Arbuscular mycorrhizal fungi (AMF) can preferentially absorb the released ammonium (NH₄⁺) over nitrate (NO₃⁻) during litter decomposition. However, the impact of AMF’s absorption of NH₄⁺ on litter nitrogen (N) decomposition is still unclear. In this study, we investigated the effects of AMF uptake for NH₄⁺ on litter N metabolic characteristics by enriching NH₄⁺ via AMF suppression and nitrification inhibition in a subtropical Cinnamomum camphora forest. The results showed that AMF suppression and nitrification inhibition significantly decelerated litter decomposition in the early stage due to the repression of NH₄⁺ in extracellular enzyme activity. In the late stage, when soil NH₄⁺ content was low, in contrast, they promoted litter decomposition by increasing the extracellular enzyme activities. Nitrification inhibition mainly promoted the utilization of plant-derived N by promoting the degradation of the amide I, amide II, and III bands by increasing protease activity, and it promoted ammonification by increasing urease activities, whereas it reduced the utilization of microbial-derived N by decreasing chitinase activity. On the contrary, AMF suppression, which significantly reduced the ammonification rate and increased the nitrification rate, only facilitated the degradation of the amide II band. Moreover, it intensified the microbial-derived N decomposition by increasing chitinase activity. The degradation of the amide I and II bands still relied on the priming effects of AMF on soil saprotrophs. This was likely driven by AMF-mediated phosphorus (P) mineralization. Nutrient acquiring, especially P by phosphatase, were the main factors in predicting litter decomposition and protein degradation. Thus, AMF could relieve the end-product repression of locally enriched NH₄⁺ in extracellular enzyme activity and promote early-stage litter decomposition. However, the promotive effects of AMF on litter protein degradation and NH₄⁺ release rely on P mineralization. Our results demonstrated that AMF could alleviate the N limitation for net primary production via accelerating litter N decomposition and reducing N loss. Moreover, they could restrict the decomposition of recalcitrant components by competing with saprotrophs for nutrients. Both pathways will contribute to C sequestration in forest ecosystems, which advances our understanding of AMF’s contribution to nutrient cycling and ecosystem processes in subtropical forests.

Clarifying the responses of microbial communities in distinct microhabitats like roots, the soil, and litter layers to secondary succession is critical for predicting the effects of global climate change on ecosystem functions. We investigated the microbial activities, compositions, and networks in these microhabitats of Fagus lucida forests ranging from 40 to 200 years. The results showed that soil physicochemical properties decreased with forest succession, except for NH₄⁺-N and available phosphorus, which decreased at the early stage. All vector angles of extracellular enzyme stoichiometry that were greater than 45° indicated that phosphorus was the key limiting element for microorganisms. The microbial community shifted from r- to K-strategists with forest succession, displaying the replacement of most bacterial phyla by Proteobacteria and Acidobacteriota, and an increase in the Acidobacteriota: Proteobacteria ratio, especially in the soil and litter layers. Soil properties, particularly NH₄⁺-N and pH, significantly affected the bacterial diversity and structure. Moreover, the bacterial network complexity increased with succession, particularly in the litter layer, and the topological properties of bacterial networks showed a stronger influence on microbial activities compared with those of fungal networks. The richness of keystone taxa in the litter layer was higher than in the soil layer and roots. However, the fungal community dominated by symbiotrophs showed lower sensitivity to soil nutrient changes and greater resilience to forest succession, displaying stable diversity and decreased network complexity, particularly in the roots. Ectomycorrhizal fungi (e.g., Russula) dominated the fungal guilds, and their abundance increased with forest succession, accompanied by a decrease in pathogenic fungi. Plant roots with significantly higher phosphatase activities played a stronger role than soils in structuring the litter microbial community, as reflected by similar carbon- and nitrogen-acquiring enzyme activities, microbial compositions, a greater share of taxa, and closer community distance. Our results revealed the increasingly important role of plant roots with forest succession in structuring the microbial community and nutrient cycling in the soil and litter layers.