Natural populations of numerous species have decreased sharply in recent years, and a number of mammalian species are also now at elevated risk of extinction globally. The long-tailed goral Naemorhedus caudatus, a vulnerable and protected species designated by IUCN (International Union for Conservation of Nature) and CITES (the Convention on International Trade in Endangered Species of Wild Fauna and Flora), is distributed in Northeast Asia including the Korean Peninsula. In South Korea, the Seoraksan National Park is known as the largest core habitat for the long-tailed goral population. In this study, phylogenetic relationships and population genetic features of the Seoraksan goral population were analyzed using fecal samples with both mitochondrial DNA and microsatellite markers. We found that Seoraksan gorals were the most closely related to Russian population, and also found that a unique Seoraksan lineage evolved there. In addition, the Seoraksan goral population showed higher genetic diversity than other South Korean populations, suggesting that this population might represent the most ecologically and evolutionarily important remnants of the long-tailed goral in South Korea. The Seoraksan goral population had a low level of genetic differentiation and a rather single genetic structure, suggesting that non-negligible levels of gene flow might have occurred across populations. Moreover, microsatellite genotype-based individual identification estimated that the population size was ≥81 in the Seoraksan National Park. Findings of our study suggest that effective conservation and restoration actions are needed for long-term conservation of N. caudatus in this protected area.

Biodiversity encompasses species diversity, which includes species richness and species evenness. High species diversity is known to contribute to community stability and the potential for maintaining healthy ecosystem functioning. However, the most commonly used species diversity indices have some limitations, as they require species-specific abundance data for each community. In contrast, phylogenetic diversity measures the evolutionary distances between species within a community, reflecting ecological and/or evolutionary divergences and niche differences, without requiring abundance data. This study assessed biodiversity by calculating phylogenetic diversity indices for freshwater fishes (19 species) and aquatic insects (49 species) at three sites within Seoraksan and Odaesan National Parks. The aquatic insects studied belong to the EPT group (Ephemeroptera, Plecoptera, and Trichoptera), which are widely used as bioindicators of aquatic ecosystems. Two mitochondrial DNA genes were used as molecular markers: COI and cyt b for fish, and COI and 16S rRNA for the EPT - group. Overall, Odaesan National Park exhibited higher phylogenetic diversity in both fishes and aquatic insects compared to Seoraksan National Park, although this difference was not statistically significant. The highest phylogenetic and species diversity were observed for fish at the OD2 site (Woljeong District) and for the EPT group at the OD1 (Gyebangsan) and SA2 (Jangsudae) sites. Correlation analysis revealed that phylogenetic diversity indices were more positively associated with species richness than species diversity indices. This study serves as a pilot project for establishing standardized methods for assessing biodiversity in national park habitats using phylogenetic diversity. It also aims to inform various policies, such as the development of biodiversity assessment systems and the prioritization of protected areas within national parks.

An unrecorded gall midge was discovered from inflorescence galls on Castanopsis sieboldii (Makino) Hatus. ex T.Yamaz. & Mashiba, which is one of major components in evergreen forests on the Korean Peninsula. The galls occurred on 20 trees out of 230 on Yokiji Island. The gall midge was identified as Schizomyia castanopsisae Elsayed & Tokuda, 2018 (Diptera: Cecidomyiidae), using morphological characters and mitochondrial DNA cytochrome oxidase subunit 1 (COI) region sequences of gall midge’s larvae. The Barcode sequences of 40 samples collected from Yokji Island were identical, and the individuals from Yokji Island formed a clade with the individuals from Kyushu, with robust bootstrap support in a maximum likelihood tree. This result suggests the gall midges may have migrated from Kyushu, Japan to Yokji Island, South Korea. However, it is too early to determine if the gall midge is truly invasive or not at present due to paucity of distribution data in the country.

River estuaries are dynamic and productive ecosystems with high regional biodiversity. Environmental DNA (eDNA) has become a useful approach to assessing biodiversity in aquatic ecosystems. This study was conducted to investigate fish community characteristics and species diversity in two river estuary ecosystems, the Taehwa River and Changwon Stream. We further compared conventional and eDNA metabarcoding analyses of the fish communities. The conventional survey was performed in May, July, and October 2022, while the eDNA analysis was conducted only in May. We observed various fish species with different life histories, including carp, goby, and marine fish. We also found that migratory fish, such as dace Tribolodon hakonensis, sweetfish Plecoglossus altivelis, and eel Auguilla japonica, occurred in the Taehwa River, suggesting high river connectivity. Marine fish species were predominant in the Changwon Stream, as this river is located close to the sea. The diversity indices showed that the Taehwa River generally had higher species richness, evenness, and diversity values than the Changwon Stream. A total of 9-19 species were detected in the conventional survey for the three sites, whereas 11-18 species were found from eDNA analysis. The findings indicate that the sensitivity of eDNA was similar to or higher than that of the conventional method. Our study findings suggest the efficiency and efficacy of eDNA-based fish community monitoring, although with some shortcomings in applying the genetic marker to Korean fish, including no clear-cut distinction for Korean endemic species and/or genetically closely related species groups.

The bitterling (Cyprinidae, Acheilongnathinae) is a temperate freshwater fish with a unique spawning symbiosis with host mussels. Female bitterlings use their extended ovipositors to lay eggs on the gills of mussels through the mussel's exhalant siphon. In the present study, in April of 2020, we investigated spawning frequencies and patterns of three bitterling fish species in host mussel species in the Nakdong River basin (Hoecheon). During field surveys, a total of four bitterling and three mussel species were found. We observed bitterling's spawning eggs/larvae in the three mussel species: Anodonta arcaeformis (proportion spawned: 45.5%), Corbicula fluminea (12.1%), and Nodularia douglasiae (45.2%). The number of bitterlings’ eggs/larvae per mussel ranged from 1 to 58. Using our developed genetic markers, we identified the eggs/ larvae of each bitterling species in each mussel species (except for A. macropterus): A. arcaeformis (spawned by Acheilognathus yamatsutae), C. fluminea (A. yamatsutae and Tanakia latimarginata), and N. douglasiae (A. yamatsutae, Rhodeus uyekii, and T. latimarginata). Approximately 57.6% of N. douglasiae mussel individuals had eggs/ larvae of more than one bitterling species, suggesting that interspecific competition for occupying spawning grounds is intense. This is the first report on bitterling’s spawning events in the Asian clam C. fluminea from Korea; however, it should be ascertained whether bitterling’s embryo undergoes successful development inside the small mussel and leaves as a free-swimming juvenile. In addition, the importance of its conservation as a new host mussel species for bitterling fishes needs to be studied further.

In recent years, macroalgal bloom occurs frequently in coastal oceans worldwide. It might be attributed to accelerating climate change. “Green tide” events caused by proliferation of green macroalgae (Ulva spp.) not only damage the local economy, but also harm coastal environments. These nuisance events have become common across several coastal regions of continents. In Korea, green tide incidences are readily seen throughout the year along the coastlines of Jeju Island, particularly the northeastern coast, since the 2000s. Ulva species are notorious to be difficult for morphology-based species identification due to their high degrees of phenotypic plasticity. In this study, to investigate temporal variation in Ulva community structure on Jeju Island between 2015 and 2020, chloroplast barcode tuf A gene was sequenced and phylogenetically analyzed for 152 specimens from 24 sites. We found that Ulva ohnoi and Ulva pertusa known to be originated from subtropical regions were the most predominant all year round, suggesting that these two species contributed the most to local green tides in this region. While U. pertusa was relatively stable in frequency during 2015 to 2020, U. ohnoi increased 16% in frequency in 2020 (36.84%), which might be associated with rising sea surface temperature from which U. ohnoi could benefit. Two species (Ulva flexuosa, Ulva procera) of origins of Europe should be continuously monitored. The findings of this study provide valuable information and molecular genetic data of genus Ulva occurring in southern coasts of Korea, which will help mitigate negative influences of green tide events on Korea coast.

본 연구는 동해유입하천 (강릉 연곡천, 양양 남대천), 한강수계 (섬강, 속사천), 낙동강수계 (길안천)에 서식하는 참갈겨니 (Zacco koreanus) 개체군을 대상으로 채집된 110개 체로부터 미토콘드리아 DNA COI 유전자 (mitochondrial DNA cytochrome oxidase I)를 분자마커로 이용하여 계통지리학적 분석을 수행하고, 추가적으로 강릉 연곡천 상·중· 하류 개체군을 대상으로 집단유전학적 분석을 수행하였다. 계통지리학 분석 결과, 동해유입하천과 한강수계의 참갈겨니 개체군은 동일한 단일계통을 나타내었고, 낙동강수계의 개체군은 상이한 계통으로 분기됨을 나타내었으며, 다른 수계 계통과의 유전적 거리 수치 범위가 평균 4.0% (3.7~4.2%)로서 동일종 이상 수준을 보여 잠재종 가능성을 시사하였다. 참갈겨니가 서식하는 수계에 따른 형태학적 차이는 연구된 바 있으나 DNA 염기서열의 변이를 이용한 분자유전학적 연구는 부족한 실정이므로 본 연구 결과는 향후 낙동강수계 참갈겨니 개체군의 계통분류학적 연구에 기초자료로 활용될 수 있을 것으로 판단된다. 추후 집단유전체학 및 생태학적 분석을 통하여 관찰된 낙동강수계 계통이 다른 종, 잠재종 혹은 단순히 큰 수준의 종내 변이를 나타내는지에 대한 추가적인 연구가 필요하다. 강릉 연곡천 상·중·하류에 서식하는 개체군의 집단유전학 분석을 통해 중류의 개체군이 상대적으로 높은 다양성을 나타냈으며 상·중·하류 개체군 간의 유전적 차이는 나타나지 않았다. 이는 상·중·하류 개체군 간 유전자 확산이 원활하게 이루어지고 있음을 의미하며 하천의 개체군 간 연결성을 판단할 수 있는 지표로 활용될 수 있다. 하지만 생태학적 시간 스케일의 연구에 더 적합한 분자마커를 이용한 추후 연구가 필요할 것으로 사료된다.

본 연구는 2015-2018년 5월 기간 동안 납자루아과 어류의 서식 집단 중 강원도 홍천 내촌천(HN), 덕치천(HD), 정선 골지천(JG) 및 조양강(JJ)을 대상으로 기 개발된 제한절편 길이 다형성(restriction fragment length polymorphism; RFLP) 분자마커를 이용하여 숙주조개 속 난 및 치어에 대한 정확한 종 동정을 수행 후 납자루아과 어류의 숙주조개 크기에 대한 산란양상을 파악하고자 하였다. 연구대상 지역 내 납자루아과 어류는 내촌천과 골지천에서 1종(묵납자루; Acheilognathus signifer), 덕치천에서 3종(각시붕어; Rhodeus uyekii, 묵납자루; A. signifer, 줄납자루; Acheilognathus yamatsutae), 조양강에서 2종(묵납자루, 줄납자루)으로 확인되었고, 네 지역에서 모두 동서하고 있는 숙주조개인 작은 말조개(Unio douglasiae sinuolatus) 982개체를 채집하였다. RFLP 분자마커를 이용하여 납자루아과 어류의 난 및 치어가 확인된 작은말조개(N=163; 16.6%)에서 총 646개체의 납자루아과 어류의 난 및 치어(묵납자루 454, 줄납자루 43, 각시붕어 149개체)를 확인하였다. 각 지역 숙주조개 크기에 따른 산란선호도를 조사하기 위해 난 및 치어가 확인된 숙주조개(mussels with [presence] eggs/fry)와 확인되지 않은 숙주조개(mussels without [absence] eggs/fry)의 각장 (shell length), 각고(shell height) 및 각폭(shell width)의 평균 크기를 비교하였다. 그 결과 3종의 납자루아과 어류가 동서하는 덕치천의 경우 난 및 치어가 확인된 숙주조개가 확인되지 않은 숙주조개보다 각장(1.98mm), 각고(0.85mm), 각폭(0.73mm)의 크기가 통계적으로 유의하게 크게 나타났으며(Mann-Whitney U tests, P=0.002, P=0.012, P=0.009), 다른 세 개의 지역에서도 난 및 치어가 확인된 조개의 각장, 각고, 각폭의 크기가 큰 결과를 보였으나, 통계적으로 유의하지는 않았다. 추가적으로 종 간 숙주조개 당 평균산란 난 및 치어의 수를 분석한 결과 각시붕어 9.31±5.94개, 묵납자루 2.86±2.45개, 줄납자루 2.50±1.32개로 각시붕어는 묵납자루와 줄납자루보다 숙주조개 당 평균 6.45~6.81개 더 많이 산란하였고, 통계적으로 유의한 결과를 나타내었다(Kruskal-Wallis test, P<0.001). 이 결과는 본 연구 대상 납자루아과 어류 3종에서 크기가 큰 작은말조개를 산란을 위한 숙주로서 선호함을 의미하고, 조개 크기에 대한 선호도 차이가 동서하는 납자루아과 종의 수가 많을수록 크게 나타났다. 또한, 묵납자루와 줄납자루의 경우 많은 숙주조개에 적은 양의 난을 추가적으로 고르게 산란하는 반면에 각시붕어는 비교적 적은 수의 숙주조개에 많은 양의 난을 산란하는 번식전략을 나타내었다. 2종 이상의 납자루아과 어류가 서식하는 덕치천(HD)과 조양강(JJ)에서 묵납자루와 줄납자루 2종이 동일한 조개에 산란하는 것이 관찰되었다(N=4). 이는 납자루아과 어류가 2종 이상 동서할 때, 동일한 자원인 작은말조개를 자신의 산란숙주로 이용하기 위한 종간경쟁(interspecific competition)이 일어나고 있는 것으로 판단된다. 본 연구에서는 기존 연구된 생태학적 연구에 유전학적인 방법을 추가하여 각 집단 간, 종 간 숙주조개 크기에 대한 산란양상을 보다 정확히 규명하여 숙주조개를 이용하는 납자루아과 어류의 생태적 적응양상을 명확히 파악하고 더 나아가 숙주조개와 납자루아과 어류의 공생(mutualism) 혹은 숙주-기생의 상호관계(host-parasite relationship)를 규명 하는데 기여할 수 있을 것으로 기대한다.

본 연구는 PCR 기반 RFLP (Restriction Fragment Length Polymorphism; 제한절편 길이 다형성) 분자기법을 활용하여 난 및 치어 대상 납자루아과 어류 3종의 동정을 좀 더 빠르고 정확하게 파악하고 납자루아과 어류의 종별 산란양상 및 번식생태 이해에 대한 기여가 목적이다. 본 연구를 위해 기존 선행된 문헌자료를 확인하고 납자루아과 어류가 2종 이상 동서하고 있는 지역을 확인하여 현지조사를 수행하였다. 현지조사 결과 확인된 납자루아과 어류는 묵납자루 (Acheilognathus signifer), 줄납자루 (A. yamatsutae) 및 각시붕어 (Rhodeus uyekii)로 총 3종이 확인되었으며, 확인된 납자루아과 어류와 동서하고 있는 숙주조개 (작은말조개; Unio douglasiae sinuolatus)를 채집하여 숙주조개 속 납자루아과 어류의 난 및 치어를 확보하였다. 현지조사 결과 확인된 납자루아과 어류 3종을 대상으로 미토콘드리아 DNA COI과 cyt b 유전자 염기서열을 비교하여 각각 종별로 특이성을 지닌 부위 (단일염기변이; Single Nucleotide Variation: SNV)에 맞는 제한효소를 선정하였고, 숙주조개 속 난 및 치어를 대상으로 genomic DNA를 추출하여 PCR-RFLP 실험을 수행한 결과 현지조사 시 확인된 납자루아과 어류 3종의 독특한 제한절편 길이 양상을 전기영동을 통하여 확인하였다. 본 연구를 통해 묵납자루, 줄납자루 및 각시붕어의 종을 판별할 수 있는 RFLP 마커를 개발하였으며, 숙주조개 난 및 치어를 대상으로 정확한 종의 동정을 보다 빠르고 효과적으로 수행하여 각각 납자루아과 종별 산란양상을 보다 정확히 규명하고 향후 이들 자연개체군의 효과적인 유지, 관리 및 보전 방법 개발에 유용하게 활용될 수 있을 것으로 판단된다.

The present study is to understand the Bongseonsa stream in the National Arboretum fish fauna variation through comparison with historical data and to evaluate the stream health situated. We performed investigations over three times from April to September 2015. In the survey, 2,960 individuals which belonging to 22 species, 8 families were collected. Dominant species by number was Zacco platypus and subdominant species was Zacco koreanus. Seven Korean endemic species (Squalidus gracilis majimae, Microphysogobio yaluensis, Zacco koreanus, Koreocobitis rotundicausata, etc) were observed and showed a ratio of 36.4%. The community analysis revealed that the structure of fish community in the study sampling sites was instability in having dominance 0.79 (±0.15), diverse 1.21 (±0.60), evenness 0.58 (±0.15) and species richness 1.49 (±0.83). The values in the Qualitative Habitat Evaluation Index (QHEI) was averagely 122.9 (±44.8) in the Bongseonsa stream and this was showed to have favorable habitat surroundings. As a result of tolerance guild analysis, the total number of sensitive species and intermediate species were higher than tolerant species. Analysis was divided into A and B two groups of fiducial 12.25% in Cluster analysis degree of similarity between study sampling sites. Fish Assessment Index (FAI) was rated A and B grade in Bongseonsa stream that stream health showed favorable. However Wangsuk stream as a urban stream rated C grade and analysed the lowest grade in the whole study sampling sites. There was high correlation beteween FAI and various indexes, dominance, diverse, evenness and sensitive species and intermediate species.

Brachymystax lenok tsinlingensis (family Salmonidae), cold freshwater fish, is endemic to Asia. This species is currently distributed throughout Russia, Mongolia, China and the Korean Peninsula. B. lenok tsinlingensis in South Korea was severely affected by anthropogenic activities such as habitat destruction, agricultural run-off and water pollution, and hence this fish has recently been dramatically decreased in its population sizes and become now critically endangered. To recover the number of individuals of B. lenok tsinlingensis, stocking or translocation programs have been conducted continuously by local governments since 1970s. However, these programs made little effort to clarify populations that may have originated from stocked, translocated or introduced fish. An understanding of genetic characteristics of endangered populations is critical to develop effective conservation and restoration plans especially because genetic diversity ensues their future fate. Therefore, we assessed the “conservation status” of this species by estimating the level of genetic diversity and genetic structure among ten geographic populations including restored populations via reinforcement and supplementation. Also, we aimed to trace the genetic origins of the newly translocated population (Chiak) through a restoration practice program. Moreover, we inferred the phylogenetic relationships among Korean lenok populations as well as across the Northeast Asia. Two hundred eighteen individuals of B. lenok tsinlingensis were sampled from ten localities (Yanggu, Injae, Seorak, Bangtae and Hongcheon: North Han River basin; Pyeongchang, Chiak and Jeongseon: South Han River basin; Taebaek and Bonghwa: Nakdong River basin in South Korea). Based on mitochondrial DNA (mtDNA) control region and eight nuclear microsatellite loci, we found extremely low levels of within-population genetic diversity, which suggests small effective population sizes (Ne) within populations. For mtDNA control region, each population housed one, or at most, two haplotypes that are restricted to the respective localities, meaning that these ‘genetically unique’ lineages will be lost permanently if the local populations undergo extinction. The overall values of haplotype diversity (h) and nucleotide diversity (π) for the entire Korean population were 0.703 ± 0.024 and 0.021 ± 0.010, respectively. In the case of microsatellites, average number of alleles across the eight loci for the entire population was 9.1 and allelic richness (AR) per population ranged from 2.375 to 4.144 (mean = 3.104). The values of observed heterozygosity (HO) and expected heterozygosity (HE) were similar to each other [HO: 0.400 ~ 0.590 (mean = 0.518); HE: 0.407 ~ 0.608 (mean = 0.504)]. The inbreeding coefficient (FIS) values were generally low, ranging from 0.048 to 0.279. Consequently, the majority of the populations (except Yanggu and Pyeongchang) were not significantly deviated from Hardy-Weinberg equilibrium (HWE), suggesting random mating at these loci tested. In addition, we found that Korean lenok populations were significantly genetically isolated from each other, with private mtDNA haplotypes and microsatellite alleles, indicating limited gene flow among populations, strong effects of genetic drift due to small Ne, or a combination of both. The Mantel test of microsatellites revealed a significant correlation (r = 0.414, P = 0.04) between genetic and geographic distances for pairwise comparisons among the ten populations, while that of mtDNA showed a lack of correlation. Given the shared identical mtDNA haplotype and similar microsatellite allelic distributions between Chiak and Hongcheon populations, we suggest that the restored (introduced) Chiak population would be inferred to be genetically originated from Hongcheon population. Phylogenetic relationships among Northeast Asian populations showed that South Korean lineages have more recently diverged from China (Yellow River), than between North Korea and Russia. Although the phylogenetic relationship would be expected to be associated with geography, South-North Korea and China populations with a similar latitude was more phylogenetically closely related. These findings may suggest a possible scenario for the historical movements of B. lenok tsinlingensis in Northeast Asia during Last Glacial Maximum (LGM). It would be supported by the line of evidence that most lenok populations migrated to southward from Northern Asia such as Russia and Mongolia during LGM because the Korean Peninsula was landlocked as inland epoch and functioned as a southern shelter with Yellow River. For this reason, the Korean Peninsula is suggested to be an important geographical region for better understanding phylogenetic relationships and evolutionary histories of B. lenok tsinlingensis across the Northeast Asia. Despite large efforts made to develop several restoration programs in South Korea for B. lenok tsinlingensis, it is still unknown whether these past restoration efforts were successful or fruitless, mainly because of little attention paid to post-restoration monitoring research. Hence, there was a lack of their published official records. In the future, conservation and restoration projects of the Korean lenok populations should consider the genetic data for a better understanding of their ecological and evolutionary trajectories. And finally, we hope that our findings here can help inform on the future effective conservation and restoration plans for B. lenok tsinlingensis populatio ns in South Korea.