Coupled RNA-SIP and metatranscriptomics of active chemolithoautotrophic communities at a deep-sea hydrothermal vent

© The International Society for Microbial Ecology, 2016. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in ISME Journal 10 (2016): 1925–1938, doi:10.1038/ismej.2015.258.The chemolithoautotrophic microbial community of the rocky subseafloor potentially provides a large amount of organic carbon to the deep ocean, yet our understanding of the activity and metabolic complexity of subseafloor organisms remains poorly described. A combination of metagenomic, metatranscriptomic, and RNA stable isotope probing (RNA-SIP) analyses were used to identify the metabolic potential, expression patterns, and active autotrophic bacteria and archaea and their pathways present in low-temperature hydrothermal fluids from Axial Seamount, an active submarine volcano. Metagenomic and metatranscriptomic results showed the presence of genes and transcripts for sulfur, hydrogen, and ammonium oxidation, oxygen respiration, denitrification, and methanogenesis, as well as multiple carbon fixation pathways. In RNA-SIP experiments across a range of temperatures under reducing conditions, the enriched 13C fractions showed differences in taxonomic and functional diversity. At 30 °C and 55 °C, Epsilonproteobacteria were dominant, oxidizing hydrogen and primarily reducing nitrate. Methanogenic archaea were also present at 55 °C, and were the only autotrophs present at 80 °C. Correspondingly, the predominant CO2 fixation pathways changed from the reductive tricarboxylic acid (rTCA) cycle to the reductive acetyl-CoA pathway with increasing temperature. By coupling RNA-SIP with meta-omics, this study demonstrates the presence and activity of distinct chemolithoautotrophic communities across a thermal gradient of a deep-sea hydrothermal vent.This work was funded by the Gordon and Betty Moore Foundation Grant GBMF3297 and NSF Center for Dark Energy Biosphere Investigations (C-DEBI) (OCE-0939564). The data collected in this study is based upon work supported by the Schmidt Ocean Institute during cruise FK010-2013 aboard R/V Falkor

Persistent currents in normal metal rings

The authors have measured the magnetic response of 33 individual cold mesoscopic gold rings, one ring at a time. The response of some sufficiently small rings has a component that is periodic in the flux through the ring and is attributed to a persistent current. Its period is close to h/e, and its sign and amplitude vary between rings. The amplitude distribution agrees well with predictions for the typical h/e current in diffusive rings. The temperature dependence of the amplitude, measured for four rings, is also consistent with theory. These results disagree with previous measurements of three individual metal rings that showed a much larger periodic response than expected. The use of a scanning SQUID microscope enabled in situ measurements of the sensor background. A paramagnetic linear susceptibility and a poorly understood anomaly around zero field are attributed to defect spins.Comment: Journal version. 4+ pages, 3 figures. See http://stanford.edu/group/moler/publications.html for the auxiliary document containing additional data and discussion (Ref. 29). Changes w.r.t. v1: Clarified some details in introduction and regarding experimental procedures, shortened abstract, added references and fixed some typo

COBRA: a research accelerator for the crustal ocean biosphere

© The Author(s), 2021. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Huber, J. A., & Orcutt, B. N. COBRA: a research accelerator for the crustal ocean biosphere. Marine Technology Society Journal, 55(3), (2021): 130–130, https://doi.org/10.4031/MTSJ.55.3.14.The deep seafloor covers two-thirds of Earth's surface area, but our understanding of the ecosystems and resources found in the deep ocean, as well as the ability of deep-sea ecosystems to withstand human perturbation, is limited. These deep-sea habitats demand urgent study as there are emergent human uses in the form of deep-sea mining and carbon sequestration that will fundamentally alter physical, chemical, and biological conditions that took millions of years to establish. We propose the international network COBRA, a research accelerator for the crustal ocean biosphere. COBRA will bring together diverse stakeholders and experts, including interdisciplinary academic and government scientists, private institutions, policy makers, data systems engineers, industry experts, and others to coordinate efforts that generate new knowledge and inform decision making about activities that could affect the deep ocean and, by extension, all of society. We will also train the next generation of leaders in ocean exploration, science, and policy through an innovative virtual program to carry this effort into future generations of ocean and earth science research. COBRA will inform policies relating to emergent industrial uses of the deep ocean, decrease the likelihood of serious harm to the environment, and maintain ecosystem services for the benefit of society

Coupled RNA-SIP and metatranscriptomics of active chemolithoautotrophic communities at a deep-sea hydrothermal vent

© The International Society for Microbial Ecology, 2016. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in ISME Journal 10 (2016): 1925–1938, doi:10.1038/ismej.2015.258.The chemolithoautotrophic microbial community of the rocky subseafloor potentially provides a large amount of organic carbon to the deep ocean, yet our understanding of the activity and metabolic complexity of subseafloor organisms remains poorly described. A combination of metagenomic, metatranscriptomic, and RNA stable isotope probing (RNA-SIP) analyses were used to identify the metabolic potential, expression patterns, and active autotrophic bacteria and archaea and their pathways present in low-temperature hydrothermal fluids from Axial Seamount, an active submarine volcano. Metagenomic and metatranscriptomic results showed the presence of genes and transcripts for sulfur, hydrogen, and ammonium oxidation, oxygen respiration, denitrification, and methanogenesis, as well as multiple carbon fixation pathways. In RNA-SIP experiments across a range of temperatures under reducing conditions, the enriched 13C fractions showed differences in taxonomic and functional diversity. At 30 °C and 55 °C, Epsilonproteobacteria were dominant, oxidizing hydrogen and primarily reducing nitrate. Methanogenic archaea were also present at 55 °C, and were the only autotrophs present at 80 °C. Correspondingly, the predominant CO2 fixation pathways changed from the reductive tricarboxylic acid (rTCA) cycle to the reductive acetyl-CoA pathway with increasing temperature. By coupling RNA-SIP with meta-omics, this study demonstrates the presence and activity of distinct chemolithoautotrophic communities across a thermal gradient of a deep-sea hydrothermal vent.This work was funded by the Gordon and Betty Moore Foundation Grant GBMF3297 and NSF Center for Dark Energy Biosphere Investigations (C-DEBI) (OCE-0939564). The data collected in this study is based upon work supported by the Schmidt Ocean Institute during cruise FK010-2013 aboard R/V Falkor

Salt marsh sediment bacterial communities maintain original population structure after transplantation across a latitudinal gradient

© The Author(s), 2018. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in PeerJ 6 (2018): e4735, doi:10.7717/peerj.4735.Dispersal and environmental selection are two of the most important factors that govern the distributions of microbial communities in nature. While dispersal rates are often inferred by measuring the degree to which community similarity diminishes with increasing geographic distance, determining the extent to which environmental selection impacts the distribution of microbes is more complex. To address this knowledge gap, we performed a large reciprocal transplant experiment to simulate the dispersal of US East Coast salt marsh Spartina alterniflora rhizome-associated microbial sediment communities across a latitudinal gradient and determined if any shifts in microbial community composition occurred as a result of the transplantation. Using bacterial 16S rRNA gene sequencing, we did not observe large-scale changes in community composition over a five-month S. alterniflora summer growing season and found that transplanted communities more closely resembled their origin sites than their destination sites. Furthermore, transplanted communities grouped predominantly by region, with two sites from the north and three sites to the south hosting distinct bacterial taxa, suggesting that sediment communities transplanted from north to south tended to retain their northern microbial distributions, and south to north maintained a southern distribution. A small number of potential indicator 16S rRNA gene sequences had distributions that were strongly correlated to both temperature and nitrogen, indicating that some organisms are more sensitive to environmental factors than others. These results provide new insight into the microbial biogeography of salt marsh sediments and suggest that established bacterial communities in frequently-inundated environments may be both highly resistant to invasion and resilient to some environmental shifts. However, the extent to which environmental selection impacts these communities is taxon specific and variable, highlighting the complex interplay between dispersal and environmental selection for microbial communities in nature.This research was conducted in the National Estuarine Research Reserve System under an award from the Estuarine Reserves Division, Office of Ocean and Coastal Resource Management, National Ocean Service, and National Oceanic and Atmospheric Administration. Support was also provided through funding to Julie Huber from a Brown-MBL Partnership SEED award, the Neal Cornell Endowed Research Fund, and the NSF Center for Dark Energy Biosphere Investigations (C-DEBI) (OCE-0939564). Additional funding was provided to Sarah Corman-Crosby by the National Park Service George Melendez Wright Climate Change Fellowship

Active subseafloor microbial communities from Mariana back-arc venting fluids share metabolic strategies across different thermal niches and taxa

© The Author(s), 2019. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Trembath-Reichert, E., Butterfield, D. A., & Huber, J. A. Active subseafloor microbial communities from Mariana back-arc venting fluids share metabolic strategies across different thermal niches and taxa. Isme Journal, 13(9), (2019): 2264-2279, doi: 10.1038/s41396-019-0431-y.There are many unknowns regarding the distribution, activity, community composition, and metabolic repertoire of microbial communities in the subseafloor of deep-sea hydrothermal vents. Here we provide the first characterization of subseafloor microbial communities from venting fluids along the central Mariana back-arc basin (15.5–18°N), where the slow-spreading rate, depth, and variable geochemistry along the back-arc distinguish it from other spreading centers. Results indicated that diverse Epsilonbacteraeota were abundant across all sites, with a population of high temperature Aquificae restricted to the northern segment. This suggests that differences in subseafloor populations along the back-arc are associated with local geologic setting and resultant geochemistry. Metatranscriptomics coupled to stable isotope probing revealed bacterial carbon fixation linked to hydrogen oxidation, denitrification, and sulfide or thiosulfate oxidation at all sites, regardless of community composition. NanoSIMS (nanoscale secondary ion mass spectrometry) incubations at 80 °C show only a small portion of the microbial community took up bicarbonate, but those autotrophs had the highest overall rates of activity detected across all experiments. By comparison, acetate was more universally utilized to sustain growth, but within a smaller range of activity. Together, results indicate that microbial communities in venting fluids from the Mariana back-arc contain active subseafloor communities reflective of their local conditions with metabolisms commonly shared across geologically disparate spreading centers throughout the ocean.This work was funded by the NOAA Ocean Exploration and Research (OER) Program, the NSF Center for Dark Energy Biosphere Investigations (C-DEBI) (OCE-0939564), and NOAA/PMEL and JISAO under NOAA Cooperative Agreement NA15OAR4320063. ETR was supported by a NASA Postdoctoral Fellowship with the NASA Astrobiology Institute and a L’Oréal USA For Women in Science Fellowship. The data collected in this study includes work supported by the Schmidt Ocean Institute during cruise FK161129 aboard R/V Falkor. We thank the captains and crews of the R/V Falkor and ROV SuBastian. Critical support in cruise planning and sampling at sea was carried out by Andra Bobbitt, Bill Chadwick, Bob Embley, Ben Larson, and Kevin Roe. Caroline Fortunato, Connor Skennerton, Rika Anderson, Karthik Anantharaman, Jaclyn Saunders, Hank Yu, Lewis Ward, Elaina Graham, and Ben Tully aided bioinformatics pipeline development and Victoria Orphan and Yunbin Guan aided with NanoSIMS analysis. This is C-DEBI Contribution 470, JISAO Contribution 2018-0173, and PMEL Contribution 4867

A Terraced Scanning Superconducting Quantum Interference Device Susceptometer with Sub-Micron Pickup Loops

Superconducting Quantum Interference Devices (SQUIDs) can have excellent spin sensitivity depending on their magnetic flux noise, pick-up loop diameter, and distance from the sample. We report a family of scanning SQUID susceptometers with terraced tips that position the pick-up loops 300 nm from the sample. The 600 nm - 2 um pickup loops, defined by focused ion beam, are integrated into a 12-layer optical lithography process allowing flux-locked feedback, in situ background subtraction and optimized flux noise. These features enable a sensitivity of ~70 electron spins per root Hertz at 4K.Comment: See http://stanford.edu/group/moler/publications.html for an auxiliary document containing additional fabrication details and discussio

Microbial Communities Are Well Adapted to Disturbances in Energy Input

Within the broader ecological context, biological communities are often viewed as stable and as only experiencing succession or replacement when subject to external perturbations, such as changes in food availability or the introduction of exotic species. Our findings indicate that microbial communities can exhibit strong internal dynamics that may be more important in shaping community succession than external drivers. Dynamic “unstable” communities may be important for ecosystem functional stability, with rare organisms playing an important role in community restructuring. Understanding the mechanisms responsible for internal community dynamics will certainly be required for understanding and manipulating microbiomes in both host-associated and natural ecosystems. Although microbial systems are well suited for studying concepts in ecological theory, little is known about how microbial communities respond to long-term periodic perturbations beyond diel oscillations. Taking advantage of an ongoing microcosm experiment, we studied how methanotrophic microbial communities adapted to disturbances in energy input over a 20-day cycle period. Sequencing of bacterial 16S rRNA genes together with quantification of microbial abundance and ecosystem function were used to explore the long-term dynamics (510 days) of methanotrophic communities under continuous versus cyclic chemical energy supply. We observed that microbial communities appeared inherently well adapted to disturbances in energy input and that changes in community structure in both treatments were more dependent on internal dynamics than on external forcing. The results also showed that the rare biosphere was critical to seeding the internal community dynamics, perhaps due to cross-feeding or other strategies. We conclude that in our experimental system, internal feedbacks were more important than external drivers in shaping the community dynamics over time, suggesting that ecosystems can maintain their function despite inherently unstable community dynamics. IMPORTANCE Within the broader ecological context, biological communities are often viewed as stable and as only experiencing succession or replacement when subject to external perturbations, such as changes in food availability or the introduction of exotic species. Our findings indicate that microbial communities can exhibit strong internal dynamics that may be more important in shaping community succession than external drivers. Dynamic “unstable” communities may be important for ecosystem functional stability, with rare organisms playing an important role in community restructuring. Understanding the mechanisms responsible for internal community dynamics will certainly be required for understanding and manipulating microbiomes in both host-associated and natural ecosystems.We are grateful for support from the National Science Foundation (grants EF-0928742 to J.J.V. and J.A.H. and OCE-1238212 to J.J.V.).S