[show abstract][hide abstract] ABSTRACT: Cytogenetic data in cartilaginous fishes are currently very inconsistent, considering that the karyotype morphology of only about seventy living species is actually known. Only in the last few years different molecular approaches, first of all physical mapping on metaphase chromosomes, have been used to investigate the cytotaxonomic relationship existing inside this interesting group of vertebrates. The aim of the work was to characterize new molecular chromosomal markers both to discriminate the different chromosome pairs and to distinguish the probable sex chromosomes in the species Torpedo torpedo, since its karyotype does not seem to exhibit heterochromosomes. Evolution of the SRY gene has received considerable attention, mainly because it has been shown to be the sex-determining locus in mammals. The gene is located in the Y chromosome where it normally occurs as a single copy. Using primers taken from the conserved SRY sequences, we characterized these regions at the molecular level and localized them on metaphase chromosomes. The PCR products revealed similar patterns in specimens of both sexes of T. torpedo, but only one fragment of the male amplification product showed a high percentage of identity with human spermatogenesis related genes, SPATA 16, SPATA 18 and UTY. Fluorescent in situ hybridization with these sequences showed the presence of spots at the subtelomeric level of two chromosome pairs in the male and of one pair in the female. Finally, these sequences are particularly useful as chromosome markers to differentiate between the male and the female karyotypes in this species.