Microbial performance under increasing nitrogen availability in a Mediterranean forest soil

In forest ecosystems, the external nitrogen (N) inputs mainly involve wet and dry depositions that potentially alter inorganic N availability in the soil and carbon (C) turnover. This study assesses the effect of a slow increase of inorganic N availability on microbial community activity and functionality in a Mediterranean forest soil. A four-month incubation experiment was performed with soil collected from the organic layer of a forest site and fertilized with a solution of ammonium nitrate. The fertilizer was supplied at an equivalent of 0, 10, 25, 50 and 75 kg N ha−1 (0, 0.3, 0.7, 1.3 and 2 mg N g−1 for control N0 and treatments N1, N2, N3 and N4, respectively). The incubation was carried out under optimal conditions, with the addition of the nutritive solution in small aliquots once a week to mimic the phenomenon of N deposition. In order to isolate the effect of N, the pH of the NH4NO3 solutions was adjusted to soil pH, and phosphorus was added in order to prevent any nutrient limitation effect. Inorganic N, C-mineralization, the activity of one oxidative enzyme (o-diphenol oxidase) and 8 hydrolitic enzymes (α-glucosidase, β-glucosidase, N-acetyl-β-d-glucosaminidase, cellulase, leucine amino-peptidase, acid phosphatase, butyric esterase and β-xylosidase) and the community level physiological profile (CLPP) were measured and analyzed during the whole incubation and at the end of the experiment as a proxy for microbial decomposition activity. In the first month, the highest N availability (N4) repressed the microbial respiration activity but stimulated microbial enzymatic activity, suggesting a change of C-pathways from spilling to enzymes and biomass investment. The treatments N1, N2 and N3 had no effect in the same period. Throughout the incubation, a general stress condition affected all the treated soils. As a consequence, treated soils exhibited higher respiration rates than the control. This was accompanied by a loss of functional diversity and an end-detected decline in biomass C. Although at the end of incubation most of the soil features showed a clear correlation with the inorganic N pool, the organic C content was strongly affected by different patterns of microbial activity during the experiment: the highest N treatment (N4) showed a lower C loss than the N3 treatment. Overall, the experiment showed how inorganic N availability can potentially alter the C cycle in a Mediterranean forest soil. The effect is non linear, depending on microbial community dynamics, on the community’s ability to adapt given the time scale of the process, and on N supply amount. Our study also revealed a common pattern in the short-term response to N addition in other, similar ecosystems with different climatic conditions.