Abstract
The capacity of several plant species or landraces to inhibit nitrification in soil (biological nitrification inhibition, BNI) has been assessed in certain tropical pastures. These assessments are commonly based on potential net nitrification rates, which do not differentiate between gross nitrification and other processes that may reduce the amount of nitrate in soil. In a greenhouse experiment using two genotypes of Urochloa humidicola with contrasting BNI capacity in vitro, we evaluated gross N transformation rates before and after (7 and 21 days) N fertilization, while periodically measuring N2O emissions. Gross nitrification rates (in fact gross nitrate production assessed by pool dilution technique) were comparable in both genotypes and were low in comparison to strong microbial NH4+ immobilization. The N2O emissions were higher in pots with low-BNI plants. The discrepancy between the potential net nitrification rates assessed in laboratory assays (higher in low-BNI plants) and gross nitrification in pot or field experiments (no differences between genotypes) can be attributed to the out-competition of ammonia oxidizers by plant N uptake and ammonia immobilizing heterotrophic microbes, resulting in low nitrification under conditions where growing plants are present. This study confirmed the capacity of certain U. humidicola genotypes to reduce N2O emissions but warrants further investigation of the underlying mechanisms. It also questions the relevance of BNI in the rhizosphere of this plant species as other mechanisms (rather than the inhibition of gross nitrification) seem to be more important in maintaining low-nitrate soil environments in soil–plant systems of U. humidicola.
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References
Anderson IC, Poth M, Homstead J, Burdige D (1993) A comparison of NO and N2O production by the autotrophic nitrifier Nitrosomonas europaea and the heterotrophic nitrifier Alcaligenes faecalis. Appl Environ Microbiol 59:3525–3533
Arango J, Moreta D, Núñez J, Hartmann K, Domínguez M, Ishitani M, Miles J, Subbarao G, Peters M, Rao I (2014) Developing methods to evaluate phenotypic variability in biological nitrification inhibition (BNI) capacity of Brachiaria grasses. Trop Grasslands - Forrajes Trop 2:6–8
Arango J, Ruden A, Martinez-Baron D, Loboguerrero AM, Berndt A, Chacón M, Torres CF, Oyhantcabal W, Gomez CA, Ricci P, Ku-Vera J, Burkart S, Moorby JM, Chirinda N (2020) Ambition meets reality: achieving GHG emission reduction targets in the livestock sector of Latin America. Front Sustain Food Syst 4:65. https://doi.org/10.3389/fsufs.2020.00065
Byrnes RC, Núñez J, Arenas L, Rao I, Trujillo C, Alvarez C, Arango J, Rasche F, Chirinda N (2017) Biological nitrification inhibition by Brachiaria grasses mitigates soil nitrous oxide emissions from bovine urine patches. Soil Biol Biochem 107:156–163. https://doi.org/10.1016/j.soilbio.2016.12.029
Cavigelli MA, Del Grosso SJ, Liebig MA, Snyder CS, Fixen PE, Venterea RT, Leytem AB, McLain JE, Watts DB (2012) US agricultural nitrous oxide emissions: context, status, and trends. Front Ecol Environ 10:537–546. https://doi.org/10.1890/120054
Chirinda N, Loaiza S, Arenas L, Ruiz V, Faverín C, Alvarez C, Savian JV, Belfon R, Zuniga K, Morales-Rincon LA, Trujillo C, Arango M, Rao I, Arango J, Peters M, Barahona R, Costa C, Rosenstock TS, Richards M, Martinez-Baron D, Cardenas L (2019) Adequate vegetative cover decreases nitrous oxide emissions from cattle urine deposited in grazed pastures under rainy season conditions. Sci Rep 9:908. https://doi.org/10.1038/s41598-018-37453-2
Dannenmann M, Gasche R, Ledebuhr A, Papen H (2006) Effects of forest management on soil N cycling in beech forests stocking on calcareous soils. Plant Soil 287:279–300. https://doi.org/10.1007/s11104-006-9077-4
De Boer W, Kowalchuk GA (2001) Nitrification in acid soils: micro-organisms and mechanisms. Soil Biol Biochem 33:853–866. https://doi.org/10.1016/S0038-0717(00)00247-9
Egenolf K, Conrad J, Schöne J, Braunberger C, Beifuß U, Walker F, Nuñez J, Arango J, Karwat H, Cadisch G, Neumann G, Rasche F (2020a) Brachialactone isomers and derivatives of Brachiaria humidicola reveal contrasting nitrification inhibiting activity. Plant Physiol Biochem 154:491–497. https://doi.org/10.1016/j.plaphy.2020.06.004
Egenolf K, Verma S, Schöne J, Klaiber I, Arango J, Cadisch G, Neumann G, Rasche F (2020b) Rhizosphere pH and cation-anion balance determine the exudation of nitrification inhibitor 3-epi-brachialactone suggesting release via secondary transport. Psysiol Plant 172:116–123. https://doi.org/10.1111/ppl.13300
Forster JC (1995) Soil Nitrogen. In: Alef K, Nannipieri P (eds) Methods in applied soil microbiology and biochemistry. Academic Press, Elsevier, Cambridge, USA, pp 79–87
Geisseler D, Horwath WR (2009) Relationship between carbon and nitrogen availability and extracellular enzyme activities in soil. Pedobiologia (jena) 53:87–98. https://doi.org/10.1016/j.pedobi.2009.06.002
Hahn J, Crutzen PJ (1982) The role of fixed nitrogen in atmospheric photochemistry. Philos Trans R Soc London b, Biol Sci 296:521–541. https://doi.org/10.1098/rstb.1982.0024
Hink L, Nicol GW, Prosser JI (2017) Archaea produce lower yields of N2O than bacteria during aerobic ammonia oxidation in soil. Environ Microbiol 19:4829–4837. https://doi.org/10.1111/1462-2920.13282
Horrocks CA, Arango J, Arevalo A, Nuñez J, Cardoso JA, Dungait JAJ (2019) Smart forage selection could significantly improve soil health in the tropics. Sci Total Environ 688:609–621. https://doi.org/10.1016/j.scitotenv.2019.06.152
Hu X, Liu C, Zheng X, Dannenmann M, Butterbach-Bahl K, Yao Z, Zhang W, Wang R, Cao G (2019) Annual dynamics of soil gross nitrogen turnover and nitrous oxide emissions in an alpine shrub meadow. Soil Biol Biochem 138:107576. https://doi.org/10.1016/j.soilbio.2019.107576
Karwat H, Moreta D, Arango J, Núñez J, Rao I, Rincón Á, Rasche F, Cadisch G (2017) Residual effect of BNI by Brachiaria humidicola pasture on nitrogen recovery and grain yield of subsequent maize. Plant Soil 420:389–406. https://doi.org/10.1007/s11104-017-3381-z
Kirkham D, Bartholomew WV (1954) Equations for following nutrient transformations in soil, utilizing tracer data1. Soil Sci Soc Am J 18:33–34. https://doi.org/10.2136/sssaj1954.03615995001800010009x
Leininger S, Urich T, Schloter M et al (2006) Archaea predominate among ammonia-oxidizing prokaryotes in soils. Nature. https://doi.org/10.1038/nature04983
Linquist B, Van Groenigen KJ, Adviento-Borbe MA, Pittelkow C, Van Kessel C (2012) An agronomic assessment of greenhouse gas emissions from major cereal crops. Glob Chang Biol 18:194–209. https://doi.org/10.1111/j.1365-2486.2011.02502.x
Liu R, Suter H, He J, Hayden H, Chen D (2015) Influence of temperature and moisture on the relative contributions of heterotrophic and autotrophic nitrification to gross nitrification in an acid cropping soil. J Soils Sediments 15:2304–2309. https://doi.org/10.1007/s11368-015-1170-y
Miranda KM, Espey MG, Wink DA (2001) A rapid, simple spectrophotometric method for simultaneous detection of nitrate and nitrite. Nitric Oxide - Biol Chem 5:62–71
Müller C, Stevens RJ, Laughlin RJ (2004) A 15N tracing model to analyse N transformations in old grassland soil. Soil Biol Biochem 36:619–632. https://doi.org/10.1016/j.soilbio.2003.12.006
Nakamura S, Saliou PS, Takahashi M et al (2020) The contribution of root turnover on biological nitrification inhibition and its impact on the ammonia-oxidizing archaea under Brachiaria cultivations. Agronomy. https://doi.org/10.3390/agronomy10071003
Nardi P, Laanbroek HJ, Nicol GW, Renella G, Cardinale M, Pietramellara G, Weckwerth W, Trinchera A, Ghatak A, Nannipieri P (2020) Biological nitrification inhibition in the rhizosphere: Determining interactions and impact on microbially mediated processes and potential applications. FEMS Microbiol Rev 44:874–908. https://doi.org/10.1093/femsre/fuaa037
Nuñez J, Arevalo A, Karwat H, Egenolf K, Miles J, Chirinda N, Cadisch G, Rasche F, Rao I, Subbarao G, Arango J (2018) Biological nitrification inhibition activity in a soil-grown biparental population of the forage grass, Brachiaria humidicola. Plant Soil 426:401–411. https://doi.org/10.1007/s11104-018-3626-5
Pandey CB, Kumar U, Kaviraj M, et al (2020) DNRA: A short-circuit in biological N-cycling to conserve nitrogen in terrestrial ecosystems. Sci. Total Environ.
Papen H, von Berg R, Hinkel I, Thoene B, Rennenberg H (1989) Heterotrophic nitrification by Alcaligenes faecalis: NO2-, NO3-, N2O, and NO production in exponentially growing cultures. Appl Environ Microbiol 55:2068–2072. https://doi.org/10.1128/aem.55.8.2068-2072.1989
Rice CW, Tiedje JM (1989) Regulation of nitrate assimilation by ammonium in soils and in isolated soil microorganisms. Soil Biol Biochem 21:597–602. https://doi.org/10.1016/0038-0717(89)90135-1
Rosswall T (1982) Microbiological regulation of the biogeochemical nitrogen cycle. Plant Soil 67:15–34. https://doi.org/10.1007/BF02182752
Rotthauwe J-H, Witzel K-P, Werner L (1997) The ammonia monooxygenase structural gene amoA as a functional marker Molecular fine-scale analysis of natural ammonia-oxidizing populations.pdf. Appl Environ Microbiol 63:4704–4712. https://doi.org/10.1128/AEM.NA
Sarr PS, Ando Y, Nakamura S, Deshpande S, Subbarao GV (2020) Sorgoleone release from sorghum roots shapes the composition of nitrifying populations, total bacteria, and archaea and determines the level of nitrification. Biol Fertil Soils 56:145–166. https://doi.org/10.1007/s00374-019-01405-3
Spohn M (2016) Element cycling as driven by stoichiometric homeostasis of soil microorganisms. Basic Appl Ecol 17:471–478. https://doi.org/10.1016/j.baae.2016.05.003
Subbarao GV, Yoshihashi T, Worthington M, Nakahara K, Ando Y, Sahrawat KL, Rao IM, Lata JC, Kishii M, Braun HJ (2015) Suppression of soil nitrification by plants. Plant Sci 233:155–164
Subbarao GV, Ishikawa T, Ito O, Nakahara K, Wang HY, Berry WL (2006) A bioluminescence assay to detect nitrification inhibitors released from plant roots: a case study with Brachiaria humidicola. Plant Soil 288:101–112. https://doi.org/10.1007/s11104-006-9094-3
Subbarao GV, Nakahara K, Hurtado MP, Ono H, Moreta DE, Salcedo AF, Yoshihashi AT, Ishikawa T, Ishitani M, Ohnishi-Kameyama M, Yoshida M, Rondon M, Rao IM, Lascano CE, Berry WL, Ito O (2009) Evidence for biological nitrification inhibition in Brachiaria pastures. Proc Natl Acad Sci U. S A 106:17302–17307. https://doi.org/10.1073/pnas.0903694106
Subbarao GV, Sahrawat KL, Nakahara K, Rao IM, Ishitani M, Hash CT, Kishii M, Bonnett DG, Berry WL, Lata JC (2013) A paradigm shift towards low-nitrifying production systems: the role of biological nitrification inhibition (BNI). Ann Bot 112:297–316. https://doi.org/10.1093/aob/mcs230
Subbarao GV, Wang HY, Ito O, Nakahara K, Berry WL (2007) NH4+ triggers the synthesis and release of biological nitrification inhibition compounds in Brachiaria humidicola roots. Plant Soil 290:245–257. https://doi.org/10.1007/s11104-006-9156-6
Syakila A, Kroeze C (2011) The global nitrous oxide budget revisited. Greenh Gas Meas Manag 1:17–26. https://doi.org/10.3763/ghgmm.2010.0007
Teutscherova N, Vazquez E, Arango J, Arevalo A, Benito M, Pulleman M (2019a) Native arbuscular mycorrhizal fungi increase the abundance of ammonia-oxidizing bacteria, but suppress nitrous oxide emissions shortly after urea application. Geoderma 338:493–501. https://doi.org/10.1016/j.geoderma.2018.09.023
Teutscherova N, Vazquez E, Arevalo A, Pulleman M, Rao I, Arango J (2019b) Differences in arbuscular mycorrhizal colonization and P acquisition between genotypes of the tropical Brachiaria grasses: is there a relation with BNI activity? Biol Fertil Soils 55:325–337. https://doi.org/10.1007/s00374-019-01353-y
Vázquez E, Teutscherova N, Dannenmann M, Töchterle P, Butterbach-Bahl K, Pulleman M, Arango J (2020) Gross nitrogen transformations in tropical pasture soils as affected by Urochloa genotypes differing in biological nitrification inhibition (BNI) capacity. Soil Biol Biochem 151:108058. https://doi.org/10.1016/j.soilbio.2020.108058
Verhagen FJM, Laanbroek HJ, Woldendrop JW (1995) Competition for ammonium between plant roots and nitrifying and heterotrophic bacteria and the effects of protozoan grazing. Plant Soil 170:241–250. https://doi.org/10.1007/BF00010477
Vitousek PM, Aber JD, Howarth RW, Likens GE, Matson PA, Schindler DW, Schlesinger WH, Tilman DG (1997) Human alteration of the global nitrogen cycle: sources and consequences. Ecol Appl 7:737–750. https://doi.org/10.1890/1051-0761(1997)007[0737:HAOTGN]2.0.CO;2
Warlo H, Machacova K, Nordstrom N et al (2018) Comparison of portable devices for sub-ambient concentration measurements of methane (CH4) and nitrous oxide (N2O) in soil research. Int J Environ Anal Chem. https://doi.org/10.1080/03067319.2018.1517871
Wrage-Mönnig N, Horn MA, Well R, et al (2018) The role of nitrifier denitrification in the production of nitrous oxide revisited. Soil Biol Biochem
Xiao R, Qiu Y, Tao J, Zhang X, Chen H, Reberg-Horton SC, Shi W, Shew HD, Zhang Y, Hu S (2020) Biological controls over the abundances of terrestrial ammonia oxidizers. Glob Ecol Biogeogr 29:384–399. https://doi.org/10.1111/geb.13030
Xiao R, Ran W, Hu S, Guo H (2021) The response of ammonia oxidizing archaea and bacteria in relation to heterotrophs under different carbon and nitrogen amendments in two agricultural soils. Appl Soil Ecol 158:103812. https://doi.org/10.1016/j.apsoil.2020.103812
Zhang J, Sun W, Zhong W, Cai Z (2014) The substrate is an important factor in controlling the significance of heterotrophic nitrification in acidic forest soils. Soil Biol Biochem 76:143–148. https://doi.org/10.1016/j.soilbio.2014.05.001
Zhang J, Zhu T, Meng T, Zhang Y, Yang J, Yang W, Müller C, Cai Z (2013) Agricultural land use affects nitrate production and conservation in humid subtropical soils in China. Soil Biol Biochem 62:107–114. https://doi.org/10.1016/j.soilbio.2013.03.006
Acknowledgements
We thank all the CIAT staff who collaborated with the soil collection, experiment establishment and maintenance at CIAT Headquarters in Palmira, Colombia. In addition, Eduardo Vázquez thanks the Spanish Ministry of Education for his FPU fellowship.
Funding
This work was implemented as part of the CGIAR Research Programs (CRP) on Climate Change, Agriculture and Food Security (CCAFS); and the Livestock CRP. We gratefully acknowledge funding from BBSRC project grants RCUK-CIAT Newton Fund—Advancing sustainable forage-based livestock production systems in Colombia (CoForLife) (BB/S01893X/1), and the UKRI Global Challenges Research Fund (GCRF) GROW Colombia grant via the UK’s BBSRC (BB/P028098/1). Financial support was also obtained from the Internal Grant Agency of Czech University of Life Sciences Prague (20213110) and from Center for Geosphere Dynamics (UNCE/SCI/006).
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Teutscherová, N., Vázquez, E., Trubač, J. et al. Gross N transformation rates in soil system with contrasting Urochloa genotypes do not confirm the relevance of BNI as previously assessed in vitro. Biol Fertil Soils 58, 321–331 (2022). https://doi.org/10.1007/s00374-021-01610-z
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DOI: https://doi.org/10.1007/s00374-021-01610-z
Keywords
- Tropical forage
- Gross nitrogen transformations
- Urochloa humidicola (Rendle) Schweick (syn. Brachiaria humidicola)
- N2O emissions
- Biological nitrification inhibition