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All sourcesNASA NTRSDOE OSTI
DOE OSTI2024

Better Living Through Biology: Studying Enzymes to Make Industrial Processes More Efficient

In biology, enzymes are the molecular machinery needed to speed up slow chemical reactions for life to occur. These molecular machines enhance chemical processes to a large degree, allowing for improbable and challenging chemical reactions to efficiently happen in water at room temperature. Many microorganisms have specialized enzymes used to tackle particularly challenging chemistry that they experience in their own environment. Our group is studying a specific enzyme using a reaction called "electron bifurcation" that energetically pairs electrons and is analogous to a trampoline, using one person's jump to propel another person higher than they could by themselves. We have learned that certain metal- and vitamin-containing pathways in this protein funnel electrons in specific directions. Our investigation into this enzyme is the foundation for future industrial applications, ranging from biological production of sustainable aviation fuel from CO2, production of nitrogen-based fertilizers, and biologically remediating environmental contamination.

BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2025

Overestimated natural biological nitrogen fixation translates to an exaggerated CO 2 fertilization effect in Earth system models

CO 2 fertilization of the terrestrial biosphere is limited by nitrogen. Biological nitrogen fixation (BNF) is the dominant natural nitrogen source to the terrestrial biosphere and can alleviate nitrogen limitation but is poorly constrained in Earth system models (ESMs). Here, in this study, we compare terrestrial BNF from an ensemble of ESMs of the 6th Coupled Model Intercomparison Project to a new global synthesis of observations across natural and agricultural biomes. We find that compared to observations, ESMs underestimate agricultural BNF but overestimate natural BNF in the present day by over 50%. Natural BNF is overestimated in the most productive ecosystems that contribute most to the terrestrial carbon sink (forests and grasslands). ESMs with different BNF representations yield a range of BNF responses to CO 2 enrichment. Some ESMs with phenomenological representations of BNF predict a natural BNF increase in response to a doubling of CO 2 that aligns with a meta-analysis of CO 2 enrichment experiments (31% increase) but fail to account for the substantial carbon cost of BNF. In contrast, ESMs with mechanistic representations of BNF account for its carbon cost as well as its regulation by nitrogen limitation but overestimate the BNF response to a doubling of CO 2 (135% increase). Overall, all current BNF representations in ESMs fall short of fully capturing its response to rising atmospheric CO 2 . Finally, we find a positive correlation between modeled present-day natural BNF and the CO 2 fertilization effect across ESMs, suggesting that overestimated natural BNF translates to an exaggerated CO 2 fertilization effect of approximately 11% in ESMs.

Biological nitrogen fixation↗
DOE OSTI2022

Whole-tree dormant season nitrogen pools for different species receiving combinations of fertilization and irrigation after one short rotation

Our understanding of the accretion and distribution of tree tissue nitrogen (N) pools across a variety of species and genotypes suitable for short-rotation woody crop (SRWC) production in response to water and N availability remains limited. We measured dormant-season, rotation-length, whole-tree N pools for five tree genotypes from four species (two eastern cottonwood, Populus deltoides Bartr., genotypes; American sycamore, Platanus occidentalis L.; American sweetgum, Liquidambar styraciflua L.; and loblolly pine, Pinus taeda L.) receiving irrigation (I), fertilization (F), their combination (IF), or no resources manipulation (C). Our results demonstrate that foliar nitrogen concentration [N] responded to fertilization but was constrained within genotype-specific ranges and varied temporally. Tree genotypes differed in their composite and component tissue N content (N C ), and these differences mostly reflected tissue mass (i.e., larger components and trees resulted in higher N C ). Resource amendments (I, F, IF) resulted in up to 3.8-fold increases in N C compared with C, which were most pronounced for sycamore and sweetgum with F and IF treatments, respectively. By the end of the rotation, forest stands accumulated 73 to 452 kg N ha –1 in tree tissues with 40 – 78 % distributed aboveground and 22 – 60 % distributed belowground. A critical difference between genotypes was that all hardwoods exhibited larger belowground N distributions than the evergreen conifer. Our results stress the importance of belowground N pools and highlight differences among genotypes. Furthermore, our study underscores valuable information about N pools across genotypes suitable for SRWC production, which can be leveraged to inform fertilization plans and devise sustainable nutrient management as production expands across marginal lands.

54 ENVIRONMENTAL SCIENCES↗
DOE OSTI2025

Soil fertility management for sustainable Miscanthus × giganteus production: Increased tiller weight from nitrogen management explains yield gains in aged miscanthus

Aging-related yield decline in Miscanthus × giganteus (miscanthus) remains a major constraint to sustainable biomass production. This study evaluated how nitrogen (N) management and soil fertility influence yield-component traits and productivity in aging miscanthus. Trials were conducted at two sites established in 2008 at the University of Illinois Energy Farm, Urbana, IL. (i) The Sun Grant trial received 0, 60, and 120 kg N ha −1 annually until 2015. Starting 2021, half of each plot received 60 or 120 kg N ha −1 , resulting in six legacy-contemporary treatments: 0N–0N, 0N–120N, 60N–0N, 60N–60N, 120N–0N, 120N–120N. (ii) The Energy Farm trial remained unfertilized until 2014, when one half of each plot received 56 kg N ha −1 , forming two treatments: 0N–0N, 0N–56N. Sun Grant trial results showed N fertilization increased tiller density (tillers m −2 ) and tiller weight (g tiller −1 ) in juvenile to early-mature miscanthus (2011–2015). After N withdrawal, both traits declined (20 % and 40 %), though legacy effects persisted in tiller weight in the aging stands (2020–2023). Contemporary N had little effect on tiller density but increased tiller weight by 34 %–77 %, resulting in 23 %–106 % higher machine-harvested biomass yield in 0–120N, 60-60N, and 120-120N plots. At the Energy Farm trial, 0N–56N plots yielded 59 %–108 % more biomass than 0N–0N. Soil total N increased (Sun Grant: 47 % by 2020; Energy Farm: 58 % by 2023), while Mehlich-3 P (42 %–44 %) and K (21 %–46 %) declined. These findings identify tiller weight as a key determinant of biomass yield in aging miscanthus and highlight the need for P and K management for long-term productivity.

09 BIOMASS FUELS↗
DOE OSTI2025

Data for Soil Fertility Management for Sustainable Miscanthus × giganteus Production: Increased Tiller Weight from Nitrogen Management Explains Yield Gains in Aged Miscanthus

Aging-related yield decline in Miscanthus × giganteus (miscanthus) remains a major constraint to sustainable biomass production. This study evaluated how nitrogen (N) management and soil fertility influence yield-component traits and productivity in aging miscanthus. Trials were conducted at two sites established in 2008 at the University of Illinois Energy Farm, Urbana, IL. (i) The Sun Grant trial received 0, 60, and 120 kg N ha−1 annually until 2015. Starting 2021, half of each plot received 60 or 120 kg N ha−1, resulting in six legacy-contemporary treatments: 0N–0N, 0N–120N, 60N–0N, 60N–60N, 120N–0N, 120N–120N. (ii) The Energy Farm trial remained unfertilized until 2014, when one half of each plot received 56 kg N ha−1, forming two treatments: 0N–0N, 0N–56N. Sun Grant trial results showed N fertilization increased tiller density (tillers m−2) and tiller weight (g tiller−1) in juvenile to early-mature miscanthus (2011–2015). After N withdrawal, both traits declined (20 % and 40 %), though legacy effects persisted in tiller weight in the aging stands (2020–2023). Contemporary N had little effect on tiller density but increased tiller weight by 34 %–77 %, resulting in 23 %–106 % higher machine-harvested biomass yield in 0–120N, 60-60N, and 120-120N plots. At the Energy Farm trial, 0N–56N plots yielded 59 %–108 % more biomass than 0N–0N. Soil total N increased (Sun Grant: 47 % by 2020; Energy Farm: 58 % by 2023), while Mehlich-3 P (42 %–44 %) and K (21 %–46 %) declined. These findings identify tiller weight as a key determinant of biomass yield in aging miscanthus and highlight the need for P and K management for long-term productivity.

miscanthus↗
DOE OSTI2021

Quantifying nitrogen loss hotspots and mitigation potential for individual fields in the US Corn Belt with a metamodeling approach

The high productivity in the US Corn Belt is largely enabled by the consumption of millions of tons of manufactured fertilizer. Excessive application of nitrogen (N) fertilizer has been pervasive in this region, and the unrecovered N eventually escaped from croplands in forms of nitrous oxide (N 2 O) emission and N leaching. Mitigating these negative impacts is hindered by a lack of practical information on where to focus and how much mitigation potential to expect. At a large scale, process-based crop models are the primary tools for predicting variables required by decision making, but their applications are prohibited by expensive computational and data storage costs. To overcome these challenges, we built a series of metamodels to learn the key mechanisms regarding the carbon (C) and N cycle from a well-validated process-based biogeochemical model, ecosys. The trained metamodel captures over 98% of the variability of the ecosys simulated outputs for 99 randomly selected counties in Iowa, Illinois, and Indiana. To identify hotspots with high mitigation potential, we introduce net societal benefit (NSB) as an indicator for synthesizing the loss in yield and social benefits through emissions and pollutants avoided. Our results show that reducing N fertilizer by 10% leads to 9.8% less N 2 O emissions and 9.6% less N leaching at the cost of 4.9% more SOC depletion and 0.6% yield reduction over the study region. The estimated total annual NSB is $\$395$ M (uncertainty ranges from $\$114$ M to $\$1271$ M), including $\$334$ from social benefits (uncertainty ranges from $\$46$ M to $\$1076$ M), $\$100$ M from saving fertilizer (uncertainty ranges from $\$13$ M to $\$455$ M), and –$\$40$ M due to yield changes (uncertainty ranges from –$\$261$ M to $\$69$ M). For the median scenario, we noted that 20% of the study area accounts for nearly 50% of the NSB, and thus represent hotspot locations for targeted mitigation. Although the uncertainty range suggests that developing such a high-resolution framework is not yet settled and the scenario based estimations are not appropriate to inform the management practices for individual farmers, our efforts shed light on the new generation of analytical tools for life cycle assessment.

54 ENVIRONMENTAL SCIENCES↗
DOE OSTI2024

Position-specific kinetic isotope effects for nitrous oxide: a new expansion of the Rayleigh model

Nitrous oxide (N 2 O) is a potent greenhouse gas and the most significant anthropogenic ozone-depleting substance currently being emitted. A major source of anthropogenic N 2 O emissions is the microbial conversion of fixed nitrogen species from fertilizers in agricultural soils. Thus, understanding the enzymatic mechanisms by which microbes produce N 2 O has environmental significance. Measurement of the 15 N/ 14 N isotope ratios of N 2 O produced by purified enzymes or axenic microbial cultures is a promising technique for studying N 2 O biosynthesis. Typically, N 2 O-producing enzymes combine nitrogen atoms from two identical substrate molecules (NO or NH 2 OH). Position-specific isotope analysis of the central (N α ) and outer (N β ) nitrogen atoms in N 2 O enables the determination of the individual kinetic isotope effects (KIEs) for N α and N β , providing mechanistic insight into the incorporation of each nitrogen atom. Previously, position-specific KIEs (and fractionation factors) were quantified using the Rayleigh distillation equation, i.e., via linear regression of δ 15 N α or δ 15 N β against [–f In f / (1 – f)], where f is the fraction of substrate remaining in a closed system. This approach, however, is inaccurate for N α and N β because it does not account for fractionation at N α affecting the isotopic composition of substrate available for incorporation into the β position (and vice versa). Therefore, we developed a new expansion of the Rayleigh model that includes specific terms for fractionation at the individual N 2 O nitrogen atoms. By applying this Expanded Rayleigh model to a variety of simulated N 2 O synthesis reactions with different combinations of normal, inverse, and/or no KIEs at N α and N β , we demonstrate that our new model is both accurate and robust. We also applied this new model to two previously published datasets describing N 2 O production from NH 2 OH oxidation in a methanotroph culture (Methylosinus trichosporium) and N 2 O production from NO by a purified Histoplasma capsulatum (fungal) P450 NOR, demonstrating that the Expanded Rayleigh model is a useful tool in calculating position-specific fractionation for N 2 O synthesis.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗
DOE OSTI2025

Observed declines in leaf nitrogen explained by photosynthetic acclimation to CO 2

Widespread evidence of decreasing leaf nutrients has raised concerns about ecosystem productivity under global change. Interpreting trends in leaf nutrients has important implications for the fate of ecosystem services, particularly the role of forests in mitigating climate change and sustaining quality food sources. Here, we challenge the common interpretation that decreasing leaf nitrogen concentration (LNC) is evidence of increasing nutrient limitations on ecosystem primary productivity. Instead, we show that declines in LNC (4% decrease per 50 ppm CO 2 increase), observed across 409 European forest plots over 22 y, can be explained by reduced photosynthetic nitrogen demand. This regional trend is consistent with leaf acclimation to increasing atmospheric CO 2 according to optimality theory. This finding suggests that enhanced photosynthetic nitrogen use efficiency due to CO 2 fertilization may lead to less nitrogen uptake and/or reallocation of nitrogen for plant growth and other functions. Our results have large implications for understanding and simulating interactions between ecosystem nitrogen and carbon cycles and suggest nitrogen requirements for terrestrial carbon uptake under elevated CO 2 may be lower than previously thought.

CO2 fertilization↗
DOE OSTI2020

Techno-Economic Analysis and Global Warming Potential of a Novel Offshore Macroalgae Biorefinery

The success of a large scale macroalgae-based biorefinery is dependent on the demonstration of favorable system economics and environmental sustainability. This study uses detailed process modeling to quantify the mass and energy flows through the various unit operations required for a novel free-floating macroalgae biorefinery concept. The modular process model served as the foundation for the techno-economic and global warming potential analyses used to quantify the sustainability of the proposed concept. This work includes detailed techno-economic results for a complete macroalgae cultivation and conversion system with multiple hatchery configurations and several emerging technologies. System optimization was achieved through the evaluation of various technology options for each unit operation. Technologies considered include traditional twine and textile substrate hatchery configurations, drone assisted seeding and biomass transport, mechanized line seeding and harvesting, adhesive spore mixtures that simplify seeding operations and improve hatchery energetics, and hydrothermal liquefaction to produce upgradable biocrude. Outputs from the system include renewable diesel (R100), naphtha, biochar, nitrogen and phosphorus fertilizers, and aqueous/solid waste streams. Three different system pathways were explored, yielding a biomass production cost ranging from $210 to $565 per dry metric ton and a minimum fuel selling price from $1.35 to $2.91 per liter of gasoline equivalent. Stochastic manipulation of the process model and sensitivity analyses support these results. The global warming potential analysis shows net greenhouse gas emissions ranging from 14 to 29 gCO2-eq MJ-1, supported by stochastic and sensitivity analyses. The recommendations from this work highlight critical areas for research and development investment such that a sustainable macroalgae cultivation and conversion system can be realized.

NOMAD seaweed cultivation, techno-economic analysi↗
DOE OSTI2020

Facile All-Optical Method for In Situ Detection of Low Amounts of Ammonia

As a key precursor for nitrogenous compounds and fertilizer, ammonia affects our lives in numerous ways. Rapid and sensitive detection of ammonia is essential, both in environmental monitoring and in process control for industrial production. Here we report a novel and nonperturbative method that allows rapid detection of ammonia at low concentrations, based on the all-optical detection of surface-enhanced Raman signals. We show that this simple and affordable approach enables ammonia probing at selected regions of interest with high spatial resolution, making in situ and operando observations possible.

09 BIOMASS FUELS↗
DOE OSTI2024

Tropical root traits in response to global changes from 1984 to 2023

This dataset is a compilation of tropical root traits data in response to different global changes in tropical sites, considering 23.50N and S as latitudinal boundaries. The global changes considered are warming, drought, flooding, cyclones, nitrogen addition, CO2 fertilization, and fire. This dataset contains 266 root trait observations from 93 studies across 24 tropical countries. The full citation from where the data was taken from is provided in the dataset, as well as the global change, the ecosystem type, location, coordinates, the root traits measured, and the direction of their response after the global change. Additional information such as the duration of the experiment, the intensity of the global change, the soil layers from where the roots were collected, the root orders, and the type of experiment are also shown. We obtained this dataset by performing a systematic literature review on Web of Science using standardized keywords in English, Spanish, and Portuguese (Yaffar, Lugli et al. in press).

54 ENVIRONMENTAL SCIENCES↗
DOE OSTI2022

Photosynthesis in aging perennial and leaf nitrogen

Perennial energy grasses are a fundamental component of the emerging bioeconomy. While their yield dynamics and environmental services are being widely studied, little attention has been given to how the physiological changes associated with perenniality (e.g., nutrient cycling) that underpin yield and environmental services, change as a function of age. Across species, perennial grasses show a photosynthetic decline as a function of stand age, however, the physiological drivers of this age-related decline are poorly understood. In this manuscript we ask: by how much does nitrogen dilution in larger older plants contribute to this photosynthetic decline? Our results support the hypothesis that nitrogen dilution did not limit photosynthesis in mature stands. Compared per unit of area-based leaf nitrogen, photosynthesis of fertilized mature stands was still 26% lower than unfertilized juvenile stands. We hypothesize that the lack of late-season carbohydrate sinks may instead drive the age-related decline in photosynthesis.

Tejera, Mauricio↗
DOE OSTI2022

Improved Moisture Management Technology for Eco-N-Control Fertilizer

Three fertilizer technologies to manage nitrogen (N) release rates in soil dominate the market today: 1) bulk controlled-release technologies, which use the intrinsic solubility properties of a N-organic co-polymer to control N release; 2) barrier-based controlled-release technologies, which use porous polymer or sulfur coatings as a physical barrier to control N release from encapsulated N nutrients; and 3) slow-release technologies, which use enzyme inhibitors to chemically slow down the transformation of N nutrients into ammonia (NH 3 ) or to temporarily reduce populations of bacteria in the soil that are responsible for converting ammonium (NH 4 + ) to highly soluble nitrates. These technologies are expensive and provide only limited control of N-release rates. Also, from another environmental perspective, synthetic polymer coatings do not decompose easily and will accumulate over time with continuous application of fertilizer. For most of the past two decades we have been developing the Eco-N-Control solid-nitride fertilizer technology, a type of bulk controlled-release technology. With Eco-N-Control, solid-nitride compounds weather like a mineral, slowly releasing N at a rate determined by their composition. Testing by an industrial collaborator, however, showed that earlier versions of Eco-N-Control released at 25% of the desired rate when deployed in unsaturated soils. The overall goal of the current project, therefore, was to strengthen our IP position and the potential for licensing of Eco-N-Control solid-nitride fertilizer technology by developing new ways to manage the level of moisture in contact with the fertilizer when applied to soil thereby ensuring the desired N-release rate. We achieved this goal by developing a biodegradable polymer cross-linking approach that substantially improved performance of our patented solid-nitride fertilizer. We found that cross-linking an environmentally friendly and biodegradable polymer with a mixture of solid nitrides, other N-nutrient compounds and other additives offers several advantages: 1) mechanical robustness of fertilizer beads, 2) chemical stability during storage, 3) maintenance of sufficient water content in the vicinity of the nitride, and 4) control of N-release rates. We also updated our N-release testing protocol to more realistically simulate the conditions found in soil. Finally, we filed an invention disclosure record to protect these new aspects of Eco-N-Control technology, which offers a potential way to lower both agricultural and industrial pollution by providing an efficient N-delivery mechanism to plants.

42 ENGINEERING↗
DOE OSTI2022

Genetic variation in nitrogen‐use efficiency and its associated traits in dryland winter wheat ( Triticum aestivum L.) cultivars released from the 1940s to the 2010s in Shaanxi Province, China

Abstract BACKGROUND Improving the nitrogen‐use efficiency (NUE) of wheat can help mitigate the problems of poor soil fertility under dryland conditions. We conducted field experiments using three nitrogen (N) fertilization levels (0, 120, and 180 kg ha −1 ) applied to eight dryland wheat cultivars to assess NUE and its associated traits. RESULTS The grain yield significantly increased with the improvement in variety, mainly as a result of a substantial increase in 1000‐grain weight and harvest index. Modern wheat varieties have stabilized at an optimal plant height and exhibited improved performance in terms of NUE, partial N productivity, N harvest index, and grain protein content compared to older varieties. The NUE of wheat gradually increased with variety replacement. The net photosynthesis rate of the flag leaves in the filling stage improved with the year of cultivar release; Increasing soil–plant analysis development (SPAD) values of flag leaves in the flowering and filling stages were observed over time, with the flag leaves of modern varieties showing a high chlorophyll content in the filling stage. Additionally, the principal component analysis showed that the SPAD value, grain number per unit area, transpiration rate, leaf area, and grain protein content positively contributed to the clustering of the N180 and modern cultivars (from the 2000s to 2010s). CONCLUSION Overall, high levels of N application did not significantly improve the NUE of wheat. However, modern wheat varieties can optimize N distribution, increase flag leaf photosynthetic capacity, and improve photosynthesis ability, thus enhancing NUE to achieve high yields under a suitable level of N supply. © 2022 Society of Chemical Industry.

Lian, Huida↗
DOE OSTI2025

Influence of fertilization on the dynamics of energy use in wheat

Plant energy use is fundamental to plant survival and growth. However, we still lack effective means to quantify plant energy use strategies. This study introduced a concept quantifying the light level at which photochemical and non-photochemical energy use in plants are in equilibrium — the photochemical compensation point (PCCP) which can be determined with chlorophyll fluorescence measurements. We used winter wheat as a test case to explore the dynamics of PCCP and its physiological and biochemical regulations. Winter wheat PCCP decreased significantly across growth stages from jointing to grain filling. Long-term nitrogen and phosphate (NP) fertilization significantly increased PCCP, whereas potassium (K) and manure (M) fertilizer supplementation had negligible effects. PCCP exhibited significant positive correlations with leaf thickness, leaf P and sulfur (S), and stomatal conductance (gs) across all growth stages. All manure-amended treatments exhibited positive correlations of PCCP with leaf N, P, K and gs, and negative correlations with leaf calcium (Ca). Random forest analysis revealed that gs was the most significant predictor of PCCP variation, followed by leaf P, iWUE, and leaf thickness across all treatments. We suggest that plant energy use strategies are strongly coupled with plant water use strategies and nutrient availability through a complex interplay of effects on physiological and biochemical traits.

energy allocation↗
DOE OSTI2021

Soil net nitrogen mineralization and leaching under Miscanthus × giganteus and Zea mays

Abstract The winter fallow period common in annual cropping systems leaves soils vulnerable to erosion and nutrient loss, especially to nitrogen (N) leaching. This vulnerability can be mitigated with perennial crops that have living roots in the ground year‐round. The mechanisms, magnitude, and consistency with which perennial crops retain N are not clear. We used an experiment to test whether a perennial crop, miscanthus ( Miscanthus × giganteus Greef et Deu.), would leach less N than continuous maize ( Zea mays L.) and how soil net N mineralization (N min ) may explain observed leaching under varied environment and management conditions. The experiment included three crossed factors: (1) cropping system (maize, juvenile miscanthus = 1–2 years old, mature miscanthus = 3–4 years old); (2) N fertilization (0 and 224 kg N ha −1 ); and (3) environment (four site‐years at two locations in Iowa, USA, that differed in climate and soil fertility). We measured N cycling dynamics, including: inorganic soil N (ammonium + nitrate), in situ N min , N leaching, crop N uptake, and calculated system N use efficiency. There were many complex interactions among factors. On average, cumulative N min under juvenile miscanthus was 111% greater than maize, but as miscanthus matured, there was no difference in N min between the perennial crop and maize. There was no difference in N leaching between juvenile miscanthus and maize, but mature miscanthus decreased N leaching by 42% and 88% compared to maize (with and without N fertilization, respectively). Across all treatments, there was no relationship between N min and N leaching, suggesting other mechanisms are regulating N leaching. Overall, mature miscanthus shows promise as a tool to reduce N losses in areas dominated by annual row‐crops.

54 ENVIRONMENTAL SCIENCES↗
DOE OSTI2025

How do Hydrological Variability and Human Activities Control the Spatiotemporal Changes of Riverine Nitrogen Export in the Upper Mississippi River Basin?

Excessive nitrogen export from agricultural watersheds remains a critical water quality challenge, with the Upper Mississippi River Basin (UMRB) significantly contributing to downstream eutrophication and hypoxia in the Gulf. This study investigates the spatiotemporal dynamics of riverine nitrate plus nitrite (NO 3 − + NO 2 − -N) export across the UMRB at high spatial resolution (12-digit Hydrologic Unit Codes or HUC12 subwatershed scale) during 2001−2020 and quantifies the effects of anthropogenic activities and hydrological variability on riverine NO 3 − + NO 2 − -N export changes in the region between 2001−2005 and 2016−2020. Our results revealed hotspots of substantial increases in NO 3 − + NO 2 − -N yields across the UMRB, with distinct regional patterns in driving factors. Over the entire UMRB, NO 3 − + NO 2 − -N yields increased by 9.7 kg/ha/yr on average from 2001−2005 to 2016−2020, with anthropogenic activities contributing 4.8 kg/ha/yr and hydrological variability contributing 4.9 kg/ha/yr. The northern and western UMRB had combined influences from both anthropogenic activities and hydrological variability, while the east-central regions had predominantly hydrologically driven changes. Agricultural sources, including fertilizer, manure, and biological nitrogen fixation, collectively contributed over 80% of NO 3 − + NO 2 − -N loading throughout the basin. Furthermore, this framework for disentangling human and hydrological impacts provides critical insights for developing effective and targeted watershed management strategies to reduce nutrient losses and improve water quality.

54 ENVIRONMENTAL SCIENCES↗
DOE OSTI2021

DIRT/3D: 3D root phenotyping for field-grown maize ( Zea mays )

The development of crops with deeper roots holds substantial promise to mitigate the consequences of climate change. Deeper roots are an essential factor to improve water uptake as a way to enhance crop resilience to drought, to increase nitrogen capture, to reduce fertilizer inputs, and to increase carbon sequestration from the atmosphere to improve soil organic fertility. A major bottleneck to achieving these improvements is high-throughput phenotyping to quantify root phenotypes of field-grown roots. We address this bottleneck with Digital Imaging of Root Traits (DIRT)/3D, an image-based 3D root phenotyping platform, which measures 18 architecture traits from mature field-grown maize (Zea mays) root crowns (RCs) excavated with the Shovelomics technique. DIRT/3D reliably computed all 18 traits, including distance between whorls and the number, angles, and diameters of nodal roots, on a test panel of 12 contrasting maize genotypes. The computed results were validated through comparison with manual measurements. Overall, we observed a coefficient of determination of r 2 > 0.84 and a high broad-sense heritability of H$_{mean}^{2}$ < 0.6 for all but one trait. The average values of the 18 traits and a developed descriptor to characterize complete root architecture distinguished all genotypes. DIRT/3D is a step toward automated quantification of highly occluded maize RCs. Therefore, DIRT/3D supports breeders and root biologists in improving carbon sequestration and food security in the face of the adverse effects of climate change.

59 BASIC BIOLOGICAL SCIENCES↗