Low molecular weight carbon (C) compounds in hotspots such as the rhizosphere can greatly affect nitrate reduction processes. Towards a better prediction of N2O emission from denitrification, we are still lacking understanding of responses to the supply of complex C compound mixtures such as rhizodeposits versus the often examined response to individually amended C compounds. In a laboratory study, we applied three C compounds, glucose, citric acid and glutamic acid, individually or as a three-compound mixture to 14NH415NO3 amended soil at 80% water-filled pore space. For the individual C compound treatments, the substrateswere enriched in 13C-C. The mixture was enriched in 13C-C either in all constituent compounds or in one of the compounds only, resulting in four different treatments. This set-up enabled quantification of the utilization of each compound for heterotrophic respiration when applied individually and in combination, and for this to be related to the dynamics of 15N-NO3- reduction to 15N-N2O. The total 15N-N2O emission from the compound mixture over 10 days was similar to the total emission predicted from the average of the individual compound treatments This could suggest potential predictability of denitrification responses to complex mixtures of C based on knowledge of its constituents. However, immediate and simultaneous peaks of 15N-N2O and 13C-CO2 fluxes from the compound mixture contrasted with observed delays in 15N-N2O and 13C-CO2 fluxes when the compounds had been applied individually. Moreover, relative contributions of the C compounds to 13C-CO2 respiration from the compound mixture were different from the predicted contributions based on their individual application. While contributions of glutamic acid-C and citric acid-C to respiration in mixture during peak 15N-N2O emission were increased, glucose utilization in the mixture treatment was significantly lower. These findings give a glimpse of the challenges we are facing when trying to predict nitrate reduction occurring in the rhizosphere where interactions between C compounds and the soil matrix, as well as within the wider heterotrophic community, determine process rates. Given that most of our understanding of the role of C in regulating nitrate reduction, is informed from single compound studies, we require more evidence on the effects and innate interactions of compound mixtures to be able to predict responses to C sources
