Comparison of genetic diversity between wild-caught broodstock and hatchery-produced offspring populations of the vulnerable Korean kelp grouper (Epinephelus bruneus) by microsatellites.

The kelp grouper Epinephelus bruneus (Perciformes: Haemulidae), is one of the most economically important fishery resources in Korea. This fish is regarded as a target for prospective aquaculture diversification; therefore, maintenance of stock quality is important. To investigate the effects of current artificial reproduction in a hatchery facility, genetic variation in wild-caught broodstock and hatchery-produced offspring of kelp grouper was analyzed using eight polymorphic nuclear microsatellite DNA loci; 77 alleles were identified. Allelic variability ranged from 2 to 22 in the broodstock and from 1 to 10 in the offspring. The average observed and expected heterozygosities were 0.620 and 0.623 in the broodstock and 0.600 and 0.513 in the offspring, respectively. The possibility of a recent genetic bottleneck was suggested in both populations of E. bruneus. The minor, but significant, genetic differentiation (FST = 0.047, P < 0.05) observed was mainly due to statistically significant reductions in the number of alleles in the offspring compared with the broodstock, suggesting that these genetic changes could be related to genetic drift. Our results demonstrate the usefulness of microsatellite markers to monitor genetic variation and raise concerns about potential harmful genetic effects of inappropriate hatchery procedures. Therefore, genetic variation between broodstock and offspring in a hatchery should be monitored in both breeding and release programs as a routine hatchery operation, and inbreeding should ideally be controlled to improve kelp grouper hatchery management. Our data provide a useful genetic basis for future planning of sustainable culture and management of E. bruneus in fisheries.

Genetic differences between the wild and hatchery-produced populations of Korean short barbeled grunter (Hapalogenys nitens) determined with microsatellite markers.

Short barbeled grunter, Hapalogenys nitens, is an economically important fishery resource. In Korea, this fish is in the early stage of domestication, and it has been regarded as the candidate marine fish species for prospective aquaculture diversification. This study presents a preliminary investigation of the future viability of sustainable fry production from short barbeled grunter. We used 12 polymorphic nuclear microsatellite DNA loci to analyze the possible genetic variability between the wild and hatchery-produced populations of short barbeled grunter from Korea and identified 91 alleles. Compared to the wild population, significant genetic changes including reduced genetic diversity (average allele number: 7.42 vs 3.75; average expected heterozygosity: 0.713 vs 0.598, Wilcoxon signed-rank test; P < 0.05) and differentiation [overall fixation index (FST) = 0.088, P < 0.01] occurred in the hatchery-produced population, as indicated by the observation of allele richness, unique allele, heterozygosity, FST, and results of molecular analysis of variance. These findings indicate that genetic drift may have promoted the differentiation between these 2 populations, which may have negative effects on sustainable fry production. Therefore, genetic variations of the wild and hatchery-produced populations should be monitored and subjected to control inbreeding through a commercial breeding program. The information presented by this paper would provide a useful genetic basis for future sustainable culturing planning and management of H. nitens.

Genetic characterization of hatchery populations of Korean spotted sea bass (Lateolabrax maculatus) using multiplex polymerase chain reaction assays.

The spotted sea bass, Lateolabrax maculatus, is an important commercial and recreational fishery resource in Korea. Aquacultural production of this species has increased because of recent resource declines, 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. In this study, the genetic diversity and structure of three cultured populations in Korea were assessed using multiplex assays with 12 highly polymorphic microsatellite loci; 144 alleles were identified. The number of alleles per locus ranged from 6 to 28, with an average of 13.1. The mean observed and expected heterozygosities were 0.724 and 0.753, respectively. Low levels of inbreeding were detected according to the inbreeding coefficient (mean FIS = 0.003-0.073). All hatchery populations were significantly differentiated from each other (overall fixation index (FST) = 0.027, P < 0.01), and no population formed a separate cluster. Pairwise multilocus FST tests, estimates of genetic distance, mantel test, and principal component analyses did not show a consistent relationship between geographic and genetic distances. These results could reflect the exchange of breeds and eggs between hatcheries and/or genetic drift due to intensive breeding practices. For optimal resource management, the genetic variation of hatchery stocks should be monitored and inbreeding controlled within the spotted sea bass stocks that are being released every year. This genetic information will be useful for the management of both L. maculatus fisheries and the aquaculture industry.

Genetic relationships of Pacific abalone (Haliotidae) species determined using universal rice primer-polymerase chain reaction fingerprinting.

Random amplified polymorphic DNA (RAPD) with universal rice primers (URP) was used to identify species and to determine phylogenetic relationships for the 6 economically important Korean Pacific abalone species: Haliotis discus hannai, H. discus discus, H. madaka, H. gigantea, H. diversicolor supertexta, and H. diversicolor diversicolor, whose morphological differentiation is difficult. Of the 12 URPs used in this study, 7 were effective in producing reproducible RAPD markers for these 6 species. Amplifications with the 7 URP primers yielded 129 reproducible amplified fragments ranging between 100 and 6000 bp in length. The dendrogram generated by the unweighted pair-group method using arithmetic averages showed that the 6 species were divided into 4 groups at 0.44 similarity level, indicating that they were genetically distant from each other and had little internal phylogenetic resolution. One group included H. discus hannai, H. discus discus, H. madaka, and H. gigantea, which were divided into 2 groups at 0.52 similarity level: one group of H. discus hannai, H. discus discus, and H. madaka, and the other of H. gigantea. H. diversicolor supertexta and H. diversicolor diversicolor belonged to the other group. Furthermore, the reproducible pattern of amplified DNA bands by URP primers indicated the possibility of using these as molecular markers for the discrimination of the 6 Pacific abalone species. These results suggest that the URP-PCR approach will be a useful tool for obtaining accurate taxonomic identification and genetic relationship of Korean Pacific abalones, which is one of the first prerequisites in effective conservation programs.

Comparative genetic variability between broodstock and offspring populations of Korean starry flounder used for stock enhancement in a hatchery by using microsatellite DNA analyses.

Korean starry flounder, Platichthys stellatus (Pleuronectidae), is one of the most economically important fishery resources in Korea. We investigated the effect of current artificial reproduction in a hatchery facility, genetic divergence between the broodstock and their offspring populations of starry flounder in a hatchery strain to be stocked into natural sea areas was accessed using 9 polymorphic nuclear microsatellite DNA loci. High levels of polymorphism were observed between the 2 populations. A total of 96 alleles were detected at the loci, with some alleles being unique in the broodstock. Allelic variability ranged from 8 to 17 in the broodstock and from 7 to 12 in the offspring population. Average observed and expected heterozygosities were estimated at 0.565 and 0.741 in the broodstock samples and 0.629 and 0.698 in the offspring population, respectively. Although no statistically significant reductions were found in heterozygosity or allelic diversity in the offspring population, a considerable loss of rare alleles was observed in the offspring population compared with that in the broodstock. Significant genetic difference was detected between the broodstock and offspring populations (FST = 0.021, P < 0.05). These results suggest that more intensive breeding practices for stock enhancement might have resulted in a further decrease of genetic diversity. Thus, genetic variations of broodstock and progeny should ideally be monitored in both breeding and release programs as a routine hatchery operation in order to improve the starry flounder hatchery management. This information might be useful for fishery management and aquaculture industry of P. stellatus.

Comparative genetic diversity of wild and hatchery-produced populations of tongue sole (Cynoglossus semilaevis) using multiplex PCR assays with polymorphic microsatellite markers.

The tongue sole, Cynoglossus semilaevis (Cynoglossidae), is one of the most economically important fishery resources in Korea. This study presents a preliminary investigation of the future viability of the complete aquaculture of tongue sole in Korea. Specifically, possible differences in genetic variability between wild populations of tongue sole from Korea and hatchery-produced populations of tongue sole from China were assessed using multiplex assays with 12 polymorphic nuclear microsatellite DNA loci. High levels of polymorphism were observed between the 2 populations. A total of 135 different alleles were found, varying from 5-15 alleles per locus, with some alleles being unique. These findings indicate a high level of genetic variability in both the wild and hatchery-produced populations. Although a considerable loss of rare alleles was observed in hatchery samples, there were no statistically significant reductions of heterozygosity or allelic diversity in the hatchery population compared to the wild population. Moreover, the inbreeding coefficient was very low (FIS = -0.010-0.052) for both populations. However, significant genetic heterogeneity was found between the 2 populations. These findings indicate that genetic drift has likely promoted differentiation between these 2 populations, and might have negative effects on the reproductive capacity of the stock, because genetic factors are important in the production of high quality seed for complete aquaculture. Therefore, aquaculture management should incorporate basic genetic principles into existing molecular monitoring protocols. The information compiled by this study is anticipated to provide a useful genetic basis for future complete culturing plans and management of C. semilaevis in fisheries.

Mitochondrial and microsatellite DNA analyses showed comparative genetic diversity between parent and offspring populations of Korean black rockfish in a hatchery facility.

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.

Population genetic structure of wild and hatchery black rockfish Sebastes inermis in Korea, assessed using cross-species microsatellite markers.

The population structure of the black rockfish, Sebastes inermis (Sebastidae), was estimated using 10 microsatellite loci developed for S. schlegeli on samples of 174 individuals collected from three wild and three hatchery populations in Korea. Reduced genetic variation was detected in hatchery strains [overall number of alleles (N(A)) = 8.07; allelic richness (A(R)) = 7.37; observed heterozygosity (H(O)) = 0.641] compared with the wild samples (overall N(A) = 8.43; A(R) = 7.83; H(O) = 0.670), but the difference was not significant. Genetic differentiation among the populations was significant (overall F(ST) = 0.0237, P < 0.05). Pairwise F(ST) tests, neighbor-joining tree, and principal component analyses showed significant genetic heterogeneity among the hatchery strains and between wild and hatchery strains, but not among the wild populations, indicating high levels of gene flow along the southern coast of Korea, even though the black rockfish is a benthic, non-migratory marine species. Genetic differentiation among the hatchery strains could reflect genetic drift due to intensive breeding practices. Thus, in the interests of optimal resource management, genetic variation should be monitored and inbreeding controlled within stocks in commercial breeding programs. Information on genetic population structure based on cross-species microsatellite markers can aid in the proper management of S. inermis populations.