INTRODUCING GENETIC MARKERS TO IDENTIFY AND DISTINGUISH FIVE SPECIES OF CYPRINIDAE IN THE CASPIAN SEA USING PCR-RFLP

Cyprinids are the main and most significant bony fishes found in the Caspian Sea. In this study, 15 specimens of Rutilus frisii kutum (Kamensky, 1901), 10 specimens of Barbus capita (Gueldenstaedtii, 1772), 10 specimens of Bream Abramis brama (Linnaeus, 1785), 10 specimens of Redlip, Aspius aspius, and 15 specimens of Kora volba, Rutilus ruti/us caspius, were collected from commercial catch stations. DNA from all specimens was extracted using the phenol-chloroform method in 500lll tubes and amplified using PCR method with a pair of primers with cytochrom b gene sequence of Kora volba. For RFLP analysis, PCR products of cytochorome gene b (1117 bp) from each species were digested with five restriction enzymes under suitable conditions of incubation. DNA bands were visualized by gel electrophoresis (polyacrylamide) and staining with silver nitrate. Five enzyme Rsa 1, Hinf I, Hhal, Hinc II, and Mbo I showed polymorphism. Genotypes obtained from digestion of enzymes Rsa I, Hinf I, Hha I showed haplotypes AAAAA for Barbus capita, BBBB for R. rutilus caspius, CCCC for Bream Abramis brama, DDDD for Aspius aspius and EEEE for R. frisii kutum. Each of these haplotypes serves as a genetic marker for the species and is of significant importance in distinguishing them.