Integrative cytogenetic and molecular analyses in Hypostomus sertanejo reveal the lowest diploid number in the genus (Siluriformes: Loricariidae)

Abstract Expanding integrative cytogenetic and molecular analyses is essential for understanding the evolutionary dynamics of chromosome number variation in fishes. This study presents the first cytogenetic characterization of Hypostomus sertanejo , an endemic species native to the semi-arid Jaguaribe River basin in Northeastern Brazil. Cytogenetic analysis revealed a diploid number of 2n = 52, which is the lowest documented for the genus, with a karyotype composed of 22m + 8sm + 16st + 6a and a fundamental number of 98. This value aligns with the ancestral diploid number suggested for the tribe Hypostomini. Fluorescence in situ hybridization identified a single 18S rDNA site coinciding with the nucleolar organizer region, while 5S rDNA was localized to an interstitial position on a unique chromosome pair. Molecular analysis based on COI gene confirmed the identity of H. sertanejo and its distinction from closely related species. These results reveal a distinctive evolutionary trajectory for H. sertanejo relative to other Hypostomus species and highlight the importance of integrating data to elucidate diversification patterns in endemic fish lineages.