Biology and Fertility of Soils

, Volume 52, Issue 5, pp 601–614 | Cite as

Dissimilatory nitrate reduction to ammonium and N2O flux: effect of soil redox potential and N fertilization in loblolly pine forests

  • K. J. MinickEmail author
  • C. B. Pandey
  • T. R. Fox
  • Santosh Subedi
Original Paper


Nitrogen (N) fertilization and soil redox potential influence N cycling processes in forested ecosystems. Gross N transformations are indicators of NH4 + and NO3 production and consumption within soil. Furthermore, dissimilatory nitrate reduction to ammonium (DNRA), a typically overlooked process in terrestrial N cycling, can conserve N within soil by reducing losses of soil N via NO3 leaching and denitrification. We tested the effects of urea fertilization and soil redox on microbial N cycling processes and N2O fluxes using a 15N tracer experiment in soils from loblolly pine plantations located in different physiographical regions (i.e., Coastal Plain of North Carolina and Piedmont of Virginia). Mineral soils (0–15 cm) from fertilized and unfertilized plots were incubated at high (Eh, 200 to 400 mV) and low redox potential (Eh, −100 to 100 mV). Site differences were limited primarily to edaphic factors, although gross N mineralization was higher in NC. Gross nitrification, DNRA, and NO3–N concentrations were higher in soils from fertilized plots. DNRA was higher at high compared to low redox potential, while N2O fluxes were higher at low redox potential. Fluxes of N2O were further enhanced in fertilized treatments incubated at low redox potential. DNRA was positively correlated with NO3 availability, but not to soil C pools. Furthermore, DNRA was negatively correlated with C/NO3 ratio, implying that NO3 pool size was the primary factor influencing DNRA. These results suggest N fertilization has alleviated limitations to nitrification, DNRA, and N2O production processes and that gaseous losses of N will prevail over N conservation pathways at low soil redox potentials.


Pinus taeda Mineralization Nitrification Anaerobic Denitrification Nitrate 



This work was conducted mainly under the India-USA Fulbright program supporting one of the authors (C.B. Pandey) in the form of a Fulbright-Nehru fellowship. Financial support from the NSF Center for Advanced Forestry Systems and the Forest Productivity Cooperative is gratefully acknowledged. Partial financial support for this work was also provided by the Virginia Agricultural Experiment Station and the McIntire-Stennis Program of the National Institute of Food and Agriculture, U.S. Department of Agriculture. We thank Chelsea Drum and Timothy Albaugh for help with sample collection and David Mitchem for assistance in the laboratory. We also thank four anonymous reviewers for their comments, which significantly improved the quality of this manuscript.


  1. Aber JD (1992) Nitrogen cycling and nitrogen saturation in temperate forest ecosystems. Trends Ecol Evol 7:220–224CrossRefPubMedGoogle Scholar
  2. Bengtsson G, Bergwall C (2000) Fate of 15N labelled nitrate and ammonium in a fertilized forest soil. Soil Biol Biochem 32:545–557CrossRefGoogle Scholar
  3. Bethke CM, Sanford RA, Kirk M, Jin Q, Flynn TM (2011) The thermodynamic ladder in geomicrobiology. Am J Sci 311:183–210CrossRefGoogle Scholar
  4. Bonin P (1996) Anaerobic nitrate reduction to ammonium in two strains isolated from coastal marine sediment: A dissimilatory pathway. FEMS Microbiol Ecol 19:27–38CrossRefGoogle Scholar
  5. Bremner JM, Blackmer AM (1978) Nitrous oxide: emission from soils during nitrification of fertilizer nitrogen. Science (New York, NY) 199:295–296CrossRefGoogle Scholar
  6. Brooks PD, Stark JM, Mclnteer BB, Preston T (1989) Diffusion method to prepare soil extracts for automated nitrogen-15 analysis. Soil Sci Soc Am J 53:1707–1711CrossRefGoogle Scholar
  7. Brunel B, Janse J, Laanbroek H, Woldendorp J (1992) Effect of transient oxic conditions on the composition of the nitrate-reducing community from the rhizosphere of Typha angustifolia. Microb Ecol 24:51–61CrossRefPubMedGoogle Scholar
  8. Caskey WH, Tiedje JM (1979) Evidence for Clostridia as agents of dissimilatory reduction of nitrate to ammonium in soils. Soil Sci Soc Am J 43:931–936CrossRefGoogle Scholar
  9. Castro MS, Peterjohn WT, Melillo JM, Steudler PA, Gholz HL, Lewis D (1994) Effects of nitrogen fertilization on the fluxes of N2O, CH4, and CO2 from soils in a Florida slash pine plantation. Can J For Res 24:9–13CrossRefGoogle Scholar
  10. Chen Z, Ding W, Xu Y, Müller C, Rütting T, Yu H, Fan J, Zhang J, Zhu T (2015) Importance of heterotrophic nitrification and dissimilatory nitrate reduction to ammonium in a cropland soil: Evidences from a 15N tracing study to literature synthesis. Soil Biol Biochem 91:65–75CrossRefGoogle Scholar
  11. Cheng Y, Wang J, Wang S, Zhang J, Cai Z (2014) Effects of soil moisture on gross N transformations and N2O emission in acid subtropical forest soils. Biol Fertil Soils 50:1099–1108CrossRefGoogle Scholar
  12. Corre MD, Beese FO, Brumme R (2003) Soil nitrogen cycle in high nitrogen deposition forest: changes under nitrogen saturation and liming. Ecol Appl 13:287–298CrossRefGoogle Scholar
  13. Fazzolari É, Nicolardot B, Germon JC (1998) Simultaneous effects of increasing levels of glucose and oxygen partial pressures on denitrification and dissimilatory nitrate reduction to ammonium in repacked soil cores. Eur J Soil Biol 34:47–52CrossRefGoogle Scholar
  14. Finzi AC, Schlesinger WH (2003) Soil-nitrogen cycling in a pine forest exposed to 5 years of elevated carbon dioxide. Ecosystems 6:444–456CrossRefGoogle Scholar
  15. Firestone MK, Davidson EA (1989) Microbial basis for NO and N2O production and consumption in soil. In: Andreae MO, Schimel DS (eds) Exchange of trace gases between terrestrial ecosystems and the atmosphere. Wiley, New York, pp 7–21Google Scholar
  16. Fisk M, Santangelo S, Minick K (2015) Carbon mineralization is promoted by phosphorus and reduced by nitrogen addition in the organic horizon of northern hardwood forests. Soil Biol Biochem 81:212–218CrossRefGoogle Scholar
  17. Fox T, Burger J, Kreh R (1986) Effects of site preparation on nitrogen dynamics in the southern Piedmont. For Ecol Manag 15:241–256CrossRefGoogle Scholar
  18. Fox TR, Allen HL, Albaugh TJ, Rubilar R, Carlson CA (2007) Tree nutrition and forest fertilization of pine plantations in the southern United States. South J Appl For 31:5–11Google Scholar
  19. Galloway JN, Aber JD, Erisman JW, Seitzinger SP, Howarth RW, Cowling EB, Cosby BJ (2003) The nitrogen cascade. Bioscience 53:341–356CrossRefGoogle Scholar
  20. Groffman PM, Tiedje JM (1989) Denitrification in north temperate forest soils: spatial and temporal patterns at the landscape and seasonal scales. Soil Biol Biochem 21:613–620CrossRefGoogle Scholar
  21. Groffman PM, Butterbach-Bahl K, Fulweiler RW, Gold AJ, Morse JL, Stander EK, Tague C, Tonitto C, Vidon P (2009) Challenges to incorporating spatially and temporally explicit phenomena (hotspots and hot moments) in denitrification models. Biogeochemistry 93:49–77CrossRefGoogle Scholar
  22. Hall SJ, Matson PA (1999) Nitrogen oxide emissions after nitrogen additions in tropical forests. Nature 400:152–154CrossRefGoogle Scholar
  23. Hart SC, Stark JM, Davidson EA, Firestone MK (1994a) Nitrogen mineralization, immobilization and nitrification. In: Weaver RW (ed) Methods of soil analysis: microbiological and biochemical properties, 3rd edn. Soil Science Society of America, Madison, pp 985–1018Google Scholar
  24. Hart SC, Nason GE, Myrold DD, Perry DA (1994b) Dynamics of gross nitrogen transformations in an old-growth forest: the carbon connection. Ecology 75:880–891CrossRefGoogle Scholar
  25. Hedin LO, VonFischer JC, Ostrom NE, Kennedy BP, Brown MG, Robertson GP (1998) Thermodynamic constraints on nitrogen transformations and other biogeochemical processes at soil-steam interfaces. Ecology 79:684–703Google Scholar
  26. Huygens D, Rütting T, Boeckx P, Van Cleemput O, Godoy R, Müller C (2007) Soil nitrogen conservation mechanisms in a pristine south Chilean Nothofagus forest ecosystem. Soil Biol Biochem 39:2448–2458CrossRefGoogle Scholar
  27. Johnson D, Edwards N, Todd D (1980) Nitrogen mineralization, immobilization, and nitrification following urea fertilization of a forest soil under field and laboratory conditions. Soil Sci Soc Am J 44:610–616CrossRefGoogle Scholar
  28. Kirkham D, Bartholomew WV (1954) Equations for following nutrient transformations in soil, utilizing tracer data. Soil Sci Soc Am Proc 18:33–34CrossRefGoogle Scholar
  29. Kraft B, Tegetmeyer HE, Sharma R, Klotz MG, Ferdelman TG, Hettich RL, Geelhoed JS, Strous M (2014) The environmental controls that govern the end product of bacterial nitrate respiration. Science (New York, NY) 345:676–679CrossRefGoogle Scholar
  30. Kuzyakov Y, Blagodatskaya E (2015) Microbial hotspots and hot moments in soil: concept & review. Soil Biol Biochem 83:184–199CrossRefGoogle Scholar
  31. Li P, Lang M (2014) Gross nitrogen transformations and related N2O emissions in uncultivated and cultivated black soil. Biol Fertil Soils 50:197–206CrossRefGoogle Scholar
  32. Linn D, Doran J (1984) Effect of water-filled pore space on carbon dioxide and nitrous oxide production in tilled and nontilled soils. Soil Sci Soc Am J 48:1267–1272CrossRefGoogle Scholar
  33. Littell RC, Milliken GA, Stroup WW, Wolfinger RD, Schabenberger O (2006) SAS for mixed models. SAS Publishing, CaryGoogle Scholar
  34. Lu W-W, Zhang H-L, Shi W-M (2013) Dissimilatory nitrate reduction to ammonium in an anaerobic agricultural soil as affected by glucose and free sulfide. Eur J Soil Biol 58:98–104CrossRefGoogle Scholar
  35. Martikainen PJ, Aarnio T, Taavitsainen VM, Päivinen L, Salonen K (1989) Mineralization of carbon and nitrogen in soil samples taken from three fertilized pine stands: long-term effects. Plant Soil 114:99–106CrossRefGoogle Scholar
  36. Matheson F, Nguyen M, Cooper A, Burt T, Bull D (2002) Fate of 15N-nitrate in unplanted, planted and harvested riparian wetland soil microcosms. Ecol Eng 19:249–264CrossRefGoogle Scholar
  37. McClain ME, Boyer EW, Dent CL, Gergel SE, Grimm NB, Groffman PM, Hart SC, Harvey JW, Johnston CA, Mayorga E (2003) Biogeochemical hot spots and hot moments at the interface of terrestrial and aquatic ecosystems. Ecosystems 6:301–312CrossRefGoogle Scholar
  38. Medinets S, Skiba U, Rennenberg H, Butterbach-Bahl K (2015) A review of soil NO transformation: associated processes and possible physiological significance on organisms. Soil Biol Biochem 80:92–117CrossRefGoogle Scholar
  39. Mehlich A (1953) Determination of P, Ca, Mg, K, Na, and NH4. Soil Testing Division Publication, p. 1–53.Google Scholar
  40. Minick KJ, Fisk MC, Groffman PM (2011) Calcium and phosphorus interact to reduce mid-growing season net nitrogen mineralization potential in organic horizons in a northern hardwood forest. Soil Biol Biochem 43:271–279CrossRefGoogle Scholar
  41. Minick KJ, Strahm BD, Fox TR, Sucre EB, Leggett ZH, Zerpa JL (2014) Switchgrass intercropping reduces soil inorganic nitrogen in a young loblolly pine plantation located in coastal North Carolina. For Ecol Manag 319:161–168CrossRefGoogle Scholar
  42. Minick KJ, Strahm BD, Fox TR, Sucre EB, Leggett ZH (2015) Microbial nitrogen cycling response to forest-based bioenergy production. Ecol Appl 25:2366–2381CrossRefPubMedGoogle Scholar
  43. Müller C, Rütting T, Abbasi MK, Laughlin RJ, Kammann C, Clough TJ, Sherlock RR, Kattge J, Jäger H, Watson CJ (2009) Effect of elevated CO2 on soil N dynamics in a temperate grassland soil. Soil Biol Biochem 41:1996–2001CrossRefGoogle Scholar
  44. Myrold DD, Tiedje JM (1985) Establishment of denitrification capacity in soil: effects of carbon, nitrate and moisture. Soil Biol Biochem 17:819–822CrossRefGoogle Scholar
  45. Nannipieri P, Eldor P (2009) The chemical and functional characterization of soil N and its biotic components. Soil Biol Biochem 41:2357–2369CrossRefGoogle Scholar
  46. Nijburg JW, Coolen M, Gerards S, Gunnewiek P, Laanbroek HJ (1997) Effects of nitrate availability and the presence of Glyceria maxima on the composition and activity of the dissimilatory nitrate-reducing bacterial community. Appl Environ Microbiol 63:931–937PubMedPubMedCentralGoogle Scholar
  47. Pandey CB, Srivastava RC, Singh RK (2009) Soil nitrogen mineralization and microbial biomass relations; and nitrogen conservation in humid—tropics. Soil Sci Soc Am J 73:1142–1149CrossRefGoogle Scholar
  48. Pett-Ridge J, Silver WL, Firestone MK (2006) Redox fluctuations frame microbial community impacts on N-cycling rates in a humid tropical forest soil. Biogeochemistry 81:95–110CrossRefGoogle Scholar
  49. Reddy K, Patrick W (1976) Effect of frequent changes in aerobic and anaerobic conditions on redox potential and nitrogen loss in a flooded soil. Soil Biol Biochem 8:491–495CrossRefGoogle Scholar
  50. Robertson GP, Vitousek PM, Matson PA, Tiedje JM (1987) Denitrification in a clearcut Loblolly pine (Pinus taeda L.) plantation in the southeastern US. Plant Soil 97:119–129CrossRefGoogle Scholar
  51. Rütting T, Huygens D, Müller C, Van Cleemput O, Godoy R, Boeckx P (2008) Functional role of DNRA and nitrite reduction in a pristine south Chilean Nothofagus forest. Biogeochemistry 90:243–258Google Scholar
  52. Rütting T, Boeckx P, Müller C, Klemedtsson L (2011) Assessment of the importance of dissimilatory nitrate reduction to ammonium for the terrestrial nitrogen cycle. Biogeosciences 8:1779–1791CrossRefGoogle Scholar
  53. SAS Institute (2013) SAS software version 9.4. SAS Institute, CaryGoogle Scholar
  54. Schmidt CS, Richardson DJ, Baggs EM (2011) Constraining the conditions conducive to dissimilatory nitrate reduction to ammonium in temperate arable soils. Soil Biol Biochem 43:1607–1611CrossRefGoogle Scholar
  55. Sexstone AJ, Parkin TB, Tiedje JM (1985a) Temporal response of soil denitrification rates to rainfall and irrigation. Soil Sci Soc Am J 49:99–103CrossRefGoogle Scholar
  56. Sexstone AJ, Revsbech NP, Parkin TB, Tiedje JM (1985b) Direct measurement of oxygen profiles and denitrification rates in soil aggregates. Soil Sci Soc Am J 49:645–651CrossRefGoogle Scholar
  57. Sgouridis F, Heppell C, Wharton G, Lansdown K, Trimmer M (2011) Denitrification and dissimilatory nitrate reduction to ammonium (DNRA) in a temperate re-connected floodplain. Water Res 45:4909–4922CrossRefPubMedGoogle Scholar
  58. Silver WL, Herman DJ, Firestone MK (2001) Dissimilatory nitrate reduction to ammonium in upland tropical forest soils. Ecology 82:2410–2416CrossRefGoogle Scholar
  59. Silver W, Thompson A, Reich A, Ewel J, Firestone M (2005) Nitrogen cycling in tropical plantation forests: potential controls on nitrogen retention. Ecol Appl 15:1604–1614CrossRefGoogle Scholar
  60. Smith MS (1982) Dissimilatory reduction of NO2− to NH4+ and N2O by a soil Citrobacter sp. Appl Environ Microb 43:854–860Google Scholar
  61. Smith RL (1991) EPA Region 3 Guidance on handling chemical concentration data near the detection limit in risk assessments. Technical guidance manual, Mid-Atlantic Risk Assessment, Environmental Protection Agency.
  62. Smith MS, Tiedje JM (1979) Phases of denitrification following oxygen depletion in soil. Soil Biol Biochem 11:261–267CrossRefGoogle Scholar
  63. Smith K, McTaggart I, Tsuruta H (1997) Emissions of N2O and NO associated with nitrogen fertilization in intensive agriculture, and the potential for mitigation. Soil Use Manag 13:296–304CrossRefGoogle Scholar
  64. Sotta ED, Corre MD, Veldkamp E (2008) Differing N status and N retention processes of soils under old-growth lowland forest in eastern Smazonia, Caxiuanã, Brazil. Soil Biol Biochem 40:740–750CrossRefGoogle Scholar
  65. Staelens J, Rütting T, Huygens D, De Schrijver A, Müller C, Verheyen K, Boeckx P (2012) In situ gross nitrogen transformations differ between temperate deciduous and coniferous forest soils. Biogeochemistry 108:259–277CrossRefGoogle Scholar
  66. Stark JM, Hart SC (1997) High rates of nitrification and nitrate turnover in undisturbed coniferous forests. Nature 385:61–64CrossRefGoogle Scholar
  67. Takaya N, Uchimura H, Lai Y, Shoun H (2002) Transcriptional control of nitric oxide reductase gene (CYP55) in the fungal denitrifier Fusarium oxysporum. Biosci Biotechnol Biochem 66:1039–1045CrossRefPubMedGoogle Scholar
  68. Templer PH, Silver WL, Pett-Ridge J, DeAngelis KM, Firestone MK (2008) Plant and microbial controls on nitrogen retention and loss in a humid tropical forest. Ecology 89:3030–3040CrossRefGoogle Scholar
  69. Tiedje JM (1988) Ecology of denitrification and dissimilatory nitrate reduction to ammonium. In: Zhender AJB (Ed) Environmental microbiology of anaerobes. Wiley, New York, pp 179–244Google Scholar
  70. Tiedje JM, Sexstone AJ, Myrold DD, Robinson JA (1982) Denitrification: ecological niches, competition and survival. A van Leeuw J Microb 48:569–583CrossRefGoogle Scholar
  71. Veldkamp E, Keller M, Nunez M (1998) Effects of pasture management on N2O and NO emissions from soils in the humid tropics of Costa Rica. Global Biogeochem Cycles 12:71–79CrossRefGoogle Scholar
  72. Venterea RT, Groffman PM, Verchot LV, Magill AH, Aber JD (2004) Gross nitrogen process rates in temperate forest soils exhibiting symptoms of nitrogen saturation. For Ecol Manag 196:129–142CrossRefGoogle Scholar
  73. Verchot LV, Holmes Z, Mulon L, Groffman PM, Lovett GM (2001) Gross vs net rates of N mineralization and nitrification as indicators of functional differences between forest types. Soil Biol Biochem 33:1889–1901CrossRefGoogle Scholar
  74. Vitousek PM, Andariese SW (1986) Microbial transformations of labelled nitrogen in a clear-cut pine plantation. Oecologia 68:601–605CrossRefGoogle Scholar
  75. Vitousek PM, Andariese SW, Matson PA, Morris L, Sanford RL (1992) Effects of harvest intensity, site preparation, and herbicide use on soil nitrogen transformations in a young loblolly pine plantation. For Ecol Manag 49:277–292CrossRefGoogle Scholar
  76. Weil RR, Islam KR, Stine MA, Gruver JB, Samson-Liebig SE (2003) Estimating active carbon for soil quality assessment: a simplified method for laboratory and field use. Am J Altern Agric 18:3–17CrossRefGoogle Scholar
  77. Yáñez MA, Fox TR, Seiler JR (2015) Early growth responses of loblolly pine varieties and families to silvicultural intensity. For Ecol Manag 356:204–215CrossRefGoogle Scholar
  78. Yin SX, Chen D, Chen LM, Edis R (2002) Dissimilatory nitrate reduction to ammonium and responsible microorganisms in two Chinese and Australian paddy soils. Soil Biol Biochem 34:1131–1137CrossRefGoogle Scholar
  79. Zak D, Grigal D (1991) Nitrogen mineralization, nitrification and denitrification in upland and wetland ecosystems. Oecologia 88:189–196CrossRefGoogle Scholar
  80. Zeller B, Recous S, Kunze M, Moukoumi J, Colin-Belgrand M, Bienaimé S, Ranger J, Dambrine E (2007) Influence of tree species on gross and net N transformations in forest soils. Ann For Sci 64:151–158CrossRefGoogle Scholar
  81. Zhou Z, Takaya N, Sakairi MAC, Shoun H (2001) Oxygen requirement for denitrification by the fungus fusarium oxysporum. Arch Microbiol 175:19–25CrossRefPubMedGoogle Scholar
  82. Zumft WG (1997) Cell biology and molecular basis of denitrification. Microbiol Mol Biol Rev 61:533–616PubMedPubMedCentralGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2016

Authors and Affiliations

  • K. J. Minick
    • 1
    Email author
  • C. B. Pandey
    • 2
  • T. R. Fox
    • 1
  • Santosh Subedi
    • 1
  1. 1.Department of Forest Resources & Environmental ConservationBlacksburgUSA
  2. 2.ICAR-Central Arid Zone Research InstituteJodhpurIndia

Personalised recommendations