Abstract
Background
The nitrogen (N) response in grass-clover is influenced by quantity and timing of N-fertilizer, but less is known about the effect of fertilizer type (animal manure vs. mineral N) and N form (ammonium vs. nitrate).
Method
A field trial was established in a mixture of white clover (Trifolium repens L.), red clover (Trifolium pratense L.) and perennial ryegrass (Lolium. perenne L.) with mineral N and slurry. DM yield in grass and clover and N2-fixation activity (%Ndfa) was determined in four cuts during the season.
Results
Fertilization resulted in a peak in soil mineral N within 1-2 weeks of application and decreased to background rate prior to plant harvest. No differences in white clover N2-fixation activity were found between fertilizer types within each N level.
Conclusions
The experiment showed that mineral N form does not affect the regulation of N2-fixation activity. The fertilizer N level was the only factor affecting white clover N2-fixation activity. Further, the results demonstrated an N threshold below which annual %Ndfa was always high (~90%). This was controlled by the companion grass’ ability to deplete the soil mineral N pool. When N fertilization exceeded this threshold (above 180 kg N ha-1 yr-1 in our case) annual %Ndfa significantly decreased as did qBNF in the harvested biomass.
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References
Allos HF, Bartholemew WV (1955) Effect on available available nitrogen on symbiotic fixation. Soil Sci Soc Am 19:182–184. https://doi.org/10.2136/sssaj1955.03615995001900020018x
Allos HF, Bartholemew WV (1959) Replacement of symbiotic fixation by available nitrogen. Soil Science 87:61–66
Anglade J, Billen G, Garnier J (2015) Relationships for estimating N-2 fixation in legumes: incidence for N balance of legume-based crop** systems in Europe. Ecosphere 6:37. https://doi.org/10.1890/ES14-00353.1
Bates D, Maechler M, Bolker B, Walker S (2015) Fitting linear mixed-effects models using lme4. J Stat Software 67:1–48. https://doi.org/10.18637/jss.v067.i01
Burchill W, James EK, Li D, Lanigan GJ, Williams M, Iannetta PPM, Humphreys J (2014) Comparison of biological nitrogen fixation in association with white clover (Trifolium repens L.) under four fertiliser nitrogen inputs as measured using 15N techniques. Plant Soil 385:287–302
Carlsson G, Huss-Danell K (2003) Nitrogen fixation in perennial forage legumes in the field. Plant Soil 253:353–372. https://doi.org/10.1023/A:1024847017371
Castle ME, Drysdale AD (1962) Liquid manure as a grassland fertilizer. I. The response to liquid manure and to dry fertilizer. J Agri Sci 58:165–171. https://doi.org/10.1017/S002185960001011X
Czaban W, Jämtgard S, Näsholm T, Rasmussen J, Nicolaisen M, Fomsgaard IS (2016) Direct acquisition of organic N by white clover even in the presence of inorganic N. Plant Soil 407:91–107. https://doi.org/10.1007/s11104-016-2896-z
Czaban W, Rasmussen J, Laursen BB, Vidkjær NH, Sapkota R, Nicolaisen M, Fomsgaard IS (2018) Multiple effects of secondary metabolites on amino acid cycling in white clover rhizosphere. Soil Biol Biochem 123:54–63. https://doi.org/10.1016/j.soilbio.2018.04.012
Dhamala NR, Rasmussen J, Carlsson G, Søegaard K, Eriksen J (2018) Effects of including forbs on N2-fixation and N yield in red clover-ryegrass mixtures. Plant Soil 424:525–537. https://doi.org/10.1007/s11104-017-3554-9
Drysdale AD (1963) Liquid manure as a grassland fertilizer: II. The response to winter applications. J Agri Sci 61:353–360. https://doi.org/10.1017/S0021859600018074
Drysdale AD (1965) Liquid manure as a grassland fertilizer. III. The effect of liquid manure on the yield and botanical composition of pasture, and its interaction with nitrogen, phosphate and potash fertilizers. J Agri Sci 65:333–340. https://doi.org/10.1017/S0021859600048899
Eriksen J, Askegaard M, Rasmussen J, Søegaard K (2015) Nitrate leaching and residual effect in dairy crop rotations with grass–clover leys as influenced by sward age, grazing, cutting and fertilizer regimes. Agri Ecosys Environ 212:75–84. https://doi.org/10.1016/j.agee.2015.07.001
Eriksen J, Askegaard M, Søegaard K (2014) Complementary effects of red clover inclusion in ryegrass–white clover swards for grazing and cutting. Grass Forage Sci 69:241–250. https://doi.org/10.1111/gfs.12025
Faurie O, Soussana JF (1993) Oxygen-induced recovery from short-term nitrate inhibition of N2 fixation in white clover plants from spaced and dense stands. Phys Plant 89:467–475. https://doi.org/10.1111/j.1399-3054.1993.tb05200.x
Fried M, Middelboe V (1977) Measurement of amount of nitrogen fixed by a legume crop. Plant Soil 47:713–715. https://doi.org/10.1007/BF00011042
Herriott JBD, Wells DA (1962) Gulle as a grassland fertilizer. Grass Forage Sci 17:167–170. https://doi.org/10.1111/j.1365-2494.1962.tb00291.x
Høgh-Jensen H, Wollenweber B, Schjoerring JK (1997) Kinetics of nitrate and ammonium absorption and accompanying H+ fluxes in roots of Lolium perenne L. and N2-fixing Trifolium repens L. Plant Cell Environ 20:1184–1192. https://doi.org/10.1046/j.1365-3040.1997.d01-145.x
Houlton BZ, Wang YP, Field CB, Vitousek PM (2008) A unifying framework for dinitrogen fixation in the terrestrial biosphere. Nature 454:327–330. https://doi.org/10.1038/nature07028
Houwaard F (1980) Influence of ammonium and nitrate nitrogen on nitrogenase activity of pea-plants as affected by light-intensity and sugar addition. Plant Soil 54:271–282. https://doi.org/10.1007/BF02181853
Johansen M (2017) Green forages for dairy cows. PhD-thesis, Aarhus University, Denmark.
Johansen M, Søegaard K, Lund P, Weisbjerg MR (2017) Digestibility and clover proportion determine milk production when silages of different grass and clover species are fed to dairy cows. J Dairy Sci 100:8861–8880. https://doi.org/10.3168/jds.2017-13401
Kusliene G, Eriksen J, Rasmussen J (2015) Leaching of dissolved organic and inorganic nitrogen from legume-based grasslands. Biol Fert Soils 51:217–230. https://doi.org/10.1007/s00374-014-0966-z
Landbrugsstyrelsen (2018) Vejledning om gødsknings- og harmoniregler In: M.-o. Fødevareministeriet (Editor), Miljø- og Fødevareministeriet Landbrugsstyrelsen Nyropsgade 30 1780 København V.
Ledgard SF, Steele KW (1992) Biological nitrogen fixation in mixed legume/grass pastures. Plant Soil 141:137–153. https://doi.org/10.1007/BF00011314
Lüscher A, Mueller-Harvey I, Soussana JF, Rees RM, Peyraud JL (2014) Potential of legume-based grassland-livestock systems in Europe: a review. Grass Forage Sci 69:206–228. https://doi.org/10.1111/gfs.12124
Macduff JH, Jarvis SC, Davidson IA (1996) Inhibition of N2 fixation by white clover (Trifolium repens L.) at low concentrations of NO3− in flowing solution culture. Plant Soil 180:287–295. https://doi.org/10.1007/BF00015312
Martin MPLD, Field RJ (1984) The nature of competition between perennial ryegrass and white clover. Grass Forage Sci 39:247–253
McAuliffe C, Chamblee DS, Uribe-Arango H, Woodhouse WW (1958) Influence of inorganic nitrogen on nitrogen fixation by legumes as revealed by N15. Agron J 50:334–337
McNeill AM, Hood RC, Wood M (1994) Direct measurement of nitrogen fixation by Trifolium repens L. and Alnus glutinosa L. using 15N2. J Exp Bot 45:749–755. https://doi.org/10.1093/jxb/45.6.749
Munoz AE, Weaver RW (1999) Competition between Subterranean Clover and Rygrass for uptake of 15N-labeled fertilizer. Plant Soil 211:173–178. https://doi.org/10.1023/A:1004646319700
Nesheim L, Boller BC, Lehmann J, Walther U (1990) The effect of nitrogen in cattle slurry and mineral fertilizers on nitrogen fixation by white clover. Grass Forage Sci 45:91–97. https://doi.org/10.1111/j.1365-2494.1990.tb02186.x
Olivares J, Bedmar EJ, Sanjuán J (2013) Biological nitrogen fixation in the context of global change. Mol Plant Microbe Interact 265:486–494. https://doi.org/10.1094/MPMI-12-12-0293-CR
Peoples MB, Herridge DF, Ladha JK (1995) Biological nitrogen fixation: An efficient source of nitrogen for sustainable agricultural production? Plant Soil 174:3–28. https://doi.org/10.1007/BF00032239
R Core Team (2018) R: A language and environment for statistical computing. R Foundation for Statistical Computing, Vienna, Austria
Rasmussen J, Søegaard K, Pirhofer-Walzl K, Eriksen J (2012) N2-fixation and residual N effect of four legume species and four companion grass species. Eur J Agron 36:66–74. https://doi.org/10.1016/j.eja.2011.09.003
Rasmussen J, Gyldfadottir T, Loges R, Eriksen J, Helgadottir A (2013) Spatial and temporal variation in N transfer in grass-white clover mixtures at three Northern European field sites. Soil Biol Biochem 57:654–662. https://doi.org/10.1016/j.soilbio.2012.07.004
Russell L (2020) emmeans: Estimated Marginal Means, aka Least-Squares Means. R package version 1(4):4
Schipanski ME, Drinkwater LE (2012) Nitrogen fixation in annual and perennial legume-grass mixtures across a fertility gradient. Plant Soil 357:147–159. https://doi.org/10.1007/s11104-012-1137-3
Silsbury JH, Catchpoole DW, Wallace W (1986) Effects of nitrate and ammonium on nitrogenase (C2H2 reduction) activity of swards of subterranean clover, Trifolium Subterraneum L. Aus J Plant Phys 13:257–273. https://doi.org/10.1071/PP9860257
Soussana JF, Tallec T (2010) Can we understand and predict the regulation of biological N2 fixation in grassland ecosystems? Nutr Cycl Agroecosyst 88:197–213. https://doi.org/10.1007/s10705-009-9335-y
Søegaard K (1998) Timing of N-application after defoliation of clover-grass. Grassland Science in Europe 3:979–982
Sørensen P (1998) Carbon mineralization, nitrogen immobilization and pH change in soil after adding volatile fatty acids. Eur J Soil Sci 49:457–462. https://doi.org/10.1046/j.1365-2389.1998.4930457.x
Walker WT, Adams FRA, Orchiston DH (1956) Fate of labeled nitrate and ammonium nitrogen when applied to grass and clover grown seperately and together. Soil Sci 81:339–352
Webb J, Sørensen P, Velthof G, Amon B, Pinto M, Rodhe L, Salomon E, Hutchings N, Burczyk P, Reid J (2013) Chapter Seven - An assessment of the variation of manure nitrogen efficiency throughout Europe and an appraisal of means to increase manure-N efficiency. DL Sparks (Editor) Adv Agron 119:371–442. https://doi.org/10.1016/B978-0-12-407247-3.00007-X
Weigelt A, Bol R, Bardgett RD (2005) Preferential uptake of soil nitrogen forms by grassland plant species. Oecologia 142:627–635. https://doi.org/10.1007/s00442-004-1765-2
Acknowledgements
We gratefully acknowledge the help of the staff at Foulumgaard and lab technicians Cecilie Kokholm, Kathrine Øster Høstgaard, Margit Paulsen and Jørgen Munksgaard Nielsen.
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The study was funded by Innovation Fund Denmark through the SmartGrass project.
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Kristensen, R.K., Rasmussen, J. & Eriksen, J. Biological N2-fixation in grass-clover ley in response to N application in cattle slurry vs. mineral fertilizer. Plant Soil 471, 629–641 (2022). https://doi.org/10.1007/s11104-021-05256-6
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DOI: https://doi.org/10.1007/s11104-021-05256-6