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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

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Abstract

A plant mixture of white clover (Trifolium repens L.), red clover (Trifolium pratense L.), and ryegrass (Lolium perenne L.) was established in the spring of 1991 under a cover-crop of barley. Treatments were two levels of nitrogen (400 and 20 kg N ha-1) and two cutting intensities (3 and 6 cuts per season). Fixation of atmospheric derived nitrogen was estimated by two 15N dilution methods, one based on application of 15N to the soil, the other utilising small differences in natural abundance of 15N.

Both methods showed that application of 400 kg N ha-1 significantly reduced dinitrogen fixation, while cutting frequency had no effect. Atmospheric derived nitrogen constituted between 50 and 64% of harvested clover nitrogen in the high-N treatment, while between 73% and 96% of the harvested clover nitrogen was derived from the atmosphere in the low-N treatment. The amounts of fixed dinitrogen varied between 31–72 kg N ha-1 and 118–161 kg N ha-1 in the high-N and low-N treatment, respectively. The highest values for biological dinitrogen fixation were estimated by the enriched 15N dilution method.

Estimates of transfer of atmospheric derived nitrogen from clover to grass obtained by the natural 15N abundance method were consistently higher than those obtained by the enriched 15N dilution method. Neither mineral nitrogen application nor defoliation frequency affected transfer of atmospheric derived nitrogen from clover to grass.

Isotopic fractionation of 14N and 15N (B value) was estimated by comparing results for nitrogen fixation obtained by the enriched 15N dilution and the natural 15N abundance method, respectively. B was on average +1.20, which was in agreement with a B value determined by growing white clover in a nitrogen free media.

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Høgh-Jensen, H., Schjoerring, J.K. 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 (1994). https://doi.org/10.1007/BF00008328

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