Reliable measurements of the input of fixed-N from forage legumes are essential in estimating the need for N fertilization in grasslands. Such measurements rely on methods that can be applied in the field for either the entire growing season or parts of it. High precision and reliability should always be aimed at when measuring N2 fixation. However, depending on the aims and availability of resources (time and equipment), methods with varying precision can be chosen. Methods with high precision are needed if the aim is to reveal factors that affect the N2-fixation rate, whereas N balances at the field or farm level may be based on N2 fixation estimates obtained with lower precision. Of the methods listed in Table 1, we only recommend 15N natural abundance (NA) and 15N isotope dilution (ID) for measurements with high precision. In addition, we propose that simple formulas based on legume biomass can be used for rapid estimates with lower precision. Nitrogen difference is not a reliable method for forage legumes because forage grasses usually are more efficient than legumes in taking up soil N (Table 1; Carlsson and Huss-Danell, 2003 and references therein).
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Carlsson, G., Huss-Danell, K. (2008). How to Quantify Biological Nitrogen Fixation in Forage Legumes in the Field. In: Dakora, F.D., Chimphango, S.B.M., Valentine, A.J., Elmerich, C., Newton, W.E. (eds) Biological Nitrogen Fixation: Towards Poverty Alleviation through Sustainable Agriculture. Current Plant Science and Biotechnology in Agriculture, vol 42. Springer, Dordrecht. https://doi.org/10.1007/978-1-4020-8252-8_14
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