Study of Kutum (Rutilus frisii kutum) population using mtDNA with PCR

Study of Mahisefid population using mtDNA with PCR Mahisefid diversity was studied in four rivers including Lamir, Sefidrood, Shirrood and Tajan. Total sample was collected from four rivers when fish migrat to river for spawning, 100 samples from Sefidrood, 98 samples from Lamir, 48 samples from Shirrood and 48 samples from Tajan. DNA was extracted with phenolcholorophorm. Samples were used for RFLP, the PCR product were digested by 20 restriction enzymes as follow: TasI, HaeIII, HinfI, HincII, SalI, DraI, AccI, AvaII, XhaI, BshNI, AvaI, BclI, BshII, MspI, PstI, RsaI, SdnI, TaqI, TruI, VspI. The four restriction enzymes including: TasI, HaeIII, HinfI, HincII showed diversity, 6 enzyme didn t have any restriction and 14 enzyme showed monomorphic. Total 20 haplotype studied that haplotype AAAA and BAAA had most frequency. The average haplotype frequency of AAAA was 29.93% and the average haplotype frequency of BAAA was 27.55%. 2- Study of population of Mahisefid with microsatellite markers 120 specimens of R.frissi kutum were used from four rivers to test thirty primers (30 samples from each river) of which 8 primers showed polymorphism. A large variation in heterozygosity average over all samples was observed among loci, that ranged from 0.13 to 0.91. For a given locus, observed heterozygosity varied greatly among the samples. For example, in Lamir 0.07 at SPY5 and 1 in Sefid Rud at CA1. Tajan had an observed heterozygosity of only 0.17, whilst the Lamir had an observed heterozygosity of 0.07 at locus SPY5. At locus SPY4, Tajan and Sefid Rud are the same and (0.53) and Shir Rud is 0.83 and Lamir is 0.47. However, despite these differences, there was clear difference in average heterozygosity observed between the samples. To investigated of Hardy-Weinberg Equilibrium in all locus and all rivers deviate showed from Hardy-Weinberg Equilibrium was significantly different ( P<0.01)except in SPY6 locus. The maximum average Ho was 0.91 in locus CA1 and minimum was 0.13 in locus SYP5 and maximum average of He was 0.89 in locus SYP4 and minimum was 0.28 in locus SYP6. Highest Ho was 1 in locus CA1 in Sefid Rud River and lowest was 0.07 in locus SYP5 in Lamir River , highest He was 0.92 in locus SYP4 in Shir Rud River and lowest was 0.24 in locus SYP6 in Tajan River. According to Nei (1978) the highest genetic distance was between Shir Rud River and Tajan River (0.07) and lowest genetic distance was between Sefid Rud River and Tajan River (0.05). The highest genetic similarity was between Sefid Rud River and Tajan River (0.95) and the lowest genetic similarity was between Shir Rud River and Tajan River (0.93). FST was significantly different (P > 0.05) between all samples except Lamir River and Sefid Rud River. The highest Fis was in Lamir River (0.84) in locus SPY5 and in Sefid Rud River in locus SPY5 (0.71), in locus CA12 (0.67) in Shir Rud River and SPY5 (0.63) in Tajan River. The lowest Fis is in locus CA1 (-0.55) in Sefid Rud River, in locus CA1 (-0.47) in Tajan River, in locus CA1 (-0.28) in Lamir River and in locus SPY6 (0.16) in Shir Rud River.

Iranian Fisheries Science Research Institute

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