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Applied and Environmental Microbiology, December 2008, p. 7321-7328, Vol. 74, No. 23
0099-2240/08/$08.00+0     doi:10.1128/AEM.01266-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Substrate-Specific Clades of Active Marine Methylotrophs Associated with a Phytoplankton Bloom in a Temperate Coastal Environment{triangledown} ,{dagger}

Josh D. Neufeld,{ddagger} Rich Boden, Hélène Moussard, Hendrik Schäfer,* and J. Colin Murrell

Department of Biological Sciences, University of Warwick, Coventry CV4 7AL, United Kingdom

Received 7 June 2008/ Accepted 1 October 2008

Marine microorganisms that consume one-carbon (C1) compounds are poorly described, despite their impact on global climate via an influence on aquatic and atmospheric chemistry. This study investigated marine bacterial communities involved in the metabolism of C1 compounds. These communities were of relevance to surface seawater and atmospheric chemistry in the context of a bloom that was dominated by phytoplankton known to produce dimethylsulfoniopropionate. In addition to using 16S rRNA gene fingerprinting and clone libraries to characterize samples taken from a bloom transect in July 2006, seawater samples from the phytoplankton bloom were incubated with 13C-labeled methanol, monomethylamine, dimethylamine, methyl bromide, and dimethyl sulfide to identify microbial populations involved in the turnover of C1 compounds, using DNA stable isotope probing. The [13C]DNA samples from a single time point were characterized and compared using denaturing gradient gel electrophoresis (DGGE), fingerprint cluster analysis, and 16S rRNA gene clone library analysis. Bacterial community DGGE fingerprints from 13C-labeled DNA were distinct from those obtained with the DNA of the nonlabeled community DNA and suggested some overlap in substrate utilization between active methylotroph populations growing on different C1 substrates. Active methylotrophs were affiliated with Methylophaga spp. and several clades of undescribed Gammaproteobacteria that utilized methanol, methylamines (both monomethylamine and dimethylamine), and dimethyl sulfide. rRNA gene sequences corresponding to populations assimilating 13C-labeled methyl bromide and other substrates were associated with members of the Alphaproteobacteria (e.g., the family Rhodobacteraceae), the Cytophaga-Flexibacter-Bacteroides group, and unknown taxa. This study expands the known diversity of marine methylotrophs in surface seawater and provides a comprehensive data set for focused cultivation and metagenomic analyses in the future.

* Corresponding author. Present address: Warwick HRI, University of Warwick, Wellesbourne CV35 9EF, United Kingdom. Phone: 44 (0)2476 575052. Fax: 44 (0)2476 574500. E-mail: H.Schaefer{at}warwick.ac.uk

{triangledown} Published ahead of print on 10 October 2008.

{dagger} Supplemental material for this article may be found at http://aem.asm.org/.

{ddagger} Present address: Department of Biology, University of Waterloo, Waterloo, Ontario N2L 3G1, Canada.

Applied and Environmental Microbiology, December 2008, p. 7321-7328, Vol. 74, No. 23
0099-2240/08/$08.00+0     doi:10.1128/AEM.01266-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.





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