English
 
User Manual Privacy Policy Disclaimer Contact us
  Advanced SearchBrowse

Item

ITEM ACTIONSEXPORT

Released

Journal Article

Identification of DNA-synthesizing bacterial cells in coastal North Sea plankton

MPS-Authors
/persons/resource/persons210667

Pernthaler,  A.
Department of Molecular Ecology, Max Planck Institute for Marine Microbiology, Max Planck Society;

/persons/resource/persons210668

Pernthaler,  J.
Department of Molecular Ecology, Max Planck Institute for Marine Microbiology, Max Planck Society;

/persons/resource/persons210747

Schattenhofer,  M.
Microbial Genomics Group, Department of Molecular Ecology, Max Planck Institute for Marine Microbiology, Max Planck Society;

/persons/resource/persons210230

Amann,  R.
Department of Molecular Ecology, Max Planck Institute for Marine Microbiology, Max Planck Society;

External Ressource
No external resources are shared
Fulltext (public)

Pernthaler2.pdf
(Publisher version), 578KB

Supplementary Material (public)
There is no public supplementary material available
Citation

Pernthaler, A., Pernthaler, J., Schattenhofer, M., & Amann, R. (2002). Identification of DNA-synthesizing bacterial cells in coastal North Sea plankton. Applied and Environmental Microbiology, 68(11), 5728-5736.


Cite as: http://hdl.handle.net/21.11116/0000-0001-D2C3-5
Abstract
We describe a method for microscopic identification of DNA- synthesizing cells in bacterioplankton samples. After incubation with the halogenated thymidine analogue bromodeoxyuridine (BrdU), environmental bacteria were identified by fluorescence in situ hybridization (FISH) with horseradish peroxidase (HRP)-linked oligonucleotide probes. Tyramide signal amplification was used to preserve the FISH staining during the subsequent immunocytochemical detection of BrdU incorporation. DNA-synthesizing cells were visualized by means of an HRP-labeled antibody Fab fragment and a second tyramide signal amplification step. We applied our protocol to samples of prefiltered (pore size, 1.2 mum) North Sea surface water collected during early autumn. After 4h of incubation, BrdU incorporation was detected in 3% of all bacterial cells. Within 20h the detectable DNA-synthesizing fraction increased to >14%. During this period, the cell numbers of members of the Roseobacter lineage remained constant, but the fraction of BrdU-incorporating Roseobacter sp. cells doubled, from 24 to 42%. In Alteromonas sp. high BrdU labeling rates after 4 to 8 h were followed by a 10-fold increase in abundance. Rapid BrdU incorporation was also observed in members of the SAR86 lineage. After 4 h of incubation, cells affiliated with this clade constituted 8% of the total bacteria but almost 50% of the visibly DNA-synthesizing bacterial fraction. Thus, this clade might be an important contributor to total bacterioplankton activity in coastal North Sea water during periods of low phytoplankton primary production. The small size and low ribosome content of SAR86 cells are probably not indications of inactivity or dormancy.