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Applied and Environmental Microbiology, August 2004, p. 4648-4657, Vol. 70, No. 8
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.8.4648-4657.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Use of Microautoradiography Combined with Fluorescence In Situ Hybridization To Determine Dimethylsulfoniopropionate Incorporation by Marine Bacterioplankton Taxa

Maria Vila,1 Rafel Simó,1* Ronald P. Kiene,2 Jarone Pinhassi,1 José M. González,3,4 Mary Ann Moran,3 and Carlos Pedrós-Alió1

Institut de Ciències del Mar-CMIMA (CSIC), 08003 Barcelona, Catalonia,1 Departamento de Microbiología, Facultad de Farmacia, Universidad de La Laguna, 38071 La Laguna, Tenerife, Spain,4 Department of Marine Sciences, University of South Alabama, Mobile, Alabama 36688,2 Department of Marine Sciences, University of Georgia, Athens, Georgia 306023

Received 19 November 2003/ Accepted 25 April 2004

The fraction of planktonic heterotrophic bacteria capable of incorporating dissolved dimethylsulfoniopropionate (DMSP) and leucine was determined at two coastal sites by microautoradioagraphy (AU). In Gulf of Mexico seawater microcosm experiments, the proportion of prokaryotes that incorporated sulfur from [35S]DMSP ranged between 27 and 51% of 4',6-diamidino-2-phenylindole (DAPI)-positive cells, similar to or slightly lower than the proportion incorporating [3H]leucine. In the northwest Mediterranean coast, the proportion of cells incorporating sulfur from [35S]DMSP increased from 5 to 42% from January to March, coinciding with the development of a phytoplankton bloom. At the same time, the proportion of cells incorporating [3H]leucine increased from 21 to 40%. The combination of AU and fluorescence in situ hybridization (FISH) revealed that the Roseobacter clade ({alpha}-proteobacteria) accounted for 13 to 43% of the microorganisms incorporating [35S]DMSP at both sampling sites. Significant uptake of sulfur from DMSP was also found among members of the {gamma}-proteobacteria and Cytophaga-Flavobacterium groups. Roseobacter and {gamma}-proteobacteria exhibited the highest percentage of DAPI-positive cells incorporating 35S from DMSP (around 50%). Altogether, the application of AU with [35S]DMSP combined with FISH indicated that utilization of S from DMSP is a widespread feature among active marine bacteria, comparable to leucine utilization. These results point toward DMSP as an important substrate for a broad and diverse fraction of marine bacterioplankton.

* Corresponding author. Mailing address: Institut de Ciències del Mar-CMIMA (CSIC), Passeig Marítim de la Barceloneta 37-49, 08003 Barcelona, Catalonia, Spain. Phone: 34 932309590. Fax: 34 932309555. E-mail: rsimo{at}icm.csic.es.

Applied and Environmental Microbiology, August 2004, p. 4648-4657, Vol. 70, No. 8
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.8.4648-4657.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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