LAUSR

Background and aims While large differences in microbial biomass and extracellular enzyme activities (EEAs) between rhizosphere and bulk soils have been demonstrated, the potentially different response of microbes and their EEAs in rhizosphere and bulk soils to nitrogen (N) deposition is still not elucidated. Methods We analyzed the microbial biomass and EEAs in the rhizosphere and bulk soils of Sibiraea angustata in an alpine shrubland on the eastern Qinghai-Tibet Plateau after chronic N application. We also analyzed the stoichiometric linkages between plants, microbes, enzymes and soils to clarify the coupled responses of aboveground plants and belowground ecological processes. Results Microbial nutrient concentrations and activities of EAAs responded differently to N addition in the rhizosphere and bulk soils. In the rhizosphere, N addition caused a significant increase in microbial biomass carbon (C), N and phosphorus (P) concentrations and greater P-degrading enzyme activity (relative to the activities of C- and N-degrading enzymes), which induced a significant reduction in enzyme C:P and N:P ratios. The rhizosphere enzyme N:P ratio was negatively correlated with the N:P ratios of plant, soil and microbe, implying that increased plant and microbial P uptake under N addition may gradually aggravate rhizosphere P limitation. However, for the bulk soil, N addition did not affect microbial biomass but significantly enhanced C-degrading enzyme activity and decreased the enzyme C:N ratio. Meanwhile, the bulk-soil enzyme C:N ratio was negatively correlated with the soil C:N ratio but independent of the plant C:N ratio, implying that N addition may enhance bulk-soil microbial C limitation. Conclusions Our study suggests that elevated N deposition may induce differential microbial nutrient limitation between the rhizosphere and bulk soils due to the plant-microbe-soil interactions in the rhizosphere. This study highlights the importance of incorporating rhizosphere microbial processes into biogeochemical models describing environmental changes.