Temporal patterns of biological dimethylsulfide (DMS) consumption during laboratory-induced phytoplankton bloom cycles

Phytoplankton bloom cycles were followed for 9 d in two 20 l carboy mesocosms filled with water from the offshore plume of Mobile Bay Alabama, USA, and incubated under fluorescent light. One of the blooms was enriched by addition of nitrate+phosphate (+nutrients), and both blooms were used to study how dimethylsulfide (DMS) concentrations and biological consumption varied over the bloom cycles. Peaks of algal biomass (15-22 µg chlorophyll a l-1) in the blooms were followed within 1 d by peaks of the DMS precursor, particulate dimethylsulfoniopropionate (DMSPp; 100-140 nM). DMS concentrations increased rapidly during the early bloom, rising from 1 nM on Day 1 up to 12 nM in the unamended carboy and up to 17 nM in the +nutrient carboy on Day 6. Maximum values for DMS concentrations, DMS consumption rates (as measured with 35S-DMS), and bacterial production were observed during the early decline of phytoplankton biomass. DMS consumption rates were initially 0.8 nM d-1 and increased to 3.1 nM d-1 in the unamended carboy and to 9.1 nM d-1 in the +nutrient carboy. Rate constants for DMS consumption (0.25-0.95 d-1) initially decreased as DMS concentrations increased, resulting in longer turnover times for DMS during the peak and early decline of the blooms. Assimilation of DMS-sulfur by bacterioplankton accounted for 4-22% of the total DMS consumption and higher rates of DMS assimilation occurred in the +nutrients bloom. Despite a bloom and decline of total heterotrophic bacterial abundances, bacterial community composition at the major phylogenetic group level remained relatively constant in both blooms, although the alpha proteobacteria showed a temporal increase in abundance in the +nutrient carboy. The concentration ratios of DMS:chlorophyll a and DMS:DMSP displayed non-linear, sigmoidal patterns over the bloom cycles and these ratios were not substantially affected by the nutrient amendment. Our results demonstrate that uncoupling of DMS production and biological consumption can occur early in a bloom cycle, causing DMS concentrations to rise significantly before biological consumption responds to draw down the DMS

Comparative genomics and mutagenesis analyses of choline metabolism in the marine Roseobacter clade

Choline is ubiquitous in marine eukaryotes and appears to be widely distributed in surface marine waters; however, its metabolism by marine bacteria is poorly understood. Here, using comparative genomics and molecular genetic approaches, we reveal that the capacity for choline catabolism is widespread in marine heterotrophs of the marine Roseobacter clade (MRC). Using the model bacterium Ruegeria pomeroyi, we confirm that the betA, betB and betC genes, encoding choline dehydrogenase, betaine aldehyde dehydrogenase and choline sulfatase, respectively, are involved in choline metabolism. The betT gene, encoding an organic solute transporter, was essential for the rapid uptake of choline but not glycine betaine (GBT). Growth of choline and GBT as a sole carbon source resulted in the re-mineralization of these nitrogen-rich compounds into ammonium. Oxidation of the methyl groups from choline requires formyltetrahydrofolate synthetase encoded by fhs in R.pomeroyi, deletion of which resulted in incomplete degradation of GBT. We demonstrate that this was due to an imbalance in the supply of reducing equivalents required for choline catabolism, which can be alleviated by the addition of formate. Together, our results demonstrate that choline metabolism is ubiquitous in the MRC and reveal the role of Fhs in methyl group oxidation in R.pomeroyi

Rapid turnover of dissolved DMS and DMSP by defined bacterioplankton communities in the stratified euphotic zone of the North Sea

Bacterioplankton-driven turnover of the algal osmolyte, dimethylsulphoniopropionate (DMSP), and its degradation product, dimethylsulphide (DMS) the major natural source of atmospheric sulphur, were studied during a Lagrangian SF6-tracer experiment in the North Sea (60°N, 3°E). The water mass sampled within the euphotic zone was characterised by a surface mixed layer (from 0 m to 13–30 m) and a subsurface layer (from 13–30 m to 45–58 m) separated by a 2°C thermocline spanning 2 m. The fluxes of dissolved DMSP (DMSPd) and DMS were determined using radioactive tracer techniques. Rates of the simultaneous incorporation of 14C-leucine and 3H-thymidine were measured to estimate bacterioplankton production. Flow cytometry was employed to discriminate subpopulations and to determine the numbers and biomass of bacterioplankton by staining for nucleic acids and proteins. Bacterioplankton subpopulations were separated by flow cytometric sorting and their composition determined using 16S ribosomal gene cloning/sequencing and fluorescence in situ hybridisation with designed group-specific oligonucleotide probes. A subpopulation, dominated by bacteria related to Roseobacter-(?-proteobacteria), constituted 26–33% of total bacterioplankton numbers and 45–48% of biomass in both surface and subsurface layers. The other abundant prokaryotes were a group within the SAR86 cluster of ?-proteobacteria and bacteria from the Cytophaga–Flavobacterium—cluster. Bacterial consumption of DMSPd was greater in the subsurface layer (41 nM d?1) than in the surface layer (20 nM d?1). Bacterioplankton tightly controlled the DMSPd pool, particularly in the subsurface layer, with a turnover time of 2 h, whereas the turnover time of DMSPd in the surface layer was 10 h. Consumed DMSP satisfied the majority of sulphur demands of bacterioplankton, even though bacterioplankton assimilated only about 2.5% and 6.0% of consumed DMSPd sulphur in the surface and subsurface layers, respectively. Bacterioplankton turnover of DMS was also faster in the subsurface layer (12 h) compared to the surface layer (24 h). However, absolute DMS consumption rates were higher in the surface layer, due to higher DMS concentrations in this layer. The majority of DMS was metabolised into dissolved non-volatile products, and bacteria could satisfy only 1–3% of their sulphur demands from DMS. Thus, structurally similar bacterioplankton communities exerted strong control over DMSPd and DMS concentrations both in the subsurface layer and surface mixed layer

An annual cycle of dimethylsulfoniopropionate-sulfur and leucine assimilating bacterioplankton in the coastal NW Mediterranean

13 pages, 6 figuresThe contribution of major phylogenetic groups to heterotrophic bacteria assimilating sulfur from dissolved dimethylsulfoniopropionate (DMSP) and assimilating leucine was analysed in surface seawaters from Blanes Bay (NW Mediterranean) over an annual study between March 2003 and April 2004. The percentage of bacteria assimilating DMSP-S showed a strong seasonal pattern, with a steady increase from winter (8 ± 5%) to summer (23 ± 3%). The same seasonal pattern was observed for the rate of DMSP-S assimilation. The annual average percentage of DMSP-S-assimilating bacteria (16 ± 8%) was lower than the corresponding percentage of leucine-assimilating cells (35 ± 16%), suggesting that not all bacteria synthesizing protein incorporated DMSP-S. Smaller differences between both percentages were recorded in summer. Members of the Alphaproteobacteria (Roseobacter and SAR11) and Gammaproteobacteria groups accounted for most of bacterial DMSP-S-assimilating cells over the year. All major bacterial groups showed an increase of the percentage of cells assimilating DMSP-S during summer, and contributed to the increase of the DMSP-S assimilation rate in this period. In these primarily P-limited waters, enrichment with P + DMSP resulted in a stimulation of bacterial heterotrophic production comparable to, or higher than, that with P + glucose in summer, while during the rest of the year P + glucose induced a stronger response. This suggested that DMSP was more important a S and C source for bacteria in the warm stratified season. Overall, our results suggest that DMSP-S assimilation is controlled by the contribution of DMSP to S (and C) sources rather than by the phylogenetic composition of the bacterioplanktonThis work was supported by the Spanish Ministry of Education and Science through a PhD studentship to M.V.-C, and through the projects MicroDIFF and MODIVUS (contracts REN2001-2120/MAR anc DTM2005-04795/MAR to J.M.G), by the Catalan government through Grant 2005SGR00021 (to R.S.) and by th EU's 5th Framework Program through project BASICs (contract EVK3-CT-2002-00078 to J.M.G.)Peer reviewe