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At least 505 records · Page 28

Representing methane emissions from wet tropical forest soils using microbial functional groups constrained by soil diffusivity

Tropical ecosystems contribute significantly to global emissions of methane (CH 4 ), and landscape topography influences the rate of CH 4 emissions from wet tropical forest soils. However, extreme events such as drought can alter normal topographic patterns of emissions. Here we explain the dynamics of CH 4 emissions during normal and drought conditions across a catena in the Luquillo Experimental Forest, Puerto Rico. Valley soils served as the major source of CH 4 emissions in a normal precipitation year (2016), but drought recovery in 2015 resulted in dramatic pulses in CH 4 emissions from all topographic positions. Geochemical parameters including (i) dissolved organic carbon (C), acetate, and soil pH and (ii) hydrological parameters like soil moisture and oxygen (O 2 ) concentrations varied across the catena. During the drought, soil moisture decreased in the slope and ridge, and O 2 concentrations increased in the valley. We simulated the dynamics of CH 4 emissions with the Microbial Model for Methane Dynamics-Dual Arrhenius and Michaelis–Menten (M3D-DAMM), which couples a microbial functional group CH 4 model with a diffusivity module for solute and gas transport within soil microsites. Contrasting patterns of soil moisture, O 2 , acetate, and associated changes in soil pH with topography regulated simulated CH 4 emissions, but emissions were also altered by rate-limited diffusion in soil microsites. Changes in simulated available substrate for CH 4 production (acetate, CO 2 , and H 2 ) and oxidation (O 2 and CH 4 ) increased the predicted biomass of methanotrophs during the drought event and methanogens during drought recovery, which in turn affected net emissions of CH 4 . A variance-based sensitivity analysis suggested that parameters related to aceticlastic methanogenesis and methanotrophy were most critical to simulate net CH 4 emissions. This study enhanced the predictive capability for CH 4 emissions associated with complex topography and drought in wet tropical forest soils.

54 ENVIRONMENTAL SCIENCES↗

Water mass age structures the auxiliary metabolic gene content of free-living and particle-attached deep ocean viral communities

Viruses play important roles in the ocean’s biogeochemical cycles. Yet, deep ocean viruses are one of the most under-explored fractions of the global biosphere. Little is known about the environmental factors that control the composition and functioning of their communities or how they interact with their free-living or particle-attached microbial hosts. We analysed 58 viral communities associated with size-fractionated free-living (0.2–0.8 μm) and particle-attached (0.8–20 μm) cellular metagenomes from bathypelagic (2150–4018 m deep) microbiomes obtained during the Malaspina expedition. These metagenomes yielded 6631 viral sequences, 91% of which were novel, and 67 represented high-quality genomes. Taxonomic classification assigned 53% of the viral sequences to families of tailed viruses from the order Caudovirales. Computational host prediction associated 886 viral sequences to dominant members of the deep ocean microbiome, such as Alphaproteobacteria (284), Gammaproteobacteria (241), SAR324 (23), Marinisomatota (39), and Chloroflexota (61). Free-living and particle-attached viral communities had markedly distinct taxonomic composition, host prevalence, and auxiliary metabolic gene content, which led to the discovery of novel viral-encoded metabolic genes involved in the folate and nucleotide metabolisms. Water mass age emerged as an important factor driving viral community composition. We postulated this was due to changes in quality and concentration of dissolved organic matter acting on the host communities, leading to an increase of viral auxiliary metabolic genes associated with energy metabolism among older water masses. These results shed light on the mechanisms by which environmental gradients of deep ocean ecosystems structure the composition and functioning of free-living and particle-attached viral communities.

59 BASIC BIOLOGICAL SCIENCES↗

PAVC Gridded 20m Alaska NGEE Tier3 PFTs v1.0

These 20-meter spatial resolution gridded products provide per-pixel fractional cover (%) of Next Generation Ecosystem Experiments (NGEE) Arctic Plant Functional Types (PFTs) Tier 3 across Alaska, north of the boreal treeline. The products were developed for the NGEE Arctic project, which is improving Arctic vegetation representation and parameterization of the E3SM Land Model. This dataset includes 8 files containing fractional cover for NGEE Tier 3 PFTs (https://data.ess-dive.lbl.gov/view/doi:10.15485/2529470): (1) bryophytes; (2) lichens; (3) non-vascular plants, i.e., the sum of lichens and bryophytes; (4) deciduous shrubs, (5) evergreen shrubs, (6) forbs, (7) graminoids, and a non-PFT class, (8) litter. Each pixel contains the percent cover (expressed as a fraction of total ground cover) that was predicted by random-forest regression models. The random-forest models were trained on cover data collected at 978 plots from 2010 to 2021, of which are archived in the Pan-Arctic Vegetation Cover (PAVC) database (https://data.ess-dive.lbl.gov/datasets/doi:10.15485/2483557). The plot cover was linked to 20-meter spatial resolution, satellite-derived predictor variables: Sentinel-2 spectra and Sentinel-1 polarizations averaged over the 2019 growing season, as well as topographical features derived from ArcticDEM. Then, spatio-temporally anomalous plot data that introduced large variability to the regression outcomes were dropped using the Cook’s distance outlier detection method, and the models were re-created using high-quality plots and their associated satellite derived explanatory variables per each PFT. The correlations between plot-observed and satellite-derived fractional cover for all PFTs were well correlated (R2 = 0.69–0.95 and 0.5 for litter) and had low RMSE bias (0.02–0.11). This research was performed as a part of the NGEE Arctic project. The NGEE Arctic project was a research effort to reduce uncertainty in Earth System Models by developing a predictive understanding of carbon-rich Arctic ecosystems and feedbacks to climate. NGEE Arctic was supported by the Department of Energy's Office of Biological and Environmental Research.

54 ENVIRONMENTAL SCIENCES↗

Using Diel Solute Signals to Assess Ecohydrological Processing in Lotic Systems

Lotic systems are a prominent ecohydrological interface within which complex, coupled interactions occur between ecological, hydrological, and geochemical processes. This chapter reviews the ecological processes driving diel signals in oxygen as well as major nutrients and other trace elements. It discusses how diel signals can be used to evaluate functioning and interactions within stream ecosystems. The chapter considers potential future research directions and outstanding questions still to be addressed by or about diel solute signals and associated processes. High-resolution sensor-based time series and associated tools and conceptual advances are being integrated into environmental management. Quantitative tools to disentangle diel biogeochemically-driven signals from other sources of variability in continuous time series are growing. Theoretical models of diel process dynamics are helping to spur hypothesis generation and new ways of approaching diel signals in lotic systems.

Kurz, Marie↗

Carbon starvation is absent regardless of season of burn in Liquidambar styraciflua L.

In the Southeastern United States, the function and stability of pine ecosystems depends on frequent low intensity fires. One of the critical effects of frequent fire is the inhibition of hardwood competition by removing above-ground stems (top-kill). Previous long-term studies have correlated early growing season burns with higher understory mortality when compared to dormant season burns. Seasonal differences in mortality from insufficient carbon reserves in roots after leaf flush were posited as the mechanism but this was never explicitly tested. In this experiment, we burned two-year-old pot-grown sweetgums (Liquidambar styraciflua L.) in the dormant season (February), growing season (May), and compared their physiological responses to plants that were unburned. By the end of the first post-burn growing season, mortality was higher after the dormant season burn than the growing season burn and unburned trees had nearly twice the mass of either the burned treatments. Tissue starch concentrations were similar across all treatments highlighting the resiliency of sweetgum carbon stocks to recover from above-ground stem removal regardless of season. We showed the critical importance of top-killing stems regardless of season of burn. Our findings further suggest that dormant season fires can be more lethal to young sweetgum, possibly due to susceptibility of tender new shoots to freezing damage.

54 ENVIRONMENTAL SCIENCES↗

Evidence-based indicator approach to guide preliminary environmental impact assessments of hydropower development

Global expansion of hydropower resources has increased in recent years to meet growing energy demands and fill worldwide gaps in electricity supply. However, hydropower induces significant environmental impacts on river ecosystems - impacts that are addressed through environmental impact assessment (EIA) processes. The need for effective EIA processes is increasing as environmental regulations are either stressed in developing countries undertaking rapid expansion of hydropower capacity or time- and resource-intensive in developed countries. Part of the challenge in implementing EIAs lies in reaching a consensus among stakeholders regarding the most important environmental factors as the focus of impact studies. To help address this gap, we developed a weight-of-evidence approach (and toolkit) as a preliminary and coarse assessment of the most relevant impacts of hydropower on primary components of the river ecosystem, as identified using river function indicators. Through a science-based questionnaire and predictive model, users identify which environmental indicators may be impacted during hydropower development as well as those indicators that have the highest levels of uncertainty and require further investigation. Furthermore, an assessment tool visualizes inter-dependent indicator relationships, which help formulate hypotheses about causal relationships explored through environmental studies. We apply these tools to four existing hydropower projects and one hypothetical new hydropower project of varying sizes and environmental contexts. We observed consistencies between the output of our tools and the Federal Energy Regulatory Commission licensing process (inclusive of EIAs) but also important differences arising from holistic scientific evaluations (our toolkit) versus regulatory policies. The tools presented herein are aimed at increasing the efficiency of the EIA processes that engender environmental studies without loss of rigor or transparency of rationale necessary for understanding, considering, and mitigating the environmental consequences of hydropower.

54 ENVIRONMENTAL SCIENCES↗

Phage-based delivery of CRISPR-associated transposases for targeted bacterial editing

Phage λ, a well-characterized temperate phage, has been recently leveraged for bacterial genome editing by selectively delivering base editors into targeted bacterial species. We extend this concept by engineering phage λ to deliver CRISPR-guided transposases, accomplishing large insertions and targeted gene disruptions. To achieve this, we engineered phage λ using homologous recombination paired with Cas13a-based counterselection for precise phage modifications. Initially, we established the utility of Cas13a in phage λ by conducting minimal recoding edits, deletions, and insertions. Subsequently, we scaled up the engineering to embed the comprehensive DNA-editing CRISPR-Cas transposase (DART) system within the phage genome, creating λ-DART phages. These modified λ-DART phages were then employed to infectEscherichia coli, generating CRISPR RNA-guided transposition events in the host genome. Applying our engineered λ-DART phages to monocultures and a mixed bacterial community comprising three genera led to efficient, precise, and specific gene knockouts and insertions in the targetedE. colicells, achieving editing efficiencies surpassing 50% of the population. This research enhances phage-mediated genome editing by enabling efficient in situ gene integrations in bacteria, offering an avenue for further application in microbial community contexts. This scalable method enables flexible microbial genome editing in situ to manipulate the function and composition of diverse ecosystems.

Science & Technology - Other Topics↗

Convergent evolution of tree hydraulic traits in Amazonian habitats: implications for community assemblage and vulnerability to drought

Amazonian droughts are increasing in frequency and severity. However, little is known about how this may influence species-specific vulnerability to drought across different ecosystem types. We measured 16 functional traits for 16 congeneric species from six families and eight genera restricted to floodplain, swamp, white-sand or plateau forests of Central Amazonia. We investigated whether habitat distributions can be explained by species hydraulic strategies, and if habitat specialists differ in their vulnerability to embolism that would make water transport difficult during drought periods. We found strong functional differences among species. Nonflooded species had higher wood specific gravity and lower stomatal density, whereas flooded species had wider vessels, and higher leaf and xylem hydraulic conductivity. The P 50 values (water potential at 50% loss of hydraulic conductivity) of nonflooded species were significantly more negative than flooded species. However, we found no differences in hydraulic safety margin among species, suggesting that all trees may be equally likely to experience hydraulic failure during severe droughts. Water availability imposes a strong selection leading to differentiation of plant hydraulic strategies among species and may underlie patterns of adaptive radiation in many tropical tree genera. Our results have important implications for modeling species distribution and resilience under future climate scenarios.

59 BASIC BIOLOGICAL SCIENCES↗

Comparing field and lab quantitative stable isotope probing for nitrogen assimilation in soil microbes

ABSTRACT Soil microbial communities play crucial roles in nutrient cycling and can help retain nitrogen in agricultural soils. Quantitative stable isotope probing (qSIP) is a useful method for investigating taxon-specific microbial growth and utilization of specific nutrients, such as nitrogen (N). Typically, qSIP is performed in a highly controlled lab setting, so the field relevance of lab qSIP studies remains unknown. We conducted and compared tandem lab and field qSIP to quantify the assimilation of 15 N by maize-associated soil prokaryotic communities at two agricultural sites. Here, we show that field qSIP with 15 N can be used to measure taxon-specific microbial N assimilation. Relative 15 N assimilation rates were generally lower in the field, and the magnitude of this difference varied by site. Rates differed by method (lab vs field) for 19% of the top N assimilating genera. The field and lab measures were more comparable when relative assimilation rates were weighted by relative abundance to estimate the proportion of N assimilated by each genus with only ~10% of taxa differing by method. Of those that differed, the taxa consistently higher in the lab were inclined to have opportunistic lifestyle strategies, whereas those higher in the field had niches reliant on plant roots or in-tact soil structure (biofilms, mycelia). This study demonstrates that 15 N-qSIP can be successfully performed using field-incubated soils to identify microbial allies in N retention and highlights the strengths and limitations of field and lab qSIP approaches. IMPORTANCE Soil microbes are responsible for critical biogeochemical processes in natural and agricultural ecosystems. Despite their importance, the functional traits of most soil organisms remain woefully under-characterized, limiting our ability to understand how microbial populations influence the transformation of elements such as nitrogen (N) in soil. Quantitative stable isotope probing (qSIP) is a powerful tool to measure the traits of individual taxa. This method has rarely been applied in the field or with 15 N to measure nitrogen assimilation. In this study, we measured genus-specific microbial nitrogen assimilation in two agricultural soils and compared field and lab 15 N qSIP methods. Our results identify taxa important for nitrogen assimilation in agricultural soils, shed light on the field relevance of lab qSIP studies, and provide guidance for the future application of qSIP to measure microbial traits in the field.

Reed, Kinsey (ORCID:0000000155178664)↗

Tunturi virus isolates and metagenome-assembled viral genomes provide insights into the virome of Acidobacteriota in Arctic tundra soils

Arctic soils are climate-critical areas, where microorganisms play crucial roles in nutrient cycling processes. Acidobacteriota are phylogenetically and physiologically diverse bacteria that are abundant and active in Arctic tundra soils. Still, surprisingly little is known about acidobacterial viruses in general and those residing in the Arctic in particular. Here, we applied both culture-dependent and -independent methods to study the virome of Acidobacteriota in Arctic soils. Five virus isolates, Tunturi 1–5, were obtained from Arctic tundra soils, Kilpisjärvi, Finland (69°N), using Tunturiibacter spp. strains originating from the same area as hosts. The new virus isolates have tailed particles with podo- (Tunturi 1, 2, 3), sipho- (Tunturi 4), or myovirus-like (Tunturi 5) morphologies. The dsDNA genomes of the viral isolates are 63–98 kbp long, except Tunturi 5, which is a jumbo phage with a 309-kbp genome. Tunturi 1 and Tunturi 2 share 88% overall nucleotide identity, while the other three are not related to one another. For over half of the open reading frames in Tunturi genomes, no functions could be predicted. To further assess the Acidobacteriota-associated viral diversity in Kilpisjärvi soils, bulk metagenomes from the same soils were explored and a total of 1881 viral operational taxonomic units (vOTUs) were bioinformatically predicted. Almost all vOTUs (98%) were assigned to the class Caudoviricetes. For 125 vOTUs, including five (near-)complete ones, Acidobacteriota hosts were predicted. Acidobacteriota-linked vOTUs were abundant across sites, especially in fens. Terriglobia-associated proviruses were observed in Kilpisjärvi soils, being related to proviruses from distant soils and other biomes. Approximately genus- or higher-level similarities were found between the Tunturi viruses, Kilpisjärvi vOTUs, and other soil vOTUs, suggesting some shared groups of Acidobacteriota viruses across soils. This study provides acidobacterial virus isolates as laboratory models for future research and adds insights into the diversity of viral communities associated with Acidobacteriota in tundra soils. Predicted virus-host links and viral gene functions suggest various interactions between viruses and their host microorganisms. Largely unknown sequences in the isolates and metagenome-assembled viral genomes highlight a need for more extensive sampling of Arctic soils to better understand viral functions and contributions to ecosystem-wide cycling processes in the Arctic.

54 ENVIRONMENTAL SCIENCES↗

AmeriFlux FLUXNET-1F US-xTL NEON Toolik (TOOL)

This is the AmeriFlux Management Project (AMP) created FLUXNET-1F version of the carbon flux data for the site US-xTL NEON Toolik (TOOL). This is the FLUXNET version of the carbon flux data for the site US-xTL NEON Toolik (TOOL) produced by applying the standard ONEFlux (1F) software. Site Description - The Toolik field site is located in a remote wilderness area at the Toolik Field Station which is on land managed by the Bureau of Land Management. The area is underlain by continuous permafrost, which exerts a major influence on hydrology and the distribution, structure, and function of terrestrial and aquatic ecosystems. Because of its location between the Brooks Range and the coastal plain, the vegetation and soils at TOOL are representative of much of the Alaskan foothills tundra. This terrestrial field site is collocated with NEON’s Toolik Lake aquatic site. The closest city, Fairbanks, Alaska is 400 miles away. The Dalton Highway (also known as the Haul Road or Pipeline Road), which traverses the middle of the state from Fairbanks to Prudhoe Bay, provides access to the area.

Network), NEON (National Ecological Observatory↗

Non-Destructive, Three-Dimensional Imaging of Processes in the Rhizosphere Utilizing High Energy Photons

Soil structure, which can be described as the aggregation and distribution of pore spaces, regulates carbon, nutrient, and water cycling across the Earth system. Yet the inability to make quantitative, dynamic, in situ measurements of soil structure and rhizosphere carbon flow has prevented meaningful incorporation of soil structural processes into Earth System Models (ESMs). The key limitations are: (i) Lack of scale integration between micron-scale soil structure and ecosystem-scale models, (ii) Poor functional linkage between soil structural properties and biogeochemical processes, and (iii) Absence of dynamic 3D measurements of rhizosphere structural changes and carbon transformations. To address these challenges, we developed a new integrated positron emission tomography (PET) - microcomputed tomography (CT) imaging platform that enables the first 4D (spatial 3D and time), non-invasive, quantitative imaging of carbon allocation and rhizosphere structural dynamics in living plants and intact soils. The system combines: • Rhizo-PET (R-PET): a high-resolution positron emission tomography scanner optimized for 11 CO 2 tracing in plant roots. • a-Se micro-CT: a high-contrast, high-spatial resolution CT system based on amorphous selenium (a-Se) direct conversion technology, enabling micron-scale visualization of soil structure and root–soil interfaces. Together, these advances allow us to quantify how carbon exudates move, transform, and stabilize within the rhizosphere, directly informing missing processes in BER-relevant carbon cycle models.

42 ENGINEERING↗

WA-IsoC_MSC1.1.0

The soil microbiome is central to the cycling of carbon and other nutrients and to the promotion of plant growth. Despite its importance, analysis of the soil microbiome is difficult due to its sheer complexity, with thousands of interacting species. Here, we reduced this complexity by developing model soil microbial consortia that are simpler and more amenable to experimental analysis but still represent important microbial functions of the native soil ecosystem. Samples were collected from an arid grassland soil and microbial communities (consisting mainly of bacterial species) were enriched on agar plates containing chitin as the main carbon source. Chitin was chosen because it is an abundant carbon and nitrogen polymer in soil that often requires the coordinated action of several microorganisms for complete metabolic degradation. Several soil consortia were derived that had tractable richness (30-50 OTUs) with diverse phyla representative of the native soil, including Actinobacteria, Bacteroidetes, Firmicutes, Proteobacteria and Verrucomicrobia. The resulting consortia can be stored as glycerol or lyophilized stocks at -80° C and revived while retaining community composition, greatly increasing their use as tools for the research community at large. One of the consortia that was particularly stable was chosen as a model soil consortium (MSC-1) for further analysis. MSC-1 species interactions were studied using both pairwise co-cultivation in liquid media and during growth in soil under several perturbations. Co-abundance analyses highlighted interspecies interactions and helped to define keystone species, including Mycobacterium, Rhodococcus, and Rhizobiales taxa. These experiments demonstrate the success of an approach based on naturally enriching a community of interacting species that can be stored, revived, and shared. The knowledge gained from querying these communities and their interactions will enable better understanding of the soil microbiome and the roles these interactions play in this environment.

54 ENVIRONMENTAL SCIENCES↗

NGEE Arctic Plant Traits: Fine Roots, Kougarok Road Mile Marker 64, Seward Peninsula, Alaska, 2016

Soil cores were collected from twelve vegetation biomass plots at the Kougarok hillslope in late July of 2016. The sampled plots were located across six ecotypes present at this site (n=2 replicates per ecotype). Soil cores were separated into depth intervals in the field and frozen for transport to Oak Ridge National Laboratory. Data package contains one *.csv and one *.pdf.Laboratory sample processing of these soils generated depth-specific data on soil properties as well as fine root biomass, length, %C, %N, delta13C, and delta15N. Only live fine roots were analyzed. Fine roots were not separated by species or plant functional type.The Next-Generation Ecosystem Experiments: Arctic (NGEE Arctic), a research effort to reduce uncertainty in Earth System Models by developing a predictive understanding of carbon-rich Arctic ecosystems and feedbacks to climate. NGEE Arctic was supported by the Department of Energy’s Office of Biological and Environmental Research.The NGEE Arctic project had two field research sites: 1) located within the Arctic polygonal tundra coastal region on the Barrow Environmental Observatory (BEO) and the North Slope near Utqiagvik (Barrow), Alaska and 2) multiple areas on the discontinuous permafrost region of the Seward Peninsula north of Nome, Alaska.Through observations, experiments, and synthesis with existing datasets, NGEE Arctic provided an enhanced knowledge base for multi-scale modeling and contributed to improved process representation at global pan-Arctic scales within the Department of Energy’s Earth system Model (the Energy Exascale Earth System Model, or E3SM), and specifically within the E3SM Land Model component (ELM).

54 ENVIRONMENTAL SCIENCES↗

Fisher Forecasting for the DESC with $\texttt{Augur}$

The Vera C. Rubin Observatory Legacy Survey of Space and Time (LSST) has begun its ten-year survey of the entire visible southern hemisphere. To ensure robust cosmological measurements, computationally inexpensive investigations of modeling choices must be made to gauge the performance of proposed cosmological analyses. In this paper, we introduce the $\texttt{Augur}$ tool of the Dark Energy Science Collaboration (DESC), which provides Fisher forecasts for cosmological inference for the LSST using software frameworks designed for DESC science. We test the pipeline by comparing it to forecasts produced by external code and direct sampling of the posterior via nested sampling methods, finding good agreement between all methods. We additionally investigate a range of modeling and hyperparameter choices for a 3$\times$2pt investigation in harmonic space, providing users with diagnostics to obtain reliable forecasts. $\texttt{Augur}$ will be continually updated to be compatible with the other tools in the DESC software ecosystem as additional probes and functionality become available.

Rogozenski, Paul [Carnegie Mellon U.; Arizona U.] ↗

Differential structure and functional gene response to geochemistry associated with the suspended and attached shallow aquifer microbiomes from the Illinois Basin, IL

Despite the clear ecological significance of the microbiomes inhabiting groundwater and connected ecosystems, our current understanding of their habitats, functionality, and the ecological processes controlling their assembly have been limited. In this study, an efficient pipeline combining geochemistry, high-throughput Fluidigm TM functional gene amplification and sequencing was developed to analyze the suspended and attached microbial communities inhabiting five groundwater monitoring wells in the Illinois Basin, USA. The dominant taxa in the suspended and the attached microbial communities exhibited significantly different spatial and temporal changes in both alpha- and beta-diversity. Further analyses of representative functional genes affiliated with N 2 fixation (nifH), methane oxidation (pmoA), and sulfate reduction (dsrB, and aprA), suggested functional redundancy within the shallow aquifer microbiomes. While more diversified functional gene taxa were observed for the suspended microbial communities than the attached ones except for pmoA, different levels of changes over time and space were observed between these functional genes. Notably, deterministic and stochastic ecological processes shaped the assembly of microbial communities and functional gene reservoirs differently. While homogenous selection was the prevailing process controlling assembly of microbial communities, the neutral processes (e.g., dispersal limitation, drift and others) were more important for the functional genes. The results suggest complex and changing shallow aquifer microbiomes, whose functionality and assembly vary even between the spatially proximate habitats and fractions. As a result, this research underscored the importance to include all the interface components for a more holistic understanding of the biogeochemical processes in aquifer ecosystems, which is also instructive for practical applications.

54 ENVIRONMENTAL SCIENCES↗

Global Diversity and Biogeography of the Zostera marina Mycobiome

Seagrasses are marine flowering plants that provide critical ecosystem services in coastal environments worldwide. Marine fungi are often overlooked in microbiome and seagrass studies, despite terrestrial fungi having critical functional roles as decomposers, pathogens, or endophytes in global ecosystems. Here, we characterize the distribution of fungi associated with the seagrass Zostera marina, using leaves, roots, and rhizosphere sediment from 16 locations across its full biogeographic range. Using high-throughput sequencing of the ribosomal internal transcribed spacer (ITS) region and 18S rRNA gene, we first measured fungal community composition and diversity. We then tested hypotheses of neutral community assembly theory and the degree to which deviations suggested that amplicon sequence variants (ASVs) were plant selected or dispersal limited. Finally, we identified a core mycobiome and investigated the global distribution of differentially abundant ASVs. We found that the fungal community is significantly different between sites and that the leaf mycobiome follows a weak but significant pattern of distance decay in the Pacific Ocean. Generally, there was evidence for both deterministic and stochastic factors contributing to community assembly of the mycobiome, with most taxa assembling through stochastic processes. The Z. marina core leaf and root mycobiomes were dominated by unclassified Sordariomycetes spp., unclassified Chytridiomycota lineages (including Lobulomycetaceae spp.), unclassified Capnodiales spp., and Saccharomyces sp. It is clear from the many unclassified fungal ASVs and fungal functional guilds that knowledge of marine fungi is still rudimentary. Further studies characterizing seagrass-associated fungi are needed to understand the roles of these microorganisms generally and when associated with seagrasses. Fungi have important functional roles when associated with land plants, yet very little is known about the roles of fungi associated with marine plants, like seagrasses. In this study, we report the results of a global effort to characterize the fungi associated with the seagrass Zostera marina across its full biogeographic range. Although we defined a putative global core fungal community, it is apparent from the many fungal sequences and predicted functional guilds that had no matches to existing databases that general knowledge of seagrass-associated fungi and marine fungi is lacking. This work serves as an important foundational step toward future work investigating the functional ramifications of fungi in the marine ecosystem.

ITS2↗

Tetranucleotide frequencies differentiate genomic boundaries and metabolic strategies across environmental microbiomes

Microbiomes are constrained by physicochemical conditions, nutrient regimes, and community interactions across diverse environments, yet genomic signatures of this adaptation remain unclear. Metagenome sequencing is a powerful technique to analyze genomic content in the context of natural environments, establishing concepts of microbial ecological trends. Here, we developed a data discovery tool-a tetranucleotide-informed metagenome stability diagram-that is publicly available in the integrated microbial genomes and microbiomes (IMG/M) platform for metagenome ecosystem analyses. We analyzed the tetranucleotide frequencies from quality-filtered and unassembled sequence data of over 12,000 metagenomes to assess ecosystem-specific microbial community composition and function. We found that tetranucleotide frequencies can differentiate communities across various natural environments and that specific functional and metabolic trends can be observed in this structuring. Our tool places metagenomes sampled from diverse environments into clusters and along gradients of tetranucleotide frequency similarity, suggesting microbiome community compositions specific to gradient conditions. Within the resulting metagenome clusters, we identify protein-coding gene identifiers that are most differentiated between ecosystem classifications. We plan for annual updates to the metagenome stability diagram in IMG/M with new data, allowing for refinement of the ecosystem classifications delineated here. This framework has the potential to inform future studies on microbiome engineering, bioremediation, and the prediction of microbial community responses to environmental change. IMPORTANCE: Microbes adapt to diverse environments influenced by factors like temperature, acidity, and nutrient availability. We developed a new tool to analyze and visualize the genetic makeup of over 12,000 microbial communities, revealing patterns linked to specific functions and metabolic processes. This tool groups similar microbial communities and identifies characteristic genes within environments. By continually updating this tool, we aim to advance our understanding of microbial ecology, enabling applications like microbial engineering, bioremediation, and predicting responses to environmental change.

Kellom, Matthew↗