eDNA (environmental DNA)란 특정 환경에 서식하는 생물로부터 유래한 DNA를 의미한다. 환경 시료로부터 추출한 eDNA를 활용하면 해당 환경에 서식하는 생물들의 효율적이고 정확한 모 니터링을 수행할 수 있다. 해수 시료로부터 얻은 eDNA를 기반으로 해양생물 다양성 연구를 수행할 수 있다. 해수 시료를 채집하고 이로부터 eDNA를 추출한 뒤, metagenome 분석을 통 해 서식하는 해양생물의 종 동정과 다양성 분석이 가능하다. 본 리뷰에서는 이처럼 해수의 eDNA를 활용하여 해양 지역의 생물 다양성 연구를 수행하는 전체적인 과정을 제시하고 있다. 아직 국내에는 해양생물 다양성 연구를 위해 eDNA를 적용하는 방법이 보편화 되어있지 않으 며, 본 리뷰를 기반으로 이와 같은 eDNA 연구 방법을 정립하는데 도움을 줄 수 있을 것이다.

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.

Freshwater jellyfish, a type of jellyfish exclusively found in freshwater, has a limited number of species but is found globally. However, their ecology and causes of occurrence are largely unknown. Therefore, understanding the distribution of polyps, which produce the larvae of freshwater jellyfish, can provide important data for comprehending their ecology. This study aims to explore the COI gene of freshwater jellyfish using environmental DNA from the microbial film in the Miho River system. Among the 12 survey points in the Miho River watershed, genetic material of freshwater jellyfish was detected in 8 points, mainly located upstream near reservoirs. These genetic materials were identified as genes of the well-known freshwater jellyfish species, Craspedacusta sowerbii. Notably, the C. sowerbii genes found in the Miho River watershed survey points were closely related to a species previously discovered in Italy. Consequently, utilizing environmental DNA to explore the genetic traces of freshwater jellyfish enables rapid screening of areas with a high likelihood of freshwater jellyfish occurrence. This approach is deemed to provide crucial information for understanding the distribution and ecology of freshwater jellyfish in Korea.

Environmental DNA (eDNA) can exist in both intracellular and extracellular forms in natural ecosystems. When targeting harmful cyanobacteria, extracellular eDNA indicates the presence of traces of cyanobacteria, while intracellular eDNA indicates the potential for cyanobacteria to occur. However, identifying the “actual” potential for harmful cyanobacteria to occur is difficult using the existing sediment eDNA analysis method, which uses silica beads and cannot distinguish between these two forms of eDNA. This study analyzes the applicability of a density gradient centrifugation method (Ludox method) that can selectively analyze intracellular eDNA in sediment to overcome the limitations of conventional sediment eDNA analysis. PCR was used to amplify the extracted eDNA based on the two different methods, and the relative amount of gene amplification was compared using electrophoresis and Image J application. While the conventional bead beating method uses sediment as it is to extract eDNA, it is unknown whether the mic gene amplified from eDNA exists in the cyanobacterial cell or only outside of the cell. However, since the Ludox method concentrates the intracellular eDNA of the sediment through filtration and density gradient, only the mic gene present in the cyanobacteria cells could be amplified. Furthermore, the bead beating method can analyze up to 1 g of sediment at a time, whereas the Ludox method can analyze 5 g to 30 g at a time. This gram of sediments makes it possible to search for even a small amount of mic gene that cannot be searched by conventional bead beating method. In this study, the Ludox method secured sufficient intracellular gene concentration and clearly distinguished intracellular and extracellular eDNA, enabling more accurate and detailed potential analysis. By using the Ludox method for environmental RNA expression and next-generation sequencing (NGS) of harmful cyanobacteria in the sediment, it will be possible to analyze the potential more realistically.

Targeting Microcystin (MC), which is most abundantly detected in the North-Han River water area, we analyzed the relationship between the MC biosynthesis gene (mcyA gene), cyanobacteria cell density, and MC concentration, derived an RNA-MC conversion formula, and derived the cyanobacteria. The concentration of MC present in cells was predicted. In the North-Han River waters, the mcyA gene was found mainly at downstream sites of the North-Han River after Muk-Hyeon Stream junction, and higher copy numbers were found on average than other sites. In the Uiam Lake waters upstream of the North-Han River, the mcyA gene copy number increased at the Kong-Ji Stream point, and after September, the mcyA gene copy number decreased throughout the North-Han River waters. The expression of the mcyA gene was concentrated in the short period of summer due to the spatio-temporal difference between upstream and downstream water bodies. The mcyA gene expression level was not only highly correlated with MC concentration, but also correlated with the cell density of Microcystis aeruginosa and Dolichospermum circinale, which are known to biosynthesize MC. Six conversion formulas derived based on the RNA-MC relationship showed statistical significance (p<0.05) and exhibited high correlation coefficients (r) of 0.9 or higher. The expression level of MC biosynthesis gene present in eRNA determines the synthesis of cyanotoxin substances in water, quickly quantifies gene activity, and can be fully utilized for early warning of MC development.

Recently, with the development of genetic technology, interest in environmental DNA (eDNA) to study biodiversity according to molecular biological approaches is increasing. Environmental DNA has many advantages over traditional research methods for biological communities distributed in the environment but highly depends on the established base sequence database. This study conducted a comprehensive analysis of the habitat status and classification at the genus level, which is mainly used in eDNA (12S rRNA, 16S rRNA, 18S rRNA, COI, and CYTB), focusing on Korean registration taxon groups (phytoplankton, zooplankton, macroinvertebrates, and fish). As a result, phytoplankton and zooplankton showed the highest taxa proportion in 18S rRNA, and macroinvertebrates observed the highest ratio in the nucleotide sequence database in COI. In fish, all genes except 18S rRNA showed a high taxon ratio. Based on the Korean registration taxon group, the gene construction of the top 20 genera according to bio density observed that most of the phytoplankton were registered in 18S rRNA, and the most significant number of COI nucleotide sequences were established in macroinvertebrates. In addition, it was confirmed that there is a nucleotide sequence for the top 20 genera in 12S rRNA, 16S rRNA, and CYTB in fish. These results provided comprehensive information on the genes suitable for eDNA research for each taxon group.

국내 침입외래종 미국가재(Procambarus clarkii)의 생물모니터링을 환경유전자 분석과 함께 2021년 2월부터 10월까 지 완주군, 함평군, 나주시, 구례군 청주시, 5지점에서 수행하였다. 조사방법은 우산형통발과 둥근뜰채를 이용하였으며, eDNA 분석을 위해 8~10 L의 물을 채수하였다. 조사지 내의 동소종인 저서성 대형무척추동물과 함께 과거 분포기록 및 국내 가재류 유통 현황도 분석하였다. 조사결과 미국가재는 총 122개체가 확인되었으며, 함평군에서 59개체 (48.36%)로 가장 많은 개체수와 높은 환경유전자(eDNA)가 검출되었고 계절상 5월달에 출현 빈도가 가장 높았다. 암컷과 수컷의 비율은 21:5로 암컷이 우세하였으며, 크기는 암컷이 72.2±21.1 mm, 수컷이 80.5±15.6 mm, 어린개체가 25.3±9.8 mm이었다. 국내에 유입된 미국가재는 남서쪽 지역을 중심으로 확산되고 있는 것으로 파악되었으며, 과거에 출현 기록이 있는 서울지역에서는 확인되지 않았다. 미국가재에 외부공생하는 끈거머리지렁이류는 확인되지 않았으며, 국내의 출현한 미국가재는 일본에서부터 수입된 개체로 추정되었다. 국내에서 유통되고 있는 외래 가재류는 8종 이상이 었으며, 이중 마블가재(Procambarus virginalis)는 2021년에 환경부에서 유입주의 생물로 지정된 종으로 파악되었다. 미국가재 조사지역 일대에 서식하는 저서성 대형무척추동물은 총 3문 5강 39과 69종이 출현하였으며, 잠자리목이 24.62%, 딱정벌레목과 노린재목이 각각 16.92%로 우세하게 나타났다. 이 중 한반도고유종 1종, 적색목록 범주의 준위협(NT)으로 구분되는 1종, 갑각류는 총 6종이 출현하였다. 섭식기능군에서는 잡아먹는무리가 서식기능군에서는 기어오르는무리가 전체적으로 우세하여 출현하였다. 조사지역 내의 미국가재는 수초와 수변부 식생이 풍부한 지역을 선호하며, 수질이 탁한 곳에서도 내성이 강한 것으로 분석되었다. 잡식성인 미국가재는 동소종에서 우세하게 출현하는 육식성 저서생물과 상호 경쟁관계를 유지할 것으로 보이며, 먹이사슬에 따른 생태계 교란이 지속적으로 일어날 것으로 판단된다.

A pump-type eDNA filtering system that can control voltage and hydraulic pressure respectively has been developed, and applied a filter case that can filter out without damaging the filter. The filtering performance of the developed system was evaluated by comparing the eDNA concentration with the conventional vacuum-pressured filtering method at the catchment conduit intake reservoir. The developed system was divided into a voltage control (manual pump system) method and a pressure control (automatic pump system) method, and the pressure was measured during filtering and the pressure change of each system was compared. The voltage control method started with 65 [KPa] at the beginning of the filtering, and as the filtering time elapsed, the amount of filtrate accumulated in the filter increased, so the pressure gradually increased. As a result of controlling the pressure control method to maintain a constant pressure according to the designed algorithm, there was a difference in the width of the hydraulic pressure fluctuation during the filtering process according to the feedback time of the hydraulic pressure sensor, and it was confirmed that the pressure was converged to the target pressure. The filtering performance of the developed system was confirmed by measuring the eDNA concentration and comparing the voltage control method and the hydraulic control method with the control group. The voltage control method obtained similar results to the control group, but the hydraulic control method showed lower results than the control group. It is considered that the low eDNA concentration in the hydraulic control method is due to the large pressure deviation during filtering and maintaining a constant pressure during the filtering process. Therefore, rather than maintaining a constant pressure during filtering, it was confirmed that a voltage control method in which the pressure is gradually increased as the filtrate increases with the lapse of filtering time is suitable for collecting eDNA. As a result of comparing the average concentration of eDNA in lentic zone and lotic zone as a control group, it was found to be 96.2 [ng μL-1] and 88.4 [ng μL-1l], respectively. The result of comparing the average concentration of eDNA by the pump method was also high in the lentic zone sample as 90.7 [ng μL-1] and 74.8 [ng μL-1] in the lentic zone and the lotic zone, respectively. The high eDNA concentration in the lentic zone is thought to be due to the influence of microorganisms including the remaining eDNA.

Recently, environmental DNA (eDNA)-based metabarcoding approaches have been proposed to evaluate the status of freshwater ecosystems owing to various advantages, including fast and easy sampling and minimal habitat disruption from sampling. Therefore, as a case study, we applied eDNA metabarcoding techniques to evaluate the effects of an abandoned mine land located near a headwater stream of Nakdonggang River, South Korea, by examining benthic macroinvertebrate diversity and compared the results with those obtained using the traditional Surber-net sampling method. The number of genera was higher in Surbernet sampling (29) than in the eDNA analysis (20). The genus richness tended to decrease from headwater to downstream in eDNA analysis, whereas richness tended to decrease at sites with acid-sulfated sediment areas using Surber-net sampling. Through cluster analysis and non-metric multidimensional scaling, the sampling sites were differentiated into two parts: acid-sulfated and other sites using Surber-net sampling, whereas they were grouped into the two lowest downstream and other sites using eDNA sampling. To evaluate freshwater ecosystems using eDNA analysis in practical applications, it is necessary to constantly upgrade the methodologies and compare the data with field survey methods.

환경유전자 (eDNA)는 다양한 환경 (수중, 토양, 대기)에 존재하는 생물체로부터 유래된 유전물질을 의미한다. eDNA는 높은 민감도, 짧은 조사시간 등 많은 장점들이 존재하며 이로 인해 생물 모니터링 및 유해생물과 멸종위기 생물을 탐색하는 분야에 다양하게 활용되고 있다. 이러한 eDNA 를 채집하기 위해서는 대상생물 및 대상유전자뿐만 아니라 현장 여과방법 및 eDNA 보존방법과 같이 매우 다양한 항목들을 고려해야 한다. 특히 환경에서 eDNA를 채집하는 방법은 eDNA 농도와 직결되는 항목으로서 적절한 채집방법을 사용하여 eDNA를 채집할 때 정확한 분석결과를 얻을 수 있다. 또한 현장에서 채집한 eDNA를 보존하고 추출하는 과정에서도 정확한 방법을 사용하였을 때 현장에 분포하는 eDNA의 농도를 정확하게 파악할 수 있다. 특히 eDNA 연구를 시작하는 연구자들에게 eDNA 분야는 초기 진입 장벽이 매우 높은 기술로서 이를 위한 기초 자료가 매우 절실하다. 본 연구에서는 본 연구는 eDNA가 수생태계를 연구하기 위한 도구로서 보다 널리 이용되며, eDNA를 이용하기 시작하는 연구자들에게 도움을 주고자 수생태계에서 eDNA를 채집하고 및 운반하는 방법과 실험실에서 eDNA를 추출하는 방법을 소개하고, 보다 간편하고 효율적인 eDNA 채집 도구와 방법을 제시하였다.

본 연구에서는 멸종위기종이 서식하는 4개 하천 (금강, 지천, 황지천, 섬진강)에서 환경유전자 (environmental DNA, eDNA)와 보편적 어구를 이용한 조사 방법을 적용하여 지점별 종 다양성을 확인하고, 이를 통해 eDNA의 활용을 고찰 하였다. eDNA 조사를 통해서 확인된 종 수는 지점 평균 (± 표준편차) 19종 (±4.4)이며, 이는 어구를 이용한 정량 조사의 10종 (±4.8)과 비교하여 높게 나타났다. 대부분의 지점에서 eDNA 조사가 어구를 이용한 조사보다 효율이 높게 나타났다. 반면 eDNA 조사 결과 고유종 및 근연종에 대해서 동정의 오류가 확인되어, universal primer (MiFish primer set)에 대한 국내 적용의 한계를 확인하였다. 또한 멸종위기종의 서식 여부도 일부 종에 대해서 eDNA 조사 결과가 현장 조사 및 문헌과의 차이를 보였다. 현재 개발된 universal primer는 국내에서 서식하는 모든 담수종의 서식을 확인 하는 데 있어서 결과의 신뢰성을 담보할 수 없기 때문에 universal primer의 보완 및 개발이 필요하며, 멸종위기종과 같은 특정종의 서식 확인을 위해서는 종특이적 마커 개발을 통한 적용이 고려되어야 한다. 마지막으로 eDNA의 현장 조사 방법에 대한 매뉴얼이 개발될 경우, 수생태계 조사에 대한 활용성이 증대될 수 있을 것이다.

에드나 오브라이언(Edna O’Brien)은 금세기 이전에는 저평가된 작가이 다. 우리나라에서는 오브라이언뿐만 아니라, 제임스 조이스(James Joyce) 이후의 아일 랜드 작가들은 심도 있는 비평적 주목을 받지 못하고 있다. 이멜다 수녀 (“Sister Imelda”)에서 오브라이언은 이멜다 수녀의 세속적 욕망을 교묘하게 숨기고 있다. 독자 는 탐정소설을 읽고 있는 것처럼 이야기 속에 감추어진 욕망을 파헤쳐야 한다. 오브라 이언은 남성중심 사회에서 정해진 성역할을 강요하는 이성애적 관계에 대해 화자가 회의적인 시각을 드러내게 하면서 동성애적 욕망을 정당시 한다.