Chromosome number, morphology and location of the nucleolus organizer region (NOR) are useful cytological characters for taxonomic and evolutionary studies. The karyotype of Upeneus moluccensis from the north-eastern Mediterranean was investigated through conventional Giemsa staining, Ag-NOR staining, C-banding and GTG-banding techniques. The results showed 2n = 44 (FN = 46), 2m + 2st + 40a chromosomes and four NORs. Two telomeric C-band regions were observed on a metacentric and subtelocentric chromosome pair. On the basis of the chromosome morphology, the GTG and C-banding pattern, it is hypothesized that Upeneus moluccensis is more primitive than the Mediterranean native fish species, Mullus barbatus and Mullus surmuletus. In this study, the cytogenetic characterization of U. moluccensis has been carried out for the first time.