Isotope 15N enrichment of soil-ammonium N as a reference to estimate N2 fixation by stem and root-inoculated S. rostrata

Abstract

A pot experiment in the greenhouse was conducted to compare the contribution of N derived from the atmosphere or from biological N2 fixation by Sesbania rostrata inoculated with Azorhizobium caulinodans, applied either to roots or to roots and stems (single or multiple stem inoculation). Two subsequent crops were grown for 50 days under flooded conditions. N derived from air was estimated by 15N dilution using 15N enrichment of soil NH inf4 sup+ -N and of Echinochloa crusgalli as the non-N2-fixing reference datum and compared with estimates obtained by the N-difference method. The first crop was grown to stabilize the 15N into the soil organic N fraction. The 15N enrichment of soil NH inf4 sup+ -N in the second crop declined slowly. The extractability ratio (15N enrichment of extractable soil N to 15N enrichment of total soil N) decreased from 4.8 to 4.1 50 days after planting. The enrichment of soil NH inf4 sup+ -N was comparable to that of E. crus-galli, resulting in similar estimates of N derived from air when either soil NH inf4 sup+ -N or enrichment of E. crus-galli was used as a non-fixing reference. The N-difference method did not always provide reliable estimates of N derived from air; percentages ranged from 75 to more than 80 by 50 days after planting in both crops and did not differ among treatments. The study demonstrates the potential of using 15N enrichment of soil NH inf4 sup+ -N as a non-N2-fixing reference for reliable BNF estimates of crops in lowland puddled soil.