Abstract:In this study, the karyotype and Ag-NORs of wild Hemibarbus labeo from Nandujiang River of Hainan were analyzed using the methods of intraperitoneal injection of phytohemagglutinin (PHA) and colchicine, and direct preparation of head kidney cells. Results showed that H. labeo had a diploid chromosome number of 50, the karyotype formula was 2n = 18m + 12sm + 12st + 8t, the number of chromosome arms (NF) was 80, no heterotypic chromosome was observed, and Ag-NORs were located on the ends of the short arms of pair No.5 and pair No.11. Comprehensive analyses of the chromosomal evolution in subfamily Gobioninae indicated that H. labeo had the primitive karyotype of Gobioninae fish. Our study will not only supplement the cytogenetic data of H. labeo and provide information on the taxonomy and evolution of the subfamily Gobioninae, genus Hemibarbus, but will also lay the theoretical foundation for germplasm protection and germplasm improvement of H. labeo.