Concentrations of dimethylsulphoniopropionate and activities of dimethylsulphide-producing enzymes in batch cultures of nine dinoflagellate species

Dinoflagellates are recognised as one of the major phytoplankton groups that produce dimethylsulphoniopropionate (DMSP), the precursor of the marine trace gas dimethylsulphide (DMS) which has climate-cooling potential. To improve the prospects for including dinoflagellates in global climate models that include DMSP-related processes, we increased the data base for this group by measuring DMSP, DMS-producing enzyme activity (DPEA), carbon, nitrogen and Chl a in nine clonal dinoflagellate cultures (1 heterotrophic and 8 phototrophic strains). Growth rates ranged from 0.11 to 1.92?day^?1 with the highest value being for the heterotroph Crypthecodinium cohnii. Overall, we observed two orders of magnitude variability in DMSP content (11–364?mM) and detected DPEA in five of the nine strains (0.61–59.73?fmol?cell^?1?h^?1). Cell volume varied between 454 and 18,439?μm³ and whilst C and N content were proportional to the cell volume, DMSP content was not. The first DMSP measurements for a dinoflagellate from Antarctic waters and a species with diatom-like plastids are included. Lower DMSP concentrations were found in three small athecate species and a dinoflagellate with haptophyte-like plastids. The highest concentrations and production rates tended to be in globally distributed dinoflagellates and the heterotroph. Photosynthetic species that are distributed in temperate to tropical waters showed low DMSP concentrations and production rates and the polar representative showed moderate concentration and a low production rate. Estuarine species had the lowest concentrations and production rates. These data should help refine the inclusion of dinoflagellates as a functional group in future global climate models.     

The quantitative role of microzooplankton grazing in dimethylsulfide (DMS) production in the NW Mediterranean

The ubiquitous, biogenic trace gas dimethylsulfide (DMS) represents the largest natural source of atmospheric sulfur. Given DMS involvement in cloud formation and climate, understanding and parameterizing the oceanic DMS source and cycling processes is a necessary challenge. We report DMS cycling rates from microzooplankton dilution grazing experiments conducted monthly during 1 year in coastal northwestern Mediterranean waters. Concentrations of DMS, its algal precursor dimethylsulfoniopropionate (DMSPt) and chlorophyll a (Chla) ranged 0.9–11 nmol L^?1, 10–71 nmol L^?1, and 0.2–1.5 µg L^?1, respectively. By comparing the growth and stock production rates of the DMSP-producing algae to those of total phytoplankton, we estimated that 3?±?4% (range 0.4–12%) of the carbon primary production was invested in DMSP biosynthesis. Microzooplankton grazing rates on DMSP-producing phytoplankton (0.46–1.45 day^?1) were generally higher than those on the bulk assemblage (0.08–0.99 day^?1), except in midsummer months. This could have been due to the smaller size of most DMSP producers. There was no indication of micrograzer selection against DMSP-containing phytoplankton, since they were not grazed at lower rates than the bulk phytoplankton assemblage. A proportion of 6–20% of the grazed DMSP was converted into DMS, and this grazing-derived production accounted for 32–96% of dark gross DMS production by the total community. Bacteria consumed daily?≤?14–100% of the gross DMS production, which resulted in biological DMS turnover times of 1 to?≥?10 days. Throughout the year, grazing-mediated DMS production explained 73% of the variance in the DMS concentration, implying that microzooplankton grazing plays a major role in controlling DMS concentration in surface waters across a broad range of environmental and productivity conditions in the Mediterranean Sea. These findings should help improve the representation of herbivore grazing in prognostic models to predict the distribution and dynamics of the global DMS emission and its feedback response to changing climate.     

Dimethylsulfoniopropionate Turnover Is Linked to the Composition and Dynamics of the Bacterioplankton Assemblage during a Microcosm Phytoplankton Bloom

Processing of the phytoplankton-derived organic sulfur compound dimethylsulfoniopropionate (DMSP) by bacteria was studied in seawater microcosms in the coastal Gulf of Mexico (Alabama). Modest phytoplankton blooms (peak chlorophyll a [Chl a] concentrations of ~2.5 μg liter⁻¹) were induced in nutrient-enriched microcosms, while phytoplankton biomass remained low in unamended controls (Chl a concentrations of ~0.34 μg liter⁻¹). Particulate DMSP concentrations reached 96 nM in the enriched microcosms but remained approximately 14 nM in the controls. Bacterial biomass production increased in parallel with the increase in particulate DMSP, and nutrient limitation bioassays in the initial water showed that enrichment with DMSP or glucose caused a similar stimulation of bacterial growth. Concomitantly, increased bacterial consumption rate constants of dissolved DMSP (up to 20 day⁻¹) and dimethylsulfide (DMS) (up to 6.5 day⁻¹) were observed. Nevertheless, higher DMSP S assimilation efficiencies and higher contribution of DMSP to bacterial S demand were found in the controls compared to the enriched microcosms. This indicated that marine bacterioplankton may rely more on DMSP as a source of S under oligotrophic conditions than under the senescence phase of phytoplankton blooms. Phylogenetic analysis of the bacterial assemblages in all microcosms showed that the DMSP-rich algal bloom favored the occurrence of various Roseobacter members, flavobacteria (Bacteroidetes phylum), and oligotrophic marine Gammaproteobacteria. Our observations suggest that the composition of the bacterial assemblage and the relative contribution of DMSP to the overall dissolved organic sulfur/organic matter pool control how efficiently bacteria assimilate DMSP S and thereby potentially divert it from DMS production.     

Spatial distributions of dimethyl sulfur compounds,DMSP-lyase activity,and phytoplankton community in the East China Sea during fall

The spatial distributions of dimethylsulfide (DMS), dimethylsulfoniopropionate (DMSP), DMSP-lyase activity (DLA) and their controlling factors including nutrients, phytoplankton community and bacterial abundance were investigated in the East China Sea (ECS) during fall from October 19 to November 2, 2015. Diatoms and dinoflagellates dominated the phytoplankton community, while other taxonomic groups were rare and mainly found in the oligotrophic open sea. Affected by the high nutrients concentrations, Chl a, DMS, DMSP and DLA showed high values in eutrophic inshore waters, and decreased from the costal zones to the open sea. Statistical analysis suggested that diatoms and dinoflagellates were the main controlling factors of DMS, DMSP, and DLA in ECS. For size-fractionated samples, a reduced contribution of the microplankton from inshore stations to offshore stations affected by the trophic conditions was noted. Meanwhile, this decrease in microplankton led to an increase in the ratio of DLA contributed by picoplankton and free-living bacteria from the estuary area to the offshore region. The DMS sea-to-air flux was calculated using the equation of Nightingale et al. (Glob Biogeochem Cy 14(1):373–387, 2000), and approximately 2.88 × 10^?2 Tg of sulfur was transferred from the sea into the atmosphere in the form of DMS in ECS during fall.     

Interacting effects of ocean acidification and warming on growth and DMS‐production in the haptophyte coccolithophore Emiliania huxleyi

The production of the marine trace gas dimethyl sulfide (DMS) provides 90% of the marine biogenic sulfur in the atmosphere where it affects cloud formation and climate. The effects of increasing anthropogenic CO₂ and the resulting warming and ocean acidification on trace gas production in the oceans are poorly understood. Here we report the first measurements of DMS‐production and data on growth, DMSP and DMS concentrations in pH‐stated cultures of the phytoplankton haptophyte Emiliania huxleyi. Four different environmental conditions were tested: ambient, elevated CO₂ (+CO₂), elevated temperature (+T) and elevated temperature and CO₂ (+TCO₂). In comparison to the ambient treatment, average DMS production was about 50% lower in the +CO₂ treatment. Importantly, temperature had a strong effect on DMS production and the impacts outweighed the effects of a decrease in pH. As a result, the +T and +TCO₂ treatments showed significantly higher DMS production of 36.2 ± 2.58 and 31.5 ± 4.66 μmol L^?1 cell volume (CV) h^?1 in comparison with the +CO₂ treatment (14.9 ± 4.20 μmol L^?1 CV h^?1). As the cultures were aerated with an air/CO₂ mixture, DMS was effectively removed from the incubation bottles so that concentration remained relatively low (3.6–6.1 mmol L^?1 CV). Intracellular DMSP has been shown to increase in E. huxleyi as a result of elevated temperature and/or elevated CO₂ and our results are in agreement with this finding: the ambient and +CO₂ treatments showed 125 ± 20.4 and 162 ± 27.7 mmol L^?1 CV, whereas +T and +TCO₂ showed significantly increased intracellular DMSP concentrations of 195 ± 15.8 and 211 ± 28.2 mmol L^?1 CV respectively. Growth was unaffected by the treatments, but cell diameter decreased significantly under elevated temperature. These results indicate that DMS production is sensitive to CO₂ and temperature in E. huxleyi. Hence, global environmental change that manifests in ocean acidification and warming may not result in decreased DMS as suggested by earlier studies investigating the effect of elevated CO₂ in isolation.     

Abundance and Distribution of Dimethylsulfoniopropionate Degradation Genes and the Corresponding Bacterial Community Structure at Dimethyl Sulfide Hot Spots in the Tropical and Subtropical Pacific Ocean

Dimethylsulfoniopropionate (DMSP) is mainly produced by marine phytoplankton but is released into the microbial food web and degraded by marine bacteria to dimethyl sulfide (DMS) and other products. To reveal the abundance and distribution of bacterial DMSP degradation genes and the corresponding bacterial communities in relation to DMS and DMSP concentrations in seawater, we collected surface seawater samples from DMS hot spot sites during a cruise across the Pacific Ocean. We analyzed the genes encoding DMSP lyase (dddP) and DMSP demethylase (dmdA), which are responsible for the transformation of DMSP to DMS and DMSP assimilation, respectively. The averaged abundance (±standard deviation) of these DMSP degradation genes relative to that of the 16S rRNA genes was 33% ± 12%. The abundances of these genes showed large spatial variations. dddP genes showed more variation in abundances than dmdA genes. Multidimensional analysis based on the abundances of DMSP degradation genes and environmental factors revealed that the distribution pattern of these genes was influenced by chlorophyll a concentrations and temperatures. dddP genes, dmdA subclade C/2 genes, and dmdA subclade D genes exhibited significant correlations with the marine Roseobacter clade, SAR11 subgroup Ib, and SAR11 subgroup Ia, respectively. SAR11 subgroups Ia and Ib, which possessed dmdA genes, were suggested to be the main potential DMSP consumers. The Roseobacter clade members possessing dddP genes in oligotrophic subtropical regions were possible DMS producers. These results suggest that DMSP degradation genes are abundant and widely distributed in the surface seawater and that the marine bacteria possessing these genes influence the degradation of DMSP and regulate the emissions of DMS in subtropical gyres of the Pacific Ocean.     

CONCENTRATIONS OF DIMETHYLSULFONIOPROPIONATE AND DIMETHYL SULFIDE ARE STRAIN‐SPECIFIC IN SYMBIOTIC DINOFLAGELLATES (SYMBIODINIUM SP., DINOPHYCEAE)1

Dimethyl sulfide (DMS) and dimethylsulfoniopropionate (DMSP) are sulfur compounds that may function as antioxidants in algae. Symbiotic dinoflagellates of the genus Symbiodinium show strain‐specific differences in their susceptibility to temperature‐induced oxidative stress and have been shown to contain high concentrations of DMSP. We investigated continuous cultures of four strains from distinct phylotypes (A1, A13, A2, and B1) that can be characterized by differential thermal tolerances. We hypothesized that strains with high thermal tolerance have higher concentrations of DMSP and DMS in comparison to strains with low thermal tolerance. DMSP concentrations were strain‐specific with highest concentrations occurring in A1 (225 ± 3.5 mmol · L^?1 cell volume [CV]) and lowest in A2 (158 ± 3.8 mmol · L^?1 CV). Both strains have high thermal tolerance. Strains with low thermal tolerance (A13 and B1) showed DMSP concentrations in between these extremes (194 ± 19.0 and 160 ± 6.1 mmol · L^?1 CV, respectively). DMS data further confirmed this general pattern with high DMS concentrations in A1 and A13 (4.1 ± 1.22 and 2.1 ± 0.37 mmol · L^?1 CV, respectively) and low DMS concentrations in A2 and B1 (0.3 ± 0.06 and 0.5 ± 0.22 mmol · L^?1 CV, respectively). Hence, the strain‐specific differences in DMSP and DMS concentrations did not match the different abilities of the four phylotypes to withstand thermal stress. Future work should quantify the possible dynamics in DMSP and DMS concentrations during periods of high oxidative stress in Symbiodinium sp. and address the role of these antioxidants in zooxanthellate cnidarians.     

Effect of PAR irradiance intensity on Phaeocystis antarctica (Prymnesiophyceae) growth and DMSP,DMSO, and acrylate concentrations

Phaeocystis antarctica forms extensive spring blooms in the Southern Ocean that coincide with high concentrations of dimethylsulfoniopropionate (DMSP), dimethylsulfoxide (DMSO), dimethylsulfide (DMS), and acrylate. We determined how concentrations of these compounds changed during the growth of axenic P. antarctica cultures exposed to light-limiting, sub-saturating, and saturating PAR irradiances. Cellular DMSP concentrations per liter cell volume (CV) ranged between 199 and 403 mmol · L_CV⁻¹, with the highest concentrations observed under light-limiting PAR. Cellular acrylate concentrations did not change appreciably with a change in irradiance level or growth, ranging between 18 and 45 mmol · L_CV⁻¹, constituting an estimated 0.2%–2.8% of cellular carbon. Both dissolved acrylate and DMSO increased substantially with irradiance during exponential growth on a per-cell basis, ranging from 0.91 to 3.15 and 0.24 to 1.39 fmol · cell⁻¹, respectively, indicating substantial export of these compounds into the dissolved phase. Average cellular DMSO:DMSP ratios increased 7.6-fold between exponential and stationary phases of batch growth, with a 3- to 13-fold increase in cellular DMSO likely formed from abiotic reactions of DMSP and DMS with reactive oxygen species (ROS). At mM levels, cellular DMSP and acrylate are proposed to serve as de facto antioxidants in P. antarctica not regulated by oxidative stress or changes in ROS. Instead, cellular DMSP concentrations are likely controlled by other physiological processes including an overflow mechanism to remove excess carbon via acrylate, DMS, and DMSO during times of unbalanced growth brought on by physical stress or nutrient limitation. Together, these compounds should aid P. antarctica in adapting to a range of PAR irradiances by maintaining cellular functions and reducing oxidative stress.     

The effect of light intensity and daylength on the β-dimethylsulphoniopropionate (DMSP) content of marine green macroalgae from Antarctica*

Abstract. The β-dimethylsulphoniopropionate (DMSP) concentrations of 5 Antarctic green algae grown at 2, 30 and 55 μmol photons m^?2 s^?1 were determined during a period of one year, cultivated under fluctuating daylengths mimicking the conditions of the natural habitat at King George Island. Antarctica. The intracellular DMSP content of all species decreased simultaneously with decreasing daylengths and vice versa. Additionally, the DMSP level was affected by the light intensity: the higher the photon fluence rate the greater the algal DMSP concentration. Under conditions of darkness, there was a degradation of the DMSP pool in members of the Acrosiphoniales, while the DMSP content in members of the Ulvales did not change. The results indicate a light-dependent DMSP accumulation in algae. Therefore, they may help to explain the seasonal variability of DMSP and its cleavage product dimethylsulphide (DMS) in coastal waters.     

New Routes for Aerobic Biodegradation of Dimethylsulfoniopropionate

Dimethylsulfoniopropionate (DMSP), an osmolyte in marine plants, is biodegraded by cleavage of dimethyl sulfide (DMS) or by demethylation to 3-methiolpropionate (MMPA) and 3-mercaptopropionate (MPA). Sequential demethylation has been observed only with anoxic slurries of coastal sediments. Bacteria that grew aerobically on MMPA and DMSP were isolated from marine environments and phytoplankton cultures. Enrichments with DMSP selected for bacteria that generated DMS, whereas MMPA enrichments selected organisms that produced methanethiol (CH₃SH) from either DMSP or MMPA. A bacterium isolated on MMPA grew on MMPA and DMSP, but rapid production of CH₃SH from DMSP occurred only with DMSP-grown cells. Low levels of MPA accumulated during growth on MMPA, indicating demethylation as well as demethiolation of MMPA. The alternative routes for DMSP biodegradation via MMPA probably impact on net DMS fluxes to the marine atmosphere.     

Macroscale patterns of the biological cycling of dimethylsulfoniopropionate (DMSP) and dimethylsulfide (DMS) in the Northwest Atlantic

The influence of the seasonal development of microplankton communities on the cycling of dimethylsulfide (DMS) and its precursor dimethylsulfoniopropionate (DMSP) was investigated along a South–North gradient (36–59^°N) in the Northwest (NW) Atlantic Ocean. Three surveys allowed the sampling of surface mixed layer (SML) waters at stations extending from the subtropical gyre to the Greenland Current during May, July and October 2003. Pools and transformation rates of DMSP and DMS were quantified and related to prevailing physical and biochemical conditions, phytoplankton abundance and taxonomic composition, as well as bacterioplankton abundance and leucine uptake. The South–North progression of the diatom bloom, a prominent feature in the NW Atlantic, did not influence the production of DMS whereas conditions in the N Atlantic Drift lead to a persistent bloom of DMSP-rich flagellate-dominated phytoplankton community and high net DMS production rates. Macroscale patterns of the observed variables were further explored using principal component analysis (PCA). The first axis of the PCA showed a strong association between the spatio-temporal distribution of DMSP and the abundance of several phytoplankton groups including dinoflagellates and prymnesiophytes, as well as with microbial-mediated DMSP_d consumption and yields and rates of the conversion of DMSP into DMS. The second axis revealed a strong association between concentrations of DMS and SML depth and photosynthetically active radiation, a result supporting the prominent role of solar radiation as a driver of DMS dynamics.     

Dynamics of Dimethyl Sulfide in a Marine Microbial Mat

Abstract The microbial mat was chosen as a model ecosystem to study dynamics of dimethyl sulfide (DMS) in marine sediments in order to gain insight into key processes and factors which determine emission rates. A practical advantage, compared to open ocean ecosystems, is that microbial mats contain high biomasses of different functional groups of bacteria involved in DMS dynamics, and that DMS concentrations are generally high enough to allow direct measurement of emission rates. Field data showed that, during the seasonal development of microbial mats, concentrations of chlorophyll a corresponded to dimethylsulfoniopropionate (DMSP). DMSP is an important precursor of DMS. It was demonstrated, with laboratory cultures, that various species of benthic diatoms produce substantial amounts of DMSP. The abundances of aerobic and anaerobic DMS- or DMSO-utilizing bacteria were estimated using the most-probable-number technique. Laboratory experiments with relatively undisturbed sediment cores showed that microbial mats act as a sink for DMS under oxic/light (day) conditions, and as a source of DMS under anoxic/dark (night) conditions. Axenic culture studies with Chromatium vinosum M2 and Thiocapsa pfennigii M8 (isolated from a microbial mat) showed that, under anoxic/light conditions, DMS was quantitatively converted to dimethylsulfoxide (DMSO). T. roseopersicina M11 converted DMSP to DMS and acrylate, apparently without use of either substrate. Received: 5 May 1997; Accepted: 21 August 1997     

Towards a systematic understanding of structure–function relationship of dimethylsulfoniopropionate‐catabolizing enzymes

Each year, several million tons of dimethylsulfoniopropionate (DMSP) are produced by marine phytoplankton and bacteria as an important osmolyte to regulate their cellular osmosis. Microbial breakdown of DMSP to the volatile gas dimethylsulfide (DMS) plays an important role in global biogeochemical cycles of the sulphur element between land and the sea. Understanding the enzymes involved in the transformation of DMSP and DMS holds the key to a better understanding of oceanic DMSP cycles. Recent work by Shao et al. (2019) has resolved the crystal structure of two important enzymes, DmdB and DmdC, involved in DMSP transformation through the demethylation pathway. Their work represents an important step towards a systematic understanding of the structure–function relationships of DMSP‐catabolizing enzymes in marine microbes.     

Recent insights into oceanic dimethylsulfoniopropionate biosynthesis and catabolism

Dimethylsulfoniopropionate (DMSP), a globally important organosulfur compound is produced in prodigious amounts (2.0 Pg sulfur) annually in the marine environment by phytoplankton, macroalgae, heterotrophic bacteria, some corals and certain higher plants. It is an important marine osmolyte and a major precursor molecule for the production of climate-active volatile gas dimethyl sulfide (DMS). DMSP synthesis take place via three pathways: a transamination ‘pathway-’ in some marine bacteria and algae, a Met-methylation ‘pathway-’ in angiosperms and bacteria and a decarboxylation ‘pathway-’ in the dinoflagellate, Crypthecodinium. The enzymes DSYB and TpMMT are involved in the DMSP biosynthesis in eukaryotes while marine heterotrophic bacteria engage key enzymes such as DsyB and MmtN. Several marine bacterial communities import DMSP and degrade it via cleavage or demethylation pathways or oxidation pathway, thereby generating DMS, methanethiol, and dimethylsulfoxonium propionate, respectively. DMSP is cleaved through diverse DMSP lyase enzymes in bacteria and via Alma1 enzyme in phytoplankton. The demethylation pathway involves four different enzymes, namely DmdA, DmdB, DmdC and DmdD/AcuH. However, enzymes involved in the oxidation pathway have not been yet identified. We reviewed the recent advances on the synthesis and catabolism of DMSP and enzymes that are involved in these processes.     

Biogenic production of DMSP and its degradation to DMS—their roles in the global sulfur cycle

Dimethyl sulfide(DMS) is the most abundant form of volatile sulfur in Earth's oceans, and is mainly produced by the enzymatic clevage of dimethylsulfoniopropionate(DMSP). DMS and DMSP play important roles in driving the global sulfur cycle and may affect climate. DMSP is proposed to serve as an osmolyte, a grazing deterrent, a signaling molecule, an antioxidant, a cryoprotectant and/or as a sink for excess sulfur. It was long believed that only marine eukaryotes such as phytoplankton produce DMSP. However, we recently discovered that marine heterotrophic bacteria can also produce DMSP, making them a potentially important source of DMSP. At present, one prokaryotic and two eukaryotic DMSP synthesis enzymes have been identified.Marine heterotrophic bacteria are likely the major degraders of DMSP, using two known pathways: demethylation and cleavage.Many phytoplankton and some fungi can also cleave DMSP. So far seven different prokaryotic and one eukaryotic DMSP lyases have been identified. This review describes the global distribution pattern of DMSP and DMS, the known genes for biosynthesis and cleavage of DMSP, and the physiological and ecological functions of these important organosulfur molecules, which will improve understanding of the mechanisms of DMSP and DMS production and their roles in the environment.     

Dimethylsulfoniopropionate turnover is linked to the composition and dynamics of the bacterioplankton assemblage during a microcosm phytoplankton bloom

Processing of the phytoplankton-derived organic sulfur compound dimethylsulfoniopropionate (DMSP) by bacteria was studied in seawater microcosms in the coastal Gulf of Mexico (Alabama). Modest phytoplankton blooms (peak chlorophyll a [Chl a] concentrations of approximately 2.5 microg liter(-1)) were induced in nutrient-enriched microcosms, while phytoplankton biomass remained low in unamended controls (Chl a concentrations of approximately 0.34 microg liter(-1)). Particulate DMSP concentrations reached 96 nM in the enriched microcosms but remained approximately 14 nM in the controls. Bacterial biomass production increased in parallel with the increase in particulate DMSP, and nutrient limitation bioassays in the initial water showed that enrichment with DMSP or glucose caused a similar stimulation of bacterial growth. Concomitantly, increased bacterial consumption rate constants of dissolved DMSP (up to 20 day(-1)) and dimethylsulfide (DMS) (up to 6.5 day(-1)) were observed. Nevertheless, higher DMSP S assimilation efficiencies and higher contribution of DMSP to bacterial S demand were found in the controls compared to the enriched microcosms. This indicated that marine bacterioplankton may rely more on DMSP as a source of S under oligotrophic conditions than under the senescence phase of phytoplankton blooms. Phylogenetic analysis of the bacterial assemblages in all microcosms showed that the DMSP-rich algal bloom favored the occurrence of various Roseobacter members, flavobacteria (Bacteroidetes phylum), and oligotrophic marine Gammaproteobacteria. Our observations suggest that the composition of the bacterial assemblage and the relative contribution of DMSP to the overall dissolved organic sulfur/organic matter pool control how efficiently bacteria assimilate DMSP S and thereby potentially divert it from DMS production.     

Denitrification kinetics and denitrifier abundances in sediments of lakes receiving atmospheric nitrogen deposition (Colorado, USA)

The transport and deposition of anthropogenic nitrogen (N) to downwind ecosystems is significant and can be a dominant source of new N to many watersheds. Bacterially mediated denitrification in lake sediments may ameliorate the effects of N loading by permanently removing such inputs. We measured denitrification in sediments collected from lakes in the Colorado Rocky Mountains (USA) receiving elevated (5–8?kg?N?ha^?1?y^?1) or low (<2?kg?N?ha^?1?y^?1) inputs of atmospheric N deposition. The nitrate (NO₃ ^?) concentration was significantly greater in high-deposition lakes (11.3?μmol?l^?1) compared to low-deposition lakes (3.3?μmol?l^?1). Background denitrification was positively related to NO₃ ^? concentrations and we estimate that the sampled lakes are capable of removing a significant portion of N inputs via sediment denitrification. We also conducted a dose–response experiment to determine whether chronic N loading has altered sediment denitrification capacity. Under Michaelis–Menten kinetics, the maximum denitrification rate and half-saturation NO₃ ^? concentration did not differ between deposition regions and were 765?μmol?N?m^?2?h^?1 and 293?μmol?l^?1?NO₃ ^?, respectively, for all lakes. We enumerated the abundances of nitrate- and nitrite-reducing bacteria and found no difference between high- and low-deposition lakes. The abundance of these bacteria was related to available light and bulk sediment resources. Our findings support a growing body of evidence that lakes play an important role in N removal and, furthermore, suggest that current levels of N deposition have not altered the abundance of denitrifying bacteria or saturated the capacity for sediment denitrification.     

Regulatory and Functional Diversity of Methylmercaptopropionate Coenzyme A Ligases from the Dimethylsulfoniopropionate Demethylation Pathway in Ruegeria pomeroyi DSS-3 and Other Proteobacteria

The organosulfur compound dimethylsulfoniopropionate (DMSP) is produced by phytoplankton and is ubiquitous in the surface ocean. Once released from phytoplankton, marine bacteria degrade DMSP by either the cleavage pathway to form the volatile gas dimethylsulfide (DMS) or the demethylation pathway, yielding methanethiol (MeSH), which is readily assimilated or oxidized. The enzyme DmdB, a methylmercaptopropionate (MMPA)-coenzyme A (CoA) ligase, catalyzes the second step in the demethylation pathway and is a major regulatory point. The two forms of DmdB present in the marine roseobacter Ruegeria pomeroyi DSS-3, RPO_DmdB1 and RPO_DmdB2, and the single form in the SAR11 clade bacterium “Candidatus Pelagibacter ubique” HTCC1062, PU_DmdB1, were characterized in detail. DmdB enzymes were also examined from Ruegeria lacuscaerulensis ITI-1157, Pseudomonas aeruginosa PAO1, and Burkholderia thailandensis E264. The DmdB enzymes separated into two phylogenetic clades. All enzymes had activity with MMPA and were sensitive to inhibition by salts, but there was no correlation between the clades and substrate specificity or salt sensitivity. All Ruegeria species enzymes were inhibited by physiological concentrations (70 mM) of DMSP. However, ADP reversed the inhibition of RPO_DmdB1, suggesting that this enzyme was responsive to cellular energy charge. MMPA reversed the inhibition of RPO_DmdB2 as well as both R. lacuscaerulensis ITI-1157 DmdB enzymes, suggesting that a complex regulatory system exists in marine bacteria. In contrast, the DmdBs of the non-DMSP-metabolizing P. aeruginosa PAO1 and B. thailandensis E264 were not inhibited by DMSP, suggesting that DMSP inhibition is a specific adaptation of DmdBs from marine bacteria.     

Microbial controls on DMSP degradation and DMS formation in the Sargasso Sea

Bacterial degradation of dimethylsulfoniopropionate (DMSP) represents one of the main sources of the climatically–active trace gas dimethylsulfide (DMS) in the upper ocean. Short-term enrichment studies to stimulate specific pathways of DMSP degradation in oligotrophic waters from the Sargasso Sea were used to explore regulatory connections between the different bacterial DMSP degradation steps and determine potential biological controls on DMS formation in the open ocean. Experiments were conducted with surface water at the BATS station in the western North Atlantic Ocean. We added selected organic substrates (25 nmol L^?1 final concentration) to induce different steps of DMSP degradation in the microbial community, and then measured DMSP dynamics (assimilation and turnover rates), DMS yields (using ³⁵sulfur-DMSP tracer), and bacterial production rates. In most treatments, the main fate of consumed S-DMSP was excretion as a non-volatile S product. ³⁵S-DMSP tracer turnover rates (accumulation + assimilation + excretion of transformed products as DMS or others) increased upon addition of DMSP and glucose, but not acrylate, methymercaptopropionate (MMPA), methanethiol, DMS or glycine betaine. DMS yields from ³⁵S-DMSP never exceeded 16 % except in a short term DMSP enrichment, for which the yield reached 45 % (±17 %). Results show that availability of non-sulfur containing labile C sources (glucose, acrylate) decreased bacterial DMS production while stimulating bacterial heterotrophic production, and suggest an influence of bacterial sulfur demand in controlling DMS-yielding pathways. However, regulatory effects on ³⁵S-DMSP fate were not consistent across all reduced sulfur compounds (i.e., methanethiol or MMPA), and may reflect alternate roles of DMSP as a bacterial energy source and osmolyte.