Quantification of Nitrogen Cycling and DMSP Genes in Svalbard, Kongsfjorden Using Real Time Quantitative PCR
실시간 정량 PCR을 이용한 스발바르, 콩스피오르덴의 질소 순환 및 DMSP 유전자의 정량화
- 주제(키워드) 도움말 Kongsfjorden , qPCR , Nitrogen cycle gene , DMSP gene Cloning , Metabarcoding
- 발행기관 강릉원주대학교 일반대학원
- 지도교수 도움말 한덕기
- 발행년도 2024
- 학위수여년월 2025. 2
- 학위명 석사
- 학과 및 전공 도움말 일반대학원 해양생명과학과
- 세부분야 해당없음
- 실제URI http://www.dcollection.net/handler/kangnung/000000012061
- UCI I804:42001-000000012061
- 본문언어 영어
초록/요약 도움말
In this study, we evaluated qPCR primer sets for nitrogen cycling (amoA and nirK) and DMSP metabolism genes (dmdA, dddP, dddL) in the surface waters of Kongsfjorden and Svalbard, quantified gene copy numbers by qPCR, and compared them to the local microbial community by metabarcoding. A total of 13 qPCR primer tests for 10 mechanisms yielded 6 gene sequences, which were subsequently cloned and compared to the UniProt database. The only available standards for the nitrogen cycle (amoA, nirK) and DMSP metabolism genes (dmdA, dddP, dddL) were the aforementioned gene sequences, with the exception of the classical nitrogen gene (nifH), for which no raw data were available in the database. Metabarcoding analysis revealed that nitrogen cycling and DMSP-related genes were detected in specific microbial communities and that the increase in cyanobacteria during summer was related to nitrogen metabolism. In particular, cyanobacteria have the ability to fix nitrogen, contributing to the nitrification process, suggesting that they may play an important role in nitrogen cycling during the summer. Furthermore, the expression of sulfur metabolism-related genes was strongly associated with microbial communities, such as Rhodobacteraceae and SAR11_Clade, which play important roles in the release of sulfate through DMSP degradation. This study provides an important basis for understanding nitrogen and sulfur metabolism gene expression patterns in the Kongsfjorden region, and may contribute to a deeper understanding of the biological productivity and functional characteristics of ecosystems in the region.
more목차 도움말
Abstract in English ································································ i
Abstract in Korean ······························································· iii
List of Tables ······································································· v
List of Figures ····································································· v
Ⅰ. Introduction ···································································· 1
Ⅱ. Materials and Methods ····················································· 5
1. Sample collection ································································· 5
2. Real Time Quantitative PCR Preparation ···································· 8
3. Real Time Quantitative PCRs ··················································· 11
4. Sequencing and Data Processing,Statistical Analyses ····················· 13
III. Results ············································································· 15
1. Physicochemical properties ······················································ 15
2. qPCR Primer Test ·································································· 17
3. qPCR measurement ······························································· 20
4. Community Composition of Metabarcoding ·································· 25
IV. Discussion ······································································· 30
V. References ········································································ 32
