The severity of drought is predicted to increase across Europe due to climate change. Droughts can substantially impact terrestrial nitrogen (N) cycling and the corresponding microbial communities. Here, we investigated how ammonia-oxidizing bacteria (AOB), archaea (AOA), and complete ammonia oxidizers (comammox) as well as inorganic N pools and N2O fluxes respond to simulated drought under different cropping systems. A rain-out shelter experiment was conducted as part of a long-term field experiment comparing cropping systems that differed mainly in fertilization strategy (organic, mineral, or mixed mineral and organic) and plant protection management (biodynamic versus conventional pesticide use). We found that the effect of drought varied depending on the specific ammonia-oxidizing (AO) groups and the type of cropping system. Drought had the greatest impact on the structure of the AOA community compared to the other AO groups. The abundance of ammonia oxidizers was also affected by drought, with comammox clade B exhibiting the highest sensitivity. Additionally, drought had, overall, a stronger impact on the AO community structure in the biodynamic cropping system than in the mixed and mineral-fertilized conventional systems. The responses of ammonia-oxidizing communities to drought were comparable between bulk soil and rhizosphere. We observed a significant increase in NH4+ and NO3− pools during the drought period, which then decreased after rewetting, indicating a strong resilience. We further found that drought altered the complex relationships between AO communities and mineral N pools, as well as N2O fluxes. These results highlight the importance of agricultural management practices in influencing the response of nitrogen cycling guilds and their processes to drought.
