities were consistent in separating the three species and attributing the unidentified individuals to V. hispanica. This result was corroborated by the assignment test. ISSRs represent an attractive tool for specimen and species identification. We recommend their use, coupled with appropriate statistical analyses, in species identification when only small portions of the organisms are available.