The creation of genetic basic population of rainbow trout (Onchorhynchus mykiss) based on study of genetic variation in brood stocks using microsatellite

In order to perform the project, 446 samples of rainbow trout (Onchorhynchus mykiss) from 24 different regions in Iran were collected. About 2-3 g of caudal fin samples was collected from each specimen and preserved in absolute ethyl alcohol and then transferred to the genetic laboratory. Genomic DNA was extracted using the phenol-chloroform method and then DNA content and quality was determined using spectrophotometry and agarose gel electrophoresis, respectively. Polymerase Chain Reaction (PCR) of genomic DNA fin samples was carried out using 10 pairs of microsatellite primers. All PCR products were electrophoresed on 6% polyacrylamide gel and stained with silver nitrate. Following the scoring of alleles, all parameters including allelic frequency, effective number of allele, observed and expected heterozygosity, shanon index, measurement of similarity and genetic distance and Hardy-Weinberg equilibrium, Fst , Rst and gene flow were calculated using AMOVA analysis in the GenAlex and Popgene programs. The results showed that 8 pairs of microsatellite primers were polymorphic. In total, 50 alleles were determined with the range size of 64-280 bp. The locus omyf had maximum number of allele (26) and loci OTSG 474 and Strurruta58 had minimum number of allele (5). The observed heterozygosity was between 0.86 and 0.964. Hardy-Weinberg departure was observed for all loci from farms 18, 15, 4, E20 and 21 and were disequilibrium (P<0.05). The farms 14, 8, 7 and 6 were equilibrium at 3 loci, but showed disequilibrium in other loci. The other farms were equilibrium at 1 or 2 loci and disequilibrium at 8 or 9 loci. The FST results showed that maximum FST (0.24) were between farms 1 and 11in which had minimum of gene flow (3.7). Minimum FST (0.04) were between farms 8 and 9 in which had maximum of gene flow (346). Based on the results of AMOVA analysis, significant differences were detected between all farms (P<0.01). Furthermore, based on Nei 's standard (1972) maximum genetic distance (0.89) were observed between farms 2 and 11 and maximum genetic similarity (0.15) were detected between farms 3 and 4. This result suggests that the unique genetic variation of rainbow trout in hatchery farms of Iran represents a highly valuable genetic resource and provide useful information for creating a based population in the future breeding programs.