Journal of Soils and Sediments

, Volume 15, Issue 5, pp 1113–1118 | Cite as

The effect of nitrification inhibitors in reducing nitrification and the ammonia oxidizer population in three contrasting soils

  • Rui LiuEmail author
  • Helen Hayden
  • Helen Suter
  • Jizheng He
  • Deli Chen
Soils, Sec2 • Global Change, Environ Risk Assess, Sustainable Land Use • Short Original Communication



3,4-Dimethylpyrazole phosphate (DMPP) and acetylene (C2H2) are widely used nitrification inhibitors. These nitrification inhibitors have shown inconsistent efficacy in different soils, demonstrating the importance of determining which soil and microbial factors cause this variability. The aim of the present study was to investigate the efficacy of DMPP and C2H2 to inhibit nitrification and the ammonia oxidizer population in three contrasting soil types from Australia.

Materials and methods

Three contrasting soils of different pHWater (4.6, 7.0, and 8.0) collected from different agriculture systems in Australia were used in a laboratory incubation experiment for 28 days to compare the efficacy of DMPP and C2H2 to inhibit nitrification. We measured mineral nitrogen (N) concentrations during the incubation. In addition, quantitative PCR was applied to quantify the ammonia oxidizer population and to investigate the population change in response to DMPP and C2H2 addition.

Results and discussion

Acetylene completely blocked nitrification in the three soils while DMPP was more effective in inhibiting nitrification in the neutral soil (93.5 %) than in the alkaline soil (85.1 %) and acid soil (70.5 %). Ammonia-oxidizing archaea (AOA) were more abundant than ammonia-oxidizing bacteria (AOB) in all three control soils, with the highest AOA abundance found in the acid soil. The addition of DMPP and C2H2 significantly decreased AOA abundance in all soils (P < 0.05) and significantly suppressed AOB abundance in the neutral soil and slightly blocked AOB growth in the alkaline soil, though it had no effect on AOB abundance in the acid soil.


Our results show C2H2 completely inhibited nitrification and performed better than DMPP in our study. DMPP was more effective in the neutral soil than other two soils. Neither DMPP nor C2H2 was a selective nitrification inhibitor in neutral and alkaline soils in which both AOA and AOB were inhibited. Neither DMPP nor C2H2 had any effect on AOB abundance in the acid soil. Soil pH plays an important role in the effectiveness of DMPP and C2H2 in inhibiting nitrification and ammonia oxidizer population.


3,4-Dimethylpyrazole phosphate (DMPP) Acetylene AmoA Archaea Bacteria Soil pH 



The authors would like to acknowledge the financial support by Incitec Pivot Limited and the Australian Government Department of Agriculture through the Grains Research and Development Corporation. Dr. John Freney, Dr. Shu Kee (Raymond) Lam, and Dr. Hangwei Hu provided their assistance during manscript preparation.


  1. Barth G, Von Tucher S, Schmidhalter U (2001) Influence of soil parameters on the effect of 3, 4-dimethylpyrazole-phosphate as a nitrification inhibitor. Biol Fertil Soils 34:98–102CrossRefGoogle Scholar
  2. Barth G, Von Tucher S, Schmidhalter U (2008) Effectiveness of 3, 4-dimethylpyrazole phosphate as nitriflcation inhibitor in soil as influenced by inhibitor concentration, application form, and soil matric potential. Pedosphere 18:378–385CrossRefGoogle Scholar
  3. Bremner J, Blackmer AM (1978) Nitrous oxide: emission from soils during nitrification of fertilizer nitrogen. Science 199:295–296CrossRefGoogle Scholar
  4. Chalk PM (1990) Effect of a nitrification inhibitor on immobilization and mineralization of soil and fertilizer nitrogen. Vol-22Google Scholar
  5. Chen D, Suter HC, Islam A, Edis R (2010) Influence of nitrification inhibitors on nitrification and nitrous oxide (N2O) emission from a clay loam soil fertilized with urea. Soil Biol Biochem 42:660–664CrossRefGoogle Scholar
  6. Crawford D, Chalk P (1993) Sources of N uptake by wheat (Triticum aestivum L.) and N transformations in soil treated with a nitrification inhibitor (nitrapyrin). PLSO 149:59–72Google Scholar
  7. De Boer W, Kowalchuk G (2001) Nitrification in acid soils: micro-organisms and mechanisms. Soil Biol Biochem 33:853–866CrossRefGoogle Scholar
  8. Di HJ, Cameron KC, Shen JP (2009) Nitrification driven by bacteria and not archaea in nitrogen-rich grassland soils. Nat Geosci 2:621–624CrossRefGoogle Scholar
  9. Di HJ, Cameron KC, Shen JP, Winefoeld CS, O'callaghan M, Bowatte S, He JZ (2010) Ammonia‐oxidizing bacteria and archaea grow under contrasting soil nitrogen conditions. FEMS Microbiol Ecol 72:386–394CrossRefGoogle Scholar
  10. Erguder TH, Boon N, Wittebolle L, Marzorati M, Verstraete W (2009) Environmental factors shaping the ecological niches of ammonia oxidizing archaea. FEMS Microbiol Rev 33:855–869CrossRefGoogle Scholar
  11. Fierer N, Jackson RB (2006) The diversity and biogeography of soil bacterial communities. Proc Natl Acad Sci U S A 103:626–631CrossRefGoogle Scholar
  12. Francis CA, Roberts KJ, Beman JM, Santoro AE, Oakley BB (2005) Ubiquity and diversity of ammonia-oxidizing archaea in water columns and sediments of the ocean. Proc Natl Acad Sci U S A 102:14683–14688CrossRefGoogle Scholar
  13. Girvan MS, Bullimore J, Pretty JN, Osborn AM, Ball AS (2003) Soil type is the primary determinant of the composition of the total and active bacterial communities in arable soils. Appl Environ Microbiol 69:1800–1809CrossRefGoogle Scholar
  14. Gubry-Rangin C, Nicol GW, Prosser JI (2010) Archaea rather than bacteria control nitrification in two agricultural acidic soils. FEMS Microbiol Ecol 74:566–574CrossRefGoogle Scholar
  15. He JZ, Shen JP, Zhang LM, Zhu YG, Zheng YM, Xu MG, Di HJ (2007) Quantitative analyses of the abundance and composition of ammonia‐oxidizing bacteria and ammonia‐oxidizing archaea of a Chinese upland red soil under long‐term fertilization practices. Environ Microbiol 9:2364–2374CrossRefGoogle Scholar
  16. Hu HW, Zhang LM, Dai Y, Di HJ, He JZ (2013) pH-dependent distribution of soil ammonia oxidizers across a large geographical scale as revealed by high-throughput pyrosequencing. J Soils Sediments 13:1439–1449CrossRefGoogle Scholar
  17. Hynes RK, Knowles R (1982) Inhibition by acetylene of ammonia oxidation in Nitrosomonas europaea. FEMS Microbiol Lett 4:319–321CrossRefGoogle Scholar
  18. Jia Z, Conrad R (2009) Bacteria rather than Archaea dominate microbial ammonia oxidation in an agricultural soil. Environ Microbiol 11:1658–1671CrossRefGoogle Scholar
  19. Joye SB, Hollibaugh JT (1995) Influence of sulfide inhibition of nitrification on nitrogen regeneration in sediments. Science 270:623–625CrossRefGoogle Scholar
  20. Juliette LY, Hyman MR, Arp DJ (1993) Mechanism-based inactivation of ammonia monooxgenase in Nitrosomonas europaea by Allysulfide. Environ Microbiol 59:3728–3735Google Scholar
  21. Kemmitt SJ, Wright DG, Keith WT, Jones DL (2006) pH regulation of carbon and nitrogen dynamics in two agricultural soils. Soil Biol Biochem 38:898–911CrossRefGoogle Scholar
  22. Kleineidam K, Kosmrlj K, Kublik S, Palmer I, Pfab H, Ruser P, Fiedler S, Schloter M (2011) Influence of the nitrification inhibitor 3, 4-dimethylpyrazole phosphate (DMPP) on ammonia-oxidizing bacteria and archaea in rhizosphere and bulk soil. Chemosphere 84:182–186CrossRefGoogle Scholar
  23. Martens-Habbena W, Berube PM, Urakawa H, Jose R, Stahl DA (2009) Ammonia oxidation kinetics determine niche separation of nitrifying Archaea and Bacteria. Nature 461:976–979CrossRefGoogle Scholar
  24. Nicol GW, Leininger S, Schleper C, Prosser JI (2008) The influence of soil pH on the diversity, abundance and transcriptional activity of ammonia oxidizing archaea and bacteria. Environ Microbiol 10:2966–2978CrossRefGoogle Scholar
  25. Offre P, Prosser JI, Nicol GW (2009) Growth of ammonia-oxidizing archaea in soil microcosms is inhibited by acetylene. FEMS Microbiol Ecol 70:99–108CrossRefGoogle Scholar
  26. Rotthauwe JH, Witzel KP, Liesack W (1997) The ammonia monooxygenase structural gene amoA as a functional marker: molecular fine-scale analysis of natural ammonia-oxidizing populations. Environ Microbiol 63:4704–4712Google Scholar
  27. Shen JP, Zhang LM, Zhu YG, Zhang JB, He JZ (2008) Abundance and composition of ammonia‐oxidizing bacteria and ammonia‐oxidizing archaea communities of an alkaline sandy loam. Environ Microbiol 10:1601–1611CrossRefGoogle Scholar
  28. Shi M, Zhang MT, Shen F, Liang DL, Dang HL (2011) Effects of nitrification inhibitors on nitrification inhibition and nitrite accumulation in calcareous soil. Sci Agric Sin 3:010Google Scholar
  29. Šimek M, Cooper J (2002) The influence of soil pH on denitrification: progress towards the understanding of this interaction over the last 50 years. Eur J Soil Sci 53:345–354CrossRefGoogle Scholar
  30. Suzuki C, Nagaoka K, Shimada A, Takenaka M (2009) Bacterial communities are more dependent on soil type than fertilizer type, but the reverse is true for fungal communities. Soil Sci Plant Nutr 55:80–90CrossRefGoogle Scholar
  31. Verhamme DT, Prosser JI, Nicol GW (2011) Ammonia concentration determines differential growth of ammonia-oxidising archaea and bacteria in soil microcosms. ISME J 5:1067–1071CrossRefGoogle Scholar
  32. Zerulla W, Barth T, Dressel J, Erhardt K, Klaus H, Pasda G, Radle M, Wissemeier A (2001) 3, 4-Dimethylpyrazole phosphate (DMPP)—a new nitrification inhibitor for agriculture and horticulture. Biol Fertil Soils 34:79–84CrossRefGoogle Scholar
  33. Zhang LM, Hu HW, Shen JP, He JZ (2011) Ammonia-oxidizing archaea have more important role than ammonia-oxidizing bacteria in ammonia oxidation of strongly acidic soils. ISME J 6:1032–1045CrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2015

Authors and Affiliations

  • Rui Liu
    • 1
    Email author
  • Helen Hayden
    • 2
  • Helen Suter
    • 1
  • Jizheng He
    • 1
  • Deli Chen
    • 1
  1. 1.Faculty of Veterinary and Agricultural SciencesThe University of MelbourneMelbourneAustralia
  2. 2.Department of Environment and Primary Industries, AgriBioMelbourneAustralia

Personalised recommendations