Diel investments in metabolite production and consumption in a model microbial system

Organic carbon transfer between surface ocean photosynthetic and heterotrophic microbes is a central but poorly understood process in the global carbon cycle. In a model community in which diatom extracellular release of organic molecules sustained growth of a co-cultured bacterium, we determined quantitative changes in the diatom endometabolome and the bacterial uptake transcriptome over two diel cycles. Of the nuclear magnetic resonance (NMR) peaks in the diatom endometabolites, 38% had diel patterns with noon or mid-afternoon maxima; the remaining either increased (36%) or decreased (26%) through time. Of the genes in the bacterial uptake transcriptome, 94% had a diel pattern with a noon maximum; the remaining decreased over time (6%). Eight diatom endometabolites identified with high confidence were matched to the bacterial genes mediating their utilization. Modeling of these coupled inventories with only diffusion-based phytoplankton extracellular release could not reproduce all the patterns. Addition of active release mechanisms for physiological balance and bacterial recognition significantly improved model performance. Estimates of phytoplankton extracellular release range from only a few percent to nearly half of annual net primary production. Improved understanding of the factors that influence metabolite release and consumption by surface ocean microbes will better constrain this globally significant carbon flux

Temporal variations in concentrations of dissolved combined and free amino acids during an autumnal phytoplankton bloom in the Chukchi Shelf, western Arctic Ocean

第6回極域科学シンポジウム分野横断セッション：[IA] 急変する北極気候システム及びその全球的な影響の総合的解明―GRENE北極気候変動研究事業研究成果報告2015―11月19日（木）　国立極地研究所１階交流アトリウ

Growth-stage-related shifts in phytoplankton endometabolome composition set the stage for bacterial heterotrophy

Phytoplankton-derived metabolites fuel a large fraction of heterotrophic bacterial production in the global ocean, yet methodological challenges have limited our understanding of the organic molecules transferred between these microbial groups. In an experimental bloom study consisting of three heterotrophic marine bacteria growing together with the diatom Thalassiosira pseudonana, we concurrently measured diatom endometabolites (i.e., potential exometabolite supply) by nuclear magnetic resonance (NMR) spectroscopy and bacterial gene expression (i.e., potential exometabolite uptake) by metatranscriptomic sequencing. Twenty-two diatom endometabolites were annotated, with nine increasing in internal concentration in the late stage of the bloom, eight decreasing, and five showing no variation through the bloom progression. Some metabolite changes could be linked to shifts in diatom gene expression, as well as to shifts in bacterial community composition and their expression of substrate uptake and catabolism genes. Yet an overall low match indicated that endometabolome concentration was not a good predictor of exometabolite availability, and that complex physiological and ecological interactions underlie metabolite exchange. Six diatom endometabolites accumulated to higher concentrations in the bacterial co-cultures compared to axenic cultures, suggesting a bacterial influence on rates of synthesis or release of glutamate, arginine, leucine, 2,3-dihydroxypropane-1-sulfonate, glucose, and glycerol-3-phosphate. Better understanding of phytoplankton metabolite production, release, and transfer to assembled bacterial communities is key to untangling this nearly invisible yet pivotal step in ocean carbon cycling

NMR and Metabolomics—A Roadmap for the Future

Metabolomics investigates global metabolic alterations associated with chemical, biological, physiological, or pathological processes. These metabolic changes are measured with various analytical platforms including liquid chromatography-mass spectrometry (LC-MS), gas chromatographymass spectrometry (GC-MS) and nuclear magnetic resonance spectroscopy (NMR). While LC-MS methods are becoming increasingly popular in the field of metabolomics (accounting for more than 70% of published metabolomics studies to date), there are considerable benefits and advantages to NMR-based methods for metabolomic studies. In fact, according to PubMed, more than 926 papers on NMR-based metabolomics were published in 2021—the most ever published in a given year. This suggests that NMR-based metabolomics continues to grow and has plenty to offer to the scientific community. This perspective outlines the growing applications of NMR in metabolomics, highlights several recent advances in NMR technologies for metabolomics, and provides a roadmap for future advancements

NMR and Metabolomics—A Roadmap for the Future

Metabolomics investigates global metabolic alterations associated with chemical, biological, physiological, or pathological processes. These metabolic changes are measured with various analytical platforms including liquid chromatography-mass spectrometry (LC-MS), gas chromatography-mass spectrometry (GC-MS) and nuclear magnetic resonance spectroscopy (NMR). While LC-MS methods are becoming increasingly popular in the field of metabolomics (accounting for more than 70% of published metabolomics studies to date), there are considerable benefits and advantages to NMR-based methods for metabolomic studies. In fact, according to PubMed, more than 926 papers on NMR-based metabolomics were published in 2021—the most ever published in a given year. This suggests that NMR-based metabolomics continues to grow and has plenty to offer to the scientific community. This perspective outlines the growing applications of NMR in metabolomics, highlights several recent advances in NMR technologies for metabolomics, and provides a roadmap for future advancements

Low production and slow turnover of heterotrophic microbes in the deep water of the Canada Basin, western Arctic

Data on microbial metabolic activities and biomass in deep oceans have begun to provide new insights into regional variability in the extent of and patterns in vertical organic carbon delivery in the ocean’s interior. However, such information is scarce in the polar region, especially in the major Arctic basins. Here we examined full-depth distributions of heterotrophic prokaryote production and biomass in the Canada Basin, western Arctic. Prokaryote production was high in the Pacific-origin water masses located in the upper-mesopelagic layer (100-200 m). In the deeper layer (300-3,000 m), the depth-integrated prokaryote production was low relative to other oceanic regions, with the estimated turnover time of 6 years in the bathypelagic layer. This turnover time was among the longest values reported in lower latitude oceans. Despite the low production and slow turnover, the estimated prokaryote carbon consumption far exceeded (38 fold) the reported sinking particulate organic carbon fluxes at depths of 120-200 m. This large apparent carbon imbalance may be partly explained by the shelf-basin exchange of organic carbon due to lateral intrusion of Pacific-origin water masses.Oral presentation abstract SS58, Association for the Sciences of Limnology and Oceanography (ASLO) 2013 Aquatic Science Meeting (17-22 February 2013, New Orleans, Louisiana

Bacterial Substrate Transformation Tracked by Stable-Isotope-Guided NMR Metabolomics: Application in a Natural Aquatic Microbial Community

The transformation of organic substrates by heterotrophic bacteria in aquatic environments constitutes one of the key processes in global material cycles. The development of procedures that would enable us to track the wide range of organic compounds transformed by aquatic bacteria would greatly improve our understanding of material cycles. In this study, we examined the applicability of nuclear magnetic resonance spectroscopy coupled with stable-isotope labeling to the investigation of metabolite transformation in a natural aquatic bacterial community. The addition of a model substrate (13C6–glucose) to a coastal seawater sample and subsequent incubation resulted in the detection of >200 peaks and the assignment of 22 metabolites from various chemical classes, including amino acids, dipeptides, organic acids, nucleosides, nucleobases, and amino alcohols, which had been identified as transformed from the 13C6–glucose. Additional experiments revealed large variability in metabolite transformation and the key compounds, showing the bacterial accumulation of glutamate over the incubation period, and that of 3-hydroxybutyrate with increasing concentrations of 13C6–glucose added. These results suggest the potential ability of our approach to track substrate transformation in aquatic bacterial communities. Further applications of this procedure may provide substantial insights into the metabolite dynamics in aquatic environments

Coupled Response of Bacterial Production to a Wind-induced Fall Phytoplankton Bloom and Sediment Resuspension in the Chukchi Sea Shelf, Western Arctic Ocean

Heterotrophic bacterial abundance and production, dissolved free amino acid (DFAA) and dissolved combined amino acid (DCAA) concentrations, and other microbial parameters were determined for seawater samples collected at a fixed station (maximum water depth, 56 m) deployed on the Chukchi Sea Shelf, in the western Arctic Ocean, during a 16-day period in September 2013. During the investigation period, the sampling station experienced strong winds and a subsequent phytoplankton bloom, which was thought to be triggered by enhanced vertical mixing and upward nutrient fluxes. In this study, we investigated whether bacterial and dissolved amino acid parameters changed in response to these physical and biogeochemical events. Bacterial abundance and production in the upper layer increased with increasing chlorophyll a concentration, despite a concomitant decrease in seawater temperature from 3.2°C to 1.5°C. The percentage of bacteria with high nucleic acid content during the bloom was significantly higher than that during the prebloom period. The ratio of the depth-integrated (0–20 m) bacterial production to primary production differed little between the prebloom and bloom period, with an overall average value of 0.14 ± 0.03 (± standard deviation, n = 8). DFAA and DCAA concentrations varied over a limited range throughout the investigation, indicating that the supply and consumption of labile dissolved amino acids were balanced. These results indicate that there was a tightly coupled, large flow of organic carbon from primary producers to heterotrophic bacteria during the fall bloom. Our data also revealed that bacterial production and abundance were high in the bottom nepheloid (low transmittance) layer during strong wind events, which was associated with sediment resuspension due to turbulence near the seafloor. The impacts of fall wind events, which are predicted to become more prominent with the extension of the ice-free period, on bacterial processes and the dynamics of organic matter in the Chukchi Sea Shelf could have far-reaching influences on biogeochemical cycles and ecosystem dynamics in broader regions of the Arctic Ocean

Ecological drivers of bacterial community assembly in synthetic phycospheres

In the nutrient-rich region surrounding marine phytoplankton cells, heterotrophic bacterioplankton transform a major fraction of recently fixed carbon through the uptake and catabolism of phytoplankton metabolites. We sought to understand the rules by which marine bacterial communities assemble in these nutrient-enhanced phycospheres, specifically addressing the role of host resources in driving community coalescence. Synthetic systems with varying combinations of known exometabolites of marine phytoplankton were inoculated with seawater bacterial assemblages, and communities were transferred daily to mimic the average duration of natural phycospheres. We found that bacterial community assembly was predictable from linear combinations of the taxa maintained on each individual metabolite in the mixture, weighted for the growth each supported. Deviations from this simple additive resource model were observed but also attributed to resource-based factors via enhanced bacterial growth when host metabolites were available concurrently. The ability of photosynthetic hosts to shape bacterial associates through excreted metabolites represents a mechanism by which microbiomes with beneficial effects on host growth could be recruited. In the surface ocean, resource-based assembly of host-associated communities may underpin the evolution and maintenance of microbial interactions and determine the fate of a substantial portion of Earth’s primary production.Simons Foundation (Grant 542385

Bacterial Substrate Transformation Tracked by Stable-Isotope-Guided NMR Metabolomics: Application in a Natural Aquatic Microbial Community

The transformation of organic substrates by heterotrophic bacteria in aquatic environments constitutes one of the key processes in global material cycles. The development of procedures that would enable us to track the wide range of organic compounds transformed by aquatic bacteria would greatly improve our understanding of material cycles. In this study, we examined the applicability of nuclear magnetic resonance spectroscopy coupled with stable-isotope labeling to the investigation of metabolite transformation in a natural aquatic bacterial community. The addition of a model substrate (13C6–glucose) to a coastal seawater sample and subsequent incubation resulted in the detection of >200 peaks and the assignment of 22 metabolites from various chemical classes, including amino acids, dipeptides, organic acids, nucleosides, nucleobases, and amino alcohols, which had been identified as transformed from the 13C6–glucose. Additional experiments revealed large variability in metabolite transformation and the key compounds, showing the bacterial accumulation of glutamate over the incubation period, and that of 3-hydroxybutyrate with increasing concentrations of 13C6–glucose added. These results suggest the potential ability of our approach to track substrate transformation in aquatic bacterial communities. Further applications of this procedure may provide substantial insights into the metabolite dynamics in aquatic environments