Tree Species Effects on Microbial Nutrient Limitation in Afforested Mine Soils Revealed by Enzyme Stoichiometry and Substrate-Induced Respiration

Microbial nutrient cycling in afforested mine soils may be affected by the plant litter quality. This study investigated how different tree species—Scots pine (Pinus sylvestris), silver birch (Betula pendula), European larch (Larix decidua), and black alder (Alnus glutinosa)—influence microbial carbon (C), nitrogen (N), and phosphorus (P) limitations in reclaimed sandy mine soils. We combined substrate-induced respiration (SIR) and ecoenzymatic stoichiometry (EES) to diagnose these metabolic constraints. The SIR analysis revealed a universal primary limitation by labile C across all tree species, with glucose addition stimulating respiration by 271%–333%, regardless of the soil organic carbon content. However, EES revealed distinct secondary nutrient constraints driven by species-specific litter quality. Alder stands exhibited severe P limitation, likely due to high P demand for symbiotic N-fixation and intense competition for P between trees and microbes. In contrast, birch stands showed stoichiometric homeostasis and a slight N deficiency. Coniferous species exhibited P limitation and low enzymatic activity, indicating a strategy focused on intensive nutrient acquisition under low-energy conditions associated with recalcitrant needle litter. These findings demonstrate that while energy limitation is a universal constraint in mine soils, tree species determine the nature and intensity of secondary nutrient limitations due to differences in litter stoichiometry.