Processes in the surface ocean regulate dissolved organic matter distributions in the deep

Marine dissolved organic matter (DOM) is a major global carbon pool, consisting of thousands of compounds with distinct lifetimes. While marine DOM persists for millennia, its molecular and isotopic composition imply that it is dynamic on shorter timescales. To determine the extent to which DOM deviates from conservative water mass mixing, we conducted a two-endmember mixing analysis on dissolved organic carbon (DOC) concentration and DOM molecular composition in the Atlantic and Pacific. Endmembers were the deep water masses near their formation sites. For DOM composition, we considered 6118 molecular formulae (MF) identified via Fourier-transform ion cyclotron resonance mass spectrometry in solid-phase extracts (SPE) of 837 samples. Bulk DOC and SPE-DOC concentrations behaved conservatively in both basins and ≥70% of the MF (14–20 μM SPE-DOC) mixed conservatively. However, a small fraction (10%–20%) of the MF (<3 μM SPE-DOC) were added or removed during mixing. These MF were more reduced and oxidized, respectively, than the conservative fraction. There were also MF absent from the endmembers; these accounted for ≤1 μM of SPE-DOC and positively correlated with DOM lability. Based on their distribution across the two basins, we conclude that the conserved MF are formed in the surface subtropical ocean and modified in overturning areas. In the deep ocean, however, these MF are solely controlled by mixing. This finding contrasts with the current paradigm of slow, continuous degradation of recalcitrant DOM in the deep ocean. Our analysis illustrates the importance of the surface ocean in controlling DOM cycling in the deep

Improved Mass Accuracy and Isotope Confirmation through Alignment of Ultrahigh-Resolution Mass Spectra of Complex Natural Mixtures

Fourier-transform ion cyclotron resonance mass spectrometry (FT-ICR-MS) is one of the state-of-the-art methods to analyze complex natural organic mixtures. The precision of detected masses is crucial for molecular formula attribution. Random errors can be reduced by averaging multiple measurements of the same mass, but because of limited availability of ultrahigh-resolution mass spectrometers, most studies cannot afford analyzing each sample multiple times. Here we show that random errors can be eliminated also by averaging mass spectral data from independent environmental samples. By averaging the spectra of 30 samples analyzed on our 15 T instrument we reach a mass precision comparable to a single spectrum of a 21 T instrument. We also show that it is possible to accurately and reproducibly determine isotope ratios with FT-ICR-MS. Intensity ratios of isotopologues were improved to a degree that measured deviations were within the range of natural isotope fractionation effects. In analogy to δ13C in environmental studies, we propose Δ13C as an analytical measure for isotope ratio deviances instead of widely employed C deviances. In conclusion, here we present a simple tool, extensible to Orbitrap-based mass spectrometers, for postdetection data processing that significantly improves mass accuracy and the precision of intensity ratios of isotopologues at no extra cost

Comparison of different solid phase extraction sorbents for the qualitative assessment of dissolved organic nitrogen in freshwater samples using FT-ICR-MS

Fate and reactivity of dissolved organic matter (DOM) is directly linked to its chemical composition. Therefore, molecular characterisation, for example using Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR-MS), is used for a better understanding of those factors. To study organic compounds in the water column, an efficient extraction method is important. The commonly used extraction method for FT-ICR-MS is solid phase extraction (SPE) using a reversed-phase sorbent (BondElut PPL). But this method, to the best of our knowledge, was not evaluated for its ability to extract organic nitrogen compounds which are important building blocks of life and therefore an important fraction of DOM. In this study, several solid phase sorbents were tested for their ability to extract organic nitrogen compounds from water samples of natural aqueous environments. Different cartridges concerning their retention mechanism and pore size were tested. Three cartridges with different extraction mechanism (reversed phase, cation exchange or a mixture of both) or different pore size were tested. Except for one sorbent type, which heavily contaminated the samples with organic molecules, the tested cartridges leached neither a significant amount of dissolved organic carbon (DOC) nor dissolved organic nitrogen (DON). The sorbents were tested with lake water to be able to investigate their functionality in real conditions. It could be shown, that the molecular composition of the sample should be considered for the choice of the sorbent material. Additionally, it was shown that a mixed-bed sorbent is a valuable complementary SPE sorbent for the molecular characterisation of lacustrine samples using FT-ICR-MS and it might also be useful for a quantitative extraction. Furthermore, it could be shown that HyperSep Retain CX sorbent allows to extract a broader range of organic nitrogen compounds leading to a more comprehensive data set for investigating organic nitrogen compounds in lakes using FT-ICR-MS

Dissolved Organic Matter Cycling in the Coastal Upwelling System Off Central Peru During an “El Niño” Year

The Peruvian upwelling system (PUS) is among the most productive regions in the ocean, with high rates of primary production and an intense oxygen minimum zone (OMZ). The main perturbation of this system is associated to “El Niño” (EN), which affects water mass distribution and reduces primary production. Previous studies in the PUS provided first insights into the dynamics of dissolved organic matter (DOM), but high-resolution studies involving the molecular characterization of the DOM pool to reveal the processes that affect the carbon cycle in this highly productive system are lacking. We characterized the molecular composition of solid-phase extractable DOM (SPE-DOM) in the coastal upwelling system off Central Peru and linked it to specific processes that affect DOM cycling. Seasonal sampling (April, August, and December) was carried out off Central Peru (12°S) during 2015, a low productivity year marked by EN conditions. The DOM molecular composition was obtained via Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR-MS). Solid-phase extractable dissolved organic carbon (SPE-DOC) concentrations showed significant differences (p &lt; 0.05) between the water masses present off central Peru. In order to explore if changes in SPE-DOC concentrations were the result of water mass mixing, we applied a conservative mixing (CM) model. The model revealed a non-conservative behavior of SPE-DOC and allowed us to identify two distinct groups of samples with increased and decreased SPE-DOC concentrations, respectively, and one group of samples inside the CM range. Differences in environmental parameters characterizing these groups were in accordance with respective processes associated to production and degradation of SPE-DOC. The trends observed for molecular parameters revealed the imprint of processes related to DOM production and DOM degradation, both biotic (microbial degradation) and abiotic (photodegradation). Our study suggests that even under low productivity conditions like EN, there is an active cycling of the DOM pool off central Peru

Biodegradability of hydrothermally altered deep-sea dissolved organic matter

Deep-sea dissolved organic matter (DOM) constitutes a huge carbon reservoir in the worlds' oceans that – despite its abundance – is virtually unused as a substrate by marine heterotrophs. Heating within hydrothermal systems induces major molecular modifications of deep-sea DOM. Here, we tested the hypothesis that hydrothermal heating of deep-sea DOM enhances bioavailability. Aliquots of DOM extracted from the deep North Pacific (North Equatorial Pacific Intermediate Water; NEqPIW) were re-dissolved in artificial seawater and subjected to temperatures of 100 and 200 °C (40 MPa) using Dickson-type reactors. In agreement with earlier findings we observed a temperature-related drop in dissolved organic carbon (DOC) concentration (−6.1% at 100 °C, −21.0% at 200 °C) that predominantly affected the solid-phase extractable (SPE-DOC) fraction (−18.2% at 100 °C, −51.4% at 200 °C). Fourier-transform ion cyclotron resonance mass spectrometric (FT-ICR-MS) analysis confirmed a temperature-related reduction of average molecular mass, O/C ratios, double bond equivalents (DBE) and a relative increase in aromaticity (AImod). This thermally altered DOM was added (25 μmol L−1 DOC) to deep-water samples from the South West Pacific (Kermadec Arc, RV Sonne / SO253, 32° 37.706′ S | 179° 38.728′ W) and incubated with the prevailing natural microbial community. After 16 days at 4 °C in the dark, prokaryotic cell counts in incubations containing the full spectrum of thermally-degraded DOM (extractable and non-extractable compounds) had increased considerably (on average 21× for DOM100°C and 27× for DOM200°C). In contrast, prokaryotic growth in incubations to which only solid-phase extractable thermally-altered DOM was added was not enhanced compared to control incubations. The experiments demonstrate that temperature-driven degradation of deep-sea recalcitrant DOM within hydrothermal systems turns fractions of it accessible to microbes. The thermally-produced DOM compounds that stimulate microbial growth are not retained on reversed-phase resins (SPE-DOM) and are likely low-molecular mass organic acids. Despite the comprehensive compositional modifications of the solid-phase extractable (SPE-DOM) fraction through heating, it remains inaccessible to microbes at the investigated concentration levels. The microbial incubation resulted in only minor and mostly insignificant overall changes in SPE-DOM molecular composition and concentration

Does the Chemodiversity of Bacterial Exometabolomes Sustain the Chemodiversity of Marine Dissolved Organic Matter?

Marine dissolved organic matter (DOM) is a complex mixture of chemical compounds. At 750 Pg C, it is one of the biggest pools of reduced carbon on Earth. It has been proposed that the diversity of DOM is responsible for its recalcitrance. We hypothesize that the chemodiversity of marine DOM is a reflection of the chemodiversity of bacterial exometabolomes. To test this, we incubated two model strains of the Roseobacter group; Phaeobacter inhibens and Dinoroseobacter shibae in pure culture using three different simple organic compounds as sole carbon sources (glutamate, glucose, and acetate and succinate for P. inhibens and D. shibae, respectively). The exometabolome of the model organisms was characterized using Fourier Transform Ion Cyclotron Resonance Mass Spectrometry (FT-ICR-MS) and ecological diversity measures. We detected thousands of molecular masses in the exometabolomes of P. inhibens and D. shibae (21,105 and 9,386, respectively), reflecting the capability of single bacterial strains to diversify simple organic compounds. The chemical composition of the exometabolomes changed with growth phase and also differed according to the strain incubated and the utilized substrate. We mimicked a higher diversity of substrates, bacterial species and heterogeneous growth (different growth phases) to approach the complexity of natural environments, by computationally creating combinations of detected exometabolomes. We compared the chemodiversity of these combinations, indicative for chemodiversity of freshly produced microbial DOM to that of refractory DOM from one of the oldest oceanic water masses (North Equatorial Pacific Intermediate Water). Some combinations of exometabolomes showed higher richness than the deep ocean refractory DOM, and all the combinations showed higher functional diversity. About 15% of the 13,509 molecular formulae detected in exometabolomes and refractory oceanic DOM were shared, i.e., occurred in Roseobacter exometabolomes and in deep water samples. This overlap provides further support for our hypothesis that marine bacteria from the Roseobacter group contribute to the sustainability of marine DOM chemodiversity and stability

Disentangling Biological Transformations and Photodegradation Processes from Marine Dissolved Organic Matter Composition in the Global Ocean

Dissolved organic matter (DOM) holds the largest amount of organic carbon in the ocean, with most of it residing in the deep for millennia. Specific mechanisms and environmental conditions responsible for its longevity are still unknown. Microbial transformations and photochemical degradation of DOM in the surface layers are two processes that shape its molecular composition. We used molecular data (via Fourier transform ion cyclotron resonance mass spectrometry) from two laboratory experiments that focused on (1) microbial processing of fresh DOM and (2) photodegradation of deep-sea DOM to derive independent process-related molecular indices for biological formation and transformation (Ibio) and photodegradation (Iphoto). Both indices were applied to a global ocean data set of DOM composition. The distributions of Iphoto and Ibio were consistent with increased photodegradation and biological reworking of DOM in sunlit surface waters, and traces of these surface processes were evident at depth. Increased Ibio values in the deep Southern Ocean and South Atlantic implied export of microbially reworked DOM. Photodegraded DOM (increased Iphoto) in the deep subtropical gyres of Atlantic and Pacific oceans suggested advective transport in warm-core eddies. The simultaneous application of Iphoto and Ibio disentangled and assessed two processes that left unique molecular signatures in the global ocean

Carbon assimilating fungi from surface ocean to subseafloor revealed by coupled phylogenetic and stable isotope analysis

Fungi are ubiquitous in the ocean and hypothesized to be important members of marine ecosystems, but their roles in the marine carbon cycle are poorly understood. Here, we use 13C DNA stable isotope probing coupled with phylogenetic analyses to investigate carbon assimilation within diverse communities of planktonic and benthic fungi in the Benguela Upwelling System (Namibia). Across the redox stratified water column and in the underlying sediments, assimilation of 13C-labeled carbon from diatom extracellular polymeric substances (13C-dEPS) by fungi correlated with the expression of fungal genes encoding carbohydrate-active enzymes. Phylogenetic analysis of genes from 13C-labeled metagenomes revealed saprotrophic lineages related to the facultative yeast Malassezia were the main fungal foragers of pelagic dEPS. In contrast, fungi living in the underlying sulfidic sediments assimilated more 13C-labeled carbon from chemosynthetic bacteria compared to dEPS. This coincided with a unique seafloor fungal community and dissolved organic matter composition compared to the water column, and a 100-fold increased fungal abundance within the subseafloor sulfide-nitrate transition zone. The subseafloor fungi feeding on 13C-labeled chemolithoautotrophs under anoxic conditions were affiliated with Chytridiomycota and Mucoromycota that encode cellulolytic and proteolytic enzymes, revealing polysaccharide and protein-degrading fungi that can anaerobically decompose chemosynthetic necromass. These subseafloor fungi, therefore, appear to be specialized in organic matter that is produced in the sediments. Our findings reveal that the phylogenetic diversity of fungi across redox stratified marine ecosystems translates into functionally relevant mechanisms helping to structure carbon flow from primary producers in marine microbiomes from the surface ocean to the subseafloor

Combined Carbohydrates Support Rich Communities of Particle-Associated Marine Bacterioplankton

Carbohydrates represent an important fraction of labile and semi-labile marine organic matter that is mainly comprised of exopolymeric substances derived from phytoplankton exudation and decay. This study investigates the composition of total combined carbohydrates (tCCHO; >1 kDa) and the community development of free-living (0.2–3 μm) and particle-associated (PA) (3–10 μm) bacterioplankton during a spring phytoplankton bloom in the southern North Sea. Furthermore, rates were determined for the extracellular enzymatic hydrolysis that catalyzes the initial step in bacterial organic matter remineralization. Concentrations of tCCHO greatly increased during bloom development, while the composition showed only minor changes over time. The combined concentration of glucose, galactose, fucose, rhamnose, galactosamine, glucosamine, and glucuronic acid in tCCHO was a significant factor shaping the community composition of the PA bacteria. The richness of PA bacteria greatly increased in the post-bloom phase. At the same time, the increase in extracellular β-glucosidase activity was sufficient to explain the observed decrease in tCCHO, indicating the efficient utilization of carbohydrates by the bacterioplankton community during the post-bloom phase. Our results suggest that carbohydrate concentration and composition are important factors in the multifactorial environmental control of bacterioplankton succession and the enzymatic hydrolysis of organic matter during phytoplankton blooms

Aerosols as a source of dissolved black carbon to the ocean

溶解黑碳（dissolved black carbon, DBC）在开阔大洋的年龄可达上万年，是海洋中到目前为止已知的年龄最老、最大的惰性溶解有机碳。因此海洋溶解黑碳的源汇问题是全球碳循环研究的重要部分。该研究团队通过测定2015年春季东、黄海以及西北太平洋的气溶胶中水溶性有机碳(water soluble organic carbon, WSOC)、水溶性黑碳 (water soluble black carbon, WSBC)的含量，并结合超高分辨质谱---傅里叶变换离子回旋共振质谱 (FT-ICR-MS)，解析了海洋气溶胶中溶解黑碳的浓度以及分子组成。进一步分析发现气溶胶中溶解黑碳的浓度与水溶性有机碳浓度显著高度相关。预测的未来生物质燃烧以及沙尘输送的变化可能增加大气沉降溶解黑碳的通量，影响区域甚至全球碳库。该研究结果将有助于把溶解黑碳纳入到海洋碳循环研究模型中。 近海海洋环境科学国家重点实验室助理教授鲍红艳为论文第一作者，高树基教授为共同通讯作者。【Abstract】Dissolved black carbon (DBC) is the largest known slow-cycling organic carbon pool in the world’s oceans. Atmospheric deposition could significantly contribute to the oceanic DBC pool, but respective information is lacking. Here we estimate that, during the dust outbreak season, the atmospheric dry deposition of water-soluble black carbon (WSBC) is～ 40% of the riverine input to the China coastal seas. The molecular composition of atmospheric WSBC determined by ultrahigh-resolution mass spectrometry, reveals similar soil-derived sources as for riverine discharge. WSBC is significantly positively correlated with water-soluble organic carbon (WSOC) in marine aerosols, and water-soluble black carbon contributes on average 2.8 ± 0.65% to the total WSOC. Based on this relationship, the global atmospheric deposition of DBC to the ocean is estimated to be 1.8 ± 0.83 Tg yr−1. Anticipated future changes in biomass burning and dust mobilization might increase these numbers, with consequences for regional ecosystems and global carbon reservoirs.This study was funded by the Major State Basic Research Development Program of China (973 program) (No. 2014CB953702) and the China Postdoctoral Science Foundation (No. 2013M540529). Hanse-Wissenschaftskolleg (the Institute for Advanced Study in Germany) is acknowledged for its support of Dr. H. Bao. 本研究受到973项目“不同营养水平下海洋氮循环关键过程及其对大气物质沉降的响应”（No.2014CB953702）以及中国博士后科学基金（No.2013M540529）的资助； 鲍红艳博士在德国的工作受到德国Hanse-Wissenschaftskolleg奖学金的资助