Individual cell DNA synthesis within natural marine bacterial assemblages as detected by 'click' chemistry

TitleIndividual cell DNA synthesis within natural marine bacterial assemblages as detected by 'click' chemistry
Publication TypeJournal Article
Year of Publication2014
AuthorsSmriga S., Samo T.J, Malfatti F., Villareal J., Azam F
JournalAquatic Microbial Ecology
Volume72
Pagination269-280
Date Published2014/07
Type of ArticleArticle
ISBN Number0948-3055
Accession NumberWOS:000340220300007
Keywordsbacterioplankton; BrdU; bromodeoxyuridine; Click chemistry; dividing cells; Epifluorescence microscopy; growth-rates; immunocytochemistry; in-situ hybridization; Marine bacteria; microautoradiography; Microradiography; ocean; segregation; Single cell growth; thymidine incorporation; waters
Abstract

Individual cell growth rates enhance our understanding of microbial roles in regulating organic matter flux in marine and other aquatic systems. We devised a protocol to microscopically detect and quantify bacteria undergoing replication in seawater using the thymidine analog 5-ethynyl-2'-deoxyuridine (EdU), which becomes incorporated into bacterial DNA and is detected with a 'click' chemistry reaction in <1 h. Distinct EdU localization patterns were observed within individual labeled cells, e.g. some displayed 2 or more distinct EdU loci within a single DAPI-stained region, which likely indicated poleward migration of nascent DNA during the early phase of replication. Cell labeling ranged from 4.4 to 49%, comparable with cell labeling in parallel incubations for H-3-thymidine microautoradiography. Meanwhile, EdU signal intensities in cells ranged >3 orders of magnitude, wherein the most intensely labeled cells comprised most of a sample's sum community EdU signal, e.g. 26% of cells comprised 80% of the sum signal. This ability to rapidly detect and quantify signals in labeled DNA is an important step toward a robust approach for the determination of single-cell growth rates in natural assemblages and for linking growth rates with microscale biogeochemical dynamics.

DOI10.3354/ame01698
Short TitleAquat. Microb. Ecol.
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