Ecosystem carbon (C) fluxes in terrestrial ecosystems are affected by varying environmental conditions (e.g., soil heterogeneity and weather) and land management. However, the interactions between soil respiration (Rs) and net ecosystem exchange (NEE) and their spatio-temporal dependence on environmental conditions and land management at field scale is not well understood. We performed repeated C flux measurement at 21 sites during the 2013 growing season in a temperate upland grassland in Germany, which was fertilized and cut three times according to the agricultural practice typical of the region. Repeated measurements included determination of NEE, Rs, leaf area index (LAI), meteorological conditions as well as physical and chemical soil properties. Temporal variability of Rs was controlled by air temperature, while LAI influenced the temporal variability of NEE. The three grass cuts reduced LAI and affected NEE markedly. More than 50% of NEE variability was explained by defoliation at field scale. Additionally, soil heterogeneity affected NEE, but to a lower extent (>30%), while Rs remained unaffected. We conclude that grassland management (i.e., repeated defoliation) and soil heterogeneity affects the spatio-temporal variability of NEE at field scale.