Arbuscular mycorrhizal fungi (AMF) represent an important route for plant carbon (C) inputs into the soil. Nonetheless, the C input via AMF as well as its impact on soil organic matter (SOM) stabilization and C sequestration remains largely unknown. A mycorrhizal wild type progenitor (MYC) and its mycorrhiza defective mutant (reduced mycorrhizal colonization: rmc) of tomato were continuously labeled with 13CO2 to trace root C inputs into the soil and quantify rhizosphere priming effects (RPE) as affected by AMF symbiosis and N fertilization. Mycorrhizal abundance and 13C incorporation into shoots, roots, soil and CO2 were measured at 8, 12 and 16 weeks after transplanting. AMF symbiosis decreased the relative C allocation (% of total assimilated C) to roots, in turn increased the net rhizodeposition. Positive RPE was recorded for both MYC and rmc plants, ranging from 16–71% and 25–101% of the unplanted control, respectively. Although net rhizodeposition was higher for MYC than rmc plants 16 weeks after transplanting, the RPE was comparatively lower. This indicated a higher potential for C sequestration by plants colonized with AMF (MYC) because the reduced nutrient availability restricts the activity of free-living decomposers. Although N fertilization decreased the relative C allocation to roots, rhizosphere and bulk soil, it had no effect on the absolute amount of rhizodeposition to the soil. The RPE and N-cycling enzyme activities decreased by N fertilization 8 and 12 weeks after transplanting, suggesting a lower microbial N demand from SOM mining. The positive relationship between enzyme activities involved in C cycling, microbial biomass C and SOM decomposition underlines the microbial activation hypothesis, which explains the RPE. We therefore concluded that AMF symbiosis and N fertilization increase C sequestration in soil not only by increasing root C inputs, but also by lowering native SOM decomposition and RPE. © 2019 Elsevier Ltd