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Selection and development of aptamers for pyoverdine

The rhizosphere is a small region surrounding plant roots that is enriched in biochemicals from root exudates and populated with fungi, nematode, and bacteria. Interaction of rhizosphere organisms with plants is mainly promoted by exudates from the roots. Root exudates contain biochemicals that come from primary and secondary metabolisms of plants. These biochemicals attract microbes, which influence plant nutrition. The rhizosphere bacteria (microbiome) are vital to plant nutrient uptake and influence biotic and abiotic stress and pathogenesis. Pseudomonas is a genus of gammaproteobacteria known for its ubiquitous presence in natural habitats and its striking ecological, metabolic, and biochemical diversity. Within the genus, members of the Pseudomonas fluorescens group are common inhabitants of soil and plant surfaces, and certain strains function in the biological control of plant disease, protecting plants from infection by soilborne and aerial plant pathogens. The soil bacterium Pseudomonas protegens Pf-5 (also known as Pseudomonas fluorescensPf-5) is a well-characterized biological strain, which is distinguished by its prolific production of the secondary metabolite, pyoverdine. Knowledge of the distribution of P. fluorescens secretory activity around plant roots is very important for understanding the interaction between P. fluorescens and plants and can be achieved by real time tracking of pyoverdine. To achieve the capability of real-time tracking in soil, we have used a structure-switching SELEX strategy to select high affinity ssDNA and 2’ F-Y-RNA aptamers with specificity for pyoverdine over other siderophores. Pyoverdines from various strains possess a conserved chromophore along with strain-specific peptide chain. Some of the isolated aptamers bind to chromophore of pyoverdine. These will identify pyoverdines from a variety of pseudomonads. Identification of aptamers that selectively bind the peptide portion of pyoverdine Pf-5 is underway. The identified aptamers that are broadly and narrowly specific for pyoverdines or Pf-5 will be optimized for integration into an electrochemical biosensor to track, in real time, P fluorescens activity and other pseudomonads in and around the rhizosphere.

59 BASIC BIOLOGICAL SCIENCES↗

Agricultural impacts on nitrogen cycling: climate and air pollution

Ratios of key soil nitrogen fluxes from two terrestrial ecosystem models (TEMs) are compared to observational values in the literature. The fluxes included nitrification, denitrification, mineralization, immobilization and plant uptake, with the latter two fluxes partitioned into NH 4 + and NO 3 -. The two TEMs, ELMv1-ECA and CLM5.0, originated from a common model but have diverged in their handling of plant/microbe competition for soil mineral N. Both models predict unrealistically low assimilation (i.e, immobilization) of NO 3 - by soil bacteria compared to literature values and unrealistic dominance of plant uptake by a single form of mineral nitrogen (NO 3 - for ELM, NH 4 + for CLM5.0). Global ratios of net mineralization (i.e., gross mineralization – gross immobilization) appear unrealistically high in both models compared to literature values while rates of nitrification/gross mineralization appear unrealistically high (ELM) and low (CLM5.0). The ratio of denitrification/nitrification is likely high in both models (3:1 ELM; 2:1 CLM5.0), but few experimental data exist to evaluate these ratios, in part because nitrification and denitrification are quantified with different techniques and generally are not measured simultaneously. The same is true of plant N uptake and microbial N assimilation. Single grid point experiments with CLM5.0 at the Hubbard Brook long-term ecological research site in New Hampshire indicate that the parameterization of nitrification is a potentially important focus area that can bring soil nitrogen flux ratios into better agreement with observations. The current flux ratios raise questions about the credibility of present and future simulations of mineral N limitation on the carbon cycle.

54 ENVIRONMENTAL SCIENCES↗

KBase Narrative - Metagenome-assembled genomes from Amazonian soil microbial consortia

We report 17 metagenome-assembled genomes (MAGs) recovered from microbial consortia of forest and pasture soils, in the Brazilian Eastern Amazon. Recovered MAGs constitute novel genomes with the potential to act in important ecological processes, including the degradation of carbohydrate and sulfur and nitrogen cycling.

Mandro, Jessica Adriele↗

Midwest Food Security & Agriculture II: Leveraging NASA Earth Observations to Analyze and Display Crop Phenology Data and Weather Conditions to Support Expansion of Small Grain Crops in the Midwest

Agriculture in the Midwest is dominated by monoculture systems that strip the soil of nutrients, decrease yields, and worsen water quality. Crop diversification and cultivating small grains is economically and ecologically advantageous, but limited in practice due to a lack of data about small-grain crop performance as well as monoculture-favoring insurance coverage, economic incentives, and sociocultural traditions. Practical Farmers of Iowa (PFI) strives to create more sustainable agriculture and supports Midwestern farmers in adding small grains into their crop rotations. The DEVELOP team partnered with the USDA Agricultural Research Service and the USA National Phenology Network to assist PFI in applying NASA Earth observation data to track small-grain crop performance. This project developed a Google Earth Engine user interface that enables PFI farmers to compare satellite-assessed vegetation production and climatic conditions of their farm with other PFI farmers. The team assessed the crop performance through Normalized Difference Vegetation Index (NDVI) values derived from Landsat 7 Enhanced Thematic Mapper Plus (ETM+) and Landsat 8 Operational Land Imager (OLI). Additionally, the team created a regression model to analyze the predictive properties of vegetation indices and crop yield, ultimately establishing a relationship with mean NDVI from the early growing season. PFI farmers can utilize these products to make informed decisions about their crop production as well as advocate for more expansive insurance policies to protect small-grain crops.

Sophie Barrowman↗

Vegetation classification map and covariates associated with NEON AOP survey, East River, CO 2018

This package includes geospatial data layers developed to investigate how environmental gradients—specifically topography and near-surface soil properties—drive the spatial arrangement of dominant plant communities in mountainous watersheds. The geospatial products, which support the analysis of these ecological relationships, are derived from airborne hyperspectral and LiDAR datasets acquired by the National Ecological Observatory Network (NEON) Airborne Observation Platform (AOP), in conjunction with an extensive ground field campaign conducted in summer 2018. This work is part of the DOE Watershed Function Science Focus Area (SFA) and features geospatial datasets developed based on observations and ground data collected at East River, Colorado, in collaboration with the National Ecological Observatory Network (NEON) Airborne Observation Platform (AOP) survey in June 2018. Classification Map: - Classification Map (PNG, GeoTIFF): Derived from hyperspectral and LiDAR airborne data using a machine learning approach. - Class Code Mapper (CSV): Associates pixel values with corresponding vegetation/non-vegetation classes. - Classification Reference Data (CSV): Reference data used in the machine learning procedure. LiDAR-Derived Products: - Topographical Metrics (GeoTIFFs): Elevation, slope, curvature, TWI, TPI, solar insolation, and canopy height model (CHM), smoothed with a 5x5 pixel window. Vegetation Indices: - GeoTIFFs of NDVI, NDNI, NDWI: Vegetation indices derived from hyperspectral data. Urban Masks: - Urban Mask (GeoTIFF): Applied to the mapping to convert bare soil classes to urban classes. Software Compatibility: GeoTIFFs: Can be visualized with GIS software or libraries that support GeoTIFF images. CSV Files: Can be opened with any software that handles comma-separated values. The FLMD file provides details and links to the source datasets used to derive the products. The manuscript (in the Method session) provides details on how each product was derived. This work was supported by the Watershed Function Science Focus Area at Lawrence Berkeley National Laboratory funded by the US Department of Energy, Office of Science, Biological and Environmental Research under Contract No. DE-AC02-05CH11231. Update on 2026-03-25: Since the original dataset publication date of 02/28/2020, this package has a new classification map derived by an improved methodology. This update also includes additional ground data that improved the representation of some of the communities. See the methods for further details on what has changed between versions.

2018 NEON and 2025 CHESS Campaigns↗

Nitrogen Deposition Weakens Soil Carbon Control of Nitrogen Dynamics Across the Contiguous United States

ABSTRACT Anthropogenic nitrogen (N) deposition is unequally distributed across space and time, with inputs to terrestrial ecosystems impacted by industry regulations and variations in human activity. Soil carbon (C) content normally controls the fraction of mineralized N that is nitrified ( ƒ nitrified ), affecting N bioavailability for plants and microbes. However, it is unknown whether N deposition has modified the relationships among soil C, net N mineralization, and net nitrification. To test whether N deposition alters the relationship between soil C and net N transformations, we collected soils from coniferous and deciduous forests, grasslands, and residential yards in 14 regions across the contiguous United States that vary in N deposition rates. We quantified rates of net nitrification and N mineralization, soil chemistry (soil C, N, and pH), and microbial biomass and function (as beta‐glucosidase (BG) and N ‐acetylglucosaminidase (NAG) activity) across these regions. Following expectations, soil C was a driver of ƒ nitrified across regions, whereby increasing soil C resulted in a decline in net nitrification and ƒ nitrified . The ƒ nitrified value increased with lower microbial enzymatic investment in N acquisition (increasing BG:NAG ratio) and lower active microbial biomass, providing some evidence that heterotrophic microbial N demand controls the ammonium pool for nitrifiers. However, higher total N deposition increased ƒ nitrified , including for high soil C sites predicted to have low ƒ nitrified , which decreased the role of soil C as a predictor of ƒ nitrified . Notably, the drop in contemporary atmospheric N deposition rates during the 2020 COVID‐19 pandemic did not weaken the effect of N deposition on relationships between soil C and ƒ nitrified . Our results suggest that N deposition can disrupt the relationship between soil C and net N transformations, with this change potentially explained by weaker microbial competition for N. Therefore, past N inputs and soil C should be used together to predict N dynamics across terrestrial ecosystems.

Nieland, Matthew A. [Stockbridge School of Agricul↗

Redox potential in Typha-dominated tidal brackish marsh, PIE LTER, Plum Island Sound, MA, June–December 2022

This dataset includes soil redox potential measurements collected at multiple depths within a tidal brackish wetland in the upper estuary of the Plum Island Ecosystems Long-Term Ecological Research site (PIE LTER), Plum Island Sound, Newbury, Massachusetts. Measurements were taken to evaluate temporal variation in redox potential in relation to hydrological events at three replicate locations. Data were recorded every 5 minutes using a Campbell Scientific Volt116 connected to a CR6 datalogger with SWAP instrument redox probes (ORP-30-4-B) and reference electrodes. Measurements were made at the AmeriFlux site US-PLo at four soil depths (5, 10, 15, and 30 cm). The file redox_soiltemp_2022.csv contains temperature-corrected redox values and soil temperature following Silva-Machado et al. (2024). Metadata files redox_soiltemp_dd.csv and redox_soiltemp_flmd.csv provide detailed descriptions of variables and site locations.

54 ENVIRONMENTAL SCIENCES↗

The Risk Assessment Information System Compendium of Ecological Screening Benchmarks for Chemicals and Radionuclides (2025) (Volume I – Text, Radionuclides for all Media, Chemicals for Air and Biota)

The Risk Assessment Information System Ecological Benchmark Tool compiles screening level benchmark values for assessing potential ecological risks posed by chemical and radionuclide contaminants. Drawing from a wide array of federal, state, and international sources, it consolidates benchmarks across environmental media, including soil, water, sediment, air, and biota, and presents values for a broad range of receptors and exposure pathways. The tool includes both effect-based and no-effect thresholds, with media- and organism-specific guidance derived from peer-reviewed literature and regulatory frameworks. By integrating diverse benchmark types into a unified reference, the tool supports consistent, scientifically grounded evaluations in ecological risk assessments and environmental management practices.

54 ENVIRONMENTAL SCIENCES↗

Restoring Soil Fertility on Degraded Lands to Meet Food, Fuel, and Climate Security Needs via Perennialization

A continuously growing pressure to increase food, fiber, and fuel production to meet worldwide demand and achieve zero hunger has put severe pressure on soil resources. Abandoned, degraded, and marginal lands with significant agricultural constraints—many still used for agricultural production—result from inappropriately intensive management, insufficient attention to soil conservation, and climate change. Continued use for agricultural production will often require ever more external inputs such as fertilizers and herbicides, further exacerbating soil degradation and impeding nutrient recycling and retention. Growing evidence suggests that degraded lands have a large potential for restoration, perhaps most effectively via perennial cropping systems that can simultaneously provide additional ecosystem services. Here we synthesize the advantages of and potentials for using perennial vegetation to restore soil fertility on degraded croplands, by summarizing the principal mechanisms underpinning soil carbon stabilization and nitrogen and phosphorus availability and retention. We illustrate restoration potentials with example systems that deliver climate mitigation (cellulosic bioenergy), animal production (intensive rotational grazing), and biodiversity conservation (natural ecological succession). Perennialization has substantial promise for restoring fertility to degraded croplands, helping to meet future food security needs.

54 ENVIRONMENTAL SCIENCES↗

Community RNA-Seq: multi-kingdom responses to living versus decaying roots in soil

Abstract Roots are a primary source of organic carbon input in most soils. The consumption of living and detrital root inputs involves multi-trophic processes and multiple kingdoms of microbial life, but typical microbial ecology studies focus on only one or two major lineages. We used Illumina shotgun RNA sequencing to conduct PCR-independent SSU rRNA community analysis (“community RNA-Seq”) and simultaneously assess the bacteria, archaea, fungi, and microfauna surrounding both living and decomposing roots of the annual grass, Avena fatua. Plants were grown in 13CO2-labeled microcosms amended with 15N-root litter to identify the preferences of rhizosphere organisms for root exudates (13C) versus decaying root biomass (15N) using NanoSIMS microarray imaging (Chip-SIP). When litter was available, rhizosphere and bulk soil had significantly more Amoebozoa, which are potentially important yet often overlooked top-down drivers of detritusphere community dynamics and nutrient cycling. Bulk soil containing litter was depleted in Actinobacteria but had significantly more Bacteroidetes and Proteobacteria. While Actinobacteria were abundant in the rhizosphere, Chip-SIP showed Actinobacteria preferentially incorporated litter relative to root exudates, indicating this group’s more prominent role in detritus elemental cycling in the rhizosphere. Our results emphasize that decomposition is a multi-trophic process involving complex interactions, and our methodology can be used to track the trajectory of carbon through multi-kingdom soil food webs.

Nuccio, Erin E. (ORCID:000000030189183X)↗

Soil disturbance and invasion magnify CO 2 effects on grassland productivity, reducing diversity

Climate change, disturbance, and plant invasion threaten grassland ecosystems, but their combined and interactive effects are poorly understood. Here, in this study, we examine how the combination of disturbance and plant invasion influences the sensitivity of mixed-grass prairie to elevated carbon dioxide (eCO 2 ) and warming. We established subplots of intact prairie and disturbed/invaded prairie within a free-air CO 2 enrichment (to 600 ppmv) by infrared warming (+1.5°C day, 3°C night) experiment and followed plant and soil responses for 5 years. Elevated CO 2 initially led to moderate increases in biomass and plant diversity in both intact and disturbed/invaded prairie, but these effects shifted due to strong eCO 2 responses of the invasive forb Centaurea diffusa. In the final 3 years, biomass responses to eCO 2 in disturbed/invaded prairie were 10 times as large as those in intact prairie (+186% vs. +18%), resulting in reduced rather than increased plant diversity (-17% vs. +10%). At the same time, warming interacted with disturbance/invasion and year, reducing the rate of topsoil carbon recovery following disturbance. The strength of these interactions demonstrates the need to incorporate disturbance into predictions of climate change effects. In contrast to expectations from studies in intact ecosystems, eCO 2 may threaten plant diversity in ecosystems subject to soil disturbance and invasion.

54 ENVIRONMENTAL SCIENCES↗

Black carbon enriches short-range-order ferrihydrite in Amazonian Dark Earth: Interplay mechanism and environmental implications

Our study underpins the mechanism of organo-mineral interaction between black carbon (BC, biochar) and associated minerals in the historical BC-rich Amazonian Dark Earth (ADE) by using synchrotron-based microscopic (TXM), microspectroscopic (μFTIR) and spectroscopic (XAS and μ-diffraction) approaches. The BC-rich ADE contained over 100% more poorly crystalline minerals than the adjacent tropical soil. Linear combination fitting of k-spacing in the X-ray Absorption Spectra (XAS) revealed that ferrihydrite contributed to 81.1% of the Fe-minerals in BC. A small but distinct peak was observed at 5.7 Å –1 in the extended X-ray absorption fine structure k oscillation of BC, revealing the presence of Fe—C (including Fe-O-C) covalent bonds. No Fe—C path was yielded by the XAS fitting when an obvious peak downshift of the first (Fe—Fe 1 ) shell was observed, suggesting that the availability of inner-sphere Fe—C complexation was limited to the BC surface and interphase region. In this study, the main minerals for organo-mineral complexation were short-range-order (SRO) ferrihydrite on BC instead of corner-sharing FeO 6 octahedra. Compared to ADE, the coordination number of the first (Fe—Fe 1 ) and second (Fe—Fe 2 ) shell was higher in BC, revealing a higher degree of order in coordination between the neighboring Fe mineral crystals. Black C limited the progressive aging of amorphous Fe phases and greatly enriched SRO ferrihydrite in the redox-fluctuating and high-leaching environment. The transformation of SRO ferrihydrite into the more crystalline Fe oxides was controlled by the local pH environment. A strong signal from the complexed phenolic group (aryl-OH, 1241 cm –1 ) and a distinct band of inner-sphere complexation (Fe-aryl C, 1380–1384 cm –1 ) were identified in the FTIR spectra. The enrichment of poorly crystalline minerals can have positive feedback on the long-term stabilization of BC. The scale-up application of biochar to agricultural and ecological systems may have a long-lasting impact on the enrichment and transformation of the SRO minerals in the soil.

54 ENVIRONMENTAL SCIENCES↗

Validating spatial structure in canopy water content using geostatistics

Heterogeneity in ecological phenomena are scale dependent and affect the hierarchical structure of image data. AVIRIS pixels average reflectance produced by complex absorption and scattering interactions between biogeochemical composition, canopy architecture, view and illumination angles, species distributions, and plant cover as well as other factors. These scales affect validation of pixel reflectance, typically performed by relating pixel spectra to ground measurements acquired at scales of 1m(exp 2) or less (e.g., field spectra, foilage and soil samples, etc.). As image analysis becomes more sophisticated, such as those for detection of canopy chemistry, better validation becomes a critical problem. This paper presents a methodology for bridging between point measurements and pixels using geostatistics. Geostatistics have been extensively used in geological or hydrogeolocial studies but have received little application in ecological studies. The key criteria for kriging estimation is that the phenomena varies in space and that an underlying controlling process produces spatial correlation between the measured data points. Ecological variation meets this requirement because communities vary along environmental gradients like soil moisture, nutrient availability, or topography.

Sanderson, E. W.↗

Myanmar Ecological Forecasting: Utilizing NASA Earth Observations to Monitor, Map, and Analyze Mangrove Forests in Myanmar for Enhanced Conservation

Mangroves supply many essential environmental amenities, such as preventing soil erosion, filtering water pollution, and protecting shorelines from harmful waves, floods, storms and winds. The Mangroves in Myanmar not only provide citizens with a food source, but they also offer firewood, charcoal, and construction materials. The depletion of mangroves is threatening more than the biodiversity however; Myanmar's fiscal livelihood is also in harm's way. Mangroves are valued at $100,000 to $277,000 per square kilometer and if managed in a sustainable fashion, can infuse constant income to the emerging Myanmarese economy. This study analyzed three coastline regions, the Ayeyarwady Delta, Rakhine and Tanintharyi, and mapped the spatial extent of mangrove forest during the dry season in 2000 and 2013. The classifications were derived from Landsat 7 Enhanced Thematic Mapper Plus (ETM+) and Landsat 8 Operation Land Imager (OLI) imagery, as well as the Terra Advanced Spaceborne Thermal Emission and Reflection Radiometer (ASTER) digital elevation model information. This data was atmospherically corrected, mosaicked, masked and classified in ENVI, followed by ArcGIS to perform raster calculations and create final products. Forest degradation collected from 2000 to 2013 was later used to forecast the density and health of Mangroves in the year 2030. These results were subsequently presented to project partners Dr. Peter Leimgruber and Ellen Aiken at the Smithsonian Conservation Biology Institute in Front Royal, VA. After the presentation of the project to the partners, these organizations formally passed on to the Myanmar Ministry of Environment, Conservation and Forestry for policy makers and forest managers to utilize in order to protect the Myanmar mangrove ecosystem while sustaining a healthy economy.

Weber, Samuel J.↗

Soil organic carbon is not just for soil scientists: measurement recommendations for diverse practitioners

Soil organic carbon (SOC) regulates terrestrial ecosystem functioning, provides diverse energy sources for soil microorganisms, governs soil structure, and regulates the availability of organically bound nutrients. Investigators in increasingly diverse disciplines recognize how quantifying SOC attributes can provide insight about ecological states and processes. Today, multiple research networks collect and provide SOC data, and robust, new technologies are available for managing, sharing, and analyzing large data sets. Here, we advocate that the scientific community capitalize on these developments to augment SOC data sets via standardized protocols. We describe why such efforts are important and the breadth of disciplines for which it will be helpful, and outline a tiered approach for standardized sampling of SOC and ancillary variables that ranges from simple to more complex. We target scientists ranging from those with little to no background in soil science to those with more soil-related expertise, and offer examples of the ways in which the resulting data can be organized, shared, and discoverable.

54 ENVIRONMENTAL SCIENCES↗

METABOLIC: high-throughput profiling of microbial genomes for functional traits, metabolism, biogeochemistry, and community-scale functional networks

Background Advances in microbiome science are being driven in large part due to our ability to study and infer microbial ecology from genomes reconstructed from mixed microbial communities using metagenomics and single-cell genomics. Such omics-based techniques allow us to read genomic blueprints of microorganisms, decipher their functional capacities and activities, and reconstruct their roles in biogeochemical processes. Currently available tools for analyses of genomic data can annotate and depict metabolic functions to some extent; however, no standardized approaches are currently available for the comprehensive characterization of metabolic predictions, metabolite exchanges, microbial interactions, and microbial contributions to biogeochemical cycling. Results We present METABOLIC (METabolic And BiogeOchemistry anaLyses In miCrobes), a scalable software to advance microbial ecology and biogeochemistry studies using genomes at the resolution of individual organisms and/or microbial communities. The genome-scale workflow includes annotation of microbial genomes, motif validation of biochemically validated conserved protein residues, metabolic pathway analyses, and calculation of contributions to individual biogeochemical transformations and cycles. The community-scale workflow supplements genome-scale analyses with determination of genome abundance in the microbiome, potential microbial metabolic handoffs and metabolite exchange, reconstruction of functional networks, and determination of microbial contributions to biogeochemical cycles. METABOLIC can take input genomes from isolates, metagenome-assembled genomes, or single-cell genomes. Results are presented in the form of tables for metabolism and a variety of visualizations including biogeochemical cycling potential, representation of sequential metabolic transformations, community-scale microbial functional networks using a newly defined metric “MW-score” (metabolic weight score), and metabolic Sankey diagrams. METABOLIC takes ~ 3 h with 40 CPU threads to process ~ 100 genomes and corresponding metagenomic reads within which the most compute-demanding part of hmmsearch takes ~ 45 min, while it takes ~ 5 h to complete hmmsearch for ~ 3600 genomes. Tests of accuracy, robustness, and consistency suggest METABOLIC provides better performance compared to other software and online servers. To highlight the utility and versatility of METABOLIC, we demonstrate its capabilities on diverse metagenomic datasets from the marine subsurface, terrestrial subsurface, meadow soil, deep sea, freshwater lakes, wastewater, and the human gut. Conclusion METABOLIC enables the consistent and reproducible study of microbial community ecology and biogeochemistry using a foundation of genome-informed microbial metabolism, and will advance the integration of uncultivated organisms into metabolic and biogeochemical models. METABOLIC is written in Perl and R and is freely available under GPLv3 at https://github.com/AnantharamanLab/METABOLIC.

59 BASIC BIOLOGICAL SCIENCES↗