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Community shifts of actively growing lake bacteria after N-acetyl-glucosamine addition: improving the BrdU-FACS method
- In aquatic environments, community dynamics of bacteria, especially actively growing bacteria (AGB), are tightly linked with dissolved organic matter (DOM) quantity and quality. We analyzed the community dynamics of DNA-synthesizing and accordingly AGB by linking an improved bromodeoxyuridine immunocytochemistry approach with fluorescence-activated cell sorting (BrdU-FACS). FACS-sorted cells of even oligotrophic ecosystems in winter were characterized by 16S rRNA gene analysis. In incubation experiments, we examined community shifts of AGB in response to the addition of N-acetyl-glucosamine (NAG), one of the most abundant aminosugars in aquatic systems. Our improved BrdU-FACS analysis revealed that AGB winter communities of oligotrophic Lake Stechlin (northeastern Germany) substantially differ from those of total bacteria and consist of Alpha-, Beta-, Gamma-, Deltaproteobacteria, Actinobacteria, Candidatus OP10 and Chloroflexi. AGB populations with different BrdU-fluorescence intensities and cell sizes represented different phylotypesIn aquatic environments, community dynamics of bacteria, especially actively growing bacteria (AGB), are tightly linked with dissolved organic matter (DOM) quantity and quality. We analyzed the community dynamics of DNA-synthesizing and accordingly AGB by linking an improved bromodeoxyuridine immunocytochemistry approach with fluorescence-activated cell sorting (BrdU-FACS). FACS-sorted cells of even oligotrophic ecosystems in winter were characterized by 16S rRNA gene analysis. In incubation experiments, we examined community shifts of AGB in response to the addition of N-acetyl-glucosamine (NAG), one of the most abundant aminosugars in aquatic systems. Our improved BrdU-FACS analysis revealed that AGB winter communities of oligotrophic Lake Stechlin (northeastern Germany) substantially differ from those of total bacteria and consist of Alpha-, Beta-, Gamma-, Deltaproteobacteria, Actinobacteria, Candidatus OP10 and Chloroflexi. AGB populations with different BrdU-fluorescence intensities and cell sizes represented different phylotypes suggesting that single-cell growth potential varies at the taxon level. NAG incubation experiments demonstrated that a variety of widespread taxa related to Alpha-, Beta-, Gammaproteobacteria, Bacteroidetes, Actinobacteria, Firmicutes, Planctomycetes, Spirochaetes, Verrucomicrobia and Chloroflexi actively grow in the presence of NAG. The BrdU-FACS approach enables detailed phylogenetic studies of AGB and, thus, to identify those phylotypes which are potential key players in aquatic DOM cycling.…
Author details: | Yuya Tada, Hans-Peter GrossartORCiDGND |
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DOI: | https://doi.org/10.1038/ismej.2013.148 |
ISSN: | 1751-7362 |
ISSN: | 1751-7370 |
Pubmed ID: | https://pubmed.ncbi.nlm.nih.gov/23985742 |
Title of parent work (English): | The ISME journal : multidisciplinary journal of microbial ecology |
Publisher: | Nature Publ. Group |
Place of publishing: | London |
Publication type: | Article |
Language: | English |
Year of first publication: | 2014 |
Publication year: | 2014 |
Release date: | 2017/03/27 |
Tag: | Lake Stechlin; N-acetyl-glucosamine (NAG); actively growing bacteria (AGB); bacterial community composition; bromodeoxyuridine (BrdU) immunocytochemistry; fluorescence-activated cell sorting (FACS) |
Volume: | 8 |
Issue: | 2 |
Number of pages: | 14 |
First page: | 441 |
Last Page: | 454 |
Funding institution: | Leibniz Institute of Freshwater Ecology and Inland Fisheries (IGB); German Science Foundation [DFG-GR1540/17-1] |
Organizational units: | Mathematisch-Naturwissenschaftliche Fakultät / Institut für Geowissenschaften |
Peer review: | Referiert |
Institution name at the time of the publication: | Mathematisch-Naturwissenschaftliche Fakultät / Institut für Erd- und Umweltwissenschaften |