scholarly journals Estimation of isotopologue variation of N<sub>2</sub>O during denitrification by <i>Pseudomonas aureofaciens</i> and <i>Pseudomonas</i> chlororaphis: Implications for N<sub>2</sub>O source apportionment

2017 ◽  
Author(s):  
Joshua A. Haslun ◽  
Nathaniel E. Ostrom ◽  
Eric L. Hegg ◽  
Peggy H. Ostrom
Keyword(s):  
Electron Donor ◽  
Isotope Effects ◽  
Microbial Processes ◽  
Site Preference ◽  
N2o Emissions ◽  
Fractionation Factor ◽  
N2o Production ◽  
Soil Microbial ◽  
Donor Source ◽  
Single Reaction

Abstract. Soil microbial processes, stimulated by agricultural fertilization, account for 90 % of anthropogenic nitrous oxide (N2O), the leading source of ozone depletion and a potent greenhouse gas. Efforts to reduce N2O flux commonly focus on reducing fertilization rates. Management of microbial processes responsible for N2O production may also be used to reduce N2O emissions, but this requires knowledge of the prevailing process. To this end, stable isotopes of N2O have been applied to differentiate N2O produced by nitrification and denitrification. To better understand the factors contributing to isotopic variation during denitrification, we characterized the δ15N, δ18O and site preference (SP; the intramolecular distribution of 15N in N2O) of N2O produced during NO3− reduction by Pseudomonas chlororaphis subsp. aureofaciens and P. c. subsp. chlororaphis. In addition to species, treatments included electron donor (citrate and succinate) and electron donor concentration (0.01 mM, 0.1 mM, 1 mM, and 10 mM) as factors. In contrast to the expectation of a Rayleigh model, all treatments exhibited curvilinear behaviour between δ15N or δ18O and [−flnf/(1−f)]. The curvilinear behaviour indicates that the fractionation factor changed over the course of the reaction, something that is not unexpected for a multi-step process such as denitrification. Using the derivative of the equation, we estimated that the net isotope effects (η) vary by as much as 100 ‰ over the course of a single reaction, placing challenges for using δ15N and δ18O as apportionment tools. In contrast, SP for denitrification was not affected by the extent of the reaction, the electron donor source, or concentration, although the mean SP of N2O produced by each species differed. Therefore, SP remains a robust indicator of the origin of N2O. To improve apportionment estimates with SP, future studies could evaluate other factors that contribute to the variation in SP.

scholarly journals Estimation of isotope variation of N<sub>2</sub>O during denitrification by <i>Pseudomonas aureofaciens</i> and <i>Pseudomonas chlororaphis</i>: implications for N<sub>2</sub>O source apportionment

2018 ◽  
Vol 15 (12) ◽  
pp. 3873-3882 ◽  
Author(s):  
Joshua A. Haslun ◽  
Nathaniel E. Ostrom ◽  
Eric L. Hegg ◽  
Peggy H. Ostrom
Keyword(s):  
Nitrous Oxide ◽  
Electron Donor ◽  
Isotope Effects ◽  
Microbial Processes ◽  
Site Preference ◽  
N2o Emissions ◽  
Pseudomonas Chlororaphis ◽  
Fractionation Factor ◽  
N2o Production ◽  
Soil Microbial

Abstract. Soil microbial processes, stimulated by agricultural fertilization, account for 90 % of anthropogenic nitrous oxide (N2O), the leading source of ozone depletion and a potent greenhouse gas. Efforts to reduce N2O flux commonly focus on reducing fertilization rates. Management of microbial processes responsible for N2O production may also be used to reduce N2O emissions, but this requires knowledge of the prevailing process. To this end, stable isotopes of N2O have been applied to differentiate N2O produced by nitrification and denitrification. To better understand the factors contributing to isotopic variation during denitrification, we characterized the δ15N, δ18O and site preference (SP; the intramolecular distribution of 15N in N2O) of N2O produced during NO3- reduction by Pseudomonas chlororaphis subsp. aureofaciens and P. c. subsp. chlororaphis. The terminal product of denitrification for these two species is N2O because they lack the gene nitrous oxide reductase, which is responsible for the reduction of N2O to N2. In addition to species, treatments included electron donor (citrate and succinate) and electron donor concentration (0.01, 0.1, 1 and 10 mM) as factors. In contrast to the expectation of a Rayleigh model, all treatments exhibited curvilinear behaviour between δ15N or δ18O and the extent of the reaction. The curvilinear behaviour indicates that the fractionation factor changed over the course of the reaction, something that is not unexpected for a multi-step process such as denitrification. Using the derivative of the equation, we estimated that the net isotope effects (η) vary by as much as 100 ‰ over the course of a single reaction, presenting challenges for using δ15N and δ18O as apportionment tools. In contrast, SP for denitrification was not affected by the extent of the reaction, the electron donor source or concentration, although the mean SP of N2O produced by each species differed. Therefore, SP remains a robust indicator of the origin of N2O. To improve apportionment estimates with SP, future studies could evaluate other factors that contribute to the variation in SP.

scholarly journals A new look at an old concept: using <sup>15</sup>N<sub>2</sub>O isotopomers to understand the relationship between soil moisture and N<sub>2</sub>O production pathways

SOIL ◽  
2019 ◽  
Vol 5 (2) ◽  
pp. 265-274 ◽  
Author(s):  
Katelyn A. Congreves ◽  
Trang Phan ◽  
Richard E. Farrell
Keyword(s):  
Soil Moisture ◽  
Mitigation Strategies ◽  
Soil Conditions ◽  
Site Preference ◽  
N2o Emissions ◽  
Spectroscopic Techniques ◽  
N2o Production ◽  
Nitrification And Denitrification ◽  
The Relationship ◽  
Source Partitioning

Abstract. Understanding the production pathways of potent greenhouse gases, such as nitrous oxide (N2O), is essential for accurate flux prediction and for developing effective adaptation and mitigation strategies in response to climate change. Yet there remain surprising gaps in our understanding and precise quantification of the underlying production pathways – such as the relationship between soil moisture and N2O production pathways. A powerful, but arguably underutilized, approach for quantifying the relative contribution of nitrification and denitrification to N2O production involves determining 15N2O isotopomers and 15N site preference (SP) via spectroscopic techniques. Using one such technique, we conducted a short-term incubation where N2O production and 15N2O isotopomers were measured 24 h after soil moisture treatments of 40 % to 105 % water-filled pore space (WFPS) were established for each of three soils that differed in nutrient levels, organic matter, and texture. Relatively low N2O fluxes and high SP values indicted nitrification during dry soil conditions, whereas at higher soil moisture, peak N2O emissions coincided with a sharp decline in SP, indicating denitrification. This pattern supports the classic N2O production curves from nitrification and denitrification as inferred by earlier research; however, our isotopomer data enabled the quantification of source partitioning for either pathway. At soil moisture levels < 53 % WFPS, the fraction of N2O attributed to nitrification (FN) predominated but thereafter decreased rapidly with increasing soil moisture (x), according to FN=3.19-0.041x, until a WFPS of 78 % was reached. Simultaneously, from WFPS of 53 % to 78 %, the fraction of N2O that was attributed to denitrification (FD) was modelled as FD=-2.19+0.041x; at moisture levels of > 78 %, denitrification completely dominated. Clearly, the soil moisture level during transition is a key regulator of N2O production pathways. The presented equations may be helpful for other researchers in estimating N2O source partitioning when soil moisture falls within the transition from nitrification to denitrification.

scholarly journals Revisiting the relationship between soil moisture and N<sub>2</sub>O production pathways by measuring <sup>15</sup>N<sub>2</sub>O isotopomers

2019 ◽  
Author(s):  
Kate A. Congreves ◽  
Trang Phan ◽  
Richard E. Farrell
Keyword(s):  
Soil Moisture ◽  
Pore Space ◽  
Mitigation Strategies ◽  
Soil Conditions ◽  
Site Preference ◽  
N2o Emissions ◽  
Spectroscopic Techniques ◽  
N2o Production ◽  
Nitrification And Denitrification ◽  
The Relationship

Abstract. Understanding the production pathways of potent greenhouse gases, such as nitrous oxide (N2O), is essential for accurate flux prediction and for developing effective adaptation and mitigation strategies in response to climate change. Yet, there remain surprising gaps in our understanding and precise quantification of the underlying production pathways – such as the relationship between soil moisture and N2O production pathways. A powerful, but arguably underutilized, approach for quantifying the relative contribution of nitrification and denitrification to N2O production involves determining 15N2O isotopomers and 15N site preference (SP) via spectroscopic techniques. Using one such technique we conducted a short-term incubation to precisely quantify the relationship between soil moisture and N2O production pathways. For each of three soils, microcosms were arranged in a complete random design with four replicates; each microcosm consisted of air-dried soil packed into plastic petri dishes wherein moisture treatments were established for water contents equivalent to 45 to 105 % water-filled pore space (WFPS). The microcosms were placed in 1-L jars and sealed; headspace samples were collected after 24-h and analyzed for total N2O concentrations using gas chromatography, and for 15N2O isotopomers using cavity ring-down spectroscopy. Relatively low N2O fluxes and high SP values indicted nitrification during dry soil conditions, whereas at higher soil moisture, peak N2O emissions coincided with a sharp decline in SP indicating denitrification. This pattern supports the classic N2O production curves from nitrification and denitrification as inferred by earlier research; however, our isotopomer data enabled the quantification of source partitioning for either pathway thereby providing clarity on N2O sources during the transition from nitrification to denitrification. At soil moisture levels  78 %, denitrification completely dominated. Clearly, the soil moisture levels during transition is a key regulation of N2O production pathways.

scholarly journals Natural Nitrogen Isotope Ratios as a Potential Indicator of N2O Production Pathways in a Floodplain Fen

Water ◽  
2020 ◽  
Vol 12 (2) ◽  
pp. 409
Author(s):  
Mohit Masta ◽  
Holar Sepp ◽  
Jaan Pärn ◽  
Kalle Kirsimäe ◽  
Ülo Mander
Keyword(s):  
Water Table ◽  
Isotope Composition ◽  
Nitrogen Isotope ◽  
Water Table Depth ◽  
Organic Soils ◽  
Site Preference ◽  
N2o Emissions ◽  
N2o Production ◽  
Nitrifier Denitrification ◽  
Anoxic Environment

Nitrous oxide (N2O), a major greenhouse gas and ozone depleter, is emitted from drained organic soils typically developed in floodplains. We investigated the effect of the water table depth and soil oxygen (O2) content on N2O fluxes and their nitrogen isotope composition in a drained floodplain fen in Estonia. Measurements were done at natural water table depth, and we created a temporary anoxic environment by experimental flooding. From the suboxic peat (0.5–6 mg O2/L) N2O emissions peaked at 6 mg O2/L and afterwards decreased with decreasing O2. From the anoxic and oxic peat (0 and >6 mg O2/L, respectively) N2O emissions were low. Under anoxic conditions the δ15N/δ14N ratio of the top 10 cm peat layer was low, gradually decreasing to 30 cm. In the suboxic peat, δ15N/δ14N ratios increased with depth. In samples of peat fluctuating between suboxic and anoxic, the elevated 15N/14N ratios (δ15N = 7–9‰ ambient N2) indicated intensive microbial processing of nitrogen. Low values of site preference (SP; difference between the central and peripheral 15N atoms) and δ18O-N2O in the captured gas samples indicate nitrifier denitrification in the floodplain fen.

scholarly journals Isotopic evidence for alteration of nitrous oxide emissions and producing pathways contribution under nitrifying conditions

2019 ◽  
Author(s):  
Guillaume Humbert ◽  
Mathieu Sébilo ◽  
Justine Fiat ◽  
Longqi Lang ◽  
Ahlem Filali ◽  
...  
Keyword(s):  
Nitrous Oxide ◽  
Biofilm Reactor ◽  
Nitrite Reduction ◽  
Nitrous Oxide Emissions ◽  
Site Preference ◽  
N2o Emissions ◽  
Ammonium Oxidation ◽  
N2o Production ◽  
Oxidation Rates ◽  
Nitrifier Denitrification

Abstract. Nitrous oxide (N2O) emissions by a nitrifying biofilm reactor were investigated with N2O isotopocules. The site preference of N2O (15N-SP) indicated the contribution of producing and consuming pathways in response to changes in oxygenation level (from 0 to 21 % O2 in the gas mix), temperature (from 13.5 to 22.3 °C), and ammonium concentrations (from 6.2 to 62.1 mg N L−1). Nitrite reduction, either nitrifier-denitrification or heterotrophic denitrification, was the main N2O producing pathway under the tested conditions. Nitrite oxidation rates decreased as compared to ammonium oxidation rates at temperatures above 20 °C and sub-optimal oxygen levels, increasing N2O production by the nitrite reduction pathway. Below 20 °C, a difference in temperature sensitivity between hydroxylamine and ammonium oxidation rates is most likely responsible for an increase in the N2O production via the hydroxylamine oxidation pathway (nitrification). A negative correlation between the reaction kinetics and the apparent isotope fractionation was additionally shown from the variations of δ15N and δ18O values of N2O produced from ammonium.

scholarly journals Isotopic evidence for alteration of nitrous oxide emissions and producing pathways' contribution under nitrifying conditions

2020 ◽  
Vol 17 (4) ◽  
pp. 979-993
Author(s):  
Guillaume Humbert ◽  
Mathieu Sébilo ◽  
Justine Fiat ◽  
Longqi Lang ◽  
Ahlem Filali ◽  
...  
Keyword(s):  
Nitrous Oxide ◽  
Biofilm Reactor ◽  
Nitrite Reduction ◽  
Nitrous Oxide Emissions ◽  
Site Preference ◽  
N2o Emissions ◽  
Ammonium Oxidation ◽  
N2o Production ◽  
Oxidation Rates ◽  
Nitrifier Denitrification

Abstract. Nitrous oxide (N2O) emissions from a nitrifying biofilm reactor were investigated with N2O isotopocules. The nitrogen isotopomer site preference of N2O (15N-SP) indicated the contribution of producing and consuming pathways in response to changes in oxygenation level (from 0 % to 21 % O2 in the gas mix), temperature (from 13.5 to 22.3 ∘C) and ammonium concentrations (from 6.2 to 62.1 mg N L−1). Nitrite reduction, either nitrifier denitrification or heterotrophic denitrification, was the main N2O-producing pathway under the tested conditions. Difference between oxidative and reductive rates of nitrite consumption was discussed in relation to NO2- concentrations and N2O emissions. Hence, nitrite oxidation rates seem to decrease as compared to ammonium oxidation rates at temperatures above 20 ∘C and under oxygen-depleted atmosphere, increasing N2O production by the nitrite reduction pathway. Below 20 ∘C, a difference in temperature sensitivity between hydroxylamine and ammonium oxidation rates is most likely responsible for an increase in N2O production via the hydroxylamine oxidation pathway (nitrification). A negative correlation between the reaction kinetics and the apparent isotope fractionation was additionally shown from the variations of δ15N and δ18O values of N2O produced from ammonium. The approach and results obtained here, for a nitrifying biofilm reactor under variable environmental conditions, should allow for application and extrapolation of N2O emissions from other systems such as lakes, soils and sediments.

scholarly journals Stable-Isotope-Aided Investigation of the Effect of Redox Potential on Nitrous Oxide Emissions as Affected by Water Status and N Fertilization

Water ◽  
2020 ◽  
Vol 12 (10) ◽  
pp. 2918
Author(s):  
Jihuan Wang ◽  
Heye R. Bogena ◽  
Harry Vereecken ◽  
Nicolas Brüggemann
Keyword(s):  
Nitrous Oxide ◽  
Redox Potential ◽  
Isotope Analysis ◽  
N Cycling ◽  
Site Preference ◽  
N2o Emissions ◽  
Soil N ◽  
N2o Production ◽  
Soil Redox Potential ◽  
Soil Redox

Soils are the dominant source of atmospheric nitrous oxide (N2O), especially agricultural soils that experience both waterlogging and intensive nitrogen fertilization. However, soil heterogeneity and the irregular occurrence of hydrological events hamper the prediction of the temporal and spatial dynamics of N2O production and transport in soils. Because soil moisture influences soil redox potential, and as soil N cycling processes are redox-sensitive, redox potential measurements could help us to better understand and predict soil N cycling and N2O emissions. Despite its importance, only a few studies have investigated the control of redox potential on N2Oemission from soils in detail. This study aimed to partition the different microbial processes involved in N2O production (nitrification and denitrification) by using redox measurements combined with isotope analysis at natural abundance and 15N-enriched. To this end, we performed long-term laboratory lysimeter experiments to mimic common agricultural irrigation and fertilization procedures. In addition, we used isotope analysis to characterize the distribution and partitioning of N2O sources and explored the 15N-N2O site preference to further constrain N2O microbial processes. We found that irrigation, saturation, and drainage induced changes in soil redox potential, which were closely related to changes in N2O emission from the soil as well as to changes in the vertical concentration profiles of dissolved N2O, nitrate (NO3−) and ammonium (NH4+). The results showed that the redox potential could be used as an indicator for NH4+, NO3−, and N2O production and consumption processes along the soil profile. For example, after a longer saturation period of unfertilized soil, the NO3− concentration was linearly correlated with the average redox values at the different depths (R2 = 0.81). During the transition from saturation to drainage, but before fertilization, the soil showed an increase in N2O emissions, which originated mainly from nitrification as indicated by the isotopic signatures of N2O (δ15N bulk, δ18O and 15N-N2O site preference). After fertilization, N2O still mainly originated from nitrification at the beginning, also indicated by high redox potential and the increase of dissolved NO3−. Denitrification mainly occurred during the last saturation period, deduced from the simultaneous 15N isotope analysis of NO3− and N2O. Our findings suggest that redox potential measurements provide suitable information for improving the prediction of soil N2O emissions and the distribution of mineral N species along the soil profile under different hydrological and fertilization regimes.

scholarly journals Nitrogen and oxygen availabilities control water column nitrous oxide production during seasonal anoxia in the Chesapeake Bay

2018 ◽  
Vol 15 (20) ◽  
pp. 6127-6138 ◽  
Author(s):  
Qixing Ji ◽  
Claudia Frey ◽  
Xin Sun ◽  
Melanie Jackson ◽  
Yea-Shine Lee ◽  
...  
Keyword(s):  
Nitrous Oxide ◽  
Chesapeake Bay ◽  
Water Column ◽  
N2o Emissions ◽  
N2o Production ◽  
Isotope Tracer ◽  
Relative Contribution ◽  
Environmental Controls ◽  
Long Run ◽  
Nitrate And Nitrite

Abstract. Nitrous oxide (N2O) is a greenhouse gas and an ozone depletion agent. Estuaries that are subject to seasonal anoxia are generally regarded as N2O sources. However, insufficient understanding of the environmental controls on N2O production results in large uncertainty about the estuarine contribution to the global N2O budget. Incubation experiments with nitrogen stable isotope tracer were used to investigate the geochemical factors controlling N2O production from denitrification in the Chesapeake Bay, the largest estuary in North America. The highest potential rates of water column N2O production via denitrification (7.5±1.2 nmol-N L−1 h−1) were detected during summer anoxia, during which oxidized nitrogen species (nitrate and nitrite) were absent from the water column. At the top of the anoxic layer, N2O production from denitrification was stimulated by addition of nitrate and nitrite. The relative contribution of nitrate and nitrite to N2O production was positively correlated with the ratio of nitrate to nitrite concentrations. Increased oxygen availability, up to 7 µmol L−1 oxygen, inhibited both N2O production and the reduction of nitrate to nitrite. In spring, high oxygen and low abundance of denitrifying microbes resulted in undetectable N2O production from denitrification. Thus, decreasing the nitrogen input into the Chesapeake Bay has two potential impacts on the N2O production: a lower availability of nitrogen substrates may mitigate short-term N2O emissions during summer anoxia; and, in the long-run (timescale of years), eutrophication will be alleviated and subsequent reoxygenation of the bay will further inhibit N2O production.

scholarly journals Spatial and temporal variability of nitrous oxide emissions in a mixed farming landscape of Denmark

2012 ◽  
Vol 9 (8) ◽  
pp. 2989-3002 ◽  
Author(s):  
K. Schelde ◽  
P. Cellier ◽  
T. Bertolini ◽  
T. Dalgaard ◽  
T. Weidinger ◽  
...  
Keyword(s):  
Land Use ◽  
Nitrous Oxide ◽  
Agricultural Land ◽  
Nitrous Oxide Emissions ◽  
Soil Conditions ◽  
Spatial And Temporal Variability ◽  
N2o Emissions ◽  
N2o Production ◽  
Mixed Farming ◽  
N2o Fluxes

Abstract. Nitrous oxide (N2O) emissions from agricultural land are variable at the landscape scale due to variability in land use, management, soil type, and topography. A field experiment was carried out in a typical mixed farming landscape in Denmark, to investigate the main drivers of variations in N2O emissions, measured using static chambers. Measurements were made over a period of 20 months, and sampling was intensified during two weeks in spring 2009 when chambers were installed at ten locations or fields to cover different crops and topography and slurry was applied to three of the fields. N2O emissions during spring 2009 were relatively low, with maximum values below 20 ng N m−2 s−1. This applied to all land use types including winter grain crops, grasslands, meadows, and wetlands. Slurry application to wheat fields resulted in short-lived two-fold increases in emissions. The moderate N2O fluxes and their moderate response to slurry application were attributed to dry soil conditions due to the absence of rain during the four previous weeks. Cumulative annual emissions from two arable fields that were both fertilized with mineral fertilizer and manure were large (17 kg N2O-N ha−1 yr−1 and 5.5 kg N2O-N ha−1 yr−1) during the previous year when soil water conditions were favourable for N2O production during the first month following fertilizer application. Our findings confirm the importance of weather conditions as well as nitrogen management on N2O fluxes.

scholarly journals Trace gas fluxes of CO<sub>2</sub>, CH<sub>4</sub> and N<sub>2</sub>O in a permanent grassland soil exposed to elevated CO<sub>2</sub> in the Giessen FACE study

2011 ◽  
Vol 11 (17) ◽  
pp. 9333-9342 ◽  
Author(s):  
M. Kaleem Abbasi ◽  
C. Müller
Keyword(s):  
Elevated Co2 ◽  
15N Tracer ◽  
Organic N ◽  
N2o Emissions ◽  
Ch4 Oxidation ◽  
N2o Production ◽  
N Application ◽  
Oxidation Rates ◽  
Controlled Conditions ◽  
N2o Fluxes

Abstract. Long-term field observations showed that N2O fluxes observed shortly after N application were not significantly affected by elevated CO2 in the Giessen Free Air Carbon dioxide Enrichment (FACE) study. To further investigate this unexpected result a 15N tracer study was carried out under controlled conditions where in parallel treatments either the NH4+ pool (15NH4NO3) or the NO3− pool (NH415NO3) was enriched with 15N. Fluxes of CO2, CH4, and N2O as well as the 15N enrichment of the N2O were measured. Denitrifying Enzyme Activity (DEA), total denitrification (N2 + N2O) and N2-to-N2O ratios were quantified in separate experiments. Over the 57 day incubation, N2O fluxes averaged 0.090 ng N2O-N g−1 h−1 under ambient and 0.083 ng N2O-N g−1 h−1 under elevated CO2 (not significantly different). The N2O production processes were identified by a two-source model. Results showed that N2O must have also been produced by a third source – possibly related to organic N transformation – which was stimulated by elevated CO2. Soil CO2 fluxes were approximately 20 % higher under elevated CO2 than soil from ambient but the differences were not significant. CH4 oxidation rates were on average −1.75 ng CH4-C g−1 h−1 in the elevated and −1.17 ng CH4-C g−1 h−1 in the ambient indicating that elevated CO2 increased the CH4 oxidation by 49 % compared to ambient CO2 under controlled conditions. N fertilization increased CH4 oxidation by 3-fold in both CO2 treatments. CO2 did not have any significant effect on DEA while total denitrification and N2-to-N2O ratios increased by 36 and 33 %, respectively. The results indicate that shortly after N application elevated CO2 must have stimulated both the N2O production and reduction to N2 to explain the increased N2-to-N2O ratio and at the same time explain the non-responsiveness of the N2O emissions. Thus, the observed variation of the CO2 effect on N2O emissions throughout the year is possibly governed by the dynamics of the N2O reductase activity.

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