Plant and Soil

, Volume 385, Issue 1–2, pp 287–302 | Cite as

Comparisons of biological nitrogen fixation in association with white clover (Trifolium repens L.) under four fertiliser nitrogen inputs as measured using two 15N techniques

  • W. Burchill
  • E. K. James
  • D. Li
  • G. J. Lanigan
  • M. Williams
  • P. P. M. Iannetta
  • J. Humphreys
Regular Article



The objectives of this study were to compare techniques for measuring biological nitrogen fixation (BNF) and to assess how fertiliser N input affects the balance between BNF and sustainable herbage production on perennial ryegrass (Lolium perenne L.)/white clover (Trifolium repens L.) grassland.


Biological N fixation and herbage production by white clover based grassland was measured in 2011 and 2012 under four nominal annual fertiliser N inputs: 0 (0N), 86 (86N), 140 (140N) and 280 kg ha−1 (280N). Biological N fixation was measured using the 15N isotope dilution and 15N natural abundance techniques under all fertiliser N inputs and also using the nitrogen difference technique under 0N.


The two 15N techniques produced similar annual estimates of above-ground BNF across the range of fertiliser N inputs. Fertiliser N input resulted in higher herbage dry matter yield, but reduced annual BNF which averaged 80, 64, 66 and 47 kg ha−1 on 0N, 86N, 140N and 280N, respectively, across both techniques and years.


The two 15N techniques were comparable in estimating BNF. Intermediate fertiliser N inputs achieved a balance between minimising detrimental impact on BNF and potential N loss while producing more herbage DM yield than clover swards receiving no fertiliser N.


15N techniques Isotope dilution Natural abundance Fertiliser nitrogen White clover N fixation 



This work was funded by the EU Framework Programme 7 project Legume Futures. The author thanks Laura Lopez del Egido and the Stable Isotope Facility at the James Hutton Institute for assistance with the B s -value analyses and Kevin McNamara, internship students and the staff of Solohead Research Farm for assistance with field sampling.

Supplementary material

11104_2014_2199_MOESM1_ESM.doc (57 kb)
Table S.1 Shoot δ15N values of perennial ryegrass and a combination of weed species in the June 2012 harvest and their effect on the proportion of N derived from the atmosphere (%Ndfa) in clover when used as reference plants using the 15N natural abundance technique on treatments receiving annual fertiliser N inputs of 0 (0N), 86 (86N), 140 (140N) and 280 (280N) kg ha−1. (DOC 57 kb)
11104_2014_2199_Fig7_ESM.gif (3 kb)
Fig S.1

Proportion of nitrogen derived from the atmosphere (%Ndfa) in white clover estimated by 15N isotope dilution (ID) data (white circle), 15N natural abundance data (black box) and 15N ID data using a B s -value (grey triangle) on the final three harvest dates and a supplementary harvest in March 2013 (mm–yy) on treatments receiving annual fertiliser N input of (a) 0, (B) 86, (c) 140 and (d) 280 kg ha−1. Error bars represents the interaction S.E.M of harvest date × technique. (GIF 3 kb)

11104_2014_2199_MOESM2_ESM.tif (32 kb)
High resolution image (TIFF 32 kb)


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Copyright information

© Springer International Publishing Switzerland 2014

Authors and Affiliations

  • W. Burchill
    • 1
    • 2
  • E. K. James
    • 3
  • D. Li
    • 4
  • G. J. Lanigan
    • 5
  • M. Williams
    • 2
  • P. P. M. Iannetta
    • 3
  • J. Humphreys
    • 1
  1. 1.Animal & Grassland Research and Innovation CentreCorkIreland
  2. 2.Department of Botany, School of Natural SciencesTrinity College DublinDublin 2Ireland
  3. 3.The James Hutton InstituteDundeeUK
  4. 4.Huanjiang Observation and Research Station for Karst Ecosystems, Key Laboratory of Agro-ecological Processes in Subtropical RegionInstitute of Subtropical Agriculture, Chinese Academy of SciencesChangshaChina
  5. 5.Johnstown Castle Environment Research CentreWexfordIreland

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