PMID- 12406771
OWN - NLM
STAT- MEDLINE
DCOM- 20021217
LR  - 20210526
IS  - 0099-2240 (Print)
IS  - 1098-5336 (Electronic)
IS  - 0099-2240 (Linking)
VI  - 68
IP  - 11
DP  - 2002 Nov
TI  - Identification of DNA-synthesizing bacterial cells in coastal North Sea plankton.
PG  - 5728-36
AB  - 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 micro m) 
      North Sea surface water collected during early autumn. After 4 h of incubation, 
      BrdU incorporation was detected in 3% of all bacterial cells. Within 20 h 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.
FAU - Pernthaler, Annelie
AU  - Pernthaler A
AD  - Max Planck Institute for Marine Microbiology, Bremen, Germany.
FAU - Pernthaler, Jakob
AU  - Pernthaler J
FAU - Schattenhofer, Martha
AU  - Schattenhofer M
FAU - Amann, Rudolf
AU  - Amann R
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Appl Environ Microbiol
JT  - Applied and environmental microbiology
JID - 7605801
RN  - 0 (Antimetabolites)
RN  - 0 (DNA, Bacterial)
RN  - G34N38R2N1 (Bromodeoxyuridine)
SB  - IM
MH  - Animals
MH  - Antimetabolites/pharmacology
MH  - Bacteria/drug effects/isolation & purification/*metabolism
MH  - Bromodeoxyuridine/pharmacology
MH  - DNA, Bacterial/*biosynthesis
MH  - Immunohistochemistry
MH  - In Situ Hybridization, Fluorescence
MH  - Marine Biology
MH  - Microbial Sensitivity Tests
MH  - Plankton
MH  - *Water Microbiology
PMC - PMC129917
EDAT- 2002/10/31 04:00
MHDA- 2002/12/18 04:00
PMCR- 2002/11/01
CRDT- 2002/10/31 04:00
PHST- 2002/10/31 04:00 [pubmed]
PHST- 2002/12/18 04:00 [medline]
PHST- 2002/10/31 04:00 [entrez]
PHST- 2002/11/01 00:00 [pmc-release]
AID - 0701 [pii]
AID - 10.1128/AEM.68.11.5728-5736.2002 [doi]
PST - ppublish
SO  - Appl Environ Microbiol. 2002 Nov;68(11):5728-36. doi: 
      10.1128/AEM.68.11.5728-5736.2002.