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

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 successful in eight loci of Kareius bicoloratus and Pleuronectes yokohamae, but in only seven loci of Microstomus achne, Pleuronectes punctatissimus, and Pleuronectes schrenki. Three to five loci could be amplified in six other species. In the three species selected, K. bicoloratus, P. punctatissimus, and P. yokohamae, seven to eight cross-amplified loci exhibited polymorphisms comprising 1-22 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 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.