Time- and age-related effects of experimentally simulated nitrogen deposition on the functioning of montane heathland ecosystems.

Ecosystems adapted to low nitrogen (N) conditions such as Calluna-heathlands are especially sensitive to enhanced atmospheric N deposition that affects many aspects of ecosystem functioning like nutrient cycling, soil properties and plant-microbial-enzyme relationships. We investigated the effects of five levels of experimentally-simulated N deposition rates (i.e., N fertilization treatments: 0, 10, 20 and 50kgNha-1yr-1 for 3years, and 56kgNha-1yr-1 for 10years) on: plant, litter, microbial biomass and soil nutrient contents, soil extracellular enzymatic activities, and plant root ericoid mycorrhizal colonization. The study was conducted in marginal montane Calluna-heathlands at different developmental stages resulting from management (young/building-phase and mature-phase). Our findings revealed that many soil properties did not show a statistically significant response to the experimental addition of N, including: total N, organic carbon (C), C:N ratio, extractable N-NO3-, available phosphorus (P), urease and β-glucosidase enzyme activities, and microbial biomass C and N. Our results also evidenced a considerable positive impact of chronic (10-year) high-N loading on soil extractable N-NH4+, acid phosphatase enzyme activity, Calluna root mycorrhizal colonization by ericoid fungi, Calluna shoot N and P contents, and litter N content and N:P ratio. The age of heathland vegetation influenced the effects of N addition on ericoid mycorrhizal colonization, resulting in higher colonized roots in young heathlands at the control, low and medium N-input rates; and in mature ones at the high and chronically high N rates. Also, young heathlands exhibited greater soil extractable N-NO3-, available P, microbial biomass N, Calluna shoot N and P contents, and litter N content, compared to mature ones. Our results highlighted that accounting for the N-input load and duration, as well as the developmental stage of the vegetation, is important for assessing the effects of added N, particularly at the heathlands' southern distribution limit.