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At least 343 records · Page 19

A meta-analysis of global cropland soil carbon changes due to cover cropping

Including cover crops within agricultural rotations may increase soil organic carbon (SOC). However, contradictory findings generated by on-site experiments make it necessary to perform a comprehensive assessment of interactions between cover crops, environmental and management factors, and changes in SOC. In this study, we collected data from studies that compared agricultural production with and without cover crops, and then analyzed those data using meta-analysis and regression. Our results showed that including cover crops into rotations significantly increased SOC, with an overall mean change of 15.5% (95% confidence interval of 13.8% to 17.3%). Whereas medium-textured soils had highest SOC stocks with (mean = 39 Mg ha-1) and without (mean = 37 Mg ha-1) cover crops, fine-textured soils showed the greatest increase in SOC after the inclusion of cover crops (mean change of 39.5%). Coarse-textured (11.4%) and medium-textured soils (10.3%) had comparatively smaller changes in SOC. Cover crop mixtures resulted in greater increases in SOC compared to mono-species cover crops, and using legumes resulted in greater SOC increases than grasses and broadleaf species. Cover cropping was associated with significant SOC increases in shallow soils (= 30 cm), but not in subsurface soils (> 30 cm). The regression analysis revealed that SOC changes from cover cropping were correlated with improvements in soil quality, specifically decreased runoff and erosion and increased mineralizable carbon, mineralizable nitrogen, and soil nitrogen. Soil carbon change was also affected by annual temperature, number of years after start of cover crop usage, latitude, and initial SOC concentrations. Finally, the mean rate of carbon sequestration from cover cropping across all studies was 0.56 Mg ha-1 yr-1. If 15% of current global cropland were to adopt cover crops, this value would translate to 0.17 Pg of carbon sequestered per year or ~1-2% of current fossil fuels emissions. Altogether, these results indicated that inclusion of cover crops into agricultural rotations can enhance soil carbon concentrations, improve many soil quality parameters, and serve as a potential sink for atmosphere CO2.

Cover crop, soil organic carbon, soil quality, soi↗

Scripting a new dialogue between diazotrophs and crops

Diazotrophs are bacteria and archaea that can reduce atmospheric dinitrogen (N 2 ) into ammonium. Plant–diazotroph interactions have been explored for over a century as a nitrogen (N) source for crops to improve agricultural productivity and sustainability. This scientific quest has generated much information about the molecular mechanisms underlying the function, assembly, and regulation of nitrogenase, ammonium assimilation, and plant–diazotroph interactions. This review presents various approaches to manipulating N fixation activity, ammonium release by diazotrophs, and plant–diazotroph interactions. Here, we discuss the research avenues explored in this area, propose potential future routes, emphasizing engineering at the metabolic level via biorthogonal signaling, and conclude by highlighting the importance of biocontrol measures and public acceptance.

59 BASIC BIOLOGICAL SCIENCES↗

Single-cell visualization and quantification of trace metals in Chlamydomonas lysosome-related organelles

Significance Transition metals are of crucial importance for primary productivity; their scarcity limits crop yield in agriculture and carbon sequestration on a global scale. Copper (Cu), iron (Fe), and manganese (Mn) are among the most important trace elements that enable the redox chemistry in oxygenic photosynthesis. The single-celled, eukaryotic green alga Chlamydomonas reinhardtii is a choice experimental system for studying trace metal homeostasis in the context of phototrophy, offering all the advantages of a classical microbial system with a well-characterized photosystem and trace metal metabolism machinery of relevance to plants. This project identifies and differentiates different trace metal storage sites in Chlamydomonas and uncovers the dynamics of trace metal storage and mobilization in situations of fluctuating resources.

54 ENVIRONMENTAL SCIENCES↗

Engineering Crassulacean Acid Metabolism in C 3 and C 4 Plants

Carbon dioxide (CO 2 ) is a major greenhouse gas contributing to changing climatic conditions, which is a grand challenge affecting the security of food, energy, and environment. Photosynthesis plays the central role in plant-based CO 2 reduction. Additionally, plants performing CAM (crassulacean acid metabolism) photosynthesis have a much higher water use efficiency than those performing C 3 or C 4 photosynthesis. Therefore, there is a great potential for engineering CAM in C 3 or C 4 crops to enhance food/biomass production and carbon sequestration on arid, semiarid, abandoned, or marginal lands. Recent progresses in CAM plant genomics and evolution research, along with new advances in plant biotechnology, have provided a solid foundation for bioengineering to convert C 3 /C 4 plants into CAM plants. Here, we first discuss the potential strategies for CAM engineering based on our current understanding of CAM evolution. Then we describe the technical approaches for engineering CAM in C 3 and C 4 plants, with a focus on an iterative four-step pipeline: (1) designing gene modules, (2) building the gene modules and transforming them into target plants, (3) testing the engineered plants through an integration of molecular biology, biochemistry, metabolism, and physiological approaches, and (4) learning to inform the next round of CAM engineering. Finally, we discuss the challenges and future opportunities for fully realizing the potential of CAM engineering.

59 BASIC BIOLOGICAL SCIENCES↗

Nitrogen acquisition and retention pathways in sustainable perennial bioenergy grass cropping systems

Abstract Perennial tall grasses show promise as bioenergy crops due to high productivity and efficient nutrient use. Ongoing research on bioenergy grasses seeks to reduce their reliance on synthetic nitrogen (N) fertilizer, the manufacture of which relies on fossil fuel combustion. Excessive use of fertilizers also causes adverse environmental consequences and leads to the evolutionary loss of plant traits beneficial to sustainable N cycle. Notably, perennial tall grasses have exhibited the potential to maintain high biomass yield without the need for N fertilizer or causing soil N depletion. Perennial grasses can be adept at interacting with their microbial partners to facilitate N acquisition and retention via mechanisms such as biological N fixation and nitrification inhibition. These inherent N management traits should be preserved and optimized at the this early stage of bioenergy grass breeding programs. This review examines the impact of external N on bioenergy grass production and explores the potential of leveraging advantageous N‐cycling attributes of perennial tall grasses, laying groundwork for future management and research efforts. With minimized dependency on external N input, the cultivation of perennial energy grasses will pave the way toward more resilient agricultural systems and play a significant role in addressing key global energy and environmental challenges.

Agriculture↗

Rational design and testing of abiotic stress‐inducible synthetic promoters from poplar cis ‐regulatory elements

Summary Abiotic stress resistance traits may be especially crucial for sustainable production of bioenergy tree crops. Here, we show the performance of a set of rationally designed osmotic‐related and salt stress‐inducible synthetic promoters for use in hybrid poplar. De novo motif‐detecting algorithms yielded 30 water‐deficit (SD) and 34 salt stress (SS) candidate DNA motifs from relevant poplar transcriptomes. We selected three conserved water‐deficit stress motifs (SD18, SD13 and SD9) found in 16 co‐expressed gene promoters, and we discovered a well‐conserved motif for salt response (SS16). We characterized several native poplar stress‐inducible promoters to enable comparisons with our synthetic promoters. Fifteen synthetic promoters were designed using various SD and SS subdomains, in which heptameric repeats of five‐to‐eight subdomain bases were fused to a common core promoter downstream, which, in turn, drove a green fluorescent protein (GFP) gene for reporter assays. These 15 synthetic promoters were screened by transient expression assays in poplar leaf mesophyll protoplasts and agroinfiltrated Nicotiana benthamiana leaves under osmotic stress conditions. Twelve synthetic promoters were induced in transient expression assays with a GFP readout. Of these, five promoters (SD18‐1, SD9‐2, SS16‐1, SS16‐2 and SS16‐3) endowed higher inducibility under osmotic stress conditions than native promoters. These five synthetic promoters were stably transformed into Arabidopsis thaliana to study inducibility in whole plants. Herein, SD18‐1 and SD9‐2 were induced by water‐deficit stress, whereas SS16‐1, SS16‐2 and SS16‐3 were induced by salt stress. The synthetic biology design pipeline resulted in five synthetic promoters that outperformed endogenous promoters in transgenic plants.

09 BIOMASS FUELS↗

Automated Sorghum Phenotyping and Trait Development Platform

There is an urgent need to accelerate energy crop development for the production of renewable transportation fuels from biomass. Our interdisciplinary team developed mobile, ground-based and aerial phenotyping platforms and advanced remote sensing data analysis tools to acquire and process imagery from different types of cameras (e.g. RGB, multispectral, hyperspectral, and thermal sensors) and LiDAR point cloud data in large-scale sorghum field trials during phase 1 of the project. Phase 2 focused on Technology to Market activities to deliver these systems to the market place coupled with targeted research programs to address specific limitations or challenges for these systems.

09 BIOMASS FUELS↗

Broad Spectrum Antifungal Pond Protection

To decrease operating costs associated with fungal infections in algal crops used for biofuel production, we developed bacterial consortia that displayed antifungal properties. These bacteria were grown in culture with algae species without any additional operating costs or need for re-inoculation with bacteria. These co-cultures maintained their antifungal properties for the entirety of the project period and increased mean time to failure (MTTF) by up to 350% when challenged with high levels of fungal pests. Multiple fungal and fungus-like pests were tested and the consortia showed efficacy against three species.

60 APPLIED LIFE SCIENCES↗

EMSL Community Science Campaign Meeting: Critical Minerals and Materials - Rhizo Critical Campaign Breakout Session Report Summary

The “Critical Minerals Biogeochemistry in the Rhizosphere – Ultramafic Soils (Rhizo Critical)” campaign breakout (BO) session was organized to identify major knowledge gaps and fundamental research needs in rhizosphere microbiology and geochemistry that, if addressed, could transform our ability to recover critical minerals from ultramafic soil systems. We sought to identify significant challenges that must be surmounted in the pursuit of deeper science knowledge. Our ultimate goal is to understand this landscape well enough to identify and prioritize opportunities for EMSL to make the greatest impact with Environmental Transformations and Interactions (ETI) science area research campaigns focused on the biogeochemical processes controlling the behavior of critical minerals and materials in the rhizosphere. The increasing demand for critical materials and minerals (CMM) in the U.S. has heightened interest in low-grade ores with much attention on ultramafic soils, which contain valuable metals such as nickel (Ni), chromium (Cr), manganese, cobalt (Co), and copper (Lee et al., 2025; DOE CMM Report, 2023) used in advanced battery, magnet, wiring and wind turbines, and stainless steel technologies. Metal hyperaccumulating plants grown in ultramafic soils can extract economically valuable concentrations of CMMs through the process of phytomining. This technology has evolved from phytoremediation, which involves using plants to cleanse contaminated environments by removing, detoxifying, or stabilizing pollutants like metals and organic compounds. Hyperaccumulator plants are capable of storing metals in their living tissues at concentrations hundreds to thousands of times higher than those found in 'normal' plants. For instance, while the average concentration of Ni in the dry matter of plants growing in typical soils is usually less than 5 µg g?¹, Ni hyperaccumulation is defined by concentrations exceeding 1,000 µg g?¹ (Corzo Remigio et al., 2020; Reeves et al., 2018). Phytomining research has primarily focused on Ni (Rylott and van der Ent, 2025), for which the U.S. has very limited conventional mines in operation. Most soils typically contain Ni concentrations ranging from 7 to 50 mg kg-1, whereas serpentine soils exhibit significantly higher levels, with Ni content often ranging between 700 and 8,000 mg kg-1 (Sobczyk et al., 2017). While more than 500 plant species in over 50 different families have been identified as Ni hyperaccumulators (Kidd et al., 2018), Ni phytomining (and phytominng in general) remains largely untested because most studies are short-term, small-scale, and conducted under simplified or artificially enriched conditions, so they fail to capture the low metal concentrations, environmental variability, and management constraints that would be needed for a field-scale demonstration. Few hyperaccumulator species have been validated as true “metal crops,” and their biomass production, stress tolerance, and rooting characteristics are usually too poor to yield economically meaningful metal outputs. Critically, the basic mechanisms of metal uptake, transport, and sequestration, especially as shaped by belowground processes such as root exudation, rhizosphere chemistry, and root–microbe interactions that control metal mobility and bioavailability (Montreemuk et al., 2023; Kidd et al., 2018; Durand et al., 2023; Alford et al., 2010), are still only partially understood, and downstream metal recovery from biomass is rarely optimized. Because these limitations stem from gaps in fundamental knowledge rather than from a failure of the concept itself (Rylott and van der Ent, 2025; van der Ent et al., 2015), there is a strong need for basic science that dissects plant metal homeostasis, rhizosphere and microbial processes, and their integration with soil chemistry and process engineering to design more robust, scalable phytomining systems.

Ahkami, Amirhossein↗

Establishment of a genome editing tool using CRISPR-Cas9 ribonucleoprotein complexes in the non-model plant pathogen Sphaerulina musiva

CRISPR-Cas9 is a versatile genome editing system widely used since 2013 to introduce site-specific modifications into the genomes of model and non-model species. This technology is used in various applications, from gene knock-outs, knock-ins, and over-expressions to more precise changes, such as the introduction of nucleotides at a targeted locus. CRISPR-Cas9 has been demonstrated to be easy to establish in new species and highly efficient and specific compared to previous gene editing strategies such as Zinc finger nucleases and transcription activator-like effector nucleases. Grand challenges for emerging CRISPR-Cas9 tools in filamentous fungi are developing efficient transformation methods for non-model organisms. In this paper, we have leveraged the establishment of CRISPR-Cas9 genome editing tool that relies on Cas9/sgRNA ribonucleoprotein complexes (RNPs) in the model species Trichoderma reesei and developed the first protocol to efficiently transform the non-model species, Sphaerulina musiva. This fungal pathogen constitutes a real threat to the genus Populus, a foundational bioenergy crop used for biofuel production. Herein, we highlight the general considerations to design sgRNAs and their computational validation. We also describe the use of isolated protoplasts to deliver the CRISPR-Cas9 RNP components in both species and the screening for targeted genome editing events. The development of engineering tools in S. musiva can be used for studying genes involved in diverse processes such as secondary metabolism, establishment, and pathogenicity, among many others, but also for developing genetic mitigation approaches. The approach described here provides guidance for potential development of transformation systems in other non-model spore-bearing ascomycetes.

59 BASIC BIOLOGICAL SCIENCES↗

Role of SaPCR2 in Zn Uptake in the Root Elongation Zone of the Zn/Cd Hyperaccumulator Sedum alfredii

Zn pollution is a potential toxicant for agriculture and the environment. Sedum alfredii is a Zn/Cd hyperaccumulator found in China and has been proven as a useful resource for the phytoremediation of Zn-contaminated sites. However, the molecular mechanism of Zn uptake in S. alfredii is limited. In this study, the function of SaPCR2 on Zn uptake in S. alfredii was identified by gene expression analysis, yeast function assays, Zn accumulation and root morphology analysis in transgenic lines to further elucidate the mechanisms of uptake and translocation of Zn in S. alfredii. The results showed that SaPCR2 was highly expressed in the root elongation zone of the hyperaccumulating ecotype (HE) S. alfredii, and high Zn exposure downregulated the expression of SaPCR2 in the HE S. alfredii root. The heterologous expression of SaPCR2 in yeast suggested that SaPCR2 was responsible for Zn influx. The overexpression of SaPCR2 in the non-hyperaccumulating ecotype (NHE) S. alfredii significantly increased the root uptake of Zn, but did not influence Mn, Cu or Fe. SR-μ-XRF technology showed that more Zn was distributed in the vascular buddle tissues, as well as in the cortex and epidermis in the transgenic lines. Root morphology was also altered after SaPCR2 overexpression, and a severe inhibition was observed. In the transgenic lines, the meristematic and elongation zones of the root were lower compared to the WT, and Zn accumulation in meristem cells was also reduced. These results indicate that SaPCR2 is responsible for Zn uptake, and mainly functions in the root elongation zone. This research on SaPCR2 could provide a theoretical basis for the use of genetic engineering technology in the modification of crops for their safe production and biological enhancement.

59 BASIC BIOLOGICAL SCIENCES↗

Janus: A Python Package for Agent-Based Modeling of Land Use and Land Cover Change

Janus is an open source Python package for agent-based modeling (ABM) of land use and land cover change (LULCC). Many ABMs of LULCC have been created across platforms, some of which are not ideal for large scale, high resolution scenarios. This model provides a simple object-oriented framework for creating ABMs specific to LULCC. The organizational philosophy of the modeling framework is to create software objects (agents) that are associated with specific and contextual attributes which are isolated from where those agents exist in the spatial setting of the model, yet provide clear linkages between the agent, their environment, and other agents in the simulation. In this way, the framework allows for assembly of LULCC ABMs with low (programmatic) overhead, making the models extensible and providing clear mechanisms for integrating them with process-oriented biophysical models. Provided with Janus is a suite of geospatial data preprocessing tools that can use arbitrary land cover products as an input. Crop choice decisions are based on potential crop prices, these can be created synthetically, or drawn from integrated human-Earth systems models such as the GCAM. Janus is publicly accessible through GitHub and provides an example dataset for testing.

54 ENVIRONMENTAL SCIENCES↗

Engineering Crassulacean Acid Metabolism in C3 and C4 Plants

Carbon dioxide (CO2) is a major greenhouse gas contributing to changing climatic conditions, which is a grand challenge affecting the security of food, energy, and environment. Photosynthesis plays the central role in plant-based CO2 reduction. Plants performing CAM (crassulacean acid metabolism) photosynthesis have a much higher water use efficiency than those performing C3 or C4 photosynthesis. Therefore, there is a great potential for engineering CAM in C3 or C4 crops to enhance food/biomass production and carbon sequestration on arid, semiarid, abandoned, or marginal lands. Recent progresses in CAM plant genomics and evolution research, along with new advances in plant biotechnology, have provided a solid foundation for bioengineering to convert C3/C4 plants into CAM plants. Here, we first discuss the potential strategies for CAM engineering based on our current understanding of CAM evolution. Then we describe the technical approaches for engineering CAM in C3 and C4 plants, with a focus on an iterative four-step pipeline: (1) designing gene modules, (2) building the gene modules and transforming them into target plants, (3) testing the engineered plants through an integration of molecular biology, biochemistry, metabolism, and physiological approaches, and (4) learning to inform the next round of CAM engineering. Finally, we discuss the challenges and future opportunities for fully realizing the potential of CAM engineering.

Yang, Xiaohan↗

Cover crops and poultry litter impact on soil structural stability in dryland soybean production in southeastern United States

Abstract This study explored the efficacy of soil aggregate indices in quantifying soil structural development, utilizing 5‐year field experiment data from the Southeastern United States. The experiment utilized a split‐plot design with cover crops (native vegetation as control, cereal rye (Secale cerealeL.), winter wheat (Triticum aestivum), hairy vetch (Vicia villosa), and mustard (Brassica rapa) plus cereal rye as the main factor and fertilizer source (no fertilizer as control, inorganic fertilizer with phosphorus, potassium, and elemental sulfur, and poultry litter) as the secondary factor. Aggregate size fractions were determined using the wet‐sieving method, and aggregate stability index (ASI), mean weight diameter (MWD), geometric mean diameter (GMD), and fractal dimension (FD) were calculated to assess soil structural stability. Main effects results indicated that cereal rye (55.11%) and poultry litter (50.97%) exhibited the highest ASI values. The highest MWD, GMD, and FD were observed under mustard plus cereal rye (1.187 mm), cereal rye (0.462 mm), and hairy vetch (2.573), respectively. Principal component analysis revealed that cover crops significantly improved soil aggregate structure and stability, overcoming limitations of sole fertilization practices. Regression analysis suggested that ASI, MWD, and GWD positively correlated with soil organic carbon, whereas FD negatively correlated with MWD, GMD, and ASI. Principal component analysis exhibited that FD decreased with increasing soil organic carbon, ASI, MWD, and GMD, demonstrating that lower FD values indicate enhanced soil aggregation and structure. Assessed indices, FD included, effectively gauged soil structural stability. These metrics should be prioritized in managerial decisions to support soil productivity and health in agricultural systems.

Agriculture↗

The Utility of Satellites and Autonomous Remote Sensing Platforms for Monitoring Offshore Aquaculture Farms: A Case Study for Canopy Forming Kelps

The emerging sector of offshore kelp aquaculture represents an opportunity to produce biofuel feedstock to help meet growing energy demand. Giant kelp represents an attractive aquaculture crop due to its rapid growth and production, however precision farming over large scales is required to make this crop economically viable. These demands necessitate high frequency monitoring to ensure outplant success, maximum production, and optimum quality of harvested biomass, while the long distance from shore and large necessary scales of production makes in person monitoring impractical. Remote sensing offers a practical monitoring solution and nascent imaging technologies could be leveraged to provide daily products of the kelp canopy and subsurface structures over unprecedented spatial scales. Here, we evaluate the efficacy of remote sensing from satellites and aerial and underwater autonomous vehicles as potential monitoring platforms for offshore kelp aquaculture farms. Decadal-scale analyses of the Southern California Bight showed that high offshore summertime cloud cover restricts the ability of satellite sensors to provide high frequency direct monitoring of these farms. By contrast, daily monitoring of offshore farms using sensors mounted to aerial and underwater drones seems promising. Small Unoccupied Aircraft Systems (sUAS) carrying lightweight optical sensors can provide estimates of canopy area, density, and tissue nitrogen content on the time and space scales necessary for observing changes in this highly dynamic species. Underwater color imagery can be rapidly classified using deep learning models to identify kelp outplants on a longline farm and high acoustic returns of kelp pneumatocysts from side scan sonar imagery signal an ability to monitor the subsurface development of kelp fronds. Current sensing technologies can be used to develop additional machine learning and spectral algorithms to monitor outplant health and canopy macromolecular content, however future developments in vehicle and infrastructure technologies are necessary to reduce costs and transcend operational limitations for continuous deployment in an offshore setting.

54 ENVIRONMENTAL SCIENCES↗

Minimizing Fraud in the Carbon Offset Market Using Blockchain Technologies

Fraud in the Environmental Benefit Credit (EBC) markets is pervasive. To make matters worse, the cost of creating EBCs is often higher than the market price. Consequently, a method to create, validate, and verify EBCs and their relevance is needed to mitigate fraud. The EBC market has focused on geologic (fossil fuel) CO 2 sequestration projects that are often over budget and behind schedule and has failed to capture the "lowest hanging fruit" EBCs - terrestrial sequestration via the agricultural industry. This project reviews a methodology to attain possibly the least costly EBCs by tracking the reduction of inputs required to grow crops. The use of bio- stimulant products, such as humate, allows a farmer to use less nitrogen without adversely affecting crop yield. Using less nitrogen qualifies for EBCs by reducing nitrous oxide emissions and nitrate runoff from a farmer's field. A blockchain that tracks the bio-stimulant material from source to application provides a link between a tangible (bio-stimulant commodity) and the associated intangible (EBCs) assets. Covert insertion of taggants in the bio-stimulant products creates a unique barcode that allows a product to be digitally tracked from beginning to end. This process (blockchain technology) is so robust, logical, and transparent that it will enhance the value of the associated EBCs by mitigating fraud. It provides a real time method for monetizing the benefits of the material. Substantial amounts of energy are required to produce, transport, and distribute agricultural inputs including fertilizer and water. Intelligent optimization of the use of agricultural inputs can drive meaningful cost savings. Tagging and verification of product application provides a valuable understanding of the dynamics in the water/food energy nexus, a major food security and sustainability issue. As technology in agriculture evolves so to must methods to verify the Enterprise Resource Planning (ERP) potential of innovative solutions. The technology reviewed provides the ability to combine blockchain and taggants ("taggant blockchains") as the engine by which to (1) mitigate fraudulent carbon credits; (2) improve food chain security, and (3) monitor and manage sustainability. The verification of product quality and application is a requirement to validate benefits. Recent upgrades to humic and fulvic quality protocols known as ISO CD 19822 TC134 offers an analytical procedure. This work has been assisted by the Humic Products Trade Association and International Humic Substance Society. In addition, providing proof of application of these products and verification of the correct application of prescriptive humic and bio-stimulant products is required. Individual sources of humate have unique and verifiable characteristics. Additionally, methods for prescription of site- specific agricultural inputs in agricultural fields are available. (See US Patents 734867B2, US 90658633B2.) Finally, a method to assure application rate is required through the use of taggants. Sensors using organic solid to liquid phase change nanoparticles of various types and melting temperatures added to the naturally occurring materials provide a barcode. Over 100 types of nanoparticles exist ensuring numerous possible barcodes to reduce industry fraud. Taggant materials can be collected from soil samples of plant material to validate a blockchain of humic, fulvic and other soil amendment products. Other non-organic materials are also available as taggants; however, the organic tags are biodegradable and safe in the environment allowing for use during differing application timeliness.

54 ENVIRONMENTAL SCIENCES↗

The Realistic Potential of Soil Carbon Sequestration in U.S. Croplands for Climate Mitigation

Existing estimates of the climate mitigation potential from cropland carbon sequestration (C-sequestration) are limited because they tend to assume constant rates of soil organic carbon change over all available cropland area, use relatively coarse land delineations, and often fail to adequately consider the agronomic and socioeconomic dimensions of agricultural land use. This results in an inflated estimate of the C-sequestration potential. We address this gap by defining a more appropriate land base for cover cropping in the United States for C-sequestration purposes: stable croplands in annual production systems that can integrate cover cropping without irrigation. Our baseline estimate of this suitable stable cropland area is 32% of current U.S. cropland extent. Even an alternative, less restrictive definition of stability results in a large reduction in area (44% of current U.S. croplands). Focusing cover crop implementation to this constrained land base would increase durability of associated C-sequestration and limit soil carbon loss from land conversion to qualify for carbon-specific incentives. Applying spatially-variable C-sequestration rates from the literature to our baseline area yields a technical potential of 19.4 Tg CO 2 e yr –1 annually, about one-fifth of previous estimates. We also find the cost of realizing about half (10 Tg CO 2 e yr –1 ) of this potential could exceed 100 USD Mg CO 2 e –1 , an order of magnitude higher than previously thought. While our economic analyses suggest that financial incentives are necessary for large-scale adoption of cover cropping in the U.S., they also imply any C-sequestration realized under such incentives is likely to be additional.

54 ENVIRONMENTAL SCIENCES↗