The use of nitrogen-15 natural abundance and nitrogen yield of non-nodulating isolines to estimate nitrogen fixation by soybeans (Glycine max L.) across three elevations

Summary Dissimilarities in soil N uptake between N₂-fixing and reference non-N₂-fixing plants can lead to inaccurate N₂ fixation estimates by N difference and ¹⁵N enrichment methods. The natural ¹⁵N abundance ( ¹⁵N) method relies on a stabilized soil ¹⁵N pool and may provide reliable estimates of N₂ fixation. Estimates based on the ¹⁵N and differences in N yield of nodulating and non-nodulating isolines of soybean were compared in this study. Five soybeans from maturity groups 00, IV, VI, and VIII and their respective non-nodulating isolines were grown at three elevations differing in ambient temperature and soil N availability. Despite large differences in phenological development and N yield between the non-nodulating isolines, the ¹⁵N values measured on seeds were relatively constant within a site. The ¹⁵N method consistently produced lower N₂ fixation estimates than the N difference method, but only in three of the 15 observations did they differ significantly. The average crop N derived from N₂ fixation across sites and maturity groups was 81% by N difference compared to 71% by ¹⁵N. The magnitude of difference between the two methods increased with increasing proportions of N derived from N₂ fixation. These differences between the two methods were not related to differences in total N across sites or genotypes. The low N₂ fixation estimates based on ¹⁵N might indicate that the nodulating isolines had assimilated more soil N than the non-nodulating ones. A lower variance indicated that the estimates by N difference using non-nodulating isolines were more precise than those by ¹⁵N. Since the differences between the estimates were large only at high N₂ fixation levels (low soil N availability), either method may be used in most situations when a non-nodulating isoline is used as the reference plant. The ¹⁵N method may have a comparative advantage over N difference and ¹⁵N enrichment methods in the absence of a suitable non-N₂-fixing reference plant such as a non-nodulating isoline.