Nitrogen fixation in chickpea. II. Comparison of ¹⁵N enrichment and ¹⁵N natural abundance methods for estimating nitrogen fixation

Abstract

The 15N enrichment and 15N natural abundance methods for estimating N2 fixation in chickpea were compared over a range of soil NO3-N levels at crop establishment varying from 10 to 326 kg N/ha (0-120 cm depth). Barley was used as a non-N2 fixing control crop. Both methods estimated reduced N2 fixation as soil NO3-N levels at crop establishment increased. Similar estimates of % N2 fixation were obtained at high values, but at low values the enrichment method gave lower estimates, some of which were negative. The 15N natural abundance method provided realistic estimates of % N2 fixation across all soil N03-N levels at crop establishment. An asymptotic curve described a close ( R2 = 0.95) relationship between these factors. Standard errors of estimates of means for the 15N natural abundance method remained acceptable and relatively stable over the full range of measurements; however, with the 15N enrichment method they became unacceptably large at low values of % N2 fixation. These large errors may have been partly due to legume and control plants assimilating mineral N of differing 15N enrichment. High mineral N levels associated with low values of % N2 fixation were also shown to reduce reliability of N2 fixation values estimated by the 15N enrichment method. These errors caused potentially greater inaccuracy at low values of % N2 fixation than at high values. To compare N2 fixation means statistically, transformations were necessary to stabilize variance and to impart lower weightings to plots with low values of % N2 fixation.