BackgroundIn farmland, microbes in soils are affected by exogenous carbon, nitrogen, and soil depth and are responsible for soil organic carbon (SOC) mineralization. The cherry industry has been evolving rapidly in northwest China and emerged as a new source of income for local farmers to overcome poverty. Accordingly, it is highly imperative to probe the effect of defoliation and nitrogen addition on carbon dioxide (CO2) emissions and microbial communities in soils of dryland cherry orchards.MethodsCO2emissions and microbial communities were determined in soil samples at three depths, including 0–10 cm, 10–30 cm, and 30–60 cm, from a 15-year-old rain-fed cherry orchard. The samples were respectively incubated with or without 1% defoliation under three input levels of nitrogen (0 mg kg−1, 90 mg kg−1, and 135 mg kg−1) at 25°C in the dark for 80 days.ResultsDefoliation and nitrogen addition affected CO2emissions and microbial communities and increased microbial biomass carbon (MBC), the activity of soil catalase, alkaline phosphatase, and cellulase in soils of the dryland cherry orchard. The culture with defoliation significantly promoted CO2emissions in soils at the three depths mainly by increasing the MBC, catalase, alkaline phosphatase, and cellulase activities, resulted in positive priming index. Nitrogen addition elevated the MBC and changed soil enzymes and reduced CO2emissions in soils at the three depths. Moreover, the priming index was higher in deep soils than in top and middle soils under the condition of defoliation and nitrogen addition. No significant differences were observed in the soil bacterial diversity (Chao1, Shannon, and Simpson) among all treatments. Meanwhile, the relative abundance ofProteobacteriawas markedly increased and that ofAcidobacteriawas substantially diminished in soils at the three depths by defoliation and nitrogen addition. The results sustained that defoliation and nitrogen can regulate SOC dynamics by directly and indirectly affecting soil microbial activities and communities. As a result, the combination of defoliation return and nitrogen fertilization management is a promising strategy to increase SOC and promote soil quality in dryland cherry orchards.