Nitrogen nutrition effects on δ13C of plant respired CO2 are mostly caused by concurrent changes in organic acid utilisation and remobilisation.

Nitrogen (N) nutrition impacts on primary carbon metabolism and can lead to changes in δ13C of respired CO2. However, uncertainty remains as to whether (1) the effect of N nutrition is observed in all species, (2) N source also impacts on respired CO2 in roots and (3) a metabolic model can be constructed to predict δ13C of respired CO2 under different N sources. Here, we carried out isotopic measurements of respired CO2 and various metabolites using two species (spinach, French bean) grown under different NH4+:NO3− ratios. Both species showed a similar pattern, with a progressive 13C‐depletion in leaf‐respired CO2 as the ammonium proportion increased, while δ13C in root‐respired CO2 showed little change. Supervised multivariate analysis showed that δ13C of respired CO2 was mostly determined by organic acid (malate, citrate) metabolism, in both leaves and roots. We then took advantage of nonstationary, two‐pool modelling that explained 73% of variance in δ13C in respired CO2. It demonstrates the critical role of the balance between the utilisation of respiratory intermediates and the remobilisation of stored organic acids, regardless of anaplerotic bicarbonate fixation by phosphoenolpyruvate carboxylase and the organ considered. Summary statement: Phaseolus vulgaris and Spinacia oleracea grown under varying NH4+:NO3− ratio show 13C‐depletion in leaf‐respired CO2 as NH4+ proportion increases. Supervised multivariate analysis shows that δ13C of respired CO2 is mostly determined by malate‐citrate metabolism in leaves and roots of both species. [ABSTRACT FROM AUTHOR]