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DEVELOPMENT OF EST-SSR MARKERS FOR ROCKFISH SEBASTES SCHLEGELII AND CROSS-SPECIES AMPLIFICATION
Author(s): 
Pages: 1096-1102
Year: Issue:  5
Journal: Oceanologia et Limnologia Sinica

Keyword:  Sebastes schlegeliiESTmicrosatellite markerstransferability;
Abstract: We designed primers with published expressed sequence tag(EST) sequences in combination with PCR amplification, polymorphic EST-SSR(simple sequence repeats) markers in rockfish Sebastes schlegelii. The results show that the discovery rate of EST-SSR on EST sequences was 9.14%; dinucleotide motif had the highest frequency at 33.03%; and TG/CA motif was the richest one. In total, primers, 46 pairs out of 56 pairs that designed proved effective, and 18 pairs expressed polymorphism. We then applied these loci to analyze 30 S. schlegelii DNA specimens in population diversity. The observed number of alleles(Na) ranged 2—9 in average of 3.89. The observed heterozygosity(Ho), expected heterozygosity(He) and their average values were 0.0333—0.8000, 0.0333—0.7927, 0.3037 and 0.3757, respectively. Polymorphic information content(PIC) ranged 0.0323—0.7522, five loci were highly polymorphic, and 4 were moderately polymorphic. After Bonferroni correction, 3 loci deviated significantly from Hardy-Weinberg Equilibrium(P-value), but no linkage disequilibrium phenomenon. The 18 loci were surveyed in S. koreanus and Sebastiscus marmoratus, the cross-species proportions and polymorphic rates were 88.89%, 100.00% and 33.33%, 88.89%, respectively. Therefore, these EST-SSR loci of S. schlegelii may facilitate studies on genetic diversity, phylogeny, and genomes mapping of related species.
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