Skip to main content
SpringerLink
Log in

Use of a 15N2 labelling technique to estimate exudation by white clover and transfer to companion ryegrass of symbiotically fixed N

  • Regular Article
  • Published:
Plant and Soil Aims and scope Submit manuscript

Abstract

Backgrounds and aims

N rhizodeposition by legumes leads to enrichment of N in soils and in companion plants. N rhizodeposition can be divided into two major components, root exudation and root senescence. Our aim was to quantify N root exudation in white clover (Trifolium repens L.) through an estimation of short-term N rhizodeposition and to assess its impact on N transfer to companion perennial ryegrass (Lolium perenne L.) grown in mixture with clover.

Method

15N2 provided in the root atmosphere for 3 days was used to estimate transfer of symbiotically fixed nitrogen (SFN) to the growing medium by clover grown in pure stand and to ryegrass by clover grown in mixture for 2 months.

Results

The proportion of N rhizodeposited over the 3 days increased from 3.5 % of SFN in pure stand to 5.3 % in mixture. The 15N-enrichment of ammonium from the adhering substrate shows that a part of the rhizodeposited N was released in the form of ammonium. 4 % of the rhizodeposited N was taken up by ryegrass during the labelling period.

Conclusions

This study showed a significant contribution of root N exudation to the total N rhizodeposition of legumes and in the transfer of N to grasses.

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

Access this article

Log in via an institution

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

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

References

  • Abdallah M, Dubousset L, Meuriot F, Etienne P, Avice JC, Ourry A (2010) Effect of mineral sulphur availability on nitrogen and sulphur uptake and remobilization during the vegetative growth of Brassica napus L. J Exp Bot 61:2635–2646

    Article  PubMed  CAS  Google Scholar 

  • Barea JM, Azcoz R, Azcon-Aguilar C (1989) Time-course of N2-fixation (15N) in the field by clover growing alone or in mixture with ryegrass to improve pasture productivity, and inoculated with vesicular-arbuscular mycorrhizal fungi. New Phytol 112:399–404

    Article  Google Scholar 

  • Bazot S (2005) Contribution à l’étude de l’allocation des photoassimilats récents dans la plante et la rhizosphère chez une graminée pérenne (Lolium perenne L.). Thèse de l’Institut National Polytechnique de Lorraine, 174 p.

  • Bingham IJ, Rees RM (2008) Senescence and N release from clover roots following permanent excision of the shoot. Plant Soil 303:229–240

    Article  CAS  Google Scholar 

  • Brophy LS, Heichel GH (1989) Nitrogen release from roots of alfalfa and soybean grown in sand culture. Plant Soil 116:77–84

    Article  CAS  Google Scholar 

  • Cliquet JB, Murray PJ, Boucaud J (1997) Effect of the arbuscular mycorrhizal fungus Glomus fasciculatum on the uptake of amino nitrogen by Lolium perenne L. New Phytol 137:345–349

    Article  CAS  Google Scholar 

  • Dahlin AS, Mårtenson AM (2008) Cutting regimes determines allocation of fixed nitrogen in white clover. Biol Fertil Soil 45:199–204

    Article  CAS  Google Scholar 

  • Dahlin AS, Stenberg M (2010) Transfer of N from red clover to perennial ryegrass in mixed stands under different cutting strategies. Eur J Agron 33:149–156

    Article  CAS  Google Scholar 

  • Dubach M, Russelle MP (1994) Forage legume roots and nodules and their role nitrogen transfer. Agron J 86:259–266

    Article  Google Scholar 

  • 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

    Article  CAS  Google Scholar 

  • Fujita K, Ofosu-Budu KG, Ogata S (1992) Biological nitrogen fixation in mixed legume-cereal cropping systems. Plant Soil 141:155–175

    Article  CAS  Google Scholar 

  • Fustec J, Lesuffleur F, Mahieu S, Cliquet JB (2009) Nitrogen rhizodeposition of legumes. Agron Sustain Dev 30:57–66

    Article  Google Scholar 

  • Gylfadóttir T, Helgadóttir A, Høgh-Jensen H (2007) Consequences of including adapted white clover in northern european grassland: transfer and deposition of nitrogen. Plant Soil 297:93–104

    Article  Google Scholar 

  • Hamel C, Barrantes-Cartin U, Furlan V, Smith DL (1991) Endomycorrhizal fungi in nitrogen transfer from soybean to maize. Plant Soil 138:33–40

    Article  CAS  Google Scholar 

  • Hartwig UA (1998) The regulation of symbiotic N2 fixation: a conceptual model of N feedback from the novel ecosystem to the gene expression level. Perspect Plant Ecol Evol System 1(1):92–120

    Article  Google Scholar 

  • Haynes RJ (1980) Competitive aspects of the grass/legume association. Adv Agron 33:227–261

    Article  Google Scholar 

  • Haystead A, Malaczuck N, Grove T (1988) Underground transfer of nitrogen between pasture plants infected with vesicular-arbuscular mycorrhizal fungi. New Phytol 108:417–423

    Article  Google Scholar 

  • He X, Xu M, Qiu GY, Zhou J (2009) Use of 15N stable isotope to quantify nitrogen transfer between mycorrhizal plants. J Plant Ecol 2:107–118

    Article  Google Scholar 

  • 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 Comm Mass Spectrom 18:2415–2425

    Article  CAS  Google Scholar 

  • Høgh-Jensen H, Schjoerring JK (1994) Measurement of biological dinitrogen fixation in grassland: Comparison of the enriched 15N dilution and the natural 15N abundance methods at different nitrogen application rates and defoliation frequencies. Plant Soil 166:153–163

    Article  Google Scholar 

  • Høgh-Jensen H, Schjoerring JK (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

    Article  Google Scholar 

  • Høgh-Jensen H, Schjoerring JK (2000) Below-ground nitrogen transfer between different grassland species: direct quantification by 15N leaf feeding compared with indirect dilution of soil 15N. Plant Soil 227:171–183

    Article  Google Scholar 

  • Høgh-Jensen H, Schjoerring JK (2001) Rhizodeposition of nitrogen by red clover, white clover and ryegrass leys. Soil Biol Biochem 33:439–448

    Article  Google Scholar 

  • Jalonen R, Nygren P, Sierra J (2009) Transfer of nitrogen from a tropical legume tree to an associated fodder grass via root exudation and common mycelial network. Plant Cell Environ 32:1366–1376

    Article  PubMed  CAS  Google Scholar 

  • Janzen HH, Gilbertson C (1994) Exchange of N-15 among plants in controlled environments studies. Can J Soil Sci 74:109–110

    Article  Google Scholar 

  • Jensen ES (1996) Grain yield, symbiotic N2 fixation and interspecific competition for inorganic N in pea-barley intercrops. Plant Soil 182:25–38

    Article  CAS  Google Scholar 

  • Jeudy C, Ruffel S, Freixes S, Tillard P, Santoni AL, Morel S, Journet EP, Duc G, Gojon A, Lepetit M, Salon C (2010) Adaptation of Medicago truncatula to nitrogen limitation is modulated via local and systemic nodule developmental responses. New Phytol 185:817–828

    Article  PubMed  CAS  Google Scholar 

  • Johansen A, Jensen ES (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

    Article  CAS  Google Scholar 

  • Jones DL, Darrah PR (1993) Influx and efflux of amino acids from Zea mays L. Roots and their implication for N nutrition and the rhizosphere. Plant Soil 155(156):87–90

    Article  Google Scholar 

  • Jørgensen FV, Jensen ES, Schjoerring JK (1999) Dinitrogen fixation in white clover grown in pure stand and mixture with ryegrass estimated by the immobilized 15N isotope dilution method. Plant Soil 208:293–305

    Article  Google Scholar 

  • Kurppa M, Leblanc HA, Nygren P (2010) Detection of nitrogen transfer from N2-fixing shade trees to cacao saplings in 15 labelled soil: ecological and experimental considerations. Agrofores Syst 80:223–229

    Article  Google Scholar 

  • Lainé P, Bigot J, Ourry A, Boucaud J (1994) Effects of low temperature on nitrate uptake, and xylem and phloem flows of nitrogen, in Secale cereale L. and Brassica napus L. New Phytol 127:675–683

    Article  Google Scholar 

  • Ledgard SF, Steele KW (1992) Biological nitrogen fixation in mixed legume/grass pastures. Plant Soil 141:137–153

    Article  CAS  Google Scholar 

  • Lesuffleur FS, Cliquet JB (2010) Characterisation of root amino acid exudation in white clover. Plant Soil 333:191–201

    Article  CAS  Google Scholar 

  • Lipson DA, Raab TK, Schmidt SK, Monson RK (1999) Variation in competitive abilities of plants and microbes for specific amino acids. Biol Fertil Soils 29:257–261

    Article  CAS  Google Scholar 

  • Macduff JH, Jackson SB (1992) Influx and efflux of nitrate and ammonium in italian ryegrass and white clover roots: comparisons between effects of darkness and defoliation. J Exp Bot 43:525–535

    Article  CAS  Google Scholar 

  • Marty C, Pornon A, Escaravage N, Winterton P, Lamaze T (2009) Complex interactions between a legume and two grasses in a subalpine meadow. Am J Bot 96:1814–1820

    Article  PubMed  Google Scholar 

  • 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

  • Moyer-Henry KA, Burton JW, Israel DW, Rufty TW (2006) Nitrogen transfer between plants: a 15N natural abundance study with crop and weed species. Plant Soil 282:7–20

    Article  CAS  Google Scholar 

  • N’Guyen C (2003) Rhizodeposition of organic C by plants: mechanisms and controls. Agron Sustain Dev 23:375–396

    Google Scholar 

  • Nyfeler D, Huguenin-Elie O, Suter M, Frossard E, Lüscher A (2011) Grass-legume mixtures can yield more nitrogen than legume pure stands due to mutual stimulation of nitrogen uptake from symbiotic and non-symbiotic sources. Agric Ecosyst Environ 140:155–163

    Article  Google Scholar 

  • Ofosu-Budu KG, Fujita K, Ogata S (1990) Excretion of ureide and other nitrogenous compounds by the root system of soybean at different growth stages. Plant Soil 128:135–142

    Article  CAS  Google Scholar 

  • Parsons R, Stanforth A, Raven JA, Sprent JI (1993) Nodule growth and activity may be regulated by a feedback mechanism involving phloem nitrogen. Plant Cell Environ 16:125–136

    Article  CAS  Google Scholar 

  • Paynel F, Cliquet JB (2003) N transfer from white clover to perennial ryegrass, via exudation of nitrogenous compounds. Agron Sustain Dev 23:503–510

    CAS  Google Scholar 

  • Paynel F, Lesuffleur F, Bigot J, Diquélou S, Cliquet JB (2008) A study of 15N transfer between legumes and grasses. Agron Sustain Dev 28:281–290

    Article  CAS  Google Scholar 

  • Paynel F, Murray PJ, Cliquet JB (2001) Root exudates: a pathway for short-term N transfer from clover and ryegrass. Plant Soil 229:235–243

    Article  CAS  Google Scholar 

  • Pirhofer-Walzl K, Rasmussen J, Høgh-Jensen H, Eriksen J, Søegaard K, Rasmussen J (2012) Nitrogen transfer from forage legumes to nine neighbouring plants in a multi-species grassland. Plant Soil 350:71–84

    Article  CAS  Google Scholar 

  • Rasmussen J, Eriksen J, Jensen ES, Esbensen KH, Høgh-Jensen H (2007) In situ carbon and nitrogen dynamics in ryegrass-clover mixtures: transfers, deposition and leaching. Soil Biol Biochem 39:804–815

    Article  CAS  Google Scholar 

  • Rogers JB, Scott Laidlaw A, Christie P (2001) The role of arbuscular mycorrhizal fungi in the transfer of nutrients between white clover and perennial ryegrass. Chemosphere 42:153–159

    Article  PubMed  CAS  Google Scholar 

  • Russelle MP, Allan DL, Gourley CJP (1994) Direct assessment of symbiotically fixed nitrogen in the rhizosphere of alfalfa. Plant Soil 159:233–243

    Article  CAS  Google Scholar 

  • Shepherd T, Davies HV (1994) Patterns of short-term amino acid accumulation and loss in the root-zone of liquid-cultured forage rape (Brassica napus L.). Plant Soil 158:99–109

    Article  CAS  Google Scholar 

  • Sierra J, Nygren P (2006) Transfer of N fixed by a legume tree to the associated grass in a tropical silvopastoral system. Soil Biol Biochem 38:1893–1903

    Article  CAS  Google Scholar 

  • Sokal RR, Rohlf FJ (1995) In: Freeman WH, Company (eds) Biometry; the principles and practice of statistics in biology research, 3rd edn. Freeman, New York, p 897

    Google Scholar 

  • Soussana JF, Hartwig UA (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

    Article  CAS  Google Scholar 

  • Sturite I, Henriksen TM, Breland TA (2007) Longevity of White Clover (Trifolium repens) leaves, stolons and roots, and consequences for nitrogen dynamics under northern temperate climatic conditions. Ann Bot 100:33–40

    Article  PubMed  Google Scholar 

  • Ta TC, Faris M (1987) Species variation in the fixation and transfer of nitrogen from legumes to associated grasses. Plant Soil 98:265–274

    Article  CAS  Google Scholar 

  • Ta TC, Macdowall FDH, Faris MA (1986) Excretion of nitrogen assimilated from N2 fixed by nodulated roots of alfalfa (Medicago sativa). Can J Bot 64:2063–2067

    Article  CAS  Google Scholar 

  • Thomas BD, Bowman WD (1998) Influence of a N2-fixing Trifolium on plant species composition and biomass production in alpine tundra. Oecologia 115:26–31

    Article  Google Scholar 

  • Voisin AS, Salon C, Jeudy C, Warembourg F (2003) Symbiotic Nitrogen Fixation in relation to C economy of Pisum sativum L. as a function of plant phenology. J Exp Bot 54:2733–2744

    Article  PubMed  CAS  Google Scholar 

  • White J, Prell J, James EK, Poole P (2007) Nutrient sharing between symbionts. Plant Physiol 144:604–614

    Article  PubMed  CAS  Google Scholar 

  • Wacquant JP, Ouknider M, Jacquart M (1989) Evidence for a periodic excretion of nitrogen by roots of grass-legume associations. Plant Soil 116:57–58

    Article  CAS  Google Scholar 

  • Wichern F, Eberhardt E, Mayer J, Joergensen RG, Müller T (2008) Nitrogen rhizodeposition in agricultural crops. Soil Biol Biochem 40:30–48

    Article  CAS  Google Scholar 

Download references

Acknowledgments

We thank Anne-Françoise Ameline for technical assistance during the labeling experiment and Marie-Paule Bataille for ammonium and isotopic analysis. We are grateful to Dr Laurence Cantrill for English improvement of the manuscript.

Author information

Authors and Affiliations

  1. INRA UMR 950, EVA Ecophysiologie Végétale, Agronomie et nutritions N,C,S, Université de Caen, Esplanade de la Paix, 14032, Caen Cedex, France

    F. Lesuffleur & J. B. Cliquet

  2. Université de Caen UMR 950, EVA Ecophysiologie Végétale, Agronomie et nutritions N,C,S, Université de Caen, Esplanade de la Paix, 14032, Caen Cedex, France

    F. Lesuffleur & J. B. Cliquet

  3. UMR1347 Agroécologie AgroSup/INRA/uB, 17 rue Sully, BP 86510, 21065, Dijon Cedex, France

    C. Salon & C. Jeudy

Authors
  1. F. Lesuffleur
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. C. Salon
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. C. Jeudy
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. J. B. Cliquet
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to J. B. Cliquet.

Additional information

Responsible Editor: Ad C. Borstlap.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Lesuffleur, F., Salon, C., Jeudy, C. et al. Use of a 15N2 labelling technique to estimate exudation by white clover and transfer to companion ryegrass of symbiotically fixed N. Plant Soil 369, 187–197 (2013). https://doi.org/10.1007/s11104-012-1562-3

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11104-012-1562-3

Keywords

Search

Navigation