This paper describes the karyotype analysis of Garra rufa samples with G-, C- banding and Ag-NOR staining techniques from four distinct localities (Mersin, Hatay, Kahramanmaras and Sivas) in Turkey. The diploid chromosomes number was found as 50 in females of both Mersin and Sivas regions samples. The karyotype was determined as 26M + 10SM + 8ST + 6A chromosome and fundamental number (NF) 94 in Mersin and 28M + 14SM + 4ST + 4A chromosome (NF=96) in Sivas regions samples. Diploid chromosomes number both Hatay and Kahramanmaras regions male and female samples were described as 46. The karyotype of Hatay region female samples was included 22M + 12SM + 8ST + 4A (NF=88) chromosome and male 22M + 12SM + 7ST + 5A chromosome (NF=87). The karyotype of Kahramanmaras region female samples was included 32M + 6SM + 6ST + 2A chromosomes (NF=90) and male 31M + 6SM + 6ST + 3A chromosomes (NF=89). The male presented a pair of differentiated heteromorphic chromosomes, characterizing as XX/XY sex chromosome system of Kahramanmaras and Hatay region samples. The comparative analysis revealed intraspecific variability of NORs and fixed differences in their number in the four populations. The results are compared with data available for populations from native geographic ranges.