Agronomy for Sustainable Development

, Volume 28, Issue 2, pp 281–290 | Cite as

A study of 15N transfer between legumes and grasses

  • Florence Paynel
  • Fabien Lesuffleur
  • Jacques Bigot
  • Sylvain Diquélou
  • Jean-Bernard Cliquet
Original Article


The overuse of classical N fertilisers contributes substantially to environmental degradation by pollution of groundwater by nitrates. This leaching of N in waters is also an economic flaw for farmers because only a part of the fertiliser is used by the plants. Here, systems involving mixtures of legumes and grasses represent a sustainable alternative because legumes can fix atmospheric N2 using symbiotic microbes. N transfer in those mixtures has been thoroughly investigated but little is known concerning the effect of N fertiliser on N transfer between N-fixing legumes and companion grasses. In white clover (Trifolium repens L.) — perennial ryegrass (Lolium perenne L.) associations, N is transferred mostly through rhizodeposition into the soil by clover followed by re-uptake by ryegrass. Rhizodeposition of N occurs through senescence and decomposition of legume tissue or through exudation of N compounds by living cells. Ammonium and amino acids are the main compounds exuded and their exudation is thought to occur by passive diffusion attributed to a concentration gradient from root to soil. In this study, we test the hypothesis that greater N transfer from clover to grass, as seen in N-rich soils or nutrient solutions, is due to greater N rhizodeposition brought about by higher ammonium and amino acid content of roots. The relations between N input, root N content, N net exudation and N transfer between legumes and grasses were investigated using 15N by growing white clover and perennial ryegrass with increasing N application in axenic microlysimeters or in pots. Ammonium and amino acid concentrations were measured in root tissues, in root bathing solutions and in soils. We found that mineral N application strongly reduced atmospheric N fixation by clover, from 3.0 to 0.9 mg per plant, and root amino acid content, from 164 to 49 nmoles per g dry weight, but had no effect on ammonium and amino acid concentrations in sterile exudates, showing for the first time that amino acid net exudation is independent of root content. In contrast, ammonium and amino acid concentrations in clover soils increased with N fixation, showing the link between N fixation and N rhizodeposition in soils. Nitrate application increased ryegrass root growth by 7–8 times, and transfer of N between clover and ryegrass (by 3 times). It is concluded that N fertiliser does not modify N exudation but decreases N fixation and ammonium rhizodeposition in soil by clover. N fertiliser increases N transfer between clover and ryegrass by increasing soil exploration by ryegrass and giving a better access to different available N sources, including the N compounds exuded from clover.


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  1. Benizri E., Courtade A., Guckert A. (1995) Fate of two microorganisms in maize simulated rhizosphere under hydroponic and sterile conditions, Soil Biol. Biochem. 27, 71–77.CrossRefGoogle Scholar
  2. Brophy L.S., Heichel G.H. (1989) Nitrogen release from roots of alfalfa and soybean grown in sand culture, Plant Soil 116, 77–84.CrossRefGoogle Scholar
  3. Boller B.C., Nösberger J. (1987) Symbiotically fixed nitrogen from field grown white and red clover mixed with ryegrass at low level of 15N fertilization, Plant Soil 104, 219–226.CrossRefGoogle Scholar
  4. Deutsch B., Kahle P., Voss M. (2006) Assessing the source of nitrate pollution in water using stable N and O isotopes, Agron. Sustain. Dev. 26, 263–267.CrossRefGoogle Scholar
  5. Elgersma A., Schlepers H., Nassiri M. (2000) Interactions between perennial ryegrass (Lolium perenne L.) and white clover (Trifolium repens L.) under contrasting nitrogen availability: productivity, seasonal patterns of species composition, N2 fixation, N transfer and N recovery, Plant Soil 221, 281–299.CrossRefGoogle Scholar
  6. Garg N., Geetanjali (2007) Symbiotic nitrogen fixation in legume nodules: process and signaling. A review, Agron. Sustain. Dev. 27, 59–68.CrossRefGoogle Scholar
  7. Grayston S.J., Wang S., Campbell C.D., Edwards A.C. (1998) Selective influence of plant species on microbial diversity in the rhizosphere, Soil Biol. Biochem. 30, 369–378.CrossRefGoogle Scholar
  8. Hertenberger G., Zampach P., Bachmann G. (2002) Plant species affect the concentration of free sugars and free amino acids in different types of soil, J. Plant Nutr. Soil Sci. 165, 557–565.CrossRefGoogle Scholar
  9. Hertenberger G., Wanek W. (2004) Evaluation of methods to measure differential 15N labeling of soil and root N pools for studies of root exudation, Rapid Commun. Mass Spectrom. 18, 2415–2425.PubMedCrossRefGoogle Scholar
  10. Høgh-Jensen H., Schjoerring J.K. (1997) Interaction between white clover and ryegrass under contrasting nitrogen availability: N2 fixation, N fertilizer recovery, N transfer and water-use efficiency, Plant Soil 197, 187–199.CrossRefGoogle Scholar
  11. Høgh-Jensen H., Schjoerring J.K. (2001) Rhizodeposition of nitrogen by red clover, white clover and ryegrass leys, Soil Biol. Biochem. 33, 439–448.CrossRefGoogle Scholar
  12. Johansen A., Jensen E.S. (1996) Transfer of N and P from intact or decomposing roots of pea to barley inter connected by an arbuscular mycorrhizal fungus, Soil Biol. Biochem. 28, 73–81.CrossRefGoogle Scholar
  13. Jones D.L, Darrah P.R. (1994) Amino acid influx at the soil-root interface of Zea mays L. and its consequences in the rhizosphere, Plant Soil 163, 1–12.Google Scholar
  14. Jones D.L., Owen A.G., Farrar J. (2002) Simple method to enable the high resolution determination of total free amino acids in soil solutions and soil extracts, Soil Biol. Biochem. 34, 1893–1902.CrossRefGoogle Scholar
  15. Jones D.L., Hodge A., Kuzyakov Y. (2004) Plant and mycorrizal regulation of rhizodeposition, New Phytol. 163, 459–480.CrossRefGoogle Scholar
  16. Kielland K. (1995) Landscape pattern of free amino acids in Artic tundra soils, Biogeochemistry 31, 85–98.CrossRefGoogle Scholar
  17. Ledgard S.F., Steele K.W. (1992) Biological nitrogen fixation in mixed legume-grass pastures, Plant Soil 141, 137–153.CrossRefGoogle Scholar
  18. Lesuffleur F., Paynel F., Bataillé M.P., Le Deunff E., Cliquet J.B. (2007) High proportions of glycine and serine in root exudates of six different species result from hight efflux rates, Plant Soil 294, 235–246.CrossRefGoogle Scholar
  19. Lipson D.A., Raab T.K., Schmidt S.K., Monson R.K. (1999) Variation in competitive abilities of plants and microbes for specific amino acids, Biol. Fert. Soils 29, 257–261.CrossRefGoogle Scholar
  20. Lipson D.A., Raab T.K., Schmidt S.K., Monson R.K. (2001) An empirical model of amino acid transformations in an alpine soil, Soil Biol. Biochem. 33, 189–198.CrossRefGoogle Scholar
  21. Murashige T., Skoog F. (1962) A revised medium for rapid growth and bio assays with tobacco tissue cultures, Physiol. Plant. 15, 473–497.CrossRefGoogle Scholar
  22. Näsholm T., Huss-Danell K., Högberg P. (2001) Uptake of glycine by field-grown wheat, New Phytol. 150, 59–63.CrossRefGoogle Scholar
  23. Nguyen C. (2003) Rhizodeposition of organic C by plants: mechanisms and control, Agronomie 23, 375–396.CrossRefGoogle Scholar
  24. Paynel F., Murray P.J., Cliquet J.B. (2001) Root exudates: a pathway for short-term N transfer from clover to ryegrass, Plant Soil 229, 235–243.CrossRefGoogle Scholar
  25. Paynel F., Cliquet J.B. (2003) N transfer from white clover to perennial ryegrass, via exudation of nitrogenous compounds, Agronomie 23, 503–510.CrossRefGoogle Scholar
  26. Phillips D.A., Fox T.C., King M.D., Bhuvaneswari T.V., Teuber L.R. (2004) Microbial products trigger amino acid exudation from plant roots, Plant Physiol. 136, 2887–2894.PubMedCrossRefGoogle Scholar
  27. Rogers J.B., Laidlaw A.S., Christie P. (2001) The role of arbuscular mycorrhizal fungi in the transfer of nutrients between white clover and perennial ryegrass, Chemosphere 42, 153–159.PubMedCrossRefGoogle Scholar
  28. Rroço E., Kosegarten H., Mengel K. (2002) Importance of plasmalemma H+-ATPase activity for N losses from intact roots of spring wheat (Triticum aestivum L.), Eur. J. Agron. 16, 187–196.CrossRefGoogle Scholar
  29. Shepherd T., Davies H.V. (1994) Pattern of short-term amino acid accumulation and loss in the root zone of liquid-cultured forage rape, Plant Soil 158, 99–109.CrossRefGoogle Scholar
  30. Soussana J.F., Hartwig U.A. (1996) The effect of elevated CO2 on symbiotic N2 fixation: a link between carbon and nitrogen cycles in grassland ecosystems, Plant Soil 187, 321–332.CrossRefGoogle Scholar
  31. Streeter T.C., Bol R., Bardgett R.D. (2000) Amino acids as a nitrogen source in temperate upland grasslands: the use of dual labelled (13C, 15N) glycine to test for direct uptake by dominant grasses, Rapid Commun. Mass Spectrum. 14, 1351–1355.CrossRefGoogle Scholar
  32. Svenning M.M., Juntilla O., Macduff J.H. (1996) Differential rates of inhibition of N2 fixation by sustained low concentrations of NH4+ and NO3 in northern ecotypes of white clover (Trifolium repens L.), J. Exp. Bot. 47, 729–738.CrossRefGoogle Scholar
  33. Ta T.C., Faris M.A. (1987) Species variation in the fixation and transfer of nitrogen from legumes to associated grasses, Plant Soil 98, 265–274.CrossRefGoogle Scholar
  34. Ta T.C., MacDowall D.H., Faris M.A. (1986) Excretion of nitrogen assimilated from N2 fixed by nodulated roots of alfalfa (Medicago sativa), Can. J. Bot. 64, 2063–2067.CrossRefGoogle Scholar
  35. Thornton B., Robinson D. (2005) Uptake and assimilation of nitrogen from solutions containing multiple N sources, Plant Cell Environ. 28, 813–821.CrossRefGoogle Scholar
  36. Thomas B.D., Bowman W.D. (1998) Influence of N2-fixing Trifolium on plant species composition and biomass production in alpine tundra, Oecologia 115, 26–31.CrossRefGoogle Scholar
  37. Umar A.S., Iqbal M. (2007) Nitrate accumulation in plants, factors affecting the process, and human health implications. A review, Agron. Sustain. Dev. 27, 45–57.CrossRefGoogle Scholar
  38. Volk R.J. (1995) 15N assays of ammonium cycling in maize roots, in: INRA (Les Colloques, n∘ 70) (Ed.), Utilisation des isotopes stables pour l’étude du fonctionnement des plantes, Paris, France, pp. 175–182.Google Scholar
  39. Weigelt A., Bol R., Bardgett R.D. (2005) Preferential uptake of soil nitrogen forms by grassland plant species, Oecologia 142, 627–635.PubMedCrossRefGoogle Scholar
  40. White J., Prell J., James E.K., Poole P. (2007) Nutrient sharing between symbionts, Plant Physiol. 144, 604–614.PubMedCrossRefGoogle Scholar

Copyright information

© Springer S+B Media B.V. 2008

Authors and Affiliations

  • Florence Paynel
    • 1
  • Fabien Lesuffleur
    • 2
  • Jacques Bigot
    • 2
  • Sylvain Diquélou
    • 2
  • Jean-Bernard Cliquet
    • 2
  1. 1.UMR CNRS/INRA/SupAgroM/UMII 5004 B&PMPBiochimie and Physiologie Moléculaire des Plantes, IBIPMontpellier Cedex 1France
  2. 2.UMR INRA-UCBN 950 EVA, Écophysiologie Végétale, Agronomie et nutritions N,C,SUniversité de CaenCaen CedexFrance

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