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The effects of elevated CO 2  (750 ppm vs. 390 ppm) were evaluated on nitrogen (N) acquisition and assimilation by three  Medicago truncatula  genotypes, including two N-fixing-deficient mutants ( dnf1-1  and  dnf1-2 ) and their wild-type (Jemalong). The proportion of N acquisition from atmosphere and soil were quantified by  15 N stable isotope, and N transportation and assimilation-related genes and enzymes were determined by qPCR and biochemical analysis. Elevated CO 2  decreased nitrate uptake from soil in all three plant genotypes by down-regulating nitrate reductase ( NR ), nitrate transporter  NRT1.1  and NR activity. Jemalong plant, however, produced more nodules, up-regulated N-fixation-related genes and enhanced percentage of N derived from fixation (%Ndf) to increase foliar N concentration and N content in whole plant (Ntotal Yield) to satisfy the requirement of larger biomass under elevated CO 2 . In contrast, both  dnf1  mutants deficient in N fixation consequently decreased activity of glutamine synthetase/glutamate synthase (GS/GOGAT) and N concentration under elevated CO 2 . Our results suggest that elevated CO 2  is likely to modify N acquisition of  M. truncatula  by simultaneously increasing N fixation and reducing nitrate uptake from soil. We propose that elevated CO 2  causes legumes to rely more on N fixation than on N uptake from soil to satisfy N requirements.

Elevated CO2 Modifies N Acquisition of Medicago truncatula by Enhancing N Fixation and Reducing Nitrate Uptake from Soil