Korean Journal of Ecology and Environment (생태와환경)

The Korean Society of Limnology (한국수생태학회)
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Application of DNA Analysis for Identification of Prey Items on Zooplankton: Selective Treatment Method

기수역 요각류 위내용물 유전자 분석: 소화기관 내외부 유전자의 선택적 처리방법

  • Chae, Yeon-Ji (Department of Environmental Science and Engineering, Kyung Hee University) ;
  • Oh, Hye-Ji (Department of Environmental Science and Engineering, Kyung Hee University) ;
  • Kim, Yong-Jae (Department of Life Science, Daejin University) ;
  • Chang, Kwang-Hyeon (Department of Environmental Science and Engineering, Kyung Hee University) ;
  • Jo, Hyunbin (Institute for Environment and Energy, Pusan National University)
  • 채연지 (경희대학교 환경학 및 환경공학과) ;
  • 오혜지 (경희대학교 환경학 및 환경공학과) ;
  • 김용재 (대진대학교 생명과학과) ;
  • 장광현 (경희대학교 환경학 및 환경공학과) ;
  • 조현빈 (부산대학교 환경.에너지연구소)
  • Received : 2021.09.13
  • Accepted : 2021.09.24
  • Published : 2021.09.30
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Abstract

Understanding the selective feeding behavior of zooplankton on phytoplankton is essential for evaluating the nutrient cycle and energy flow in the food web. Although many studies have been conducted regarding the feeding behaviors of zooplankton through gut content analyses, there are limitations in the visual identification of digested contents using a microscope. DNA techniques have been applied to overcome these limitations since they can detect and amplify small amounts of prey DNA remaining in the gut contents. We designed a method to extract prey DNA from the gut contents of the whole body of the copepod specimen and tested the resolution of DNA identification for the prey phytoplankton. The common brackish species, Sinocalanus tenellus, were collected from Saemangeum Reservoir in different sites and seasons, and gut content DNA was extracted using 2.5% bleach treatment for 2 min for removal of potential contamination sources existing in preserved specimens without dissolution of the body. The sequences of the extracted gut contents were confirmed using BLASTn suite based on the NCBI database. The phytoplankton species detected in the gut showed temporal and spatial differences. Although DNA analysis of small copepod gut contents has been suggested as an effective method to examine the dynamics of primary prey sources at the genus or species level, uncertainties such as misidentification and limitations in the detailed information of the composition still exist.

동물플랑크톤이 식물플랑크톤을 선택적으로 섭식하는 특성에 대한 이해는 수생태계 먹이사슬 내의 물질 이동에 중요하다. 하지만 해부를 통한 위내용물 추출 방법은 소형 요각류를 대상으로 적용하기에는 적절하지 않고, 유전자가 유실되거나 위내용물이 아닌 개체 외부의 유전자로 인해 오염될 가능성이 존재한다. 본 연구에서 호 내 식물 플랑크톤 조성 및 기타 환경이 상이한 두 지점을 선정하여 모든 지점에서 지속적으로 출현하는 기수성 요각류인 Sinocalanus tenellus를 대상으로 위내용물의 유전자 분석을 수행하였다. 요각류 개체 외부의 DNA를 제거하는 데 2.5%로 희석한 시판용 표백제(차아염소산나트륨 5.4%)에 2분간 처리하여 증류수로 2회 세척한 뒤 유전자를 추출하였다. 추출된 유전자는 23S rRNA을 증폭하여 서열분석을 실시하였다. Capillary sequencing 분석 결과, 원수와 처리수 및 요각류 위내용물에서 다양한 분류군(규조강, 녹조강, 남조강, 와편모조강, 은편모조강, 황갈조강)에 속하는 식물플랑크톤이 검출되었으며, 새만금호 내 시공간 차이에 따라 상이한 경향을 보였다. 현미경을 이용하여 동정한 식물플랑크톤 군집 조성의 경우 규조강이 우점한 반면, 동일한 원수의 유전자 분석(capillary sequencing) 결과에서는 주로 녹조강, 남조강 및 와편모조강이 우점하여 다소 상반된 경향을 나타냈다. 본 연구에서 적용한 위내용물 분석에 특화된 외부 유전자 제거 전처리 방법은 농도와 처리시간 조절 등의 응용방법에 따라 다양한 동물플랑크톤 분류군에 적용이 가능할 것으로 사료된다.

Keywords

Acknowledgement

이 논문은 2020년도 정부(교육부)의 재원으로 한국연구재단의 지원을 받아 수행된 기초연구사업임(NRF-2020R1C1C1009066).

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