분류군별 외래생물 탐지를 위한 환경 DNA 메타바코딩 활용 가능성 = Feasibility of Environmental DNA Metabarcoding for Invasive Species Detection According to Taxa

다국어 초록 (Multilingual Abstract)

In orderto establish an effective management strategy forinvasive species early detection and regular monitoring are required to assess their introduction or dispersal. Environmental DNA (eDNA) is actively applied to evaluate the fauna including the presence of invasive species as it has high detection sensitivity and can detect multiple species simultaneously. In Korea, the applicability evaluation of metabarcoding is being conducted mainly on fish, and research on other taxa is insufficient. Therefore, this study identified the feasibility of detecting invasive species in Korea using eDNA metabarcoding. In addition, to confirm the possibility of detection by taxa, the detection of target species was evaluated using four universal primers (MiFish, MiMammal, Mibird, Amp16S) designed for fish, mammals, birds, and amphibians. As a result, target species (Trachemys scripta, 3 sites; Cervus nippon, 3 sites; Micropterus salmoides, 7 sites; Rana catesbeiana, 4 sites) were detected in 17 of the total 55 sites. Even in the selection of dense sampling sites within the study area, there was a difference in the detection result by reflecting the ecological characteristics of the target species.
A comparison of community structures (species richness, abundance and diversity) based on the presence of invasive species focused on M.salmoides and T.scripta, showed higher diversity at the point where invasive species were detected. Also, 1 to 4 more species were detected and abundance was also up to 1.7 times higher. The results of invasive species detection through metabarcoding and the comparison of community structures indicate that the accumulation of large amounts of monitoring data through eDNA can be efficiently utilized for multidimensional ecosystem evaluation. In addition, it suggested that eDNA can be used as major data for evaluation and prediction, such as tracking biological changes caused by artificial and natural factors and environmental impact assessment.

국문 초록 (Abstract)

효과적인 외래생물 관리 전략 수립을 위해서는 도입 및 확산 여부 평가를 위한 정기 모니터링이 요구된다. 환경 DNA (eDNA, environmental DNA) 메타바코딩은 높은 검출 민감도를 가지고 다수의 종을동시에 검출할 수 있어 외래생물의 출현 여부와 그 영향을 평가하는데 활발히 활용되고 있다. 국내에서는어류를 중심으로 메타바코딩의 적용 가능성 평가가 이루어지고 있으며 타 분류군에 대한 연구는 부족한 실정이다. 따라서 본 연구에서는 환경 DNA 메타바코딩을 활용한 국내 외래생물 탐지 가능성을 확인하고자했다. 분류군별 검출 가능성을 확인하기 위해 어류, 포유류, 조류, 양서류를 목표로 디자인 된 4가지 범용 프라이머(MiFish, MiMammal, Mibird, Amp16S)를 활용하여 대상종 검출 여부를 평가하였다. 그 결과, 총55개 지점 중 17개 지점(Trachemys scripta, 3개 지점; Cervus nippon, 3개 지점; Micropterus salmoides, 7개 지점; Rana catesbeiana, 4개 지점)에서 대상종의 서식이 확인되었다. 대상지 내 조밀한 지점 선정에도 생태적 특성을 반영한 검출 지점에 차이가 나타났다. 큰입배스와 붉은귀거북을 중심으로 외래생물이 출현이 생물 군집구조(종 풍부도, 풍부도, 다양도)에 미치는 영향을 비교한 결과, 외래생물이 서식하는 지점에서의 다양도가 더 높게 나타났다. 또한 외래생물 출현 지점에서 출현 종 수가 1~4종 추가 검출되었으며풍부도 또한 1.7배 높게 나타났다. 메타바코딩을 통한 외래생물 검출 결과 및 군집구조 비교는 eDNA를 통한 다량의 모니터링 데이터 구축이 다차원적 생태계 평가에 효율적으로 활용될 수 있음을 나타냈다. 또한환경의 인위적, 자연적 변화에 따른 생물상 변화를 관찰하고 자연생태 분야의 환경영향평가 등 현황 평가및 예측을 위한 주요한 기초자료로 활용 가능성을 제시하였다.

참고문헌 (Reference)

1 김정희 ; 조현빈 ; 장민호 ; 우승현 ; 조영호 ; 윤주덕, "환경유전자의 국내 담수어류 모니터링 적용 연구 김정희" 한국수생태학회 53 (53): 63-72, 2020

2 송영근 ; 김종희 ; 원수연 ; 박찬, "환경DNA 메타바코딩 기술을 활용한 수생태계 어류종 군집조사의 가능성 – 도시 생태하천 초기분석 자료를 중심으로" 한국환경복원기술학회 22 (22): 125-138, 2019

3 김가우 ; 송영근, "환경DNA 기술을 이용한 국내 담수어류종 탐지 가능성 - 경기도 민물고기생태학습관 중심으로 -" 한국환경영향평가학회 30 (30): 1-12, 2021

4 Robert M. Dorazio, "ednaoccupancy:An r package for multiscale occupancy modelling of environmental DNA data" Wiley 18 (18): 368-380, 2017

5 Hashemzadeh Segherloo I, "eDNA metabarcoding as a means to assess distribution of subterranean fish communities:Iranian blind cave fishes as a case study" 4 (4): 402-416, 2022

6 Mizumoto H, "Utilizing environmental DNA for wide-range distributions of reproductive area of an invasive terrestrial toad in Ishikari river basin in Japan" 24 (24): 1199-1211, 2022

7 Muha TP, "Using environmental DNA to improve species distribution models for freshwater invaders" 5 : 158-, 2017

8 "University"

9 Stewart KA, "Understanding the effects of biotic and abiotic factors on sources of aquatic environmental DNA" 28 (28): 983-1001, 2019

10 Deiner K, "Transport Distance of Invertebrate Environmental DNA in a Natural River" 9 : e88786-, 2014

11 Ministry of Environment, "The 2nd Alien Speices Management Plan (2019-2023)"

12 Toshifumi Minamoto, "Surveillance of fish species composition using environmental DNA" Springer Science and Business Media LLC 13 (13): 193-197, 2011

13 Di MC, "Read counts from environmental DNA (eDNA) metabarcoding reflect fish abundance and biomass in drained ponds" 4 : 97-112, 2020

14 Valentini A, "Next-generation monitoring of aquatic biodiversity using environmental DNA metabarcoding" 25 (25): 929-942, 2016

15 Saenz-Agudelo P, "Monitoring vertebrate biodiversity of a protected coastal wetland using eDNA metabarcoding" 4 (4): 77-92, 2022

16 Thomsen PF, "Monitoring endangered freshwater biodiversity using environmental DNA" 21 (21): 2565-2573, 2012

17 Sato Y, "MitoFish and MiFish Pipeline: A Mitochondrial Genome Database of Fish with an Analysis Pipeline for Environmental DNA Metabarcoding" 35 (35): 1553-1555, 2018

18 Miya M, "MiFish, a set of universal PCR primers for metabarcoding environmental DNA from fishes: detection of more than 230 subtropical marine species" 2 : 150088-, 2015

19 Yates MC, "Meta-analysis supports further refinement of eDNA for monitoring aquatic species-specific abundance in nature" 1 (1): 5-13, 2019

20 "Information of Korean Alien Specie" National Institute of Ecology

21 Stoeckle BC, "Influence of stream characteristics and population size on downstream transport of freshwater mollusk environmental dna" 40 : 191-201, 2021

22 Nevers MB, "Influence of sediment and stream transport on detecting a source of environmental DNA" 15 : e0244086-, 2020

23 Vila M, "Handbook of Alien Species in Europe" 265-268, 2009

24 Lintermans M, "Finding the needle in the haystack : Comparing sampling methods for detecting an endangered freshwater fish" 67 (67): 1740-1749, 2016

25 Takahara T, "Estimation of Fish Biomass Using Environmental DNA" 7 : e35868-, 2012

26 Rourke ML, "Environmental DNA(eDNA)as a tool for assessing fish biomass : A review of approaches and future considerations for resource surveys" 4 (4): 9-33, 2022

27 Thomsen PF, "Environmental DNA – An emerging tool in conservation for monitoring past and present biodiversity" 183 : 4-18, 2015

28 Smart AS, "Environmental DNA sampling is more sensitive than a traditional survey technique for detecting an aquatic invader" 25 (25): 1944-1952, 2015

29 Yamamoto S, "Environmental DNA metabarcoding reveals local fish communities in a species-rich coastal sea" 7 (7): 1-12, 2017

30 Ushio M, "Environmental DNA enables detection of terrestrial mammals from forest pond water" 17 (17): e63-e75, 2017

31 Aozora Kakuda, "Environmental DNA detection and quantification of invasive red-eared sliders, Trachemy scripta elegans, in ponds and the influence of water quality" PeerJ 7 : e8155-, 2019

32 Murakami H, "Dispersion and degradation of environmental DNA from caged fish in a marine environment" 85 : 327-337, 2019

33 Ministry of Environment, "Developing a real-time web-based positioning surveillance system customized for introduced exotic species" Seoul National 2022

34 Williams KE, "Detection and persistence of environmental DNA from an invasive, terrestrial mammal" 8 (8): 688-695, 2018

35 Ushio M, "Demonstration of the potential of environmental DNA as a tool for the detection of avian species" 8 (8): 1-10, 2018

36 Czeglédi I, "Congruency between two traditional and eDNA-based sampling methods in characterising taxonomic and trait-based structure of fish communities and community-environment relationships in lentic environment" 129 : 107952-, 2021

37 Yu Z, "Comparison of species-specific qPCR and metabarcoding methods to detect small pelagic fish distribution from open ocean environmental DNA" 17 (17): 0273670-, 2022

38 Sard NM, "Comparison of fish detections, community diversity, and relative abundance using environmental DNA metabarcoding and traditional gears" 1 (1): 368-384, 2019

39 Juan Diego Lozano Mojica, "Applications of eDNA Metabarcoding for Vertebrate Diversity Studies in Northern Colombian Water Bodies" Frontiers Media SA 8 : 2021

40 Feilong Li, "Application of Environmental DNA Metabarcoding for Predicting Anthropogenic Pollution in Rivers" American Chemical Society (ACS) 2018

41 Rubenson ES, "An invader in salmonid rearing habitat: Current and future distributions of smallmouth bass (micropterus dolomieu) in the Columbia river basin" 77 (77): 314-325, 2020

42 Ministry of Environment, "Act On the Conservation And Use of Biological Diversity"

43 Leempoel K, "A comparison of eDNA to camera trapping for assessment of terrestrial mammal diversity" 287 (287): 2020

44 Gang HS, "A Study on the Management and Utilization of the Saetgang Stream in the Han River" Korea Environment Institute 2015

45 Yeoungdeungpo cultural foundation, "A Study on the Basic Environment of Waterfront Ecological Culture for the Development of Waterfront Culture Revitalization Plan" 2020

46 Ministry of Environment, "4th National Natural Environment Survey: Ichthyofauna from midstream catchment of Anyang stream"

47 Ministry of Environment, "4th National Natural Environment Survey: Ichthyofauna from midstream catchment of Anyang stream"