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Lectin is a carbohydrate-binding protein that recognizes specific cells by binding to cell-surface polysaccharides. Tumor cells generally show various glycosylation patterns, making them distinguishable from non-cancerous cells. Consequently, lectin has been suggested as a good anticancer agent. Herein, the anticancer activity of

Bacterial extracellular polymeric substances (EPS) are promising materials that have a role in enhancing growth, metabolite production, and harvesting efficiency. However, the validity of the EPS effectiveness in scale-up cultivation of microalgae is still unknown. Therefore, in order to verify whether the bacterial metabolites work in the scale-up fermentation of microalgae, we conducted a bioreactor fermentation following the addition of bacterial EPS derived from the marine bacterium, Pseudoalteromonas sp., to Euglena gracilis. Various culture strategies (i.e., batch, glucose fed-batch, and glucose and EPS fed-batch) were conducted to maximize metabolite production of E. gracilis in scale-up cultivation. Consequently, biomass and paramylon concentrations in the continuous glucose and EPS-treated culture were enhanced by 3.0-fold and 4.2-fold (36.1 ± 1.4 g L-1 and 25.6 ± 0.1 g L-1), respectively, compared to the non-treated control (12.0 ± 0.3 g L-1 and 6.1 ± 0.1 g L-1). Also, the supplementation led to the enhanced concentrations of α-tocopherols and total fatty acids by 3.7-fold and 2.8-fold, respectively. The harvesting efficiency was enhanced in EPS-supplemented cultivation due to the flocculation of E. gracilis. To the best of our knowledge, this is the first study that verifies the effect of bacterial EPS in scale-up cultivation of microalgae. Also, our results showed the highest paramylon productivity than any other previous reports. The results obtained in this study showed that the scale-up cultivation of E. gracilis using bacterial EPS has the potential to be used as a platform to guide further increases in scale and in the industrial environment. KEY POINTS: Effect of EPS on Euglena gracilis fermentation was tested in bioreactor scale. EPS supplement was effective for the paramylon, α-tocopherol, and lipid production. EPS supplement induced the flocculation of E. gracilis.

Mytilus coruscus Gould, 1861 is a mussel species in the family Mytilidae, native to the Northwest Pacific Ocean, ranging from the East China Sea, the Yellow Sea, and as far as the Peter the Great Gulf in the East Sea. In Korea, this species has been heavily exploited for nutrient-rich food resources and experienced severe reduction in their population. The aim of this study was to investigate the genetic diversity and population structure and to provide baseline data to facilitate the conservation and sustainable use of the vulnerable species M. coruscus in South Korea. Mitochondrial DNA (mtDNA) cytochrome c oxidase I (COI) sequences of 91 adult individuals from four islands and one coastal localities in South Korea were sequenced. We then compared genetic diversity and haplotype data with previously published Chinese wild populations. Mytilus coruscus populations on Korean coasts were found to exhibit high genetic diversity despite concerns regarding recent population reduction. A total of 42 haplotypes were defined by 56 polymorphic sites. High-level genetic diversity was observed on four Island sites (Hd = 0.906–0.955, π = 0.0068–0.0090). The other seashore site represented relatively lower genetic diversity (Hd = 0.529, π = 0.0011) and was genetically differentiated from the others. In a previous study, wild populations on the East China Sea exhibited similarly high genetic diversity as that observed in our study. Additionally, Chinese M. coruscus populations exhibit a distinct regional haplotype distribution pattern while sharing six haplotypes with Korean populations. The results of this study provide insights that further the current understanding regarding the evolution of M. coruscus species and provides comprehensive genetic data to facilitate the development of an effective conservation strategy.

Lectin is a carbohydrate-binding protein that recognizes specific cells by binding to cell-surface polysaccharides. Tumor cells generally show various glycosylation patterns, making them distinguishable from non-cancerous cells. Consequently, lectin has been suggested as a good anticancer agent. Herein, the anticancer activity of Bryopsis plumosa lectins (BPL1, BPL2, and BPL3) was screened and tested against lung cancer cell lines (A549, H460, and H1299). BPL2 showed high anticancer activity compared to BPL1 and BPL3. Cell viability was dependent on BPL2 concentration and incubation time. The IC50 value for lung cancer cells was 50 μg/mL after 24 h of incubation in BPL2 containing medium; however, BPL2 (50 μg/mL) showed weak toxicity in non-cancerous cells (MRC5). BPL2 affected cancer cell growth while non-cancerous cells were less affected. Further, BPL2 (20 μg/mL) inhibited cancer cell invasion and migration (rates were ˂20%). BPL2 induced the downregulation of epithelial-to-mesenchymal transition-related genes (Zeb1, vimentin, and Twist). Co-treatment with BPL2 and gefitinib (10 μg/mL and 10 μM, respectively) showed a synergistic effect compared with monotherapy. BPL2 or gefitinib monotherapy resulted in approximately 90% and 70% cell viability, respectively, with concomitant treatment showing 40% cell viability. Overall, BPL2 can be considered a good candidate for development into an anticancer agent.

Upon gene inactivation in animal models, the zebrafish (Danio rerio) has become a useful model organism for many reasons, including the fact that it is amenable to various forms of genetic manipulation. Genome editing is a type of genetic engineering in which DNA is inserted, deleted, modified, or replaced in the genome of a living organism. Mainly, CRISPR (clustered regularly interspaced short palindromic repeats) Cas9 (CRISPR-associated protein 9) is a technology that enables geneticists to edit parts of the genome. In this study, we utilized this technology to generate an mmp15b mutant by using zebrafish as an animal model. MMP15 is the membrane-type MMP (MT-MMP) which is a recently identified matrix metalloproteinase (MMP) capable of degrading all kinds of extracellular matrix proteins as well as numerous bioactive molecules. Although the newly-established mmp15b zebrafish mutant didn’t exhibit morphological phenotypes in the developing embryos, it might be further utilized to understand the role of MMP15 in liver-related diseases, such as liver fibrosis, and associated pathogeneses in humans.

Parupeneus barberinus is a tropical/subtropical reef-dwelling marine fish belonging to the family Mullidae. Herein, we report the first sequencing and assembly of the complete mitochondrial genome of P. barberinus. The complete mitochondrial genome is 16,560 bp long and has the typical vertebrate mitochondrial gene arrangement, consisting of 13 protein-coding genes, 22 tRNA genes, two rRNA genes, and a control region. Phylogenetic analysis using mitochondrial genomes of 18 species showed that P. barberinus is clustered with P. multifasciatus and P. chrysopleuron and rooted with other Mullidae species. This mitochondrial genome provides potentially important resources for addressing taxonomic issues and studying molecular evolution.

Gymnogobius heptacanthus is a small intertidal species belonging to the family Gobiidae. Herein, we report the first sequencing and assembly of the complete mitochondrial genome of G. heptacanthus. The complete mitochondrial genome is 16,529 bp long and has the typical vertebrate mitochondrial gene arrangement, consisting of 13 protein-coding genes, 22 tRNA genes, 2 rRNA genes, and a control region. Phylogenetic analysis using mitochondrial genomes of 12 species showed that G. heptacanthus is clustered with G. urotaenia and G. petschiliensis and rooted with other Gobiidae species. This mitochondrial genome provides potentially important resources for addressing taxonomic issues and studying molecular evolution.

A small and air-breathing snail, Ellobium chinense (Ellobiidae), is a vulnerable species by International Union for Conservation of Nature (IUCN). To protect and manage habitat and population of E. chinense, microsatellite markers were developed using 454 pyrosequencing and 20 polymorphic microsatellite markers were identified. A total of 146,704 sequences containing a minimum of four repeat motifs (mean, 631 base pairs) were identified from 499,505 reads. Among 80 loci containing more than nine repeat units, 34 primer sets (42.5 %) produced strong PCR products, of which 20 were polymorphic among 48 samples of E. chinense. All loci exhibited high genetic variability, with an average of 18.9 alleles per locus, and the mean observed and expected heterozygosities were 0.65 and 0.90, respectively. In addition, cross-amplification was tested for all 20 loci in the same family species, Melampus sincaporensis. None of the primer pairs resulted in effective amplification, which might be due to their high mutation rates. Our work demonstrated the utility of next-generation 454 sequencing as a method for the rapid and cost-effective identification of microsatellites. The high degree of polymorphism exhibited by the 20 newly developed microsatellites will be useful in future conservation genetic studies of this species.

Psenes arafurensis, belonging to the family Nomeidae, is described based on four specimens (184.8–199.0 mm in standard length) collected from Korea. The species is most similar to the congeneric species P. maculatus and P. pellucidus, but differs from them in the number of lateral–line scales. (44–47 in P. arafurensis vs 67–70 in P. maculatus and 120 in P. pellucidus) and the number of vertebrae (31 in P. arafurensis vs 34–38 in P. maculatus and 40–42 in P. pellucidus). The present study is a new record of P. arafurensis with voucher specimens from Korean waters.

Unfortunately, one of the co-author's family name has been incorrectly published in the original online publication. The correct family name should be Tsai.

Background The common long-arm octopus (Octopus minor) is found in mudflats of subtidal zones and faces numerous environmental challenges. The ability to adapt its morphology and behavioral repertoire to diverse environmental conditions makes the species a promising model for understanding genomic adaptation and evolution in cephalopods. Findings The final genome assembly of O. minor is 5.09 Gb, with a contig N50 size of 197 kb and longest size of 3.027 Mb, from a total of 419 Gb raw reads generated using the Pacific Biosciences RS II platform. We identified 30,010 genes; 44.43% of the genome is composed of repeat elements. The genome-wide phylogenetic tree indicated the divergence time between O. minor and Octopus bimaculoides was estimated to be 43 million years ago based on single-copy orthologous genes. In total, 178 gene families are expanded in O. minor in the 14 bilaterian species. Conclusions We found that the O. minor genome was larger than that of closely related O. bimaculoides, and this difference could be explained by enlarged introns and recently diversified transposable elements. The high-quality O. minor genome assembly provides a valuable resource for understanding octopus genome evolution and the molecular basis of adaptations to mudflats.

Stethojulis strigiventer is a tropical reef-associated marine fish belonging to the family Labridae. Herein, we report the first sequencing and assembly of the complete mitochondrial genome of S. s...

The genus Laticauda (Reptilia: Elapidae), commonly known as sea kraits, is venomous marine amphibious snakes distributed throughout the south and southeast Asian islands and mostly found in coastal waters. To facilitate genetic studies, we have developed microsatellite loci for L. semifasciata using the 454 GS-FLX pyrosequencing technique. A total of 65,680 sequences containing a minimum of five repeat motifs were identified from 451,659 reads. Among 80 loci containing more than nine repeat units, 34 primer sets (42.5%) produced strong PCR products, of which 21 were polymorphic among 36 samples of L. semifasciata. All loci exhibited high genetic variability, with an average of 7.38 alleles per locus, and the mean observed and expected heterozygosities were 0.73 and 0.76, respectively. The cross-species amplification of these loci in two laticaudine species, L. colubrina and L. laticaudata, revealed a high transferability (78.6%) and polymorphism (59.5%) of the loci. Our work demonstrated the utility of next-generation 454 sequencing as the rapid and cost-effective method for development of microsatellite markers. The high level of polymorphism in these microsatellite loci will be useful for the detection of population subdivision and the study of migration, gene flow, relatedness and philopatry of L. semifasciata and other laticaudine species.

The neritid species Clithon retropictum (Gastropoda), an oviparous snail, is distributed in freshwater and brackish water in southern Korea, south-east Japan, southern China and Taiwan. This species is listed as a class II endangered species in Korea because of its decreasing population size. It has led to an increase in the need for genetic information about this species. For this aim, in this study, we developed microsatellite markers for C. retropictum by using nextgeneration sequencing. A total of 185,972 sequences containing motifs with a minimum of five repeats motifs were identified from 3,392,119 reads. Of the 46 loci screened among 49 individuals, 36 were successfully amplified and 21 were polymorphic among 49 individuals, with 14 tri-nucleotide repeats and 7 tetra-nucleotide repeats. All loci exhibited relatively high genetic variability except one locus (CR08), with an average of 10.33 alleles per locus, and the mean observed and expected heterozygosities were 0.661 and 0.758, respectively. Our results demonstrated the utility of next-generation sequencing as a method for the rapid and cost-effective identification of microsatellites. These 21 newly developed microsatellite markers will be informative tools for investigating the genetic structure and diversity among populations of this endangered species and will help facilitate effective strategies for its conservation.

The seven-band grouper Epinephelus septemfasciatus is of considerable economic value and a candidate species for aquaculture in East Asia. Genetic characterization of hatchery populations is needed to maintain their genetic diversity and to develop more effective aquaculture practices. We analyzed the genetic structure of four cultured populations of E. septemfasciatus in Korea using 13 microsatellite markers. The number of alleles per locus ranged from 5 to 16, with an average of 9.6. The mean observed and expected heterozygosities were 0.699 and 0.679, respectively. Pairwise multilocus FST tests suggested the existence of two populations, although the isolation-by-distance model was rejected. An analysis of molecular variance (AMOVA) also revealed two genetic clusters. These results could reflect current aquacultural practices, with breeds and eggs exchanged between hatcheries, and/or genetic drift due to intensive breeding practices. Therefore, for the optimal management of resources, the genetic variation of hatchery stocks should be monitored and inbreeding controlled for the seven-band grouper stocks. This genetic information will be useful for the management of E. septemfasciatus fisheries and the aquaculture industry.

The seven-band grouper (Epinephelus septemfasciatus) is an important fishery resource of a target for prospective aquaculture diversification and maintenance of stock quality is thus important. To explore the sustainability of fry production, genetic variations in 83 seven-band groupers from two broodstock and offspring populations of a hatchery strain were analyzed using 13 polymorphic nuclear microsatellite DNA loci; 133 alleles were identified. Allelic variability ranged from 4 to 18 in the broodstock and from 3 to 11 in the offspring. The average observed and expected heterozygosities were 0.669 and 0.734 in broodstock and 0.674 and 0.649 in offspring, respectively. Although no statistically significant reductions in heterozygosity or allelic diversity were evident in offspring, considerable loss of rare alleles was apparent. The broodstock and offspring populations exhibited significant genetic differences (F ST = 0.033, P

The common squid, Todarodes pacificus, is an important fisheries species, but fluctuations in its abundance and its reproductive biology are poorly understood. To enable sustainable management of this resource, 21 novel polymorphic microsatellite loci were isolated using an enrichment method based on the magnetic/biotin capture of microsatellite sequences from a size-selected genomic library. To characterize each locus, 48 individuals from a natural T. pacificus population from the western coastal waters of Korea were genotyped. All of the microsatellite loci were highly polymorphic with 10–32 alleles per locus. The observed and expected heterozygosities ranged from 0.417 to 0.917 and 0.791 to 0.965, respectively. The high variability revealed in this study suggests that the microsatellite markers could be a useful tool for future conservation genetics studies and for the sustainable exploitation of this resource.

The Pacific oyster, Crassostrea gigas, is the main oyster species cultivated in Korea. Despite a long history of hatchery practices, little is known about the population genetics of native populations of C. gigas in Korea. This study examined the population structure and genetic diversity of 297 individuals of C. gigas in six populations, representing two major geographically separated habitats along the coast of Korea, using multiplex assays with nine highly polymorphic loci. All populations showed very high genetic diversity, with a deficit of heterozygotes and a recent bottleneck. No significant differences were found in the genetic diversity of the two geographically separated oyster groups, although significant, but minimal, genetic differentiation was observed (FST = 0.003, p = 0.002). This was mainly due to genetic differences between the oyster populations from Taean and Gaduk, suggesting that the oyster is not panmictic. This result was corroborated by an analysis of molecular variance (AMOVA) and multidimensional scaling analysis. It was inconsistent with a previous finding of no population differentiation using mitochondrial sequence analysis, implying rather recent separation of the two geographic populations. This genetic differentiation might have resulted from the combined of various factors, such as historical dispersal, the local environment, and ocean currents. Nonetheless, these results mean that the two geographical groups should be treated as independent units for proper resource management. Our results provide critical information on the ecological and economic potential of the Korean Pacific oyster.

Large amounts of the manila clam, Ruditapes philippinarum, have been imported from Dalian (China) recently to meet the increasing demand in Korea. To ensure the quality and safety of the domestic marine product, it is necessary to be able to distinguish between domestic manila clams and clams that are imported from Dalian, China. We developed expressed sequence tags (ESTs)-derived single nucleotide polymorphism markers using 454 pyrosequencing. A total of 780,000 ESTs were assembled, and 49,540 putative SNPs were identified from 46,405 contigs. Twenty-one polymorphic SNPs from 11 primers were finally selected as diagnostic markers. Genotype analysis showed that 21 SNP markers had the opposite alleles (transitions or transversions) between domestic and imported samples, which were useful for distinguishing clam origins. The GENECLASS 2.0 program was used to estimate the ability of the markers in the discrimination between domestic and imported populations. The 21 SNP markers had 98.96% ability to discriminate domestic manila clams and 100% ability to identify those imported. These markers could be useful for discriminating between domestic and imported manila clams and can contribute to the prevention of falsified labeling of this species.

The tongue sole, Cynoglossus semilaevis, is an important fishery resource in Korea. About 100 tongue sole sampled from three major habitats along the western coast of Korea were assessed using multiplex assays with 12 highly polymorphic microsatellite loci to explore the population genetic structure of the species; 151 alleles and similar high levels of gene diversity (mean number of alleles (NA) = 10.42, mean expected heterozygosity (He) = 0.78) were detected. Three populations showed significant Hardy–Weinberg equilibrium deviations at four loci. Although a significant difference in the number of unique alleles was observed among populations, genetic population subdivision was low by F-statistics (overall F ST = 0.007, p

Short barbeled grunter, Hapalogenys nitens, is an important fishery resource and it has been regarded as a target marine fish species for prospective aquaculture diversification in Korea, but sustainable fry production is not yet possible. This delay has led to an increase in the need for genetic information about this species. Furthermore, it is necessary to conserve the wild stocks for the sustainable use of the species. In this study, we report 26 novel polymorphic microsatellites with a mean of 4.9 alleles per locus. The mean observed and expected heterozygosity per locus was 0.45 and 0.50, respectively. Cross-amplification revealed variability in others two Haemulidae species. These markers developed can be a valuable tool for conservation genetics and further genetic improvement of these species.

Two species of mitten crab, Eriocheir sinensis and Eriocheir japonica, are commercially valuable in Korea. Millions of their juveniles have been released in attempts to restore ecological habitat and enhance fishery resources. Due to their habitat's singularity, exact species identification should be conducted before juveniles are released. In this study, we used a highly conserved region of the mitochondrial cytochrome b (cyt b) gene to distinguish E. sinensis from E. japonica. We designed a primer set to amplify part of the mitochondrial cyt b gene of both species and obtained 1024 base pairs (bp) of polymerase chain reaction (PCR) product. These PCR products were digested with the restriction enzyme MboII and the species-specific restriction fragment length polymorphism (RFLP) patterns were analyzed. Also, we designed primers for the rapid identification of cyt b gene (498 bp) to distinguish E. sinensis. PCR-RFLP and PCR using new primers are a fast and inexpensive method to access species identification. This simple and rapid method can provide useful information for E. sinensis and E. japonica species identification.

In the present report, we describe the first sequencing and assembly of the complete mitochondrial genome of Cheilio inermis. The mitochondrial genome of C. inermis, with 16,494 bp in length, has the typical vertebrate mitochondrial gene arrangement. It contains 13 protein-coding genes, 22 tRNA genes, two rRNA genes, and a control region. All the tRNA genes typically formed a cloverleaf secondary structure. Phylogenetic analysis using mitochondrial genomes of 11 species showed that C. inermis formed monophyletic group with other Labridae species.

Population genetics has been recognized as a key component of policy development for fisheries and conservation management. In this study, natural sea bass (Lateolabrax japonicus) populations in three ocean basins in Korea were assessed using multiplex assays with 12 highly polymorphic microsatellite loci; 203 alleles and similarly high levels of genetic diversity [mean number of alleles (NA) = 14.43, mean expected heterozygosity (He) = 0.84] were detected. All populations showed significant heterozygote deficiency at four loci, which could be explained by the presence of null alleles. The genetic population subdivision was low and was significantly different according to F-statistics (overall F ST = 0.003, R ST = 0.005). However, this substructure was not supported by an analysis of molecular variance test, analyses of isolation by distance or Bayesian analysis. The passive dispersal of eggs/larvae via the main currents appears to facilitate gene flow. The possibility of a recent genetic bottleneck was observed in all three populations of L. japonicus, indicating that overfishing and degradation of the environment in recent years has led to a decline in the sea bass populations in Korea. Our study demonstrates that sea bass in Korea do not appear to be genetically partitioned and should be managed as a single unit; however, the potential for a rapid loss of genetic diversity remains. Information regarding the genetic characteristics of Korean sea bass populations has important implications for fishery management and conservation efforts and will aid in the sustainable exploitation of fishing resources and the preservation of biodiversity.

The threadsail filefish Stephanolepis cirrhifer is one of the most important commercial fish resources in Korea. However, its natural populations have declined drastically over the last several decades. To investigate the genetic characteristics of the threadsail filefish for conservation and management purposes, its genetic variation was analyzed in Korean fishing grounds using 11 microsatellite loci. All populations showed moderate genetic diversity (mean number of alleles (NA) = 8.6, expected heterozygosity (He) = 0.73), representing a slightly lower level of genetic variability than other congeneric species. The presence of a weak genetic population substructure was detected (FST = 0.023, RST = 0.030, P

Red sea cucumber Stichopus japonicus is the most important and valuable commercial sea cucumber species in Korea. Its farming and stock enhancement started in the early 2000s and is still rapid expansion in Korea. Therefore, the analyses of genetic status of wild and hatchery populations are necessary to maintain the genetic diversity of this valuable marine resource. In this study, possible genetic similarity and differences between the wild population and hatchery population in Jeju, Korea were accessed using multiplex assays with eight highly polymorphic microsatellite loci. High levels of polymorphism were observed between the two populations. A total of 93 different alleles were found. Although a considerable loss of unique alleles and relatively high inbreeding coefficient value were observed in the hatchery samples, no statistically significant reductions were found in heterozygosity or allelic diversity in the hatchery population, compared with the wild population. However, significant genetic heterogeneity was found between two populations. These results suggest that genetic drift has probably promoted differentiation between populations, and stocking intensity in wild populations may correlate with loss of genetic integrity. Therefore, the sustainable exploitation plans of the fishery resource should be developed by applying basic genetic principles combined with molecular monitoring. This genetic baseline information of Korean red sea cucumber has important implications for designing of genetically sustainable restocking programs and more efficient conservation management guidelines for these valuable genetic materials.

The spotted sea bass, Lateolabrax maculatus, is popular in recreational fishing and aquaculture in Korea. Its natural population has declined during the past two decades; thus, beginning in the early 2000s stock-enhancement programs were introduced throughout western and southern coastal areas. In this study, genetic similarities and differences between wild and hatchery populations were assessed using multiplex assays with 12 highly polymorphic microsatellite loci; 96 alleles were identified. Although many unique alleles were lost in the hatchery samples, no significant reductions were found in heterozygosity or allelic diversity in the hatchery compared to the wild population. High genetic diversity (He = 0.724–0.761 and Ho = 0.723–0.743), low inbreeding coefficient (FIS = 0.003–0.024) and Hardy–Weinberg equilibrium were observed in both wild and hatchery populations. However, the genetic heterogeneity between the populations was significant. Therefore, genetic drift likely promoted inter-population differentiation, and rapid loss of genetic diversity remains possible. Regarding conservation, genetic variation should be monitored and inbreeding controlled in a commercial breeding program.

The Pacific abalone Haliotis diversicolor supertexta, of the family Haliotidae, is one of the most important shellfish species in eastern Asia. In Korea, it is intensively caught in Jeju Island, where it is highly appreciated as seafood. However, its natural population has declined drastically in the past several decades. To investigate its genetic characteristics for conservation and management purposes, the genetic variation in four populations from Jeju Island, Korea was analyzed using 20 microsatellite loci. All populations showed very high genetic diversity and a deficit of heterozygotes. The minor but significant genetic differentiations (both FST and RST) observed were mainly due to genetic differences between populations from Pyoson and other localities, suggesting that abalone may not be panmictic. The Pyoson abalone population showed a significant difference in allelic richness that could be related to harvesting intensity and stock enhancement activity. This result was also corroborated by an analysis of molecular variance (AMOVA). Overall, this report presents novel observations of the wild genetic population structure in H. diversicolor supertexta from Korea using microsatellite markers and provides critical information for supporting conservation strategies for this species, namely that the Pyoson abalone population should have a higher conservation priority due to its unique genetic characteristics.

The black rockfish, Sebastes inermis (Sebastidae), is an important commercial fishery resource in Korea. As a preliminary investigation into the effect of artificial reproduction in a hatchery facility, the genetic divergence between parent and offspring populations of black rockfish was accessed using 10 polymorphic nuclear microsatellite DNA loci and a mitochondrial (mt) control gene. All loci that were screened showed marked polymorphisms. mtDNA control region sequences were also highly variable. Of approximately 350 base pairs (bp) sequenced, 52 variable sites, comprising 56 base substitutions, were found among 233 individuals. Offspring populations showed less genetic variability than the parent population in terms of numbers of microsatellite alleles and mtDNA haplotypes, as well as mtDNA haplotype diversity. Statistical analysis of the fixation index (ΦST and F(ST)) and analysis of molecular variance using both DNA markers showed significant genetic differences between the parent and offspring populations. These results suggest that random genetic drift and/or inbreeding events, as well as artificial selection and founder effects, occurred when the offspring strain was reproduced in a hatchery facility despite thousands of males and females from different hatcheries being maintained for artificial reproduction. Therefore, it is necessary to improve current hatchery programs by monitoring genetic variation in both the broodstock and progeny and controlling inbreeding within stocks in commercial breeding facilities to maintain the production of high-quality black rockfish. This information will be useful for determining suitable guidelines for establishing and maintaining cultured stocks and the aquaculture industry of S. inermis.

The Korean pen shell Atrina pectinata is a commercially valuable species in Korea. The commercial catch of this fish has decreased continuously since 1990. However, its genetic characteristics have never been studied. In order to explore the population genetic structure of this species, 125 pen shells sampled from three major habitats along the western coast of Korea were genotyped at 21 microsatellite loci. Relatively high levels of genetic variability (mean number of allelic richness (AR) = 14.74; mean hetrerozygosity (He) = 0.849) was found among localities. However, despite the potential of the Korean pen shell to exhibit genetic structure (adults are sedentary), none of the genetic tests applied in this study detected significant genetic differentiation among the samples. The lack of genetic differentiation among samples may be due to high levels of larval dispersal, through passive drift with ocean currents. Alternatively, populations may have diverged too recently for significant genetic differentiation to have become evident. Furthermore, small sample sizes and the limited number of samples may have hampered the detection of genetic structure. The results of this study therefore suggested a lack of genetic structure among the populations of pen shell of Korean waters and should be managed as a single unit. This information on the genetic characteristics of Korean pen shell populations has important implications for the sustainable exploitation of the fishing resources and the preservation of biodiversity.

Eleven species of catfish (Siluriformes) found in Korean freshwater are economically important resources and include five endemic species. However, there are no studies on phylogenetic analysis of all catfish species in Korea at a molecular level. The species-level analysis of catfish species is usually carried out through morphological characters and controversial due to phenotypic variation. In this study, the partial sequences of 16S rRNA and cytochrome b mitochondrial genes were analyzed for species identification and phylogenetic relationships among 11 species of catfish from 10 different rivers in Korea. The nucleotide sequences of 16S rRNA and cytochrome b consisted of 587 and 441 nucleotide base pairs, respectively. Sequence analysis of both genes revealed that the 11 species fell into three distinct groups, which were genetically distinct from each other and exhibited identical phylogenetic resolution. Sequence divergences between congeneric species averaged 1.78% and 7.39% for 16S rRNA and cytochrome b, respectively. The phylogenetic relationships forming well-differentiated clades in the NJ, ML and BI trees were identical for both fragments. This research demonstrates that partial sequences of both the genes can efficiently identify the 11 species of catfish in Korea, indicating the usefulness of mtDNA-based approach in species identification.

The Korean littleneck clam, Ruditapes philippinarum, is a valuable and intensively exploited species in Korea. Attention to genetics can enhance strategies for using fishery resources sustainably, and a potentially important application is the use of molecular markers to assess the genetic stock structure of a harvested species. In this study, seven microsatellite loci were analyzed in five littleneck clam samples from the western and southern coasts of Korea to investigate the genetic characteristics of these clams. Very high genetic diversity was present in all five of the populations (overall He = 0.813), but no significant differences among the localities were detected (Wilcoxon signed-rank test, P > 0.05). The results showed a significant global FST (0.045), indicating genetic heterogeneity among the populations. We found three genetically distinct regions: a western coastal region (FST range 0.0137–0.0218, P > 0.005) and two regions along the southern coast of Korea (FST range 0.0473–0.1121, P < 0.005). Neighbor-joining, principle coordinate analysis, and Bayesian clustering revealed the existence of these three groups. This result was also corroborated by an analysis of molecular variance (AMOVA) and an assignment test. This report presents novel observations of the wild genetic population structure in littleneck clams along the coast of Korea and provides critical information to support conservation strategies for this species. The three groups mentioned above could be considered independent management units, and the southern groups would have a higher conservation priority due to their unique genetic characteristics.

Mytilus coruscus (family Mytilidae) is one of the most important marine shellfish species in Korea. During the past few decades, this species has become endangered due to the loss of habitats and overfishing. Despite this species’ importance, information on its genetic background is scarce. In this study, we developed microsatellite markers for M. coruscus using next-generation sequencing. A total of 263,900 raw reads were obtained from a quarter-plate run on the 454 GS-FLX titanium platform, and 176,327 unique sequences were generated with an average length of 381 bp; 2569 (1.45%) sequences contained a minimum of five di- to tetra-nucleotide repeat motifs. Of the 51 loci screened, 46 were amplified successfully, and 22 were polymorphic among 30 individuals, with seven of trinucleotide repeats and three of tetranucleotide repeats. All loci exhibited high genetic variability, with an average of 17.32 alleles per locus, and the mean observed and expected heterozygosities were 0.67 and 0.90, respectively. In addition, cross-amplification was tested for all 22 loci in another congener species, M. galloprovincialis. None of the primer pairs resulted in effective amplification, which might be due to their high mutation rates. Our work demonstrated the utility of next-generation 454 sequencing as a method for the rapid and cost-effective identification of microsatellites. The high degree of polymorphism exhibited by the 22 newly developed microsatellites will be useful in future conservation genetic studies of this species.

Eighteen new polymorphic microsatellite markers were developed for the Korean mi-iuy croaker (Miichthys miiuy, Perciformes, Sciaenidae), and allelic variability was compared between a wild population in Mokpo, Korea, and a hatchery population in Tongyeong, Korea. All loci were amplified readily and demonstrated allelic variability, with the number of alleles ranging from 5 to 37 in the wild population, and from 4 to 12 in the farmed population. The average observed and expected heterozygosities were estimated, respectively, to be 0.74 and 0.78 in the hatchery population samples, and 0.79 and 0.86 in the wild samples. These results indicate lower genetic variability in the hatchery population compared with the wild population, and significant genetic differentiation between the wild population and the hatchery samples (F ST=0.058, P < 0.001). These microsatellite loci may be valuable for future population genetic studies, monitoring changes in the genetic variation within stocks in a commercial breeding pr...

The Korean rockfish, Sebastes schlegeli, is a valuable and intensively exploited species in Korea. We discuss the genetic diversity and genetic structure of four Korean rockfish populations using eight microsatellite loci. In total, 161 different alleles from 138 individuals were observed. Average allele number per locus ranged from 2.5 to 23 and allelic richness varied from 13.38 to 14.63 within a population. Despite a long history of stocking practices, we found very high levels of polymorphism (mean heterozygosity = 0.810), which is comparable to other congeneric species. No significant difference in genetic diversity and molecular genetic variance (FST and RST) was observed among four local samples (P > 0.1). Little indication of contemporary inbreeding (FIS= 0.051) or population structure (K = 1) was detected. This absence of differentiation may reflect high levels of gene flow along the coast of Korea. Our study demonstrates that rockfish in Korea should be managed as a single unit. Currently, the species does not appear to be genetically threatened, but the potential for a rapid loss of genetic diversity remains. This information on the genetic characteristics of Korean rockfish populations has important implications for fisheries management and conservation efforts, and will aid in the sustainable exploitation of the fishing resources and the preservation of biodiversity.

Kelp grouper (Epinephelus bruneus Bloch 1793) is a commercially important fish in Korea. In recent years, the catch of kelp grouper in the coastal waters of Korea has significantly declined. Despite its importance, little is known about its genetic diversity and conservation efforts are hampered. In this study, we isolated and characterized 12 microsatellite loci using an enrichment method based on magnetic/biotin capture of microsatellite sequences from a size-selected genomic library. All loci were readily amplified and contained TG/CA denucleotide repeats. To characterize each locus, 30 individuals from a natural E. bruneus population in the coastal waters of Jeju Island, Korea, were genotyped. All loci except three, KEm118, KEm154, and KEm219, were polymorphic, with an average of 8.1 alleles per locus (range 2–18). The mean observed and expected heterozygosities were 0.47 (range 0.19–1.00) and 0.61 (range 0.29–0.92), respectively. A significant deviation from Hardy-Weinberg equilibrium was observed at...

Populations of the Pacific abalone, Haliotis discus hannai, have been severely overexploited over the past few decades in Korea. Information regarding the levels of genetic variability and structure within populations is insufficient for the development of effective strategies for conservation of genetic diversity of this species. To assess the genetic status of this species, we examined variation at six microsatellite loci in 842 individuals of Pacific abalone collected from three hatchery stocks of the main aquaculture areas and eight wild populations, which were two from the East Sea, two from the West Sea and three from the South Sea. High levels of polymorphism at these microsatellite loci were found in both the wild and hatchery populations. The genetic variation in the hatchery stocks [overall number of alleles (N(A)) = 24.00; allelic richness (A(R)) = 19.71; observed heterozygosity (H(O)) = 0.733] was similar to that of the wild (overall N(A) = 28.13; A(R) = 22.62; H(O) = 0.775) populations. Low levels of inbreeding and significant Hardy-Weinberg equilibrium deviations were detected in both the wild and hatchery populations. Significant F(ST) values were observed for the hatchery stocks and in most cases between the wild and hatchery populations (overall F(ST) = 0.017, P < 0.01); however, only a minor portion of the genetic diversity was distributed between the wild and hatchery populations. These results reflect intensive seedling and stocking practices. This preliminary study showed genetic separation between the eastern and pooled western and southern wild populations in Korea, which was based on the F(ST) value, phylogenetic tree clustering, PCA and MDS analyses, structure analysis, and AMOVA. This strong biogeographic structure of H. d. hannai in Korea may be considered to be independent management units. This study demonstrates the feasibility of microsatellite analyses for the monitoring of genetic diversity and for revealing the population structure of the wild Pacific abalone. This information will be useful for the proper management and conservation of H. d. hannai in Korea.

Parupeneus barberinus is a tropical/subtropical reef-dwelling marine fish belonging to the family Mullidae. Herein, we report the first sequencing and assembly of the complete mitochondrial genome ...

The threadsail filefish Stephanolepis cirrhifer is a highly commercial fisheries resource in Korea that suffers intensive anthropogenic pressure across much of its range. For basic information about its current genetic status in relation to stock enhancement, the level and distribution of genetic variation between a wild and a hatchery-bred population were investigated using 10 microsatellite markers developed for Thamnaconus modestus. High levels of polymorphism were observed between the two populations. A total of 95 different alleles were found at all loci, with some alleles being unique. The allelic variability ranged from six to 13 in the wild population and from five to 13 in the hatchery one. The average observed and expected heterozygosities were estimated to be 0.72 and 0.80 in the wild sample and 0.70 and 0.79 in the hatchery one, respectively. These results showed similar genetic variability in the hatchery population, as compared with the wild population and significant genetic differentiation between the wild population and the hatchery samples (F ST = 0.016, P < 0.05). Genetic drift in the intensive breeding practices for stock enhancement has probably promoted differentiation between populations. Significant deviations from Hardy-Weinberg equilibrium were detected in both populations. Our results indicate that further studies using species-specific microsatellite markers will be necessary for a more reliable assessment of genetic diversity of the species.

Black scraper is a commercially important oceanodromous fish in Korea. The commercial catch of this fish has decreased continuously since 1990. However, its genetic characteristics have never been studied. For population genetic analysis to assist effective aquaculture and stock management efforts, we isolated and characterized 19 microsatellite loci using an microsatellite-enrichment method based on magnetic/biotin capture of microsatellite sequences from a size-selected genomic library. To characterize each locus, 30 individuals from a natural T. modestus population in southern Korea were genotyped. All loci except two, KTm213 and KTm284, were polymorphic, with an average of 15.18 alleles per locus (range, 2–41). The mean observed and expected heterozygosities were 0.67 (range, 0.33–0.97) and 0.79 (range, 0.40–1.00), respectively. A significant deviation from Hardy-Weinberg equilibrium was observed at six loci (KTm142, KTm145, KTm158, KTm24, KTm221, and KTm250). This high variability indicates that these microsatellites may be useful for high-resolution studies of population genetics.

Pen shell (Atrina pectinata) is a popular food source with a high commercial value in a number of Asian Pacific areas. The natural A. pectinata population has been declining continuously over the past several decades. Microsatellite DNA markers are a useful DNA-based tool for monitoring the genetic variation of pen shell populations. In this study, 20 polymorphic microsatellite (MS) DNA markers were identified from a partial genomic pen shell DNA library enriched in CA repeats, and used to compare allelic variation between wild and hatchery pen shell populations in Korea. A total of 438 alleles were detected at the 20 MS loci in the two populations. All loci were easily amplified and demonstrated allelic variability, with the number of alleles ranging from 5 to 35 in the wild population and from 5 to 22 in the farmed population. The average observed and expected heterozygosities were 0.69 and 0.82, respectively, in the hatchery samples and 0.69 and 0.83, respectively, in the wild samples. Statistical analysis of fixation index (F(ST)) and analysis of molecular variance (AMOVA) showed minor, but significant, genetic differences between the wild and hatchery populations (F(ST) = 0.0106, CI(95%) = 0.003-0.017). These microsatellite loci may be valuable for future aquaculture and population genetic studies for developing conservation and management plans. Further studies with additional pen shell samples are needed to conclusively determine the genetic diversity between the wild and hatchery populations.

The Pacific abalone, Haliotis discus hannai, is a popular food in Eastern Asia. Aquacultural production of this species has increased because of recent resource declines, the growing consumption, and ongoing government-operated stock release programs. Therefore, the genetic characterization of hatchery populations is necessary to maintain the genetic diversity of this species and to develop more effective aquaculture practices. We analyzed the genetic structures of five cultured populations in Korea using six microsatellite markers. The number of alleles per locus ranged from 15 to 64, with an average of 23.5. The mean observed and expected heterozygosities were 0.797 and 0.904, respectively. The inbreeding coefficient F(IS) ranged from 0.054 to 0.184 (mean F(IS) = 0.121 ± 0.056). The genetic differentiation across all populations was low but significant (overall F(ST) = 0.009, P0.01). Pairwise multilocus F(ST) tests, estimates of genetic distance, and phylogenetic and principal component analyses did not show a consistent relationship between geographic and genetic distances. These results could reflect extensive aquaculture, the exchange of breeds and eggs between hatcheries and/or genetic drift due to intensive breeding practices. Thus, for optimal resource management, the genetic variation of hatchery stocks should be monitored and inbreeding controlled within the abalone stocks that are being released every year. This genetic information will be useful for the management of both H. discus hannai fisheries and the aquaculture industry.

In this study, we developed 20 polymorphic microsatellite markers for the Korean black scraper, Thamnaconus modestus (Günther, 1877), Monacanthidae, and used them to compare allelic variation between wild and hatchery populations in Korea. All loci were readily amplified and demonstrated allelic variability, with the number of alleles ranging from 5–35 in the wild population and 5–22 in the farmed population. The average observed and expected heterozygosities were estimated, respectively, as 0.74 and 0.80 in the hatchery samples and 0.78 and 0.81 in the wild ones. These results indicate lower genetic variability in the hatchery population than in the wild population and minor, but significant, genetic differentiation between the two populations (FST = 0.005, P < 0.01). Additionally, cross-amplification was tested in another monacanthid species, Stephanolepis cirrhifer; many loci were found that yielded useful information. The high degree of polymorphism exhibited by the 20 microsatellites will be useful in future aquaculture and population genetic studies for developing conservation and management plans.

Seven-band grouper, Epinephelus septemfasciatus, is of considerable economic value and a candidate species of aquaculture in Eastern Asia. It is necessary to monitor the stocks for the sustainable use and aquaculture of the species. Monitoring genetic variation among marine resources necessitates the development of genetic markers. Here, we report 22 polymorphic microsatellite markers with 2–16 alleles per locus. The mean observed and expected heterozygosity per locus was 0.53 and 0.57, respectively. Cross-amplification revealed variability in Epinephelus bruneus and Epinephelus akaara. The 22 markers developed can be a valuable tool for genetic conservation of this species.

Pacific abalone Haliotis discus discus is an important fisheries resource in Jeju, Korea. For basic information about its current genetic status in relation to stock enhancement, the level and distribution of genetic variation between wild and released stocks of Pacific abalone in Jeju were examined at nine cross-species microsatellite markers including the use of two novel primers. High levels of polymorphism were observed between the two populations. A total of 146 different alleles were found at all loci, with some alleles being unique. The allelic variability ranged from five to 27 in the wild population and from four to 16 in the released sample. The average observed and expected heterozygosities were estimated to be 0.74 and 0.84 in the wild sample and 0.70 and 0.78 in the released sample, respectively. Although a considerable loss of rare alleles was observed in the released sample, no statistically significant reductions were found in heterozygosity or allelic diversity in the released sa...

To estimate the effect of selective breeding on the improvement of growth in olive flounder Paralichthys olivaceus, we compared the growth of a strain selected for rapid growth to that of an unselected strain from a commercial hatchery. The fish strains were fed with either moist pellets (MP) (dry matter, 59.8% crude protein; 14.1% lipid) or extruded pellets (EP) (dry matter, 50.4% crude protein; 13.8% lipid) for 190 days and were reared under similar conditions. The mortality rates were less than 2% and were not significantly different among the experimental groups. The growth rate of the selected fish was significantly greater than that of the unselected fish regardless of the diet type, and both strains fed MP grew significantly faster than those fed EP. The selected fish consumed more feed than the unselected fish. However, there was no significant difference in the feed efficiency between the selected and unselected fish. These results demonstrate that the selected fish exhibited superior growth rates, and that this was associated with a greater intake of food. Thus, selective breeding may be useful for improving the growth of commercial olive flounder.

Seven new microsatellite markers were developed for the Pacific abalone (Haliotis discus hannai, Haliotidae), and allelic variability was compared between a wild population and a hatchery population in Yeosu, Korea. All loci amplified readily and demonstrated allelic variability, with the number of alleles ranging from 6 to 15 in the wild population and from 3 to 12 in farmed populations. Average observed and expected heterozygosities were estimated at 0.65 and 0.77 in the hatchery samples, and 0.79 and 0.87 in the wild samples. These results indicated lower genetic variability in the hatchery population, as compared with the wild population and significant genetic differentiation between the wild population and the hatchery samples (F ST=0.055, p