Interactive effects of elevated atmospheric CO# and soil N availability on N # fixation and biomass production were examined using Gliricidia sepium, a tropical leguminous tree species. Our objective was to determine if elevated CO # alters the inhibitory effects of soil N on N # fixation, and whether the response of Gliricidia to elevated CO # was a function of N source originating from either substrate N fertilizer or N # fixation. We hypothesized that CO # enrichment would ameliorate the inhibitory effects of N fertilization on seedling nodulation and N # fixation through increased C partitioning to nodules. Seedlings were grown from seed for 100 d in growth chambers at either 350 or 700 µmol mol −" CO # . Seedlings were inoculated with Rhizobium spp. and grown either with 0, 1 or 10 mM N fertilizer. The δ"&N isotope-dilution technique was used to determine N source partitioning between N # fixation and inorganic N fertilizer uptake. The addition of 10 mM N fertilizer significantly reduced nodule number and mass, specific nitrogenase activity, the specific rate of N # fixation, and the proportion of plant N derived from N # fixation. Elevated CO # , however, strongly ameliorated the inhibitory effects of N fertilization, indicating that increased C availability for nodule activity may partially offset the inhibition of N # fixation caused by substrate N, as nodule sugar concentrations were stimulated with CO # enrichment. This study clearly shows that elevated CO # enhanced plant productivity and net N content of Gliricidia tree seedlings by stimulating N # fixation. In addition, seedling biomass production was greatly enhanced by elevated CO # , regardless of whether plant N was derived from the substrate or from the atmosphere. We conclude from this study that CO # enrichment mitigates the inhibitory effects of substrate N on nodule initiation and development and specific N # fixation, either through increased C allocation to nodule production and activity, or through increased N demand by the plant for biomass production. This experiment with Gliricidia provides evidence for a positive feedback between increased atmospheric CO # concentrations, C allocation to symbiotic N # -fixing bacteria, and plant C and N accumulation that may occur when N # -fixing plants are grown under conditions where substrate N may typically inhibit N # fixation.