DOI: 10.1016/j.watres.2018.11.017
Scopus记录号: 2-s2.0-85056752399
论文题名: AFISsys - An autonomous instrument for the preservation of brackish water samples for microbial metatranscriptome analysis
作者: Charvet S. ; Riemann L. ; Alneberg J. ; Andersson A.F. ; von Borries J. ; Fischer U. ; Labrenz M.
刊名: Water Research
ISSN: 431354
出版年: 2019
起始页码: 351
结束页码: 361
语种: 英语
英文关键词: AFISsys
; Gene expression
; In situ fixation
; Metatranscriptomics
; RNA
; Sampling
Scopus关键词: Biogeochemistry
; Microorganisms
; Phenols
; RNA
; Sampling
; Seawater
; AFISsys
; Autonomous instruments
; Biogeochemical cycling
; Environmental Monitoring
; Local transformations
; Long-term preservation
; Metatranscriptomics
; Microbial communities
; Gene expression
; alcohol
; brackish water
; messenger RNA
; phenol
; ribosome RNA
; sea water
; small subunit ribosomal RNA
; surface water
; transcriptome
; bioassay
; brackish water
; design method
; environmental change
; environmental monitoring
; gene expression
; genetic analysis
; in situ measurement
; instrumentation
; microbial activity
; microbial community
; molecular analysis
; RNA
; sampling
; time series analysis
; turnover
; Actinobacteria
; Alveolata
; aquatic environment
; Article
; bacterial gene
; Bacteroidetes
; biogeochemical cycling
; cell count
; Chlamydiae
; circadian rhythm
; comparative study
; controlled study
; cyanobacterium
; environmental monitoring
; experimental therapy
; Firmicutes
; gene expression
; green alga
; Haptophyta
; long term exposure
; manB gene
; microbial community
; nonhuman
; nth gene
; Planctomycetes
; priA gene
; priority journal
; Proteobacteria
; RNA analysis
; RNA sequence
; spoU gene
; time series analysis
; transcriptomics
; water sampling
; Bacteria (microorganisms)
英文摘要: Microbial communities are the main drivers of biogeochemical cycling of multiple elements sustaining life in the ocean. The rapidity of their response to stressors and abrupt environmental changes implies that even fast and infrequent events can affect local transformations of organic matter and nutrients. Modern molecular techniques now allow for monitoring of microbial activities and functions in the environment through the analysis of genes and expressed genes contained in natural microbial assemblages. However, messenger RNA turnover in cells can be as short as 30 seconds and stability varies greatly between transcripts. Sampling of in situ communities involves an inevitable delay between the collection of seawater and the extraction of its RNA, leaving the bacterial communities plenty of time to alter their gene expression. The characteristics of microbial RNA turnover make time-series very difficult because samples need to be processed immediately to limit alterations to the metatranscriptomes. To address these challenges we designed an autonomous in situ fixation multi-sampler (AFISsys) for the reliable sampling of microbial metatranscriptomes at frequent intervals, for refined temporal resolution. To advance the development of this instrument, we examined the minimal seawater volume necessary for adequate coverage of community gene expression, and the suitability of phenol/ethanol fixation for immediate and long-term preservation of transcripts from a microbial community. We then evaluated the field eligibility of the instrument itself, with two case studies in a brackish system. AFISsys is able to collect, fix, and store water samples independently at a predefined temporal resolution. Phenol/ethanol fixation can conserve metatranscriptomes directly in the environment for up to a week, for later analysis in the laboratory. Thus, the AFISsys constitutes an invaluable tool for the integration of molecular functional analyses in environmental monitoring in brackish waters and in aquatic environments in general. © 2018 Elsevier Ltd Citation statistics:
资源类型: 期刊论文
标识符: http://119.78.100.158/handle/2HF3EXSE/122147
Appears in Collections: 气候变化事实与影响
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作者单位: IOW - Leibniz Institute for Baltic Sea Research, Warnemuende, Germany; Marine Biological Section, Department of Biology, University of Copenhagen, Denmark; KTH - Royal Institute of Technology, Science for Life Laboratory, Sweden; HYDRO-BIOS Apparatebau GmbH, Altenholz, Germany
Recommended Citation:
Charvet S.,Riemann L.,Alneberg J.,et al. AFISsys - An autonomous instrument for the preservation of brackish water samples for microbial metatranscriptome analysis[J]. Water Research,2019-01-01