Skip to main content
Log in

Comparative effects of 3,4-dimethylpyrazole phosphate (DMPP) and dicyandiamide (DCD) on ammonia-oxidizing bacteria and archaea in a vegetable soil

  • Environmental biotechnology
  • Published:
Applied Microbiology and Biotechnology Aims and scope Submit manuscript

Abstract

Nitrification inhibitors (NIs) 3,4-dimethylpyrazole phosphate (DMPP) and dicyandiamide (DCD) have been used extensively to improve nitrogen fertilizer utilization in farmland. However, their comparative effects on ammonia-oxidizing bacteria (AOB) and ammonia-oxidizing archaea (AOA) in agricultural soils are still unclear. Here, we compared the impacts of these two inhibitors on soil nitrification, AOA and AOB abundance as well as their community structure in a vegetable soil by using real-time PCR and terminal restriction fragment length polymorphism (T-RFLP). Our results showed that urea application significantly increased the net nitrification rates, but were significantly inhibited by both NIs, and the inhibitory effect of DMPP was significantly greater than that of DCD. AOB growth was more greatly inhibited by DMPP than by DCD, and the net nitrification rate was significantly related to AOB abundance, but not to AOA abundance. Application of urea and NIs to soil did not change the diversity of the AOA community, with the T-RFs remaining in proportions that were similar to control soils, while the community structure of AOB exhibited obvious shifts within all different treatments compared to the control. Phylogenetic analysis showed that all AOA sequences fell within group 1.1a and group 1.1b, and the AOB community consisted of Nitrosospira cluster 3, cluster 0, and unidentified species. These results suggest that DMPP exhibited a stronger inhibitory effect on nitrification than DCD by inhibiting AOB rather than AOA.

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

Access this article

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

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

References

  • Benckiser G, Christ E, Herbert T, Weiske A, Blome J, Hardt M (2013) The nitrification inhibitor 3,4-dimethylpyrazole-phosphat (DMPP)—quantification and effects on soil metabolism. Plant Soil 371:257–266

    Article  CAS  Google Scholar 

  • Chen T, Liu XM, Zhu MZ, Zhao KL, Wu JJ, Xu JM, Huang PM (2008a) Identification of trace element sources and associated risk assessment in vegetable soils of the urban-rural transitional area of Hangzhou, China. Environ Pollut 151:67–78

    Article  CAS  PubMed  Google Scholar 

  • Chen XP, Zhu YG, Xia Y, Shen JP, He JZ (2008b) Ammonia-oxidizing archaea: important players in paddy rhizosphere soil? Environ Microbiol 10:1978–1987

    Article  CAS  PubMed  Google Scholar 

  • Chen X, Zhang LM, Shen JP, Xu Z, He JZ (2010) Soil type determines the abundance and community structure of ammonia-oxidizing bacteria and archaea in flooded paddy soils. J Soils Sediments 10:1510–1516

    Article  CAS  Google Scholar 

  • Cui M, Sun XC, Hu CX, Di HJ, Tan QL, Zhao CS (2011) Effective mitigation of nitrate leaching and nitrous oxide emissions in intensive vegetable production systems using a nitrification inhibitor, dicyandiamide. J Soils Sediments 11:722–730

    Article  CAS  Google Scholar 

  • Cui PY, Fan FL, Yin C, Li ZJ, Song AL, Wan YF, Liang YC (2013) Urea- and nitrapyrin-affected N2O emission is coupled mainly with ammonia oxidizing bacteria growth in microcosms of three typical Chinese arable soils. Soil Biol Biochem 66:214–221

    Article  CAS  Google Scholar 

  • Di HJ, Cameron K (2011) Inhibition of ammonium oxidation by a liquid formulation of 3,4-dimethylpyrazole phosphate (DMPP) compared with a dicyandiamide (DCD) solution in six new Zealand grazed grassland soils. J Soils Sediments 11:1032–1039

    Article  CAS  Google Scholar 

  • Di HJ, Cameron K (2012) How does the application of different nitrification inhibitors affect nitrous oxide emissions and nitrate leaching from cow urine in grazed pastures? Soil Use Manag 28:54–61

    Article  Google Scholar 

  • Di HJ, Cameron K, Shen JP, Winefield C, O’Callaghan M, Bowatte S, He JZ (2009) Nitrification driven by bacteria and not archaea in nitrogen-rich grassland soils. Nat Geosci 2:621–624

    Article  CAS  Google Scholar 

  • Fangueiro D, Fernandes A, Coutinho J, Moreira N, Trindade H (2009) Influence of two nitrification inhibitors (DCD and DMPP) on annual ryegrass yield and soil mineral N dynamics after incorporation with cattle slurry. Commun Soil Sci Plant Anal 40:3387–3398

    Article  CAS  Google Scholar 

  • Florio A, Clark IM, Hirsch PR, Jhurreea D, Benedetti A (2014) Effects of the nitrification inhibitor 3,4-dimethylpyrazole phosphate (DMPP) on abundance and activity of ammonia oxidizers in soil. Biol Fertil Soils 50:1–13

    Article  Google Scholar 

  • 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 102:14683–14688

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Gong P, Zhang LL, Wu ZJ, Li D (2012) Laboratory study of the effects of nitrification inhibitors on the abundance of ammonia-oxidizing bacteria and archaea in a Hap-Ustic Luvisol. Afr J Microbiol Res 6:7428–7434

    CAS  Google Scholar 

  • Gong P, Zhang LL, Wu ZJ, Chen ZH, Chen LJ (2013) Responses of ammonia-oxidizing bacteria and archaea in two agricultural soils to nitrification inhibitors DCD and DMPP: a pot experiment. Pedosphere 23:729–739

    Article  CAS  Google Scholar 

  • Hauser M, Haselwandter K (1990) Degradation of dicyandiamide by soil bacteria. Soil Biol Biochem 22:113–114

  • 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 9:2364–2374

    Article  CAS  PubMed  Google Scholar 

  • Höfferle Š, Nicol GW, Pal L, Hacin J, Prosser JI, Mandić-Mulec I (2010) Ammonium supply rate influences archaeal and bacterial ammonia oxidizers in a wetland soil vertical profile. FEMS Microbiol Ecol 74:302–315

    Article  PubMed  Google Scholar 

  • Hu HW, Xu ZH, He JZ (2014) Ammonia-oxidizing archaea play a predominant role in acid soil nitrification. Adv Agron 125:261–302

    Article  Google Scholar 

  • Irigoyen I, Muro J, Azpilikueta M, Aparicio-Tejo P, Lamsfus C (2003) Ammonium oxidation kinetics in the presence of nitrification inhibitors DCD and DMPP at various temperatures. Soil Res 41:1177–1183

    Article  CAS  Google Scholar 

  • Kelliher FM, Clough TJ, Clark H, Rys G, Sedcole JR (2008) The temperature dependence of dicyandiamide (DCD) degradation in soils: a data synthesis. Soil Biol Biochem 40:1878–1882

    Article  CAS  Google Scholar 

  • Kim JG, Jung MY, Park SJ, Rijpstra WIC, Sinninghe Damsté JS, Madsen EL, Min D, Kim JS, Kim GJ, Rhee SK (2012) Cultivation of a highly enriched ammonia-oxidizing archaeon of thaumarchaeotal group I.1b from an agricultural soil. Environ Microbiol 14:1528–1543

    Article  CAS  PubMed  Google Scholar 

  • Kleineidam K, Košmrlj K, Kublik S, Palmer I, Pfab H, Ruser R, 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 84:182–186

    Article  CAS  PubMed  Google Scholar 

  • Lehtovirta-Morley LE, Verhamme D, Nicol GW, Prosser JI (2013) Effect of nitrification inhibitors on the growth and activity of Nitrosotalea devanaterra in culture and soil. Soil Biol Biochem 62:129–133

    Article  CAS  Google Scholar 

  • Leininger S, Urich T, Schloter M, Schwark L, Qi J, Nicol GW, Prosser JI, Schuster SC, Schleper C (2006) Archaea predominate among ammonia-oxidizing prokaryotes in soils. Nature 442:806–809

    Article  CAS  PubMed  Google Scholar 

  • Lin XG, Yin R, Zhang HY, Huang JF, Chen RR, Cao ZH (2004) Changes of soil microbiological properties caused by land use changing from rice-wheat rotation to vegetable cultivation. Environ Geochem Health 26:119–128

    Article  CAS  PubMed  Google Scholar 

  • Liu C, Wang K, Zheng X (2013) Effects of nitrification inhibitors (DCD and DMPP) on nitrous oxide emission, crop yield and nitrogen uptake in a wheat-maize cropping system. Biogeosci Discuss 10:711–737

    Article  Google Scholar 

  • Liu Y, Yang Y, Qin HL, Zhu YJ, Wei WX (2014a) Differential responses of nitrifier and denitrifier to dicyandiamide in short- and long-term intensive vegetable cultivation soils. J Integr Agric 13:1090–1098

    Article  Google Scholar 

  • Liu Y, Zhang JX, Zhang XL, Xie SG (2014b) Depth-related changes of sediment ammonia-oxidizing microorganisms in a high-altitude freshwater wetland. Appl Microbiol Biotechnol 98:5697–5707

    Article  CAS  PubMed  Google Scholar 

  • McCarty G (1999) Modes of action of nitrification inhibitors. Biol Fertil Soils 29:1–9

    Article  CAS  Google Scholar 

  • Morimoto S, Hayatsu M, Takada HY, Nagaoka K, Yamazaki M, Karasawa T, Takenaka M, Akiyama H (2011) Quantitative analyses of ammonia-oxidizing archaea (AOA) and ammonia-oxidizing bacteria (AOB) in fields with different soil types. Microbes Environ 26:248–253

    Article  PubMed  Google Scholar 

  • 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 10:2966–2978

    Article  CAS  PubMed  Google Scholar 

  • O’Callaghan M, Gerard EM, Carter PE, Lardner R, Sarathchandra U, Burch G, Ghani A, Bell N (2010) Effect of the nitrification inhibitor dicyandiamide (DCD) on microbial communities in a pasture soil amended with bovine urine. Soil Biol Biochem 42:1425–1436

    Article  Google Scholar 

  • Paranychianakis NV, Tsiknia M, Giannakis G, Nikolaidis NP, Kalogerakis N (2013) Nitrogen cycling and relationships between ammonia oxidizers and denitrifiers in a clay-loam soil. Appl Microbiol Biotechnol 97:5507–5515

    Article  CAS  PubMed  Google Scholar 

  • Persson T, Wirén A (1995) Nitrogen mineralization and potential nitrification at different depths in acid forest soils. Plant Soil 168:55–65

    Article  Google Scholar 

  • Rajbanshi S, Benckiser G, Ottow J (1992) Mineralization kinetics and utilization as an N source of dicyandiamide (DCD) in soil. Naturwissenschaften 79:26–27

    Article  CAS  Google Scholar 

  • 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. Appl Environ Microbiol 63:4704–4712

    CAS  PubMed Central  PubMed  Google Scholar 

  • Schauss K, Focks A, Leininger S, Kotzerke A, Heuer H, Thiele-Bruhn S, Sharma S, Wilke BM, Matthies M, Smalla K (2009) Dynamics and functional relevance of ammonia-oxidizing archaea in two agricultural soils. Environ Microbiol 11:446–456

    Article  CAS  PubMed  Google Scholar 

  • 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 10:1601–1611

    Article  CAS  PubMed  Google Scholar 

  • Shen WS, Lin XG, Gao N, Shi WM, Min J, He XH (2011) Nitrogen fertilization changes abundance and community composition of ammonia-oxidizing bacteria. Soil Sci Soc Am J 75:2198–2205

    Article  CAS  Google Scholar 

  • Shen TL, Stieglmeier M, Dai JL, Urich T, Schleper C (2013) Responses of the terrestrial ammonia-oxidizing archaeon Ca. Nitrososphaera viennensis and the ammonia-oxidizing bacterium Nitrosospira multiformis to nitrification inhibitors. FEMS Microbiol Lett 344:121–129

    Article  CAS  PubMed  Google Scholar 

  • Sheng R, Meng DL, Wu MM, Di HJ, Qin HL, Wei WX (2013) Effect of agricultural land use change on community composition of bacteria and ammonia oxidizers. J Soils Sediments 13:1246–1256

    Article  Google Scholar 

  • Singh BK, Nazaries L, Munro S, Anderson IC, Campbell CD (2006) Use of multiplex terminal restriction fragment length polymorphism for rapid and simultaneous analysis of different components of the soil microbial community. Appl Environ Microbiol 72:7278–7285

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Subbarao GV, Ito O, Sahrawat KL, Berry WL, Nakahara K, Ishikawa T, Watanabe T, Suenaga K, Rondon M, Rao IM (2006) Scope and strategies for regulation of nitrification in agricultural systems—challenges and opportunities. Crit Rev Plant Sci 25:303–335

    Article  CAS  Google Scholar 

  • Tourna M, Stieglmeier M, Spang A, Könneke M, Schintlmeister A, Urich T, Engel M, Schloter M, Wagner M, Richter A (2011) Nitrososphaera viennensis, an ammonia oxidizing archaeon from soil. Proc Natl Acad Sci 108:8420–8425

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Walker CB, De La Torre JR, Klotz MG, Urakawa H, Pinel N, Arp DJ, Brochier AC, Chain PSG, Chan PP, Gollabgir A (2010) Nitrosopumilus maritimus genome reveals unique mechanisms for nitrification and autotrophy in globally distributed marine crenarchaea. Proc Natl Acad Sci 107:8818–8823

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Wang YN, Ke XB, Wu LQ, Lu YH (2009) Community composition of ammonia-oxidizing bacteria and archaea in rice field soil as affected by nitrogen fertilization. Syst Appl Microbiol 32:27–36

    Article  CAS  PubMed  Google Scholar 

  • Weiske A, Benckiser G, Herbert T, Ottow J (2001) Influence of the nitrification inhibitor 3,4-dimethylpyrazole phosphate (DMPP) in comparison to dicyandiamide (DCD) on nitrous oxide emissions, carbon dioxide fluxes and methane oxidation during 3 years of repeated application in field experiments. Biol Fertil Soils 34:109–117

    Article  CAS  Google Scholar 

  • Wissemeier AH, Linzmeier W, Gutser R, Weigelt W, Schmidhalter U (2001) The new nitrification inhibitor DMPP (ENTEC®)—comparisons with DCD in model studies and field applications. In: Horst WJ, Schenk MK, Bürkert A, Claassen N, Flessa H, Fommer WB, Goldbach H, Olfs HW, Römheld V, Sattelmacher B, Schmidhalter U, Schubert S, von Wirén N, Wittenmayer L (eds) Plant Nutrition, 1st edn. Springer, Netherlands, pp 702–703

    Chapter  Google Scholar 

  • Xia WW, Zhang CX, Zeng XW, Feng YZ, Weng JH, Lin XG, Zhu JG, Xiong ZQ, Xu J, Cai ZC (2011) Autotrophic growth of nitrifying community in an agricultural soil. ISME J 5:1226–1236

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Xu C, Wu LH, Ju XT, Zhang FS (2005) Role of nitrification inhibitor DMPP (3,4-dimethylpyrazole phosphate) in NO3 -N accumulation in greengrocery (Brassica campestris L. ssp. Chinensis) and vegetable soil. J Environ Sci 17:81–83

  • Yang JB, Li XC, Xu L, Hu F, Li HX, Liu MQ (2013) Influence of the nitrification inhibitor DMPP on the community composition of ammonia-oxidizing bacteria at microsites with increasing distance from the fertilizer zone. Biol Fertil Soils 49:23–30

    Article  CAS  Google Scholar 

  • Yao HY, Gao YM, Nicol GW, Campbell CD, Prosser JI, Zhang LM, Han WY, Singh BK (2011) Links between ammonia oxidizer community structure, abundance, and nitrification potential in acidic soils. Appl Environ Microbiol 77:4618–4625

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Zacherl B, Amberger A (1990) Effect of the nitrification inhibitors dicyandiamide, nitrapyrin and thiourea on Nitrosomonas europaea. Fertil Res 22:37–44

    Article  CAS  Google Scholar 

  • Zerulla W, Barth T, Dressel J, Erhardt K, von Locquenghien KH, Pasda G, Rädle M, Wissemeier A (2001) 3,4-dimethylpyrazole phosphate (DMPP)—a new nitrification inhibitor for agriculture and horticulture. Biol Fertil Soils 34:79–84

    Article  CAS  Google Scholar 

  • Zhang LM, Hu HW, Shen JP, He JZ (2012) Ammonia-oxidizing archaea have more important role than ammonia-oxidizing bacteria in ammonia oxidation of strongly acidic soils. ISME J 6:1032–1045

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Zhao CS, Hu CX, Huang W, Sun XC, Tan QL, Di HJ (2010) A lysimeter study of nitrate leaching and optimum nitrogen application rates for intensively irrigated vegetable production systems in Central China. J Soils Sediments 10:9–17

    Article  CAS  Google Scholar 

  • Zheng YL, Hou LJ, Liu M, Lu M, Zhao H, Yin GY, Zhou JL (2013) Diversity, abundance, and activity of ammonia-oxidizing bacteria and archaea in Chongming eastern intertidal sediments. Appl Microbiol Biotechnol 97:8351–8363

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

This study was supported in part by the National Basic Research Program (973 Program) of China (2013CB127403), the National Project of Scientific and Technical Supporting Programs (2012BAC17B02), the Natural Science Foundation of China (31272237, 30971859), the Foundation for University PhD Granting Discipline of the Ministry of Education (20120101110130), and International Plant Nutrition Institute (IPNI).

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Xianyong Lin.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Chen, Q., Qi, L., Bi, Q. et al. Comparative effects of 3,4-dimethylpyrazole phosphate (DMPP) and dicyandiamide (DCD) on ammonia-oxidizing bacteria and archaea in a vegetable soil. Appl Microbiol Biotechnol 99, 477–487 (2015). https://doi.org/10.1007/s00253-014-6026-7

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00253-014-6026-7

Keywords

Navigation