Soil Enzymes and Stable Isotopes as Suitable Soil–Plant Indicators of Ecosystem Functionality in Mediterranean Forests

Monitoring the soil–plant system in forest ecosystems is crucial for preserving their ecological functions and services. This study assessed carbon and nitrogen stable isotopes and ecoenzymatic stoichiometry as suitable indicators for characterizing the soil–plant system as a functional unit of ecological processes. To this end, in June 2021 six plots (1 m2 each) were selected in two typical Mediterranean forest ecotypes: a coastal stone pine forest (Pinus pinea L., PF) and a meso-hygrophilous broadleaf forest (RV). Soil samples (0–15 and 15–30 cm depth) and litter samples (40 × 40 cm) were collected and characterized in terms of physical, chemical and biochemical properties. t-tests revealed significant differences between RV and PF, indicating distinct microbial nutrient acquisition strategies. The higher C:N ratio in PF suggested lower litter quality and greater recalcitrance to microbial decomposition. Consistently, RV showed a more pronounced 13C and 15N enrichment from litter to SOM down to a 30 cm depth, confirming faster organic matter decomposition and mineralization. Enzyme activity patterns supported these findings. The higher β-glucosidase and butyrate esterase activities in RV reflected its greater microbial potential to activate biogeochemical cycles. Both forests exhibited a higher microbial demand for C and P than for N to maintain ecological stoichiometric balance, with stronger C limitation at the surface and P limitation in the subsoil, particularly in RV soil. This integrated monitoring approach provides insights into nutrient cycling and ecosystem resilience and offers tools to evaluate ecosystem functionality under changing environmental conditions, supporting sustainable forest management.