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At least 1,063 records · Page 59

Elucidating algal-bacterial community interactions by tracking volatile biomarkers (Final Report)

This feasibility study project aimed to lay further groundwork for studying and understanding how microorganisms interact with each other at the molecular level using model algal-bacterial co-cultures. A better understanding of such interactions is critical to optimizing and operating biotic platforms for a number of applications, including bioenergy, algal bioproducts and agriculture, and carbon capture. Here we explored the feasibility of characterizing and following these interactions by identifying and tracking volatile metabolites and biomarkers in the complex gaseous headspace of such cultures. We set up an experimental platform using a co-culture of Phaeodactylum tricornutum and Marinobacter subspecies 3-2 as a model system, used solid-phase microextraction fibers for volatile collection at various time points throughout culture growth, and employed gas chromatography mass spectrometry-based instrumentation available at LLNL for compound identification and quantification. We found that it is feasible to determine volatile profiles from the growth of Phaeodactylum tricornutum and Marinobacter subspecies 3-2 using our methodology and monitor profile changes over time. We also observed that volatiles distinguishing between cultures of the individual species and the co-cultures could be detected. We expect that results of this project will feed into the ongoing LLNL’s DOE BER funded Biofuels Scientific Focus Area (SFA) on algal-bacterial interactions and pave the way for future expanded volatile studies supported by DOE. The approach and technologies developed here would also be applicable and transferable to other microbial communities under study, such as cyanobacteria, rhizosphere communities and biofilms.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Microbial astronauts: assembling microbial communities for advanced life support systems

Extension of human habitation into space requires that humans carry with them many of the microorganisms with which they coexist on Earth. The ubiquity of microorganisms in close association with all living things and biogeochemical processes on Earth predicates that they must also play a critical role in maintaining the viability of human life in space. Even though bacterial populations exist as locally adapted ecotypes, the abundance of individuals in microbial species is so large that dispersal is unlikely to be limited by geographical barriers on Earth (i.e., for most environments "everything is everywhere" given enough time). This will not be true for microbial communities in space where local species richness will be relatively low because of sterilization protocols prior to launch and physical barriers between Earth and spacecraft after launch. Although community diversity will be sufficient to sustain ecosystem function at the onset, richness and evenness may decline over time such that biological systems either lose functional potential (e.g., bioreactors may fail to reduce BOD or nitrogen load) or become susceptible to invasion by human-associated microorganisms (pathogens) over time. Research at the John F. Kennedy Space Center has evaluated fundamental properties of microbial diversity and community assembly in prototype bioregenerative systems for NASA Advanced Life Support. Successional trends related to increased niche specialization, including an apparent increase in the proportion of nonculturable types of organisms, have been consistently observed. In addition, the stability of the microbial communities, as defined by their resistance to invasion by human-associated microorganisms, has been correlated to their diversity. Overall, these results reflect the significant challenges ahead for the assembly of stable, functional communities using gnotobiotic approaches, and the need to better define the basic biological principles that define ecosystem processes in the space environment. Copyright 2004 Springer-Verlag.

Review↗

Spaceflight Biospecimen and Data Sharing in Support of Science Discovery and Exploration

For decades, NASA and international partners have conducted biological experiments in space to understand effects of spaceflight and address potential hazards. To enable spaceflight back to the Moon, and then to Mars and beyond, it is imperative to further understand basic science and health risks associated with spaceflight, along with developing countermeasures. The sending of experiments and organisms into space is a costly endeavor. To maximize scientific return, sharing with the scientific community both space-flown biospecimens and data from completed experiments is essential. New fundamental, applied, and bioinformatic science insights can be gained from specimen and data sharing efforts. Data reuse enables spaceflight health risk modeling, analyzing adverse outcomes across spaceflight hazards, and deep space autonomous support for the flight medical officer. Space-flown biospecimens not required by mission Principal Investigators are regularly archived and made available for scientific request. The largest biorepository of these samples are found within NASA’s Institutional Scientific Collection at Ames Research Center (ISC-ARC), which stores over 32,000 specimens mostly from Shuttle and International Space Station (ISS) missions, but also some ground-based analog samples. The Ames Life Sciences Data Archive manages the ISC-ARC. Tissues are predominantly from mice and rats, though samples are also available from bacteria and quail. Only a handful of other similar collections exist worldwide. Rodent biospecimens exposed to simulated space radiation at Brookhaven National Laboratory are archived under the purview of NASA HRP Space Radiation Element. Microbial collection and analyses from 20 years of routine environmental monitoring of air, surfaces, and water systems of the ISS were performed to ensure a safe environment for astronauts. Samples from the ISC-ARC, space radiation and microbial collections are searchable and requestable through the NASA Life Sciences Data Archive (LSDA). Decades of planetary protection microbial isolates derived from spacecraft bioburden are archived in JPL’s microbial collection. Rodent biospecimens from spaceflight investigations conducted by the Japan Aerospace Exploration Agency (JAXA) are archived and available at the JAXA Biorepository at Tsukuba Space Center. The Russian Institute of Biomedical Problems also has a collection of animal, microbial, cellular, and fungi available for research from ground analog experiments. Several data repositories exist for scientists to utilize. The LSDA is the primary NASA source of life sciences research data and information. It contains decades of spaceflight and ground-analog research involving human, microbial, cellular, plant, and animal subjects. Data is collected from NASA-funded investigations through the Human Research Program and the Space Biology Program. The NASA Lifetime Surveillance of Astronaut Health collects and grants access to clinical and occupational health monitoring data from astronauts, with a list and description of data collected available for request through the LSDA. NASA GeneLab at ARC collects genomic, transcriptomic, proteomic, and metabolomic data from any species. It is a repository and platform for collaborative open-science bioinformatic approaches. JAXA is establishing an ‘omics-based repository in collaboration with the Tohoku Medical Megabank (ToMMo), called the JAXA-ToMMo Integrated Biobank for Space Life Science. Overall, the sharing of these biospecimen and data resources can assist researchers worldwide in understanding spaceflight effects on biology, along with enabling next generation data science applications for space exploration platforms. Websites: https://lsda.jsc.nasa.gov/ ; https://www.nasa.gov/ames/research/space-biosciences/isc-bsp ; https://www.nasa.gov/ames/research/space-biosciences/alsda

Ryan T. Scott↗

Reduce Revenue Versus Increase Expenditure: Fires and Plant Invasion Drive Soil Carbon Loss With Different Mechanisms in a Mediterranean Shrubland

Fires and plant invasions threaten Mediterranean ecosystems substantially, particularly in the context of changing climate. Our study utilized a data-model integration approach to assess the response of soil organic carbon (SOC) to fires and plant invasion under three Shared Socio-Economic Pathway (SSP) scenarios (SSP1-26, SSP2-45, and SSP5-85). We parameterized the CLM-Microbe model and then investigated the individual and interactive impacts of fires and plant invasion on soil C by comparing factorial simulations of initialization (fire/no wildfire in 2021), fire module on/off, and with and without plant invasion during 2023–2100 in a Mediterranean ecosystem. The simulations indicated a marked C loss due to the 2021 wildfire, projected fires, and plant invasion across all future climate scenarios. Specifically, the 2021 wildfire, projected fires, and plant invasion reduced the SOC (0–30 cm) by 0.12, 0.26, and 0.15 kg C m −2 under SSP1-26, 0.12, 0.30, and 0.12 kg C m −2 under SSP2-45, and 0.12, 0.24, and 0.13 kg C m −2 under SSP5-85, respectively. However, fires and plant invasion decreased SOC through distinct mechanisms. The effects of the 2021 wildfire occurred due to its negative legacy on soil microbial community and, thus, litter accumulation, suppressing the formation of soil carbon via decomposition. Influences of projected fires happen via consuming fuel and suppressing carbon input to soils. In contrast, the impacts of plant invasions were due to enhanced microbial respiration, leading to C loss. In conclusion, these findings emphasize the need for tailored C sequestration strategies considering the disparate effects of fires and plant invasions in the Mediterranean climate.

microbe↗

Nitrogen-Fixing Gamma Proteobacteria Azotobacter vinelandii—A Blueprint for Nitrogen-Fixing Plants?

The availability of fixed nitrogen limits overall agricultural crop production worldwide. The so-called modern “green revolution” catalyzed by the widespread application of nitrogenous fertilizer has propelled global population growth. It has led to imbalances in global biogeochemical nitrogen cycling, resulting in a “nitrogen problem” that is growing at a similar trajectory to the “carbon problem”. As a result of the increasing imbalances in nitrogen cycling and additional environmental problems such as soil acidification, there is renewed and increasing interest in increasing the contributions of biological nitrogen fixation to reduce the inputs of nitrogenous fertilizers in agriculture. Interestingly, biological nitrogen fixation, or life’s ability to convert atmospheric dinitrogen to ammonia, is restricted to microbial life and not associated with any known eukaryotes. It is not clear why plants never evolved the ability to fix nitrogen and rather form associations with nitrogen-fixing microorganisms. Perhaps it is because of the large energy demand of the process, the oxygen sensitivity of the enzymatic apparatus, or simply failure to encounter the appropriate selective pressure. Whatever the reason, it is clear that this ability of crop plants, especially cereals, would transform modern agriculture once again. Successfully engineering plants will require creating an oxygen-free niche that can supply ample energy in a tightly regulated manner to minimize energy waste and ensure the ammonia produced is assimilated. Nitrogen-fixing aerobic bacteria can perhaps provide a blueprint for engineering nitrogen-fixing plants. This short review discusses the key features of robust nitrogen fixation in the model nitrogen-fixing aerobe, gamma proteobacteria Azotobacter vinelandii, in the context of the basic requirements for engineering nitrogen-fixing plants.

Microbiology↗

Metabolic Potential and Activity in Fluids of the Coast Range Ophiolite Microbial Observatory, California, USA

Metabolic potential and activity associated with hydrogen and carbon monoxide were characterized in fluids sampled from the the Coast Range Ophiolite Microbial Observatory (CROMO). CROMO consists of two clusters of science-dedicated wells drilled to varying depths up to 35m in the actively serpentinizing, Jurassic-age Coast Range Ophiolite of Northern California, along with a suite of pre-existing monitoring wells at the same site. Consistent with the fluid chemistry observed in other serpentinizing systems, CROMO fluids are highly alkaline, with pH up to 12.5, high in methane, with concentrations up 1600 micromolar, and low in dissolved inorganic carbon (DIC), with concentrations of 10's to 100's of micromolar. CROMO is conspicuous for fluid H2 concentrations that are consistently sub-micromolar, orders of magnitude lower than is typical of other systems. However, higher H2 concentrations (10's -100's of micromolar) at an earlier stage of fluid chemical evolution are predicted by, or consistent with: thermodynamic models for fluid chemistry based on parent rock composition equivalent to local peridotite and with water:rock ratio constrained by observed pH; the presence of magnetite at several wt% in CROMO drill cores; and concentrations of formate and carbon monoxide that would require elevated H2 if formed in equilibrium with H2 and DIC. Calculated Gibbs energy changes for reaction of H2 and CO in each of several metabolisms, across the range of fluid composition encompassed by the CROMO wells, range from bioenergetically feasible (capable of driving ATP synthesis) to thermodynamically unfavorable. Active consumption relative to killed controls was observed for both CO and H2 during incubation of fluids from the pre-existing monitoring wells; in incubations of freshly cored solids, consumption was only observed in one sample set (corresponding to the lowest pH) out of three. The specific metabolisms by which H2 and CO are consumed remain to be determined.

Hydrogen↗

Interplay of microbial communities with mineral environments in coralline algae

Coralline algae are worldwide carbonate builders, considered to be foundational species and biodiversity hotspots. Coralline habitats face increasing pressure from human activities and effects related to Global Change, yet their ecological properties and adaptive responses remain poorly understood. The relationships of the algal microbiota with the mineral bioconstructions, as well as plasticity and resilience of coralline holobionts in a changing environment, are of particular interest. In the Gulf of California, Neogoniolithon trichotomum (Rhodophyta) is the main carbonate builder in tidal pools. We performed a multi-disciplinary assessment of the N. trichotomum microstructure using XRD, SEM microscopy and SR-FTIR spectromicroscopy. In the algal perithallus, magnesium-calcite and aragonite were spatially segregated and embedded in a polysaccharide matrix (rich in sulfated polysaccharides). Mg-calcites (18-19 mol% Mg) were the main mineral components of the thallus overall, followed by iron carbonates related to dolomite (ankerite) and siderite. Minerals of late evaporitic sequences (sylvite and bischofite) were also present, suggesting potential halophilic microenvironments within the algal thalli. The diverse set of abundant halophilic, halotolerant and oligotrophic taxa, whose abundance increase in the summer, further suggests this condition. We created an integrated model, based on environmental parameters and the microbiota distribution, that identified temperature and nutrient availability (particularly nitrate and silicate) as the main parameters related to specific taxa patterns. Among these, Hahella, Granulossicoccus, Ferrimonas, Spongiibacteraceae and cyanobacterial Xenococcaceae and Nostocaceae change significantly between seasons. These bacterial components might play relevant roles in algal plasticity and adaptive responses to a changing environment. This study contributes to the understanding of the interplay of the prokaryotic microbiota with the mineral microenvironments of coralline algae. Because of their carbonates with potential resistance to dissolution in a higher pCO 2 world and their seasonally dynamic bacteria, coralline algae are relevant targets to study coastal resilience and carbonated systems responses to changing environments.

54 ENVIRONMENTAL SCIENCES↗

Heterotrophic respiration by soil microbes in a changing climate

Soil microbes strongly influence the soil organic carbon (SOC) pool, which globally stores ~2,000 PgC. Specifically, the balance between microbial heterotrophic respiration (R H ), which degrades SOC, and plant–microbe interactions that stabilize SOC determines whether terrestrial ecosystems are a net source or sink of CO 2 to the atmosphere. Here, in this Review, we evaluate how climate change alters these competing processes. R H is approximately half of total soil respiration, at ~50 PgC yr −1 , with 70% occurring in topsoils. Warming accelerates microbial metabolism, with a 10 °C temperature increase estimated to raise R H by ~50%, an effect that is particularly strong in Arctic soils. Warming also reduces soil moisture, further modulating R H , which responds nonlinearly to soil moisture, being limited by saturation and desiccation and meeting a maximum at intermediate levels. Consequently, R H is highly sensitive to future precipitation changes and drought. However, soil management strategies could enhance SOC stocks and persistence under climate change. Bacterial and fungal inoculants can promote SOC production and stabilization, while deep-rooting plants increase SOC inputs to deeper layers that experience lower R H . Agricultural practices and biochar amendments can also enhance SOC and reduce R H . Expanding field trials across regions, climates and soil types would improve empirical understanding of these responses and support better representation of R H in predictive models, enabling more accurate assessments of climate impacts on SOC storage.

Jansson, Janet K. [Pacific Northwest National Labo↗

Disentangling direct from indirect relationships in association networks

Networks are vital tools for understanding and modeling interactions in complex systems in science and engineering, and direct and indirect interactions are pervasive in all types of networks. However, quantitatively disentangling direct and indirect relationships in networks remains a formidable task. Here, we present a framework, called iDIRECT (Inference of Direct and Indirect Relationships with Effective Copula-based Transitivity), for quantitatively inferring direct dependencies in association networks. Using copula-based transitivity, iDIRECT eliminates/ameliorates several challenging mathematical problems, including ill-conditioning, self-looping, and interaction strength overflow. With simulation data as benchmark examples, iDIRECT showed high prediction accuracies. Application of iDIRECT to reconstruct gene regulatory networks in Escherichia coli also revealed considerably higher prediction power than the best-performing approaches in the DREAM5 (Dialogue on Reverse Engineering Assessment and Methods project, #5) Network Inference Challenge. In addition, applying iDIRECT to highly diverse grassland soil microbial communities in response to climate warming showed that the iDIRECT-processed networks were significantly different from the original networks, with considerably fewer nodes, links, and connectivity, but higher relative modularity. Further analysis revealed that the iDIRECT-processed network was more complex under warming than the control and more robust to both random and target species removal ( P < 0.001). As a general approach, iDIRECT has great advantages for network inference, and it should be widely applicable to infer direct relationships in association networks across diverse disciplines in science and engineering.

59 BASIC BIOLOGICAL SCIENCES↗

A Science-Focused Artificial Intelligence (AI) Responding in Real-Time to New Information: Capability Demonstration for Ocean World Missions

Introduction: Artificial intelligence (AI) has long been considered a potential mechanism to explore increasingly challenging environments, including those with extreme temperatures and pressures, limited communication capabilities, or those with demanding terrain. We posit that missions in extreme environments could deploy an onboard AI focused on science observations and goals in order to augment a traditional concept(s) of operations (ConOps). An onboard AI capability could perform functions such as data analysis in order to make high-level decisions, including prioritized data transmission for analysis by ground-based teams or autonomously-guided follow-on analyses that maximize science return. Such a capability would empower missions to respond to scientific data of interest in real-time; a mission could make observations and perform a preliminary analysis to alert ground-based scientists to an observation of interest, enabling an informed, rapid response from Earth-based teams. Enceladus Case Study for Onboard AI: We are developing an onboard AI capability for real-time telemetry response that formulates and carries-out informed decisions in service to established mission goals, enabling increased science return of a mission. We focus our AI development for use on a constellation of SmallSats orbiting Enceladus. Our Enceladus case study tests autonomous decision-making capabilities in scenarios with complex orbital dynamics, plume ejecta, extreme cold environments, power restrictions, and a requirement to maximize science return for a potential positive detection of life, while critically evaluating the potential for false positives. Telemetry includes simulated scientific data, spacecraft onboard operational data (e.g., position, velocity, and rotation), and engineering hardware performance data. Enceladus SmallSat Constellation. Our constellation includes eight SmallSat spacecraft in an 8:35 resonant orbit-based formation, leveraging Saturn’s gravitational forces to maintain stable orbits with global coverage around Enceladus. To our knowledge, we simulate the first stable configuration of multiple spacecraft in closed orbits around Enceladus, using a full ephemeris force model (Russell and Lara, 2009). Each spacecraft’s orbit will precess, causing an eastward ground track shift (from an orbiter’s perspective) of each spacecraft for each orbit. However, all spacecraft return to their original positions relative to Enceladus after eight Enceladus revolutions around Saturn. We model communication pathways between SmallSats to understand how information would need to be transmitted across the constellation to enable AI-driven decision-making and resource allocation across the fleet. Capability Demonstration. Our simulated capability demonstration inputs position, velocity, and rotation telemetry from our Enceladus-focused constellation simulations, and mass spectrometry data collected from abiotic and biotic laboratory-analog ocean world experiments (Theiling et al., 2018; Theiling, 2021; Da Poian et al., 2023). Data from these experiments are used to simulate MS measurements and different scenarios of science observations for onboard analysis performed on each of the eight spacecraft. For these demonstrations, we integrate 24 machine learning (ML) algorithms into an onboard intelligence as a ‘knowledge base’, including algorithms evaluating data quality and those predicting (with % confidence) gas composition, ocean aqueous chemistry, and whether the sample was influenced by microbial life. The onboard AI capability is designed to use the knowledge base to come to a consensus-based decision in the interpretation of the observed data in order to request additional action outside of a pre-defined ConOps. Requested actions could include e.g., prioritized downlink to Earth (for analysis by ground-based teams) or follow-on analyses performed across the constellation. The spacecraft’s intelligent onboard planner must then determine whether sufficient resources (e.g., time, power, etc.) are available and weigh the request with mission priorities. In our simulation, the constellation is able to identify potential biosignatures using onboard ML algorithms, evaluate the confidence of that prediction, and perform follow-on analyses across the fleet to confirm the detection, in order to best prepare a transmission of these data to Earth-based teams.

astrobiology↗

Quantitative Analysis of The High‐Yield Hydrolysis of Kelp by Laminarinase and Alginate Lyase

Abstract Kelp is an abundant, farmable biomass‐containing laminarin and alginate as major polysaccharides, providing an excellent model substrate to study their deconstruction by simple enzyme mixtures. Our previous study showed strong reactivity of the glycoside hydrolase family 55 during hydrolysis of purified laminarin, raising the question of its reactivity with intact kelp. In this study, we determined that a combination of a single glycoside hydrolase family 55 β‐1,3‐exoglucanase with a broad‐specificity alginate lyase from the polysaccharide lyase family 18 gives efficient hydrolysis of untreated kelp to a mixture of simple sugars, that is, glucose, gentiobiose, mannitol‐end glucose, and mannuronic and guluronic acids and their soluble oligomers. Quantitative assignments from nanostructure initiator mass spectrometry (NIMS) and 2D HSQC NMR spectroscopy and analysis of the reaction time‐course are provided. The data suggest that binary combinations of enzymes targeted to the unique polysaccharide composition of marine biomass are sufficient to deconstruct kelp into soluble sugars for microbial fermentation.

2D HSQC NMR↗

AdenPredictor: accurate prediction of the adenylation domain specificity of nonribosomal peptide biosynthetic gene clusters in microbial genomes

Microbial natural products represent a major source of bioactive compounds for drug discovery. Among these molecules, nonribosomal peptides (NRPs) represent a diverse class that include antibiotics, immunosuppressants, anticancer agents, toxins, siderophores, pigments, and cytostatics. The discovery of novel NRPs remains a laborious process because many NRPs consist of nonstandard amino acids that are assembled by nonribosomal peptide synthetases (NRPSs). Adenylation domains (A-domains) in NRPSs are responsible for selection and activation of monomers appearing in NRPs. During the past decade, several support vector machine-based algorithms have been developed for predicting the specificity of the monomers present in NRPs. These algorithms utilize physiochemical features of the amino acids present in the A-domains of NRPSs. In this article, we benchmarked the performance of various machine learning algorithms and features for predicting specificities of NRPSs and we showed that the extra trees model paired with one-hot encoding features outperforms the existing approaches. Moreover, we show that unsupervised clustering of 453560 A-domains reveals many clusters that correspond to potentially novel amino acids. While it is challenging to predict the chemical structure of these amino acids, we developed novel techniques to predict their various properties, including polarity, hydrophobicity, charge, and presence of aromatic rings, carboxyl, and hydroxyl groups.

59 BASIC BIOLOGICAL SCIENCES↗

Did surface temperatures constrain microbial evolution?

The proposition that glaciation may not have occurred before the Cenozoic--albeit not yet a consensus position--nevertheless raises for reconsideration the surface temperature history of the earth. Glacial episodes, from the Huronian (2.3 billion years ago; BYA) through the late Paleozoic (320 to 250 million years ago; MYA) have been critical constraints on estimation of the upper bounds of temperature (Crowley 1983, Kasting and Toon 1989). Once removed, few if any constraints on the upper temperature limit other than life remain. Walker (1982) recognized that life provides an upper limit to temperature in the Precambrian. We propose a more radical concept: the upper temperature limit for viable growth of a given microbial group corresponds to the actual surface temperature at the time of the group's first appearance. In particular, we propose here that two major evolutionary developments--the emergence of cyanobacteria and aerobic eukaryotes--can be used to determine surface temperature in the Precambrian, and that only subsequent cooling mediated by higher plants and then angiosperms permitted what may possibly be the earth's first glaciation in the late Cenozoic.

NASA Program Exobiology↗

The Habitability of Proxima Centauri b: Environmental States and Observational Discriminants

Proxima Centauri b provides an unprecedented opportunity to understand the evolution and nature of terrestrial planets orbiting M dwarfs. Although Proxima Cen b orbits within its star’s habitable zone, multiple plausible evolutionary paths could have generated different environments that may or may not be habitable. Here, we use 1-D coupled climate-photochemical models to generate self-consistent atmospheres for several evolutionary scenarios, including high-O2, high-CO2, and more Earth-like atmospheres, with both oxic and anoxic compositions. We show that these modeled environments can be habitable or uninhabitable at Proxima Cen b's position in the habitable zone. We use radiative transfer models to generate synthetic spectra and thermal phase curves for these simulated environments, and use instrument models to explore our ability to discriminate between possible planetary states. These results are applicable not only to Proxima Cen b but to other terrestrial planets orbiting M dwarfs. Thermal phase curves may provide the first constraint on the existence of an atmosphere. We find that James Webb Space Telescope ( JWST) observations longward of 10 mm could characterize atmospheric heat transport and molecular composition. Detection of ocean glint is unlikely with JWST but may be within the reach of larger-aperture telescopes. Direct imaging spectra may detect O4 absorption, which is diagnostic of massive water loss and O2 retention, rather than a photosynthetic biosphere. Similarly, strong CO2 and CO bands at wavelengths shortward of 2.5 mm would indicate a CO2-dominated atmosphere. If the planet is habitable and volatile-rich, direct imaging will be the best means of detecting habitability. Earth-like planets with microbial biospheres may be identified by the presence of CH4-which has a longer atmospheric lifetime under Proxima Centauri's incident UV-and either photosynthetically produced O2 or a hydrocarbon haze layer.

Meadows, Victoria S.↗

The Emergence and Promise of Functional Chemogeography of Organic Matter

Organisms in ecosystems continuously release a myriad of organic matter molecules that undergo microbial and abiotic transformation, processes that critically influence carbon storage and climate feedbacks. Yet, a systematic understanding of what determines the transformation and persistence of organic matter across spatiotemporal scales remains elusive. We propose an emerging framework, termed “functional chemogeography,” to understand transformation and persistence of organic matter based on the chemical traits of molecules. This framework extends beyond a sole focus on intrinsic traits, which remain relatively constant across spatiotemporal scales, to emphasize extrinsic traits such as biochemical transformations and environmental responses, which vary spatiotemporally and are shaped by both intrinsic traits and the environment. When upscaled to the assemblage level using functional diversity indices, these extrinsic traits reveal a significant, and in some cases superior, capacity than intrinsic traits to explain biogeochemical processes, as demonstrated through a case study of dissolved organic matter in China's lakes. In conclusion, by integrating trait-based perspectives into predictive models, this framework helps bridge chemical complexity with ecosystem biogeochemistry, thereby advancing our ability to predict the fate of global organic carbon under environmental change.

chemical traits↗

Effects of hurricane disturbance and increased temperature on carbon cycling and storage of a Puerto Rican forest: a mechanistic investigation of above- and belowground processes (Final Technical Report)

The overall goal of the Tropical Responses for Altered Climate Experiment (TRACE) is to assess the effects of increasing temperature on tropical plant and soil carbon fluxes and storage as the forest recovers from major hurricane disturbance that occurred in September 2017. Ultimately, we aimed to reduce uncertainty and increase confidence with which tropical forests are represented in Earth System models to make more accurate global forecasts of future climate. We focused on both above- and belowground processes and explored temperature controls over critical aspects of carbon and nutrient cycling for tropical plants, soil, and microbes. TRACE is located in a wet tropical forest in the Luquillo Experimental Forest close to the USDA Forest Service Sabana Field Research Station in Luquillo, Puerto Rico. The warming treatment consists of six 4.7 m diameter plots. Three of the plots receive infrared warming and three have the same infrastructure but are not warmed (using ‘dummy’ heaters). Each plot was monitored from 2018-2023 to investigate two major questions: 1) Are there legacy effects of prior warming on forest recovery following hurricane disturbance? 2) Will the trajectory of forest recovery following disturbance be affected by warmer temperatures? Concurrent soil incubation experiments were conducted to enable more controlled mechanistic investigations of temperature response on microbial function. In sum, our goal was to use this novel climate manipulation experiment (the only of its kind in any tropical forest) and once-in-a-lifetime chance to assess how temperature and hurricane disturbance interact to affect coupled biogeochemical cycling in a tropical forest.

54 ENVIRONMENTAL SCIENCES↗

Increased salinity decreases annual gross primary productivity at a Northern California brackish tidal marsh

Tidal marshes sequester 11.4–87.0 Tg C yr –1 globally, but climate change impacts can threaten the carbon capture potential of these ecosystems. Tidal marshes occur across a wide range of salinity, with brackish marshes (0.5–18 ppt (parts per thousand)) dominating global tidal marsh extents. A diverse mix of freshwater- and saltwater-tolerant plant and microbial communities has led researchers to predict that carbon cycling in brackish wetlands may be less sensitive to changes in salinity than fresh- or saltwater wetlands. Rush Ranch, a well-monitored brackish tidal wetland of the San Francisco Bay National Estuarine Research Reserve, experiences highly variable annual salinity regimes. Within a five-year period (2014–2018), Rush Ranch experienced particularly extreme drought-induced salinization during the 2014 and 2015 growing seasons. During drought years, tidal channel salinity rose from a 15 year baseline of 4.7 ppt to growing season peaks of 10.3 ppt and 12.5 ppt. Continuous eddy covariance data from 2014 to 2018 demonstrate that during drought summers, gross primary productivity (GPP) decreased by 24%, whereas ecosystem respiration remained similar among all five years. Stepwise linear regression revealed that salinity, not air temperature or tidal height, was the dominant driver of annual GPP. A random forest model trained to predict GPP based on environmental data from low salinity years (i.e. naive to salinization) significantly over predicted GPP in drought years. When growing season salinities were doubled, annual estimates of net ecosystem exchange of CO 2 decreased by up to 30%. These results provide ecosystem-scale evidence that increased salinity influences CO 2 fluxes dominantly through reductions in GPP. This relationship provides a starting point for incorporating the effect of changes in salinity in wetland carbon models, which could improve wetland carbon forecasting and management for climate resilience.

54 ENVIRONMENTAL SCIENCES↗

Non-canonical d-xylose and l-arabinose metabolism via d-arabitol in the oleaginous yeast Rhodosporidium toruloides

Abstract R. toruloides is an oleaginous yeast, with diverse metabolic capacities and high tolerance for inhibitory compounds abundant in plant biomass hydrolysates. While R. toruloides grows on several pentose sugars and alcohols, further engineering of the native pathway is required for efficient conversion of biomass-derived sugars to higher value bioproducts. A previous high-throughput study inferred that R. toruloides possesses a non-canonical l -arabinose and d -xylose metabolism proceeding through d -arabitol and d -ribulose. In this study, we present a combination of genetic and metabolite data that refine and extend that model. Chiral separations definitively illustrate that d -arabitol is the enantiomer that accumulates under pentose metabolism. Deletion of putative d -arabitol-2-dehydrogenase (RTO4_9990) results in > 75% conversion of d -xylose to d -arabitol, and is growth-complemented on pentoses by heterologous xylulose kinase expression. Deletion of putative d -ribulose kinase (RTO4_14368) arrests all growth on any pentose tested. Analysis of several pentose dehydrogenase mutants elucidates a complex pathway with multiple enzymes mediating multiple different reactions in differing combinations, from which we also inferred a putative l -ribulose utilization pathway. Our results suggest that we have identified enzymes responsible for the majority of pathway flux, with additional unknown enzymes providing accessory activity at multiple steps. Further biochemical characterization of the enzymes described here will enable a more complete and quantitative understanding of R. toruloides pentose metabolism. These findings add to a growing understanding of the diversity and complexity of microbial pentose metabolism.

59 BASIC BIOLOGICAL SCIENCES↗