Cross-species amplification of microsatellite markers for the brown sole in the family Pleuronectidae

The amplification of eight microsatellite loci previously developed and characterized in brown sole Pleuronectes herzensteini was attempted in 11 other flatfish species. Two loci Phz6 and Phz12 were amplified in all the species examined. Cross-species amplification was succeeded in eight loci of Kareius bicoloratus and Pleuronectes yokohamae, but in seven loci of Microstomus achne, Pleuronectes punctatissimus and Pleuronectes schrenki. Five to three loci could be amplified in other six species. In the three species selected, K. bicoloratus, P. punctatissimus and P. yokohamae, cross-amplified seven to eight loci exhibited polymorphisms comprising one to 22 alleles. Expected heterozygosity (He) ranged from 0.66 to 0.96 in K. bicoloratus, from 0.62 to 0.96 in P. punctatissimus and from 0.43 to 0.91 in P. yokohamae. Observed heterozygosity (Ho) ranged from 0.65 to 1.00 in K. bicoloratus, from 0.55 to 0.95 in P. punctatissimus and from 0.40 to 0.95 in P. yokohamae. The Phz2, Phz3 and Phz12 loci significantly deviated in certain or all the three species from Hardy-Weinberg equilibrium. The mean values of homology to the flanking-region sequences of brown sole were 93.7% in K. bicoloratus, 91.2% in P. punctatissimus, and 93.9% in P. yokohamae. These results suggest that microsatellite markers for brown sole are applicable for genetic studies in flatfish species including at least these three species