Development of microsatellite markers for a hard-shelled mussel, Mytilus coruscus, and cross-species transfer

Abstract

ABSTRACT. The Korean mussel Mytilus coruscus, an endemic marine bivalve mollusk, is economically important. Its population is currently decreasing due to overexploitation and invasion of a more competitive species, Mytilus galloprovincialis. In this study, microsatellite markers for M. coruscus were developed using a cost-effective pyrosequencing technique. Among the 33,859 dinucleotide microsatellite sequences identified, 176 loci that contained more than 8 CA, CT, or AT repeats were selected for primer synthesis. Sixty-four (36.4%) primer sets were produced from the 100-to 200-bp polymerase chain reaction products obtained from 2 M. coruscus individuals. Twenty of these were chosen to amplify DNA from 82 M. coruscus individuals, and 18 polymorphic loci and 2 monomorphic loci were selected as microsatellite markers. The number of alleles and the allele richness of the polymorphic loci ranged from 2 to 22 and from 2.0 to 19.7 with means of 10.8 and 10.1, respectively. Null alleles were detected for all but three loci, which resulted in an observed heterozygosity lower than the expected heterozygosity and therefore an excess of homozygotes. In a cross-species transfer analysis of these markers using 7 Mytilidae species, the locus Mc65 was amplified from all species tested and was found to be polymorphic in all of them. Among the species, M. galloprovincialis, Lithophaga curta, and Hormomya mutabilis showed the same transferability of 25%, but the five amplified loci were polymorphic only in M. galloprovincialis and H. mutabilis. These microsatellite markers may be useful for future resource management and artificial production of juveniles for aquaculture.

Population Genetic Analysis of the Wild Hard-shelled Mussel, Mytilus Unguiculatus (Valenciennes 1858) in South Korea Using a Microsatellite Multiplex Assay

“…Sixteen microsatellite markers available in previous studies (Xu et al 2010;An et al 2013;Kang et al 2013a;Fu et al 2018) tested in this study were characterized across the seven M. unguiculatus populations and summarized in Table S1. All of them successfully produced PCR products for all samples (size ranging from 64 to 376 bp) of the seven populations with no evidence of scoring errors or large allelic dropout indicated by Micro-Checker v.2.2.3.…”

“…In this study, we conducted comprehensive genetic diversity and structure analyses of seven M. unguiculatus populations along the coastal areas of the mainland and islands of South Korea with two sets of a multiplex PCR assay (consisting of eight polymorphic microsatellite markers each) from previous studies (Xu et al 2010;Kang et al 2013a;Guo et al 2013;Fu et al 2018). The findings of this study will provide the crucial basis for developing effective conservation and management strategies for this economically and ecologically important species in South Korea.…”

Population genetic analysis of the wild hard-shelled mussel, Mytilus unguiculatus (Valenciennes 1858) in South Korea using a microsatellite multiplex assay

“…In this study, we conducted comprehensive genetic diversity and structure analyses of seven M. unguiculatus populations along the coastal areas of the mainland and islands of South Korea with two sets of multiplex PCR assays (consisting of eight polymorphic microsatellite markers each) from previous studies (Xu et al 2010; An and Lee 2012; Kang et al 2013a;Guo et al 2013;Fu et al 2018). The ndings of this study will provide the crucial basis for developing effective conservation and management strategies for this economically and ecologically important species in South Korea.…”

“…A total of 16 primer pairs developed for M. unguiculatus(Xu et al 2010; An and Lee 2012;Kang et al 2013a;Guo et al 2013;Fu et al 2018) were used in this study (TableS1). Two sets of multiplex PCR assays (Set A and Set B) were carried out in a 20µl volume using AccuPower PCR PreMix (Bioneer Inc., Daejeon, South Korea) including 1 µl gDNA and 1 µl primer pairs (the forward primer was uorescently labeled with FAM, HEX, TAMRA, and ATTO).…”

A molecular and population genetics study of the Bluff Oyster (Ostrea chilensis)

“…Cross-species transferability has had variable levels of success (Zhang and Hewitt, 2003; reviewed by Barbara et al 2007). In bivalves, Kang et al (2013) successfully used microsatellite loci in a range of mussel species, although the levels of success varied depending on the species and the loci tested. There are many other examples of successful cross amplification in other bivalves including for example: lampsilines (Galbraith et al 2011) and scallops (Marín et al 2012).…”