Hyuk Je Lee

Exploring the Impacts of Anthropogenic Disturbance on Seawater and Sediment Microbial Communities in Korean Coastal Waters Using Metagenomics Analysis

The coastal ecosystems are considered as one of the most dynamic and vulnerable environments under various anthropogenic developments and the effects of climate change. Variations in the composition and diversity of microbial communities may be a good indicator for determining whether the marine ecosystems are affected by complex forcing stressors. DNA sequence-based metagenomics has recently emerged as a promising tool for analyzing the structure and diversity of microbial communities based on environmental DNA (eDNA). However, few studies have so far been performed using this approach to assess the impacts of human activities on the microbial communities in marine systems. In this study, using metagenomic DNA sequencing (16S ribosomal RNA gene), we analyzed and compared seawater and sediment communities between sand mining and control (natural) sites in southern coastal waters of Korea to assess whether anthropogenic activities have significantly affected the microbial communities. The sand mining sites harbored considerably lower levels of microbial diversities in the surface seawater community during spring compared with control sites. Moreover, the sand mining areas had distinct microbial taxonomic group compositions, particularly during spring season. The microbial groups detected solely in the sediment load/dredging areas (e.g., Marinobacter, Alcanivorax, Novosphingobium) are known to be involved in degradation of toxic chemicals such as hydrocarbon, oil, and aromatic compounds, and they also contain potential pathogens. This study highlights the versatility of metagenomics in monitoring and diagnosing the impacts of human disturbance on the environmental health of marine ecosystems from eDNA.

Genetic diversity and genetic structure of the endangered Manchurian trout, Brachymystax lenok tsinlingensis, at its southern range margin: conservation implications for future restoration

The Manchurian trout, Brachymystax lenok tsinlingensis (family: Salmonidae), is a cold freshwater fish endemic to Northeast Asia. South Korean populations of this species, which comprise its southern range limit, have recently decreased markedly in size and are now becoming critically endangered. We assessed the current population status of this species in South Korea by estimating the levels of genetic diversity and genetic structure of five natural and four restored populations using mitochondrial DNA (mtDNA) control region sequences and eight nuclear microsatellite loci. Levels of within-population genetic diversity were low, suggesting that past effective population sizes (Ne) have been small. Each population had one or a maximum of two mtDNA haplotypes. Microsatellite allelic richness (AR) was significantly higher for natural populations (mean AR = 3.51; 95% confidence interval, 3.00–4.03) than for restored populations (mean AR = 2.61; 2.38–2.98). South Korean populations were significantly genetically isolated from one another, with private mtDNA haplotypes and microsatellite alleles, suggesting that limited gene flow has been occurring among populations. A mtDNA phylogeny revealed that South Korean lineages were more closely related to those of China than to those of North Korea and Russia. Overall, we suggest that future restoration efforts aimed at South Korean populations should consider the genetic characteristics reported here, which should help to fulfil effective conservation strategies for this highly cherished species. Our results will inform other conservation efforts, including assisted migration of freshwater fish populations at the equatorial end of the geographical range limit of the species.

Identification of Single Nucleotide Polymorphism Markers in the Laccase Gene of Shiitake Mushrooms (Lentinula edodes)

Abstract We identified single nucleotide polymorphism (SNP) markers in the laccase gene to establish a line-diagnostic system for shiitake mushrooms. A total of 89 fungal isolates representing four lines, including Korean registered, Korean wild type, Chinese, and Japanese lines, were analyzed. The results suggest that SNP markers in the laccase gene can be useful for line typing in shiitake mushrooms.

Effects of intensity and seasonal timing of disturbances on a rocky intertidal benthic community on the southern coast of Korea

The effects of intensity and timing of disturbances on recovery of marine benthic organisms were investigated on a rocky intertidal shore in Gwangyang Bay, Korea. We hypothesized that the recovery pattern of the benthic community structure would be affected by disturbance intensity and season. Twenty-eight permanent plots were set up, with disturbance intensity (cleared plots and sterile plots) and seasonal disturbance (fall 1999 and spring 2000) incorporated into the experimental design. To monitor natural seasonal variation in benthic community abundances, we established seven permanent unmanipulated plots. Turf-forming algae were observed in the unmanipulated plots throughout the experimental period, whereas green algae and invertebrate presence varied with season. In the disturbance-intensity experiment, turf-forming and green algae were dominant in cleared plots. The highest coverage of sessile organisms was observed in sterile plots, which exhibited the highest species richness because of their relatively low macroalgal coverage. Seasonal effects of disturbance were an important factor in the recovery pattern of benthic organisms under high disturbance intensity. Coverage of green algae was higher in sterile spring plots than in sterile fall plots; this result was attributed to low spatial competition, as the disturbances occurred just before green algal blooms. On the other hand, the abundances of barnacles and bivalves were highest on sterile fall plots, as these organisms were suppressed by green algal blooms in other periods. These results indicate that the effects of disturbance intensity on benthic community recovery patterns can be influenced by season of disturbance.

Stipe Length as an Indicator of Reproductive Maturity in the Kelp Ecklonia cava

We conducted testing to determine whether stipe length is a useful indicator of reproductive maturity in the kelp Ecklonia cava, a species that plays a pivotal role in ecosystem functioning and services in subtidal areas. Approximately 100 sporophytes with stipes of various lengths were collected during the fertile period (July–November, 2013). We investigated the relationships between stipe length and other morphological characteristics to determine whether stipe length could indicate the age of reproductive maturity. Primary blade length, longest blade length, thallus height, and total length showed significant relationships with stipe length. The length of the primary and longest blades gradually declined as stipe length increased above 125 mm. Zoosporangial sori were found on the blades of more than 70% of individuals with stipes longer than 125 mm, but on only 8% of individuals with stipes less than 125 mm long. Stipe length therefore seems to be an acceptable proxy for reproductive maturity. Another factor to consider, however, is that all specimens with zoosporangial sori, regardless of stipe length, had a dry weight of 80 g or more; thus, individual biomass may also be an important parameter influencing the initiation of reproduction.

Contrasting life histories contribute to divergent patterns of genetic diversity and population connectivity in freshwater sculpin fishes

BackgroundLife history characteristics are considered important factors influencing the evolutionary processes of natural populations, including the patterns of population genetic structure of a species. The sister species Cottus hangiongensis and C. koreanus are small bottom-dwelling freshwater sculpin fishes from South Korea that display marked life history divergence but are morphologically nearly indistinguishable. Cottus hangiongensis evolved an ‘amphidromous’ life history with a post-hatching pelagic larval phase. They spawn many small eggs in the low reaches of rivers, and hatched larvae migrate to the sea before returning to grow to maturity in the river mouth. In contrast, C. koreanus evolved a ‘fluvial’ landlocked type with benthic larvae. They release a smaller number of larger eggs, and the larvae undergo direct development, remaining benthic in the upstream rivers throughout their entire lives. We tested whether there were differences in patterns and levels of within-population genetic diversities and spatial population structure between the two closely related Korean sculpins using mitochondrial DNA control region sequences and seven nuclear microsatellite loci.ResultsThe combined analyses of both marker sets revealed that C. hangiongensis harboured considerably higher levels of within-population genetic diversities (e.g. haplotype/allelic richness, heterozygosities) than C. koreanus. In contrast, the fluvial sculpin exhibited noticeably more spatial population structure than did the amphidromous sculpin, as suggested by pairwise FST statistics. The finding that C. hangiongensis individuals comprised a single random mating population across the east-flowing river basins in the Korean Peninsula, whereas C. koreanus individuals comprised genetically discrete individual populations, was further supported by an individual-based Bayesian population assignment and also factorial correspondence analyses.ConclusionsThe higher genetic diversity, but lower population structure, of the amphidromous sculpin relative to the fluvial sculpin may have resulted from its greater larval dispersal and also possibly, higher fecundity accompanied by an amphidromous life history. Hence, we conclude that contrasting early life histories – including the presence or absence of the pelagic larval phase – may have led to divergent patterns of within-population genetic diversities and spatial population structure between the sister Cottus species following speciation from a common ancestor of marine sculpin.

Development of twelve novel microsatellite markers in the medicinal mushroom Cordyceps militaris (Ascomycota: Clavicipitaceae) for its strain identification and genetic analysis

Cordyceps militaris (a caterpillar fungus), which belongs to the class Ascomycetes, has extensively been used for medicinal purposes in East Asia. Here, we isolated and characterized 12 microsatellite loci from the medicinal mushroom, C. militaris. Twenty‐nine individual samples were taken from a single locality in southwestern Korea and used to characterize the developed markers. The number of alleles of these loci ranged from two to 13 (mean allelic richness = 6.44). Observed heterozygosity and expected heterozygosity in the population ranged from 0.035 to 0.880 and from 0.035 to 0.886, respectively. Five of 12 loci significantly deviated from Hardy–Weinberg Equilibrium, most likely due to heterozygote deficiency caused by inbreeding. Tests of genotypic linkage disequilibrium between the 12 loci showed no significant association of alleles. These microsatellite markers will provide valuable tools for genetic analyses for the strain identification of C. militaris as well as for its resource conservation.

Genetic structure of Korean populations of bumblebees Bombus ignitus (Hymenoptera: Apidae) as revealed by microsatellite markers

Bumblebee Bombus ignitus, which is indigenous to Korea, Japan, and China, has been recognized as a valuable pollinator for both crops and wild plants. Bombus ignitus has now become commercially important as a pollinator because of its use in the agricultural industry, particularly for greenhouse pollination. For long‐term management and effective conservation of B. ignitus, an understanding of the genetic structure of its naturally occurring populations is practically important. In this study, the genetic structure among the five populations of B. ignitus in South Korea was assessed using nine microsatellite loci. These markers showed high levels of genetic variability, with the number of alleles ranging from 6 to 22 (mean = 13.4) and the frequency of the most common allele ranging from 0.11 to 0.66. Only the Sabuk (SB) population showed a genetic signature of a recent bottleneck, which was further supported by the lowest level of allelic richness (AR) (mean AR = 3.944). Genetic differentiation was highly significant among all pairs of populations (P < 0.001) across the nine microsatellite markers, suggesting a lack of gene flow among those populations. Interestingly, FST (and RST and Dest) values were always greater for the Taebaek population than for the four remaining populations. The phylogenetic analysis showed evidence supporting our hypothesis that the Taebaek population is genetically more divergent than the other populations. Overall, our results suggest that the Korean populations of B. ignitus might have undergone geographic isolation and have become highly separated spatially from one another, thereby having a limited range of migration among their local habitats.