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Utilizing Novel Field and Data Exploration Methods to Explore Hot Moments in High-Frequency Soil Nitrous Oxide Emissions Data: Opportunities and Challenges

Soil nitrous oxide (N 2 O) emissions are an important driver of climate change and are a major mechanism of labile nitrogen (N) loss from terrestrial ecosystems. Evidence increasingly suggests that locations on the landscape that experience biogeochemical fluxes disproportionate to the surrounding matrix (hot spots) and time periods that show disproportionately high fluxes relative to the background (hot moments) strongly influence landscape-scale soil N 2 O emissions. However, substantial uncertainties remain regarding how to measure and model where and when these extreme soil N 2 O fluxes occur. High-frequency datasets of soil N 2 O fluxes are newly possible due to advancements in field-ready instrumentation that uses cavity ring-down spectroscopy (CRDS). Here, we outline the opportunities and challenges that are provided by the deployment of this field-based instrumentation and the collection of high-frequency soil N 2 O flux datasets. While there are substantial challenges associated with automated CRDS systems, there are also opportunities to utilize these near-continuous data to constrain our understanding of dynamics of the terrestrial N cycle across space and time. Finally, we propose future research directions exploring the influence of hot moments of N 2 O emissions on the N cycle, particularly considering the gaps surrounding how global change forces are likely to alter N dynamics in the future.

54 ENVIRONMENTAL SCIENCES↗

Process-based modeling of soil nitrous oxide emissions from United States corn fields under different management and climate scenarios coupled with evaluation using regional estimates

Direct emissions of soil nitrous oxide during a growing season (N 2 O gs ) can be quantified with process-based models considering interactions between management, climate, and soil moisture when key data are available. We used an adapted “parameterized CENTURY/DAYCENT-model” ( p CENTURY) calibrated with crop growth and soil organic matter decay coefficients at the county-level for the estimation of N 2 O gs in the United States Corn Belt. Model estimated N 2 O-emissions from corn-based biofuels scenarios considering crop rotation, fertilizer inputs, tillage, and weather were compared against meta-summary of field observations from 55 studies. Both model and meta-summary ranked N 2 O gs -emissions to be corn > wheat > soybean phase while model likely underestimated cover crop N 2 O gs -emissions. The N 2 O gs -emissions and the associated emission factors (EFs) were modeled and summarized to be greater after anhydrous ammonia than urea application and from conventional tilled than non-tilled fields. Modeled and observed N 2 O gs -emissions after organic and inorganic fertilizer amendment did not differ due to high variability associated with the treatments. However, the organic fertilizer associated EFs were greater according to meta-summary data because of N input rates. Regionalized weather scenarios indicate hotspots for N 2 O gs -emissions can occur where crop N uptake is limited during dry years and in eastern states also during normal or wet seasons. The p CENTURY-derived N 2 O gs EFs (0.91 ± 0.19%) for counties investigated were only slightly lower than literature (1.07 ± 0.57%) or Tier-1 (1%) values. Our preliminary evaluation of regional soil moisture estimates showed reasonable agreement between monthly soil moisture estimates and the North American Soil Moisture Dataset during the growing season, but overestimation of soil moisture in winter-spring can influence the estimates of annual N 2 O emissions so future work is needed to calibrate soil moisture-associated model parameters. Our work provided scenario-based estimates of climate and management impacts on soil N 2 O gs -emissions together with valuable spatial insights into EFs that will be improved by more accurate information of fertilizer inputs and more temporally refined model evaluation.

54 ENVIRONMENTAL SCIENCES↗

Precipitation and nitrogen application stimulate soil nitrous oxide emission

Precipitation and nitrogen (N) fertilization are the two most important drivers for soil nitrous oxide (N 2 O) emission. However, the effects of changes in N fertilization and precipitation patterns (i.e., precipitation intensity and frequency) on N 2 O emissions in agricultural fields are still unclear. In this study, we simulated soil N 2 O emission under different precipitation patterns (6 precipitation intensities, and 12 precipitation frequencies by either merging or splitting precipitation events) and N fertilization rates (low, typical, and high N fertilization) in a cornfield using the DeNitrification-DeComposition model. The model was parameterized and validated using meteorological data and N experimental measurements in Nashville, Tennessee, USA. Results showed that soil water filled pore space (WFPS) and simulated soil N 2 O emission increased as precipitation intensity increased. Less frequent but high intensity precipitation treatments reduced the soil WFPS by 25.2% and stimulated soil N 2 O emission by 45.3%, while more frequent but low intensity precipitation treatments increased soil WFPS by 9.0% and reduced soil N 2 O emission by 23.9%. Compared to typical N fertilization, the sensitivity of soil N 2 O emission to precipitation was higher under high N than low N fertilization treatments, and the response ratios were 50.0% and 40.1%, respectively. There was significant interactive effect of precipitation intensity and N fertilization on soil N 2 O emission. Furthermore, these findings improved our understanding of precipitation and N impacts on soil N 2 O emissions and provided useful knowledge for irrigation and N fertilizer management in agriculture to mitigate greenhouse gas emissions.

54 ENVIRONMENTAL SCIENCES↗

Response patterns of simulated corn yield and soil nitrous oxide emission to precipitation change

Abstract Background Precipitation plays an important role in crop production and soil greenhouse gas emissions. However, how crop yield and soil nitrous oxide (N 2 O) emission respond to precipitation change, particularly with different background precipitations (dry, normal, and wet years), has not been well investigated. In this study, we examined the impacts of precipitation changes on corn yield and soil N 2 O emission using a long-term (1981–2020, 40 years) climate dataset as well as seven manipulated precipitation treatments with different background precipitations using the DeNitrification-DeComposition (DNDC) model. Results Results showed large variations of corn yield and precipitation but small variation of soil N 2 O emission among 40 years. Both corn yield and soil N 2 O emission showed near linear relationships with precipitation based on the long-term precipitation data, but with different response patters of corn yield and soil N 2 O emission to precipitation manipulations. Corn yield showed a positive linear response to precipitation manipulations in the dry year, but no response to increases in precipitation in the normal year, and a trend of decrease in the wet year. The extreme drought treatments reduced corn yield sharply in both normal and wet years. In contrast, soil N 2 O emission mostly responded linearly to precipitation manipulations. Decreases in precipitation in the dry year reduced more soil N 2 O emission than those in the normal and wet years, while increases in precipitation increased more soil N 2 O emission in the normal and wet years than in the dry year. Conclusions This study revealed different response patterns of corn yield and soil N 2 O emission to precipitation and highlights that mitigation strategy for soil N 2 O emission reduction should consider different background climate conditions.

54 ENVIRONMENTAL SCIENCES↗

Beyond denitrification: The role of microbial diversity in controlling nitrous oxide reduction and soil nitrous oxide emissions

Many biotic and abiotic processes contribute to nitrous oxide (N 2 O) production in the biosphere, but N 2 O consumption in the environment has heretofore been attributed primarily to canonical denitrifying microorganisms. The nosZ genes encoding the N 2 O reductase enzyme, NosZ, responsible for N 2 O reduction to dinitrogen are now known to include two distinct groups: the well-studied Clade I which denitrifiers typically possess, and the novel Clade II possessed by diverse groups of microorganisms, most of which are non-denitrifiers. Clade II N 2 O reducers could play an important, previously unrecognized role in controlling N 2 O emissions for several reasons, including: (1) the consumption of N 2 O produced by processes other than denitrification, (2) hypothesized non-respiratory functions of NosZ as an electron sink or for N 2 O detoxification, (3) possible differing enzyme kinetics of Clade II NosZ compared to Clade I NosZ, and (4) greater nosZ gene abundance for Clade II compared to Clade I in soils of many ecosystems. Despite the potential ecological significance of Clade II NosZ, a census of 800 peer-reviewed original research articles discussing nosZ and published from 2013 to 2019 showed that the percentage of articles evaluating or mentioning Clade II nosZ increased from 5% in 2013 to only 22% in 2019. The census revealed that the slowly spreading awareness of Clade II nosZ may result in part from disciplinary silos, with the percentage of nosZ articles mentioning Clade II nosZ ranging from 0% in Agriculture and Agronomy journals to 32% in Multidisciplinary Sciences journals. In addition, inconsistent nomenclature for Clade I nosZ and Clade II nosZ, with 17 different terminologies used in the literature, may have created confusion about the two distinct groups of N 2 O reducers. Furthermore, we provide recommendations to accelerate advances in understanding the role of the diversity of N 2 O reducers in regulating soil N 2 O emissions.

59 BASIC BIOLOGICAL SCIENCES↗

N 2 Onet: a global collaborative network facilitating advances in measurement, modeling, and mitigation of agricultural soil nitrous oxide emissions

Nitrogen (N) fertilizer supports global food production, but its use and overuse drive emissions of nitrous oxide (N 2 O), a potent and long-lived greenhouse gas. Understanding the drivers of N 2 O fluxes remains elusive, making it difficult to predict emissions in time and space and to develop and evaluate ways to lower emissions through management. Major scientific uncertainties underlying the understanding of the drivers of N 2 O fluxes identified in a workshop of N 2 O emissions experts include poor process-based understanding of controls on soil N 2 O emissions in the field; insufficient data to reduce uncertainty in N 2 O budgets from the field to regional scales, including N 2 O emission measurements and importantly, field-scale N balances; and high uncertainty in model predictions of soil N 2 O emissions across environmental and management conditions. To reduce these uncertainties, we present the concept of N 2 Onet, a global collaborative initiative to accelerate advances in N 2 O measurement, analyses, and mitigation. N 2 Onet will serve as an observational network of supersites with multi-scale measurements; a database hub for N 2 O flux and ancillary data; and a catalyst for community building, information sharing, and training. By coalescing and coordinating the global community of researchers, N 2 Onet will provide a roadmap for reducing N 2 O emissions from agriculture worldwide.

54 ENVIRONMENTAL SCIENCES↗

Hot or Not? An Evaluation of Methods for Identifying Hot Moments of Nitrous Oxide Emissions From Soils

Abstract Effectively quantifying hot moments of nitrous oxide (N 2 O) emissions from agricultural soils is critical for managing this potent greenhouse gas. However, we are challenged by a lack of standard approaches for identifying hot moments, including (a) determining thresholds above which emissions are considered hot moments, and (b) considering seasonal variation in the magnitude and frequency distribution of net N 2 O fluxes. We used one year of hourly N 2 O flux measurements from 16 autochambers that varied in flux magnitude and frequency distribution in a conventionally tilled maize field in central Illinois, USA, to compare three approaches to identify hot moment thresholds: standard deviations (SD) above the mean, 1.5x the interquartile range (IQR), and isolation forest (IF) identification of anomalous values. We also compared these approaches on seasonally subdivided data (early, late, and non‐growing seasons) versus the whole year. Our analyses revealed that 1.5x IQR method best identified N 2 O hot moments. In contrast, using 2 or 4 SD both yielded hot moment threshold values too high, and IF yielded threshold values too low, leading to missed N 2 O hot moments or low net N 2 O fluxes mischaracterized as hot moments, respectively. Furthermore, seasonally subdividing the data set not only facilitated identification of smaller hot moments in the late‐ and non‐growing seasons when N 2 O hot moments were generally smaller but it also increased hot moment threshold values in the early growing season when N 2 O hot moments were larger. Consequently, of the methods evaluated here, we recommend using the 1.5x IQR method on whole year data sets to identify N 2 O hot moments.

Stuchiner, Emily R. [Institute for Sustainability,↗

Legacy Effects of Intercropping and Nitrogen Fertilization on Soil N Cycling, Nitrous Oxide Emissions, and the Soil Microbial Community in Tropical Maize Production

Maize-forage grasses intercropping systems have been increasingly adopted by farmers because of their capacity to recycle nutrients, provide mulch, and add C to soil. However, grasses have been shown to increase nitrous oxide (N 2 O) emissions. Some tropical grasses cause biological nitrification inhibition (BNI) which could mitigate N 2 O emissions in the maize cycle but the reactions of the N cycle and the microbial changes that explain the N 2 O emissions are little known in such intercropping systems. With this in mind, we explored intercropping of forage grasses ( Brachiaria brizantha and Brachiaria humidicola ) with distinct BNI and yield potential to increase N cycling in no-till maize production systems compared to monocrop with two N rates (0 and 150 kg ha −1 ) applied during the maize season. These grasses did not strongly compete with maize during the period of maize cycle and did not have a negative effect on grain yield. We observed a legacy of these grasses on N mineralization and nitrification through the soil microbiome during maize growth. We observed that B. humidicola , genotype with higher BNI potential, increased net N mineralization by 0.4 mg N kg −1 day −1 and potential nitrification rates by 1.86 mg NO 3 -N kg −1 day −1 , while B. brizantha increased the soil moisture, fungi diversity, mycorrhizal fungi, and bacterial nitrifiers, and reduced saprotrophs prior to maize growth. Their legacy on soil moisture and cumulative organic inputs (i.e., grass biomass) was strongly associated with enhanced mineralization and nitrification rates at early maize season. These effects contributed to increase cumulative N 2 O emission by 12.8 and 4.8 mg N 2 O-N m −2 for maize growing after B. brizantha and B. humidicola , respectively, regardless of the N fertilization rate. Thus, the nitrification inhibition potential of tropical grasses can be outweighed by their impacts on soil moisture, N recycling, and the soil microbiome that together dictate soil N 2 O fluxes.

Canisares, Lucas P.↗

Enhancement of Nitrous Oxide Emissions in Soil Microbial Consortia via Copper Competition between Proteobacterial Methanotrophs and Denitrifiers

Unique means of copper scavenging have been identified in proteobacterial methanotrophs, particularly the use of methanobactin, a novel ribosomally synthesized, post-translationally modified polypeptide that binds copper with very high affinity. The possibility that copper sequestration strategies of methanotrophs may interfere with copper uptake of denitrifiers in situ and thereby enhance N 2 O emissions was examined using a suite of laboratory experiments performed with rice paddy microbial consortia. Addition of purified methanobactin from Methylosinus trichosporium OB3b to denitrifying rice paddy soil microbial consortia resulted in substantially increased N 2 O production, with more pronounced responses observed for soils with lower copper content.

59 BASIC BIOLOGICAL SCIENCES↗

Nature-based climate solutions can help mitigate the radiative forcing that follows deforestation

Widespread expansion of agriculture and forestry has altered the surface of the Earth, the composition of the atmosphere, and as a result, the climate. Here we quantify the radiative forcing caused by the historical deforestation of an ecoregion in the U.S. Upper Midwest and the adoption of eight nature-based climate solutions. We combined regional forest inventory data with over three decades of remote sensing and in situ data from a replicated land use change experiment. Deforestation of the region caused net global warming (1626 ± 44 µW m -2 ), mainly from the 76 % reduction of ecosystem carbon stocks, but also from the 84 % reduction of the soil methane sink and the 115 % increase in soil nitrous oxide emissions. The associated albedo increase offset 24 % of this greenhouse gas induced warming. For the adoption of nature-based climate solutions, we found that conservation agriculture can provide -39 to -76 ± 31 µW m -2 of climate mitigation over a 100-year time period while short/medium length forestry rotations can provide more at -296 to -881 ± 44 µW m -2 and natural forest regeneration can provide the most at -1555 ± 44 µW m -2 . As the impacts of climate change on nature and society intensify, consideration should be given to the climate mitigation, habitat, and ecosystem services that nature-based climate solutions can provide.

54 ENVIRONMENTAL SCIENCES↗

Intermediate soil acidification induces highest nitrous oxide emissions

Global potent greenhouse gas nitrous oxide (N 2 O) emissions from soil are accelerating, with increases in the proportion of reactive nitrogen emitted as N 2 O, i.e., N 2 O emission factor (EF). Yet, the primary controls and underlying mechanisms of EFs remain unresolved. Based on two independent but complementary global syntheses, and three field studies determining effects of acidity on N 2 O EFs and soil denitrifying microorganisms, we show that soil pH predominantly controls N 2 O EFs and emissions by affecting the denitrifier community composition. Analysis of 5438 paired data points of N 2 O emission fluxes revealed a hump-shaped relationship between soil pH and EFs, with the highest EFs occurring in moderately acidic soils that favored N 2 O-producing over N 2 O-consuming microorganisms, and induced high N 2 O emissions. Our results illustrate that soil pH has a unimodal relationship with soil denitrifiers and EFs, and the net N 2 O emission depends on both the N 2 O/(N 2 O + N 2 ) ratio and overall denitrification rate. These findings can inform strategies to predict and mitigate soil N 2 O emissions under future nitrogen input scenarios.

54 ENVIRONMENTAL SCIENCES↗

Post-fire soil emissions of nitric oxide (NO) and nitrous oxide (N 2 O) across global ecosystems: a review

Wildfires may increase soil emissions of trace nitrogen (N) gases like nitric oxide (NO) and nitrous oxide (N 2 O) by changing soil physicochemical conditions and altering microbial processes like nitrification and denitrification. When 34 studies were synthesized, we found a significant increase in both NO and N 2 O emissions up to 1 year post-fire across studies spanning ecosystems globally. However, when fluxes were separated by ecosystem type, we found that individual ecosystem types responded uniquely to fire. Forest soils tended to emit more N 2 O after fire, but there was no significant effect on NO. Shrubland soils showed significant increases in both NO and N 2 O emissions after fires; often with extremely large but short-lived NO pulses occurring immediately after fire. Grassland NO emissions increased after fire, but the size of this effect was small relative to shrublands. N 2 O emissions from burned grasslands were highly variable with no significant effect. To better understand the variation in responses to fire across global ecosystems, more consistent measurements of variables recognized as important controls on soil fluxes of NO and N 2 O (e.g., N cycling rates, soil water content, pH, and substrate availability) are needed across studies. We also suggest that fire-specific elements like burn severity, microbial community succession, and the presence of char be considered by future studies. Our synthesis suggests that fires can exacerbate ecosystem N loss long after they burn, increasing soil emissions of NO and N 2 O with implications for ecosystem N loss, climate, and regional air quality as wildfires increase globally.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Microbial functional genes commonly respond to elevated carbon dioxide

Atmospheric CO 2 concentration is increasing, largely due to anthropogenic activities. Previous studies of individual free-air CO 2 enrichment (FACE) experimental sites have shown significant impacts of elevated CO 2 (eCO 2 ) on soil microbial communities; however, no common microbial response patterns have yet emerged, challenging our ability to predict ecosystem functioning and sustainability in the future eCO 2 environment. Here we analyzed 66 soil microbial communities from five FACE sites, and showed common microbial response patterns to eCO 2 , especially for key functional genes involved in carbon and nitrogen fixation (e.g., pcc/acc for carbon fixation, nifH for nitrogen fixation), carbon decomposition (e.g., amyA and pulA for labile carbon decomposition, mnp and lcc for recalcitrant carbon decomposition), and greenhouse gas emissions (e.g., mcrA for methane production, norB for nitrous oxide production) across five FACE sites. Also, the relative abundance of those key genes was generally increased and directionally associated with increased biomass, soil carbon decomposition, and soil moisture. In addition, a further literature survey of more disparate FACE experimental sites indicated increased biomass, soil carbon decay, nitrogen fixation, methane and nitrous oxide emissions, plant and soil carbon and nitrogen under eCO 2 . A conceptual framework was developed to link commonly responsive functional genes with ecosystem processes, such as pcc/acc vs. soil carbon storage, amyA/pulA/mnp/lcc vs. soil carbon decomposition, and nifH vs. nitrogen availability, suggesting that such common responses of microbial functional genes may have the potential to predict ecosystem functioning and sustainability in the future eCO 2 environment.

59 BASIC BIOLOGICAL SCIENCES↗

Depth-dependent links between microbial taxa and nitrous oxide emissions in a long-term cotton cropping system employing soil health practices

Long-term management practices can shape soil microbial communities in ways that influence nitrogen (N) dynamics and nitrous oxide (N 2 O) emissions. We leverage a 41-year continuous cotton cropping experiment with contrasting tillage, cover cropping, and N fertilization regimes to investigate how these long-term strategies influence soil microbial communities and their associations with N 2 O fluxes during the cotton growing season. Using 16S rRNA gene metabarcoding, we assessed microbial composition in surface and subsurface soils and evaluated its relationship with temporal N 2 O emissions. Among the management practices, N fertilization – a known driver of N 2 O emissions – had the strongest effect on microbial community composition and was linked to a greater number of taxa correlated to N 2 O emissions, particularly in surface soils. Soil pH emerged as a key variable influencing microbial structure across depth and was negatively associated with both N 2 O emissions and microbial composition in the surface layers of fertilized soils. In total, 57 archaeal/bacterial taxa were correlated with N 2 O fluxes, but only seven were shared across depths, suggesting distinct microbial contributors in surface and subsurface soils. Several of these taxa have been previously reported to be associated with N and C cycling processes such as nitrate respiration or carbon turnover, indicating functional context to their correlation with N 2 O fluxes. Temporal shifts in the abundance of key taxa aligned with seasonal peaks in N 2 O emissions, notably in early and late August, and were most pronounced under conventional tillage, hairy vetch cover cropping, and N fertilization. While 16S-based associations cannot confirm functional gene presence or activity, these findings demonstrate that long-term fertilization and associated soil acidification are dominant drivers of microbial shifts linked to N 2 O emissions and highlight the importance of accounting for depth-specific and seasonal microbial dynamics when evaluating management impacts on greenhouse gas emissions.

16S rRNA gene sequencing↗