Biology and Fertility of Soils

, Volume 41, Issue 6, pp 379–388 | Cite as

Contributions of nitrification and denitrification to N2O emissions from soils at different water-filled pore space

  • E. J. Bateman
  • E. M. BaggsEmail author
Original Paper


A combination of stable isotope and acetylene (0.01% v/v) inhibition techniques were used for the first time to determine N2O production during denitrification, autotrophic nitrification and heterotrophic nitrification in a fertilised (200 kg N ha−1) silt loam soil at contrasting (20–70%) water-filled pore space (WFPS). 15N-N2O emissions from 14NH415NO3 replicates were attributed to denitrification and 15N-N2O from 15NH415NO3 minus that from 14NH415NO3 replicates was attributed to nitrification and heterotrophic nitrification in the presence of acetylene, as there was no dissimilatory nitrate reduction to ammonium or immobilisation and remineralisation of 15N-NO3. All of the N2O emitted at 70% WFPS (31.6 mg N2O-N m−2 over 24 days; 1.12 μg N2O-N g dry soil−1; 0.16% of N applied) was produced during denitrification, but at 35–60% WFPS nitrification was the main process producing N2O, accounting for 81% of 15N-N2O emitted at 60% WFPS, and 7.9 μg 15N-N2O m−2 (0.28 ng 15N-N2O g dry soil−1) was estimated to be emitted over 7 days during heterotrophic nitrification in the 50% WFPS treatment and accounted for 20% of 15N-N2O from this treatment. Denitrification was the predominant N2O-producing process at 20% WFPS (2.6 μg 15N-N2O m−2 over 7 days; 0.09 ng 15N-N2O g dry soil−1; 85% of 15N-N2O from this treatment) and may have been due to the occurrence of aerobic denitrification at this WFPS. Our results demonstrate the usefulness of a combined stable isotope and acetylene approach to quantify N2O emissions from different processes and to show that several processes may contribute to N2O emission from agricultural soils depending on soil WFPS.


Denitrification Nitrification Nitrous oxide Soil water-filled pore space 



This work was funded by a Research Committee studentship awarded by the Biotechnology and Biological Sciences Research Council, UK. We thank Jon Fear for the stable isotope analyses and Trudi Krol for assisting with the soil mineral N analyses.


  1. Abbasi MK, Adams WA (2000) Gaseous N emission during simultaneous nitrification–denitrification associated with mineral N fertilisation to a grassland soil under field conditions. Soil Biol Biochem 32:1251–1259Google Scholar
  2. Anderson IC, Poth M, Homstead J, Burdige D (1993) A comparison of NO and N2O production by the autotrophic nitrifier Nitrosomonas europaea and the heterotrophic nitrifier Alcaligenes faecalis. Appl Environ Microbiol 59:3525–3533Google Scholar
  3. Badr O, Probert SD (1993) Environmental impacts of atmospheric nitrous oxide. Appl Energy 44:197–231Google Scholar
  4. Baggs EM, Richter M, Cadisch G, Hartwig UA (2003) Denitrification in grass swards is increased under elevated atmospheric CO2. Soil Biol Biochem 35:729–732CrossRefGoogle Scholar
  5. Berg P, Klemedtsson L, Rosswall T (1982) Inhibitory effect of low partial pressures of acetylene on nitrification. Soil Biol Biochem 14:301–303Google Scholar
  6. Blackmer AM, Bremner JM (1978) Inhibitory effect of nitrate on reduction of N2O to N2 by soil microorganisms. Soil Biol Biochem 10:187–191Google Scholar
  7. Bollmann A, Conrad R (1998) Influence of O2 availability on NO and N2O release by nitrification and denitrification in soils. Glob Chang Biol 4:387–396Google Scholar
  8. Bouwman AF (1996) Direct emission of nitrous oxide from agricultural soils. Nutr Cycl Agroecosyst 46:53–70Google Scholar
  9. Brookes PD, Stark JM, McInteer BB (1989) Diffusion method to prepare soil extracts for automated nitrogen-15 analysis. Soil Sci Soc Am J 53:1707–1711Google Scholar
  10. Carter JP, Hsiao YH, Spiro S, Richardson DJ (1995) Soil and sediment bacteria capable of aerobic nitrate respiration. Appl Environ Microbiol 61:2852–2858Google Scholar
  11. Clayton H, McTaggart IP, Parker J, Swan L, Smith KA (1997) Nitrous oxide emissions from fertilised grassland: a 2 year study of the effects of N fertiliser form and environmental conditions. Biol Fertil Soils 25:252–260Google Scholar
  12. Davidson EA (1991) Fluxes of nitrous oxide and nitric oxide from terrestrial ecosystems. A global inventory of nitric oxide emissions from soils. In: Rogers JE, Whitman WB (eds) Microbial production and consumption of greenhouse gases: methane, nitrogen oxides, and halomethanes. American Society for Microbiology, Washington, DC, pp 219–235Google Scholar
  13. Davidson EA, Hart SC, Shanks CA, Firestone MK (1991) Measuring gross nitrogen mineralization, immobilization, and nitrification by 15N isotopic pool dilution in intact soil cores. J Soil Sci 42:335–349Google Scholar
  14. Daum M, Zimmer W, Papen H, Kloos K, Nawrath K, Bothe H (1998) Physiological and molecular biological characterization of ammonia oxidation of the heterotrophic nitrifier Pseudomonas putida. Curr Microbiol 37:281–288Google Scholar
  15. Dobbie KE, Smith KA (2001) The effects of temperature, water-filled pore space and land use on N2O emissions from an imperfectly drained gleysol. Eur J Soil Sci 52:667–673Google Scholar
  16. Dobbie KE, McTaggart IP, Smith KA (1999) Nitrous oxide emissions from intensive agricultural systems: variations between crops and seasons, key driving variables, and mean emission factors. J Geophys Res D21:26891–26899Google Scholar
  17. Garrido F, Hénault C, Gaillard H, Pérez S, Germon JC (2002) N2O and NO emissions by agricultural soils with low hydraulic potentials. Soil Biol Biochem 34:559–575Google Scholar
  18. Goreau TJ, Kaplan WA, Wofsy SC, McElroy MB, Valois FW, Watson SW (1980) Production of NO2 and N2O by nitrifying bacteria at reduced concentrations of oxygen. Appl Environ Microbiol 40:526–532Google Scholar
  19. Houghton JT, Ding Y, Griggs DJ, Noguer M, van der Linden PJ, Dai X, Maskell K, Johnson CA (2001) Climate change 2001: the scientific basis. Contribution of Working Group I to the third assessment report of the intergovernmental panel on climate change. Cambridge University Press, CambridgeGoogle Scholar
  20. Hynes RK, Knowles R (1982) Effect of acetylene on autotrophic and heterotrophic nitrification. Can J Microbiol 28:334–340Google Scholar
  21. Kester RA, de Boer W, Laanbroek HJ (1997) Production of NO and N2O by pure cultures of nitrifying and denitrifying bacteria during changes in aeration. Appl Environ Microbiol 63:3872–3877Google Scholar
  22. Killham K (1986) Heterotrophic nitrification. In: Prosser JI (ed) Nitrification. IRL, Oxford, pp 117–126Google Scholar
  23. Knowles R (1982) Denitrification. Microbiol Rev 46:43–70Google Scholar
  24. Kroeze C, Mosier A, Bouwman AF (1999) Closing the global N2O budget: a retrospective analysis 1500–1994. Glob Biogeochem Cycles 13:1–8Google Scholar
  25. Laverman AM, Zoomer HR, Engelbrecht D, Berg MP, van Straalen NM, van Versveld HW, Verhoef HA (2000) Soil layer-specific variability in net nitrification and denitrification in an acid coniferous forest. Biol Fertil Soils 32:427–434Google Scholar
  26. Linn DM, Doran JW (1984) Effect of water-filled pore space on carbon dioxide and nitrous oxide production in tilled and nontilled soils. Soil Sci Soc Am J 48:1267–1272Google Scholar
  27. Moir JWB, Crossman LC, Spiro S, Richardson DJ (1996) The purification of ammonia monooxygenase from Paracoccus denitrificans. FEMS Lett 387:71–74Google Scholar
  28. Papen H, Vonberg R, Hinkel I, Thoene B, Rennenberg H (1989) Heterotrophic nitrification by Alcaligenes faecalis—NO2, NO3, N2O and NO production in exponentially growing cultures. Appl Environ Microbiol 55:2068–2072Google Scholar
  29. Parton WJ, Mosier AR, Ojima DS, Valentine DW, Schimel DS, Weier K, Kulmala AE (1996) Generalised model for N2 and N2O production from nitrification and denitrification. Glob Biogeochem Cycles 10:401–412Google Scholar
  30. Patureau D, Zumstein E, Delgenes JP, Moletta R (2000) Aerobic denitrifiers isolated from diverse natural and managed ecosystems. Microb Ecol 39:145–152Google Scholar
  31. Pedersen H, Dunkin KA, Firestone MK (1999) The relative importance of autotrophic and heterotrophic nitrification in a conifer forest soil as measured by 15N tracer and pool dilution techniques. Biogeochemistry 44:135–150Google Scholar
  32. Poth M, Focht DD (1985) 15N kinetic analysis of N2O production by Nitrosomonas europaea—an examination of nitrifier denitrification. Appl Environ Microbiol 49:1134–1141Google Scholar
  33. Renault P, Sierra J (1994) Modelling oxygen diffusion in aggregated soils. 2. Anaerobiosis in topsoil layers. Soil Sci Soc Am J 58:1023–1030Google Scholar
  34. Robertson LA, Kuenen JG (1990) Combined heterotrophic nitrification and aerobic denitrification in Thiosphaera pantotropha and other bacteria. Antonie van Leeuwenhoek 57:139–152Google Scholar
  35. Robertson GP, Tiedje JM (1987) Nitrous oxide sources in aerobic soils: nitrification, denitrification and other biological processes. Soil Biol Biochem 19:187–193Google Scholar
  36. Sexstone AJ, Parkin TB, Tiedje JM (1988) Denitrification response to soil wetting in aggregated and unaggregated soil. Soil Biol Biochem 20:767–769Google Scholar
  37. Shoun H, Kim D-H, Uchiyama H, Sugiyama J (1992) Denitrification by fungi. FEMS Microbiol Lett 94:277–281Google Scholar
  38. Skiba U, Ball B (2002) The effect of soil texture and soil drainage on emissions of nitric oxide and nitrous oxide. Soil Use Manage 18:56–60Google Scholar
  39. Smith KA (1980) A model of the extent of anaerobic zones in aggregated soils, and its potential application to estimates of denitrification. J Soil Sci 31:263–277Google Scholar
  40. Smith KA, Arah JRM (1990) Losses of nitrogen by denitrification and emissions of nitrogen oxides from soils. In: The Fertiliser Society Proceedings 299, LondonGoogle Scholar
  41. Stark JM, Firestone MK (1995) Mechanisms for soil-moisture effects on the activity of nitrifying bacteria. Appl Environ Microbiol 61:218–221Google Scholar
  42. Stevens RJ, Laughlin RJ, Burns LC, Arah JRM, Hood RC (1997) Measuring the contributions of nitrification and denitrification to the flux of nitrous oxide from soil. Soil Biol Biochem 29:139–151Google Scholar
  43. Webster EA, Hopkins DW (1996) Contributions from different microbial processes to N2O emissions from soil under different moisture regimes. Biol Fertil Soils 22:331–335Google Scholar
  44. Wolf I, Russow R (2000) Different pathways of formation of N2O, N2 and NO in black earth soil. Soil Biol Biochem 32:229–239Google Scholar
  45. Wrage N, Velthof GL, van Beusichem ML, Oenema O (2001) Role of nitrifier denitrification in the production of nitrous oxide. Soil Biol Biochem 33:1723–1732CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2005

Authors and Affiliations

  1. 1.Department of Agricultural Sciences, Wye CampusImperial College LondonAshfordUK
  2. 2.School of Biological SciencesUniversity of AberdeenAberdeenUK
  3. 3.Department of Biology 3University of YorkYorkUK

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