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Importance of correct B value determination to quantify biological N2 fixation and N balances of faba beans (Vicia faba L.) via 15N natural abundance

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Abstract

Quantifying biologically fixed nitrogen (BNF) by legumes through the 15N natural abundance techniques requires correct determination of a so-called B value. We hypothesized that significant variations in B values exist between faba bean (Vicia faba L.) varieties having consequences for BNF and N balance calculations. We experimentally determined B values for a range of faba bean varieties and quantified to what extent variety has an effect on B values and hence BNF quantification. Seeds of six faba bean varieties released in Ethiopia were inoculated with Rhizobium fabae strain LMG 23997-19 and grown in vermiculite with an N-free nutrient solution in a growth room until full flowering. Total N and 15N content of nodules, roots, and shoot components was analyzed separately to determine the weighted whole plant 15N fractionation during N2 fixation, i.e., the B value. Owing to its large seed size and high N content, a correction for seed N was carried out. We then calculated the percentage of N derived from air (%Ndfa), BNF, and N balance for faba beans grown in the field using three B value scenarios (variety specific B value corrected for seed N, variety specific B value without seed N correction, and a literature derived B value). Whole plant seed N corrected B values were significantly different (P < 0.05) between varieties and varied between +0.5 ± 0.4 and −1.9 ± 1.4‰ suggesting a variable isotope fractionation during N2 fixation. The %Ndfa was significantly (P < 0.05) different between varieties (59 ± 4.2–84 ± 4.5 %) using seed N corrected B values. BNF (218 ± 26.2–362 ± 34.7 kg N ha−1) was significantly (P < 0.05) different between varieties for corrected and uncorrected B values. Soil N balance did not result in statistically significant (P > 0.05) difference between varieties for all three B value scenarios. Use of inappropriate B values masked the difference between varieties and affected their ranking in terms of BNF, resulting from an over- to underestimation of 15 and 19 %, respectively. When applying the 15N natural abundance technique to compare BNF of legume accessions, we recommend determining a B value for each accession. For legumes with large seeds such as faba beans, it is moreover essential to account for seed N when determining the B value.

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Acknowledgments

The authors would like to thank financial support from VLIR-UOS in the framework of an Inter University Cooperation program with Jimma University, Ethiopia. Katja Van Nieuland is duly acknowledged for 15N analysis. Mr. Daniel Damtew and Mr. Alemseged Amanuel are duly acknowledged for kindly assisting in data collection in the field.

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Authors and Affiliations

  1. College of Agriculture and Veterinary Medicine, Jimma University, POB 1316, Jimma, Ethiopia

    Amsalu Nebiyu

  2. Isotope Bioscience Laboratory—ISOFYS, Faculty of Biosciences Engineering, Ghent University, Coupure links 653, 9000, Gent, Belgium

    Dries Huygens, Hari Ram Upadhayay & Pascal Boeckx

  3. Department of Earth and Environmental Sciences, KU Leuven, Celestijnenlaan 200E, box 2411, 3001, Leuven, Belgium

    Jan Diels

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  1. Amsalu Nebiyu
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  4. Jan Diels
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  5. Pascal Boeckx
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Correspondence to Amsalu Nebiyu.

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Nebiyu, A., Huygens, D., Upadhayay, H.R. et al. Importance of correct B value determination to quantify biological N2 fixation and N balances of faba beans (Vicia faba L.) via 15N natural abundance. Biol Fertil Soils 50, 517–525 (2014). https://doi.org/10.1007/s00374-013-0874-7

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