Abstract
Purpose
The nitrification inhibitor 3,4-dimethylpyrazol-phosphate (DMPP) and the urease inhibitor N-(n-butyl) thiophosphoric triamide (nBTPT) can mitigate N losses through reducing nitrification and ammonia volatilization, respectively. However, the impact of repeated applications of these inhibitors on nitrogen cycling microorganisms is not well documented. This study aimed to investigate the changes in the abundance and community structure of the functional microorganisms involved in nitrification and denitrification in Australian pasture soils after repeated applications of DMPP and nBTPT.
Materials and methods
Soil was collected in autumn and spring, 2014 from two pasture sites where control, urea, urea ammonium nitrate, and urea-coated inhibitors had been repeatedly applied over 2 year. Soil samples were analyzed to determine the potential nitrification rates (PNRs), the abundances of amoA, narG, nirK and bacterial 16S rRNA genes, and the community structure of ammonia oxidizers.
Results and discussion
Two years of urea application resulted in a significantly lower soil pH at Terang and a significant decrease in total bacterial 16S rRNA gene abundance at Glenormiston and led to significantly higher PNRs and abundances of ammonia oxidizers compared to the control. Amendment with either DMPP or nBTPT significantly decreased PNRs and the abundance of amoA and narG genes. However, there was no fertilizer- or inhibitor-induced change in the community structure of ammonia oxidizers.
Conclusions
These results suggest that there were inhibitory effects of DMPP and nBTPT on the functional groups mediating nitrification and denitrification, while no significant impact on the community structure of ammonia oxidizers was observed. The application of nitrification or urease inhibitor appears to be an effective approach targeting specific microbial groups with minimal effects on soil pH and the total bacterial abundance.
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References
Allison SM, Prosser JI (1991) Urease activity in neutrophilic autotrophic ammonia-oxidizing bacteria isolated from acid soils. Soil Biol Biochem 23:45–51
Bolan NS, Hedley MJ, White RE (1991) Processes of soil acidification during nitrogen cycling with emphasis on legume based pastures. Plant Soil 134:53–63
Bremner JM, McCarty GW, Yeomans JC, Chai HS (1986) Effects of phosphoroamides on nitrification, denitrification, and mineralization of organic nitrogen in soil1. Commun Soil Sci Plant Anal 17:369–384
Brochier-Armanet C, Boussau B, Gribaldo S, Forterre P (2008) Mesophilic Crenarchaeota: proposal for a third archaeal phylum, the thaumarchaeota. Nature reviews Microbiol 6:245–252
Bronson KFM, Mosier AR (1994) Suppression of methane oxidation in aerobic soil by nitrogen fertilizers, nitrification inhibitors, and urease inhibitors. Biol Fertil Soils 17:263–268
Bru D, Sarr A, Philippot L (2007) Relative abundances of proteobacterial membrane-bound and periplasmic nitrate reductases in selected environments. Appl Environ Microbiol 73:5971–5974
Burton SA, Prosser JI (2001) Autotrophic ammonia oxidation at low pH through urea hydrolysis. Appl Environ Microbiol 67:2952–2957
Chaves B, Opoku A, De Neve S, Boeckx P, Van Cleemput O, Hofman G (2006) Influence of DCD and DMPP on soil N dynamics after incorporation of vegetable crop residues. Biol Fertil Soils 43:62–68
Chen DL, Suter H, Islam A, Edis AR, Freney JR, Walker CN (2008) Prospects of improving efficiency of fertiliser nitrogen in Australian agriculture: a review of enhanced efficiency fertilisers. Aust J Soil Res 46:289–301
De Boer W, Laanbroek HJ (1989) Ureolytic nitrification at low pH by Nitrosospira spec. Arch Microbiol 152:178–181
Di HJ, Cameron KC (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
Di HJ, Cameron KC (2012) How does the application of different nitrification inhibitors affect nitrous oxide emissions and nitrate leaching from cow urine in grazed pastures? Soil use. Manage 28:54–61
Di HJ, Cameron KC (2016) Inhibition of nitrification to mitigate nitrate leaching and nitrous oxide emissions in grazed grassland: a review. J Soils Sediments 16:1401–1420
Di HJ, Cameron KC, Shen JP, Winefield CS, O’Callaghan M, Bowatte S, He JZ (2010) Ammonia-oxidizing bacteria and archaea grow under contrasting soil nitrogen conditions. FEMS Microbiol Ecol 72:386–394
Dong XX, Zhang LL, Wu ZJ, Gong P (2013) The response of nitrifier, N-fixer and denitrifier gene copy numbers to the nitrification inhibitor 3,4-dimethylpyrazole phosphate. Plant Soil Environ 59:398–403
Ernfors M, Brennan FP, Richards KG, McGeough KL, Griffiths BS, Laughlin RJ, Watson CJ, Philippot L, Grant J, Minet EP, Moynihan E, Müller C (2014) The nitrification inhibitor dicyandiamide increases mineralization–immobilization turnover in slurry-amended grassland soil. J Agr Sci 152:137–149
Giovannini C, Garcia-Mina JM, Ciavatta C, Marzadori C (2009) Ureic nitrogen transformation in multi-layer soil columns treated with urease and nitrification inhibitors. J Agric Food Chem 57:4883–4887
Hallin S, Lindgren P (1999) PCR detection of genes encoding nitrite reductase in denitrifying bacteria. Appl Soil Ecol 65:1652–1657
Hatch D, Trindade H, Cardenas L, Carneiro J, Hawkins J, Scholefield D, Chadwick D (2005) Laboratory study of the effects of two nitrification inhibitors on greenhouse gas emissions from a slurry-treated arable soil: impact of diurnal temperature cycle. Biol Fertil Soils 41:225–232
Hu HW, Macdonald CA, Trivedi P, Holmes B, Bodrossy L, He JZ, Singh BK (2015) Water addition regulates the metabolic activity of ammonia oxidizers responding to environmental perturbations in dry subhumid ecosystems. Environ Microbiol 17:444–461
Hu HW, Xu ZH, He JZ (2014) Ammonia-Oxidizing Archaea play a predominant role in acid soil nitrification. Adv Agron 125:261–294
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 13:1439–1449
Hyman MR, Wood PM (1983) Methane oxidation by Nitrosomonas europaea. Biochem J 212:31–37
Isbell RF (2002) The Australian soil classification. CSIRO Publishing, Melbourne
Kong X, Duan Y, Schramm A, Eriksen J, Petersen SO (2016) 3,4-dimethylpyrazole phosphate (DMPP) reduces activity of ammonia oxidizers without adverse effects on non-target soil microorganisms and functions. Appl Soil Ecol 105:67–75
Koper TE, El-Sheikh AF, Norton JM, Klotz MG (2004) Urease-encoding genes in ammonia-oxidizing bacteria. Appl Soil Ecol 70:2342–2348
Kou YP, Wei K, Chen GX, Wang ZY, Xu H (2015) Effects of 3,4-dimethylpyrazole phosphate and dicyandiamide on nitrous oxide emission in a greenhouse vegetable soil. Plant Soil Environ 61:29–35
Lu L, Jia Z (2013) Urease gene-containing archaea dominate autotrophic ammonia oxidation in two acid soils. Environ Microbiol 15:1795–1809
Macadam XMB, Ad P, Merino P, Estavillo JM, Pinto M, González-Murua C (2003) Dicyandiamide and 3,4-dimethyl pyrazole phosphate decrease N2O emissions from grassland but dicyandiamide produces deleterious effects in clover. J Plant Physiol 160:1517–1523
Manunza B, Deiana S, Pintore M, Gessa C (1999) The binding mechanism of urea, hydroxamic acid and N-(N-butyl)-phosphoric triamide to the urease active site. A comparative molecular dynamics study. Soil Biol Biochem 31:789–796
Menéndez S, Merino P, Pinto M, González-Murua C, Estavillo JM (2006) 3,4-dimethylpyrazol phosphate effect on nitrous oxide, nitric oxide, ammonia, and carbon dioxide emissions from grasslands. J Environ Qual 35:973–981
Menéndez S, Barrena I, Setien I, González-Murua C, Estavillo JM (2012) Efficiency of nitrification inhibitor DMPP to reduce nitrous oxide emissions under different temperature and moisture conditions. Soil Biol Biochem 53:82–89
Merino P, Menéndez S, Pinto M, González-Murua C, Estavillo JM (2005) 3, 4-dimethylpyrazole phosphate reduces nitrous oxide emissions from grassland after slurry application. Soil Use Manage 21:53–57
Osborne BB, Baron JS, Wallenstein MD (2016) Moisture and temperature controls on nitrification differ among ammonia oxidizer communities from three alpine soil habitats. Front Earth Sci Prc 10:1–12
O’Sullivan CA, Wakelin SA, Fillery IRP, Roper MM (2013) Factors affecting ammonia-oxidising microorganisms and potential nitrification rates in southern Australian agricultural soils. Soil Research 51:240–252
Philippot L, Hallin S, Schloter M (2007) Ecology of denitrifying prokaryotes in agricultural soil. Adv Agron 96:249–305
Prosser JI, Nicol GW (2012) Archaeal and bacterial ammonia-oxidisers in soil: the quest for niche specialisation and differentiation. Trends Microbiol 20:523–531
Purkhold U, Pommerening-Roser A, Juretschko S, Schmid MC, Koops HP, Wagner M (2000) Phylogeny of all recognized species of ammonia oxidizers based on comparative 16S rRNA and amoA sequence analysis implications for molecular diversity surveys. Appl Soil Ecol 66:5368–5382
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 Soil Ecol 63:4704–4712
Rowlings DW, Scheer C, Liu S, Grace PR (2016) Annual nitrogen dynamics and urea fertilizer recoveries from a dairy pasture using 15N; effect of nitrification inhibitor DMPP and reduced application rates. Agric Ecosyst Environ 216:216–225
Ruser R, Schulz R (2015) The effect of nitrification inhibitors on the nitrous oxide (N2O) release from agricultural soils-a review. J Plant Nut Soil Sci 178:171–188
Selbie DR, Buckthought LE, Shepherd MA (2015) The challenge of the urine patch for managing nitrogen in grazed pasture systems. Adv Agron 129:229–292
Shi XZ, Hu HW, Müller C, He JZ, Chen DL, Suter H (2016a) Effects of the nitrification inhibitor 3,4-dimethylpyrazole phosphate (DMPP) on nitrification and nitrifiers in two contrasting agricultural soils . doi:10.1128/AEM.01031-16Appl Environ Microbiol
Shi XZ, Hu HW, He JZ, Chen DL, Suter H (2016b) Effects of 3,4-dimethylpyrazole phosphate (DMPP) on nitrification and the abundance and community structure of soil ammonia oxidizers in three land uses . doi:10.1007/s00374-016-1131-7Biol Fertil Soils
Singh J, Kunhikrishnan A, Bolan NS, Saggar S (2013) Impact of urease inhibitor on ammonia and nitrous oxide emissions from temperate pasture soil cores receiving urea fertilizer and cattle urine. Sci Total Environ 465:56–63
Singh BK, Thomas N (2006) Multiplex-terminal restriction fragment length polymorphism. Nat Protoc 1:2428–2433
Spang A, Poehlein A, Offre P, Zumbragel S, Haider S, Rychlik N, Nowka B, Schmeisser C, Lebedeva EV, Rattei T, Bohm C, Schmid M, Galushko A, Hatzenpichler R, Weinmaier T, Daniel R, Schleper C, Spieck E, Streit W, Wagner M (2012) The genome of the ammonia-oxidizing Candidatus Nitrososphaera gargensis: insights into metabolic versatility and environmental adaptations. Environ Microbiol 14:3122–3145
Suter HC, Sultana H, Davies R, Walker C, Chen D (2016) Influence of enhanced efficiency fertilisation techniques on nitrous oxide emissions and productivity response from urea in a temperate Australian ryegrass pasture. Soil Res 54:523–532
Suzuki MT, Taylor LT, Delong EF (2000) Quantitative analysis of small-subunit rRNA genes in mixed microbial populations via 5′-nuclease assays. Appl Soil Ecol 66:4605–4614
Tolar BB, Wallsgrove NJ, Popp BN, Hollibaugh JT (2016) Oxidation of urea-derived nitrogen by thaumarchaeota-dominated marine nitrifying communities. Environ microbiol doi. doi:10.1111/1462-2920.13457
Tourna M, Freitag TE, Nicol GW, Prosser JI (2008) Growth, activity and temperature responses of ammonia-oxidizing archaea and bacteria in soil microcosms. Environ Microbiol 10:1357–1364
Tourna M, Stieglmeier M, Spang A, Könneke M, Schintlmeister A, Urich T, Engel M, Schloter M, Wagner M, Richter A, Schleper C (2011) Nitrososphaera viennensis, an ammonia oxidizing archaeon from soil. Proc Natl Acad Sci U S A 108:8420–8425
Verhamme DT, Prosser JI, Nicol GW (2011) Ammonia concentration determines differential growth of ammonia-oxidising archaea and bacteria in soil microcosms. ISME J 6:1067–1071
Walkley A, Black IA (1934) An examination of the Degtjareff method for determining soil organic matter, and a proposed modification of the chromic acid titration method. Soil Sci 37:29–38
Wang Z, Kong T, Hu S, Sun H, Yang W, Kou Y, Mandlaa XH (2015) Nitrification inhibitors mitigate earthworm-induced N2O emission—a mesocosm study. Biol Fertil Soils 51:1005–1011
Wu SF, Wu LH, Shi QW, Wang ZQ, Chen XY, Li YH (2007) Effects of a new nitrification inhibitor 3,4-dimethylpyrazole phosphate (DMPP) on nitrate and potassium leaching in two soils. J Environ Sci 19:841–847
Yu Q, Ye X, Chen Y, Zhang Z, Tian G (2008) Influences of nitrification inhibitor 3,4-dimethyl pyrazole phosphate on nitrogen and soil salt-ion leaching. J Environ Sci 20:304–308
Zaman M, Nguyen ML, Blennerhassett JD, Quin BF (2008) Reducing NH3, N2O and NO3 −-N losses from a pasture soil with urease or nitrification inhibitors and elemental S-amended nitrogenous fertilizers. Biol Fertil Soils 44:693–705
Zaman M, Saggar S, Blennerhassett JD, Singh J (2009) Effect of urease and nitrification inhibitors on N transformation, gaseous emissions of ammonia and nitrous oxide, pasture yield and N uptake in grazed pasture system. Soil Biol Biochem 41:1270–1280
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This work was financially supported by the Australian Research Council (DE150100870, LP160101134).
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Shi, X., Hu, HW., Kelly, K. et al. Response of ammonia oxidizers and denitrifiers to repeated applications of a nitrification inhibitor and a urease inhibitor in two pasture soils. J Soils Sediments 17, 974–984 (2017). https://doi.org/10.1007/s11368-016-1588-x
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DOI: https://doi.org/10.1007/s11368-016-1588-x