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Applied and Environmental Microbiology, July 2000, p. 3044-3051, Vol. 66, No. 7
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
Culturability and In Situ Abundance of Pelagic
Bacteria from the North Sea
Heike
Eilers,
Jakob
Pernthaler,*
Frank Oliver
Glöckner, and
Rudolf
Amann
Max-Planck-Institut für marine
Mikrobiologie, D-28359 Bremen, Germany
Received 9 March 2000/Accepted 28 April 2000
The culturability of abundant members of the domain
Bacteria in North Sea bacterioplankton was investigated by
a combination of various cultivation strategies and
cultivation-independent 16S rRNA-based techniques. We retrieved 16S
rRNA gene (rDNA) clones from environmental DNAs and determined the
in situ abundance of different groups and genera by fluorescence in
situ hybridization (FISH). A culture collection of 145 strains was
established by plating on oligotrophic medium. Isolates were screened
by FISH, amplified ribosomal DNA restriction analysis (ARDRA), and
sequencing of representative 16S rDNAs. The majority of isolates
were members of the genera Pseudoalteromonas,
Alteromonas, and Vibrio. Despite being readily
culturable, they constituted only a minor fraction of the
bacterioplankton community. They were not detected in the 16S rDNA
library, and FISH indicated rare (<1% of total cell counts) occurrence as large, rRNA-rich, particle-associated bacteria. Conversely, abundant members of the Cytophaga-Flavobacteria
and gamma proteobacterial SAR86 clusters, identified by FISH as 17 to
30% and up to 10% of total cells in the North Sea bacterioplankton, respectively, were cultured rarely or not at all. Whereas
SAR86-affiliated clones dominated the 16S rDNA library (44 of 53 clones), no clone affiliated to the Cytophaga-Flavobacterum
cluster was retrieved. The only readily culturable abundant group of
marine bacteria was related to the genus Roseobacter. The
group made up 10% of the total cells in the summer, and the
corresponding sequences were also present in our clone library.
Rarefaction analysis of the ARDRA patterns of all of the isolates
suggested that the total culturable diversity by our method was high
and still not covered by the numbers of isolated strains but was almost
saturated for the gamma proteobacteria. This predicts a limit to the
isolation of unculturable marine bacteria, particularly the
gamma-proteobacterial SAR86 cluster, as long as no new techniques for
isolation are available and thus contrasts with more optimistic
accounts of the culturability of marine bacterioplankton.
*
Corresponding author. Mailing address:
Max-Planck-Institut für marine Mikrobiologie, Celsiusstrasse 1, D-28359 Bremen, Germany. Phone: 49 421 2028 940. Fax: 49 421 2028 580. E-mail: jperntha{at}mpi-bremen.de.
Applied and Environmental Microbiology, July 2000, p. 3044-3051, Vol. 66, No. 7
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
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