The karyotype of Pseudophoxinus antalyae was analyzed using G-banding, C-banding, Ag-NOR staining, and Q-banding. Chromosome analysis of this species, which lives in the Berdan River and Berdan Dam Lake, was performed with the air-drying technique, using gill and fin epithelium cells. The diploid chromosome number of P antalyae was determined to be 2n = 50, comprising 16 metacentric, 14 submetacentric, 12 subtelocentric, and 8 acrocentric (NF = 92) chromosomes. The largest chromosome pair of the complements was characteristically a submetacentric. The nucleolar organizer regions (NORs) were apparently located in subtelocentric chromosome pairs. The karyotype of P. antalyae is reported for the first time.