Skip to main content
SpringerLink
Log in

Perspectives on measurement of denitrification in the field including recommended protocols for acetylene based methods

  • Published:
Plant and Soil Aims and scope Submit manuscript

Abstract

Of the biogeochemical processes, denitrification has perhaps been the most difficult to study in the field because of the inability to measure the product of the process. The last decade of research, however, has provided both acetylene and15N based methods as well as undisturbed soil core andin situ soil cover sampling approaches to implementing these methods. All of these methods, if used appropriately, give comparable results. Thus, we now have several methods, each with advantages for particular sites or objectives, that accurately measure denitrification in nature. Because of the general usefulness of the acetylene methods, updated protocols for the following three methods are given: gas-phase recirculation soil cores; static soil cores; and the denitrifying enzyme assay also known as the phase 1 assay. Despite the availability of these and other methods, denitrification budgets remain difficult to accurately establish in most environments because of the high spatial and temporal variability inherent in denitrification. Appropriate analysis of those data includes a distribution analysis of the data, and if highly skewed as is typically the case, the most accurate method to estimate the mean and the population variance is the UMVUE method (uniformly minimum variance unbiased estimator). Geostatistical methods have also been employed to improve spatial and temporal estimates of denitrification. These have occasionally been successful for spatial analysis but in the attempt described here for temporal analysis the approach was not useful.

Discussions of the importance of denitrification have always focused on quantifying the process and whether particular measured quantities are judged to be a significant amount of nitrogen. A second line of evidence discussed here is the extant genetic record that results from natural selection. These analysis lead to the conclusion that strong selection for denitrification must currently be occurring, which implies that the process is of general significance in soils.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Abd-el-Malek Y, Hosny I and Emam N F 1974 Evaluation of media used for enumeration of denitrifying bacteria. Zentralbl. Bakteriol. Parasitenkd. Infektionski Hyg. 2 Abt. 415–421.

    Google Scholar 

  • Aulakh M S, Rennie D A and Paul E A 1982 Gaseous nitrogen from cropped and summer-fallowed soils. Can. J. Soil Sci. 62, 187–196.

    Google Scholar 

  • Balderston W L, Sherr B and Payne W J 1976 Blockage by acetylene of nitrous oxide reduction inPseudomonas perfectomarinus. Appl. Environ. Microbiol. 31, 504–508.

    Google Scholar 

  • Betlach M R 1982 Evolution of bacterial denitrification and denitrifier diversity. Antonie van Leeuwenhock J. Microbiol. 48, 585–607.

    Google Scholar 

  • Blackmer A M, Robbins S G and Bremner J M 1982 Diurnal variability in rate of emission of nitrous oxide from soils. Soil Sci. Soc. Am. J. 46, 937–942.

    Google Scholar 

  • Burton D L and Beauchamp E C 1984 Field techniques using the acetylene blockage of nitrous oxide reduction to measure denitrification. Can. J. Soil Sci. 64, 555–562.

    Google Scholar 

  • Christensen S and Tiedje J M 1988 Denitrification in the field: Analysis of spatial and temporal variability.In Nitrogen Efficiency in Agricultural Soils. Eds D S Jenkinson and K A Smith. pp 295–301. Elsevier Appl. Sci. London, N.Y., USA.

    Google Scholar 

  • Duxbury J M and McCounnaughey P K 1986 Effect of fertilizer source on denitrification and nitrous oxide emissions in a maize-field. Soil Sci. Soc. Am. J. 50, 644–648.

    Google Scholar 

  • Duxbury J M 1986 Advantages of the acetylene method for measuring denitrification.In Field Measurement of Dinitrogen Fixation and Denitrification. Eds. R D Hauck and R W Weaver, pp 73–91. Soil Science Society of America, Madison, Wisconsin, USA.

    Google Scholar 

  • Federova R I, Milekhina E I and Ilkukhina N I 1973 Possiblity of using the ‘gas-exchange’ method to detect extraterrestrial life: Identification of nitrogen-fixing organisms. Izv. Akad. Nauk Arm. SSR Biol. Nauki 6, 797–806.

    Google Scholar 

  • Folorunso O A and Rolston D E 1984 Spatial variability of field-measured denitrification gas fluxes. Soil Sci. Soc. Am. J. 48, 1214–1219.

    Google Scholar 

  • Folorunso O A and Rolston D E 1985 Spatial and spectral relationships between field-measured denitrification gas fluxes and soil properties. Soil. Sci. Soc. Am. J. 49, 1087–1093.

    Google Scholar 

  • Gamble T N, Betlach M R and Tiedje J M 1977 Numerically dominant denitrifying bacteria from world soils. Appl. Environ. Microbiol. 33, 926–939.

    Google Scholar 

  • Goodroad L L and Keeney D R 1984 Nitrous oxide emissions from soils during thawing. Can. J. Soil Sci. 64, 187–194.

    Google Scholar 

  • Groffman P M 1985 Nitrification and denitrification in conventional and no-tillage soils. Soil Sci. Soc. Am. J. 49, 329–334.

    Google Scholar 

  • Groffman P M and Tiedje J M 1988 Denitrification hysteresis during wetting and drying cycles in soil. Soil Sci. Soc. Am. J. 52 1626–1629.

    Google Scholar 

  • Groffman P M and Tiedje J M 1989a Denitrification in north temperate forest soils: Spatial and temporal patterns at the landscape and seasonal scales. Soil Biol. Biochem. (Accepted).

  • Groffman P M and Tiedje J M 1989b Denitrification in north temperate forest soils: Relationships between denitrification and environmental parameters at the landscape scale. Soil Biol. Biochem. (Accepted).

  • Groffman P M, Tiedje J M, Robertson G P and Christensen S 1988 Denitrification at different temporal and geographical scales: Proximal and distal controls.in Adv. in Nitrogen Cycling in Agricultural Ecosystems. Ed. J R Wilson. pp 174–192. CAB International, Wallingford, U.K.

    Google Scholar 

  • Hallmark S L and Terry R E 1985 Field measurement of denitrification in irrigated soils. Soil Sci. 140, 35–44.

    Google Scholar 

  • Hauck R D 1986 Field measurement of denitrification—an overview.In Field Measurement of Dinitrogen Fixation and Denitrification. Eds. R D Hauck and R W Weaver. pp 59–72. Soil Science Society of America, Madison, Wisconsin, USA.

    Google Scholar 

  • Hauck R D, Meldsted S W and Yankwich P E 1958 Use of N-isotope distribution in nitrogen gas in the study of denitrification. Soil Sci. 86, 287–291.

    Google Scholar 

  • Heinemeyer O, Haider K and Mosier A 1988 Phytotron studies to compare nitrogen losses from corn planted soil by the 15-N balance or direct dinitrogen and nitrous oxide measurements. Biol. Fert. Soils 6, 73–77.

    Google Scholar 

  • Hyman M R and Arp D J 1987 Quantification and removal of some contaminating gases from acetylene used to study gas-utilizing enzymes and microorganisms. Appl. Environ. Microbiol. 53, 298–303.

    Google Scholar 

  • Journel A G and Hujbregts C J 1978 Mining Geostatistics. Academic Press, New York.

    Google Scholar 

  • Juma N G and Paul E A 1981 Use of tracers and computer simulation techniques to assess mineralization and immobilization of soil nitrogen.In Simulation of Nitrogen Behavior of Soil-Plant Systems. Eds. M J Frissel and J A Van Veen. pp 145–154. Center for Agricultural Publishing and Documentation, Wageningen.

    Google Scholar 

  • Jury W A, Letey J and Collins T 1982 Analysis of chamber methods used for measuring nitrous oxide production in the field. Soil Sci. Soc. Am. J. 46, 250–255.

    Google Scholar 

  • Kaspar H F 1984 A simple method for the measurement of N2O and CO2 flux rates across undisturbed soil surfaces. N.Z. J. Sci. 27, 243–246.

    Google Scholar 

  • Kaspar H F and Tiedje J M 1980 Response of electron-capture detector to hydrogen, oxygen, nitrogen, carbon dioxide, nitric oxide and nitrous oxide. J. Chromatography 193, 142–147.

    Google Scholar 

  • Kaspar H F, Tiedje J M and Firestone R B 1981 Denitrification and dissimilatory nitrate reduction to ammonium in digested sludge. Can. J. Microbiol. 27, 878–885.

    Google Scholar 

  • Keeney D R 1986 Critique of the acetylene blockage technique for field measurement of denitrification.In Field Measurement of Dinitrogen Fixation and Denitrification. Eds. R D Hauck and R W Weaver. pp 103–115. Soil Science Society of America, Madison, Wisconsin, USA.

    Google Scholar 

  • Knowles R 1981 Denitrification.In Soil Biochemistry, Vol. 5, p 240. Eds. E A Paul and J N Ladd. Marcel Dekker, Inc. New York.

    Google Scholar 

  • Koike I and Hattori A 1975 Growth yield of a denitrifying bacteriumPseudomonas denitrificans under aerobic and denitrifying conditions. J. Gen. Microbiol. 88, 1–10.

    Google Scholar 

  • Korner H, Runzke K, Dohler K and Zumft W G 1987 Immunochemical patterns of distribution of nitrous oxide reductase and nitrite reductase (cytochrome cd1) among denitrifying pseudomonads. Arch. Microbiol. 148, 20–24.

    Google Scholar 

  • Klemedtsson L 1986 Denitrification in arable soil with special emphasis on the influence of plant roots. Report 32, Dept. of Microbiology. Swedish University of Agricultural Sciences, Uppsala, Sweden.

    Google Scholar 

  • Klemedtsson L, Svensson B H and Rosswal T 1977 The use of acetylene inhibition of nitrous oxide reductase in quantifying denitrification in soils. Swedish J. Agric. Res. 7, 179–185.

    Google Scholar 

  • Limmer A W, Steele K W and Wilson A T 1982 Direct field measurement of N2 and N2O evolution from soil. J. Soil Sci. 33, 499–507.

    Google Scholar 

  • Lowrance R and Smittle D 1988 Nitrogen cycling in a multiplecrop vegetable production system. J. Environ. Qual. 17, 158–162.

    Google Scholar 

  • McConnaughey P K and Duxbury J M 1986 Introduction of acetylene into soil for measurement of denitrification. Soil Sci. Soc. Am. J. 50, 260–263.

    Google Scholar 

  • Matson P A and Vitousek P M 1987 Cross-system comparisons of soil nitrogen transformations and nitrous oxide flux in tropical forest ecosystems. Global Biogeochemical Cycles 1, 163–170.

    Google Scholar 

  • Matthias A D, Blackmer A M and Bremner J M 1980 A simple chamber technique for field measurement of emissions of nitrous oxide from soils. J. Environ. Qual. 9, 251–256.

    Google Scholar 

  • Michalski W P and Nicholas D J D 1988 Immunological patterns of distribution of bacterial denitrifying enzymes. Phytochemistry 27, 2451–2456.

    Google Scholar 

  • Mosier A R, Guenzi W D and Schweizer E E 1986 Field denitrification estimation by nitrogen-15 and acetylene inhibition techniques. Soil Sci. Soc. Am. J. 50, 831–833.

    Google Scholar 

  • Mosier A R, Guenzi W D and Schweizer E E 1986 Soil losses of dinitrogen and nitrous oxide from irrigated crops in north eastern Colorado. Soil. Sci. Soc. Am. J. 50, 344–348.

    Google Scholar 

  • Myrold D D 1988 Denitrification in ryegrass and winter wheat cropping systems of western Oregon. Soil Sci. Soc. Am. J. 52, 412–415.

    Google Scholar 

  • Myrold D D and Tiedje J M 1986 Simultaneous estimation of several nitrogen cycle rates using15N: Theory and application. Soil Biol. Biochem. 6, 559–568.

    Google Scholar 

  • Myrold D D and Tiedje J M 1985 Diffusional constraints on denitrification. Soil Sci. Soc. Am. J. 49, 652–657.

    Google Scholar 

  • Nommik H 1956 Investigations on denitrification in soil. Acta Agric. Scan. 6, 195–228.

    Google Scholar 

  • Parkin T B 1985 Automated analysis of nitrous oxide. Soil Sci. Soc. Am. J. 49, 273–276.

    Google Scholar 

  • Parkin T B 1987 Soil microsites as a source of denitrification variability. Soil Sci. Am. J. 51, 1194–1199.

    Google Scholar 

  • Parkin T B, Kaspar H F, Sexstone A J and Tiedje J M 1984 A gas-flow soil core method to measure field denitrification rates. Soil Biol. Biochem. 16, 323–330.

    Google Scholar 

  • Parkin T B, Mesinger J J, Chester S T, Starr J L and Robinson J A 1988 Evaluation of statistical estimation methods for lognormally distributed variables. Soil Sci. Soc. Am. J. 52, 323–329.

    Google Scholar 

  • Parkin T B and Robinson J A 1989 Stochastic models of soil denitrification. Appl. Environ. Microbiol 55, 72–77.

    Google Scholar 

  • Parkin T B, Sexstone A J and Tiedje J M 1985 Comparison of field denitrification rates determined by acetylene-based soil core and nitrogen-15 methods. Soil Sci. Soc. Am. J. 49, 94–99.

    Google Scholar 

  • Parkin T B, Starr J L and Meisinger J J 1987 Influence of sample size on measurement of soil denitrification. Soil Sci. Soc. Am. J. 51, 1492–1501.

    Google Scholar 

  • Parkin T B and Tiedje J M 1984 Application of a soil core method to investigate the effect of oxygen concentration on denitrification. Soil Biol. Biochem. 4, 331–334.

    Google Scholar 

  • Pearson E S and Hartley H O 1958 Biometrika Tables for Statisticians, Volume 1. Cambridge Univ. Press, London.

    Google Scholar 

  • Rice C W, Sierzega P E, Tiedje J M and Jacobs L W 1988 Stimulated denitrification in the microenvironment of a biodegradable organic waste injected into soil. Soil Sci. Soc. Am. J. 52, 102–108.

    Google Scholar 

  • Robertson G P, Huston M A, Evans F C and Tiedje J M 1988 Spatial variability in a successional plant community: Patterns of nitrogen mineralization, nitrification, and denitrification. Ecology 69, 1517–1524.

    Google Scholar 

  • Robertson G P and Tiedje J M 1987 Nitrous oxide sources in aerobic soils. Nitrification, denitrification and other biological processes. Soil Biol. Biochem. 19, 187–193.

    Google Scholar 

  • Robertson G P and Tiedje J M 1985 An automated technique for sampling the contents of stoppered gas-collection vials. Plant and Soil 83, 453–457.

    Google Scholar 

  • Robertson G P and Tiedje J M 1984 Denitrification and nitrous oxide production in successional and old-growth Michigan forests. Soil Sci. Soc. Am. J. 383–389.

  • Robertson G P, Vitousek P M, Matson P A and Tiedje J M 1987 Denitrification in a clearcut Loblolly pine (Pinus taeda L) plantation in the southeastern. U.S. Plant and Soil 97, 119–129.

    Google Scholar 

  • Rolston D E 1986 Limitations of the acetylene blockage technique for field measurement of denitrification.In Field Measurement of Dinitrogen Fixation and Denitrification. Eds. R D Hauck and R W Weaver. pp 93–101. Soil Science Society of America, Madison, Wisconsin, USA.

    Google Scholar 

  • Rolston D E, Broadbent F E and Goldhamer D A 1979 Field measurements of denitrification. II. Mass balance and sampling uncertainty. Soil Sci. Am. J. 43, 703–708.

    Google Scholar 

  • Rolston D E, Fried M and Goldhamer D A 1976 Denitrification measured directly from nitrogen and nitrous oxide gas fluxes. Soil Sci. Soc. Am. J. 40, 259–266.

    Google Scholar 

  • Rolston D E, Hoffman D L and Toy D W 1978 Field measurement of denitrification. I. Flux of N2 and N2O. Soil Sci. Soc. Am. J. 42, 863–869.

    Google Scholar 

  • Rolston D E, Rao P S C, Davidson J M and Jessup R E 1984 Simulation of denitrification losses of nitrate fertilizer applied to uncropped, cropped and manure-amended field plots. Soil Sci. 137, 270–279.

    Google Scholar 

  • Rolston D E, Sharpley A N, Toy D W and Broadbent F E 1982 Field measurement of denitrification. III. Rates during irrigation cycles. Soil Sci. Soc. Am. J. 46, 289–296.

    Google Scholar 

  • Romermann D and Friedrich B 1985 Denitrification byAlcaligenes entrophus is plasmid dependent. J. Bacteriol. 162, 852–854.

    Google Scholar 

  • Ryden J C, Lund L J, Letey J and Focht D D 1979 Direct measurement of denitrification loss from soils. II. Development and application of field methods. Soil Sci. Soc. Am. J. 43, 110–118.

    Google Scholar 

  • Ryden J C and Dawson K P 1982 Evaluation of the acetylene-inhibition technique for the measurement of denitrification in grassland soils. J. Sci. Food Agric. 33, 1197–1206.

    Google Scholar 

  • Ryden J C, Skinner J H and Nixon D J 1987 Soil core incubation system for the field measurement of denitrification using acetylene-inhibition. Soil Biol. Biochem. 19, 753–757.

    Google Scholar 

  • Schmidt J, Seiler W and Conrad R 1988 Emission of nitrous oxide from temperate forest soils into the atmosphere. J. Atmos. Chem. 6, 95–115.

    Google Scholar 

  • Sexstone A J, Parkin T B and Tiedje J M 1988 Denitrification response to soil wetting in aggregated and unaggregated soil. Soil Biol. Biochem. 20, 767–769.

    Google Scholar 

  • Sexstone A J, Parkin T B and Tiedje J M 1985 Temporal response of soil denitrification rates to rainfall and irrigation. Soil Sci. Soc. Am. J. 48, 99–103.

    Google Scholar 

  • Siegel R S, Hauck R D and Kurtz L T 1982 Determination of30N2 and application to measurement of N2 evolution during denitrification. Soil Sci. Soc. Am. J. 46, 68–74.

    Google Scholar 

  • Smith C J 1988 Denitrification in the field.In Advances in Nitrogen Cycling in Agricultural Ecosystems. Ed J R Wilson. pp 387–398. C.A.B. International, Wallingford U.K.

    Google Scholar 

  • Smith M S, Firestone M K and Tiedje J M 1978 The acetylene inhibition method for short-term measurement of soil denitrification and its evaluation using nitrogen-13. Soil Sci. Soc. Am J. 42, 611–615.

    Google Scholar 

  • Smith M S and Tiedje J M 1979 Phases of denitrification following oxygen depletion in soil. Soil Biol. Biochem. 11, 261–267.

    Google Scholar 

  • Svensson B H, Klemedtsson L and Rosswall T 1985 Preliminary field denitrification studies of nitrate-fertilized and nitrogen-fixing crops.In Denitrification and the Nitrogen Cycle. Ed. H L Golterman. pp 157–169. NATO Conference Series I: Ecology Vol. 9. Plenum Press, London.

    Google Scholar 

  • Terry R E and Duxbury J M 1985 Acetylene decomposition in soils. Soil Sci. Soc. Am. J. 49, 90–94.

    Google Scholar 

  • Terry R E, Jellen E N and Breakwell D P 1986 Effect of irrigation and acetylene exposure on field denitrification measurements. Soil Sci. Soc. Am. J. 50, 115–120.

    Google Scholar 

  • Tiedje J M 1982 Denitrification.In Methods of Soil Analysis, Park 2nd ed. Ed. A L Page. Agronomy Monogr. 9, 1011–1026. Amer. Soc. Agron., Madison, Wisc.

    Google Scholar 

  • Tiedje J M 1988 Ecology of denitrification and dissimilatory nitrate reduction to ammonium.In Biology of Anaerobic Microorganisms. Ed. A J B Zehnder. pp 179–244. John Wiley & Sons, New York.

    Google Scholar 

  • Tiedje J M, Firestone R B, Firestone M K, Betlach M R, Kaspar H F and Sørenson J 1981 Use of13N in studies of denitrification.In Short-lived radionuclides in Chemistry and Biology. Eds. J W Root and K A Krohn. pp 295–315. American Chemical Society.

  • Tiedje J M, Sexstone A J, Parkin T B and Revsbech N P 1984 Anaerobic processes in soil. Plant and Soil 76, 197–212.

    Google Scholar 

  • Topp E and Germon J C 1986 Acetylene metabolism and stimulation of denitrification in an agricultural soil. Appl. Environ. Microbiol. 52, 802–806.

    Google Scholar 

  • Vieira S R, Hatfield J L, Nielsen D R and Biggar J W 1983 Geostatistical theory and application to variability of some agronomical properties. Hilgardia 51, 1–75.

    Google Scholar 

  • Webster 1985 Quantitative spatial analysis of soil in the field, Adv. Soil Sci. 3, 1–70.

    Google Scholar 

  • Yeomans J C and Beauchamp E G 1982 Acetylene as a possible substrate in the denitrification process. Can. J. Soil Sci. 62, 139–146.

    Google Scholar 

  • Yeomans J C and Beauchamp E G 1978 Limited inhibition of nitrous oxide reduction in soil in the presence of acetylene. Soil Biol. Biochem. 10, 517–519.

    Google Scholar 

  • Yoshinari T and Knowles R 1976 Acetylene inhibition of nitrous oxide reduction by denitrifying bacteria. Biochem. Biophys. Res. Commun. 69, 705–710.

    Google Scholar 

  • Zar J H 1974 Biostatistical Analysis. Prentice-Hall, Inc., Englewood Cliffs, N. J.

    Google Scholar 

Download references

Author information

Author notes

  1. Stephen Simkins

    Present address: Dept. Plant and Soil Sciences, Univ. of Massachusetts, 01003, Amherst, MA, USA

  2. Peter M. Groffman

    Present address: Dept. of Natural Resources Science, Univ. of Rhode Island, 02881, Kingston, RI, USA

Authors and Affiliations

  1. Dept. of Crop and Soil Sciences and of Microbiology and Public Health, Michigan State University, 48824, E. Lansing, MI, USA

    James M. Tiedje, Stephen Simkins & Peter M. Groffman

Authors
  1. James M. Tiedje
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Stephen Simkins
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Peter M. Groffman
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Tiedje, J.M., Simkins, S. & Groffman, P.M. Perspectives on measurement of denitrification in the field including recommended protocols for acetylene based methods. Plant Soil 115, 261–284 (1989). https://doi.org/10.1007/BF02202594

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02202594

Key words

Search

Navigation