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Nitrogen (N) deposition is a component of global change that has considerable impact on belowground carbon (C) dynamics. Plant growth stimulation and alterations of fungal community composition and functions are the main mechanisms driving soil C gains following N deposition in N-limited temperate forests. In N-rich tropical forests, however, N deposition generally has minor effects on plant growth; consequently, C storage in soil may strongly depend on the microbial processes that drive litter and soil organic matter decomposition. Here, we investigated how microbial functions in old-growth tropical forest soil responded to 13 years of N addition at four rates: 0 (Control), 50 (Low-N), 100 (Medium-N), and 150 (High-N) kg N ha-1  year-1 . Soil organic carbon (SOC) content increased under High-N, corresponding to a 33% decrease in CO2 efflux, and reductions in relative abundances of bacteria as well as genes responsible for cellulose and chitin degradation. A 113% increase in N2 O emission was positively correlated with soil acidification and an increase in the relative abundances of denitrification genes (narG and norB). Soil acidification induced by N addition decreased available P concentrations, and was associated with reductions in the relative abundance of phytase. The decreased relative abundance of bacteria and key functional gene groups for C degradation were related to slower SOC decomposition, indicating the key mechanisms driving SOC accumulation in the tropical forest soil subjected to High-N addition. However, changes in microbial functional groups associated with N and P cycling led to coincidentally large increases in N2 O emissions, and exacerbated soil P deficiency. These two factors partially offset the perceived beneficial effects of N addition on SOC storage in tropical forest soils. These findings suggest a potential to incorporate microbial community and functions into Earth system models considering their effects on greenhouse gas emission, biogeochemical processes, and biodiversity of tropical ecosystems.

Nitrogen (N) deposition is a component of global change that has considerable impact on belowground carbon (C) dynamics. Plant growth stimulation and alterations of fungal community composition and functions are the main mechanisms driving soil C gains following N deposition in N-limited temperate forests. In N-rich tropical forests, however, N deposition generally has minor effects on plant growth; consequently, C storage in soil may strongly depend on the microbial processes that drive litter and soil organic matter decomposition. Here, we investigated how microbial functions in old-growth tropical forest soil responded to 13 years of N addition at four rates: 0 (Control), 50 (Low-N), 100 (Medium-N), and 150 (High-N) kg N ha-1  year-1 . Soil organic carbon (SOC) content increased under High-N, corresponding to a 33% decrease in CO2 efflux, and reductions in relative abundances of bacteria as well as genes responsible for cellulose and chitin degradation. A 113% increase in N2 O emission was positively correlated with soil acidification and an increase in the relative abundances of denitrification genes (narG and norB). Soil acidification induced by N addition decreased available P concentrations, and was associated with reductions in the relative abundance of phytase. The decreased relative abundance of bacteria and key functional gene groups for C degradation were related to slower SOC decomposition, indicating the key mechanisms driving SOC accumulation in the tropical forest soil subjected to High-N addition. However, changes in microbial functional groups associated with N and P cycling led to coincidentally large increases in N2 O emissions, and exacerbated soil P deficiency. These two factors partially offset the perceived beneficial effects of N addition on SOC storage in tropical forest soils. These findings suggest a potential to incorporate microbial community and functions into Earth system models considering their effects on greenhouse gas emission, biogeochemical processes, and biodiversity of tropical ecosystems.

Nitrogen (N) availability limits plant growth and soil carbon (C) sequestration in N‐limited ecosystems, however, plant and soil C feedback on the free‐living nitrogen fixation (FLNF) process is poorly understood. Moreover, whether this feedback is influenced by initial N availability in leguminous and non‐leguminous forest soils has not been clarified. Here, we found that the addition of plant‐ and microbial‐derived C significantly enhanced soil nitrogenase activity (13∼28%) and that microbial‐derived C had a more positive impact. These positive effects were attributed to the direct C‐energy supply (0.49∼0.84) rather than variations in soil microbial activity (−0.01∼0.21) and substrate resources (−0.45∼0.27). Long‐term N addition did not inhibit FLNF. C addition promoted FLNF in soils of the two forests, but the response rate was higher in the leguminous forest soils. Our study reveals that increased soil C availability can drive FLNF in tropical forests, enhancing our understanding of the soil C‐N coupling mechanism. Plain Language Summary: Explicit consideration of controlling factors for free‐living nitrogen fixation (an important process of nitrogen biogeochemistry), is critical to understanding the response of ecosystems to environmental change. Our study reveals a previously overlooked but important process in which increased soil carbon availability exerts positive feedback on soil nitrogen inputs via free‐living nitrogen fixation. This insight enhances our understanding of soil carbon‐nitrogen coupling mechanisms and highlights an important nitrogen source in tropical forests based on increased vegetation and soil carbon sink conditions under global environmental change scenario. Key Points: Carbon (C) input affected free‐living nitrogen fixation (FLNF) more than nitrogen (N) application in tropical forest soilsBoth plant‐ and microbial‐derived C enhanced FLNF, but microbial‐derived C had a more positive impactC addition promoted FLNF in soils of the two forests, but the response rate was higher in the leguminous forest soils [ABSTRACT FROM AUTHOR]

Highly weathered lowland (sub)tropical forests are widely recognized as nitrogen (N)‐rich and phosphorus (P)‐poor, and the input of N and P affects soil carbon (C) cycling and storage in these ecosystems. Microbial residual C (MRC) plays a crucial role in regulating soil organic C (SOC) stability in forest soils. However, the effects of long‐term N and P addition on soil MRC across different soil layers remain unclear. This study conducted a 12‐year N and P addition experiment in two typical subtropical plantation forests dominated by Acacia auriculiformis and Eucalyptus urophylla trees, respectively. We measured plant C input (fine root biomass, fine root C, and litter C), microbial community structure, enzyme activity (C/N/P‐cycling enzymes), mineral properties, and MRC. Our results showed that continuous P addition reduced MRC in the subsoil (20–40 cm) of both plantations (A. auriculiformis: 28.44% and E. urophylla: 28.29%), whereas no significant changes occurred in the topsoil (0–20 cm). N addition decreased MRC in the subsoil of E. urophylla (25.44%), but had no significant effects on A. auriculiformis. Combined N and P addition reduced MRC (34.63%) in the subsoil of A. auriculiformis but not in that of E. urophylla. The factors regulating MRC varied across soil layers. In the topsoil (0–10 cm), plant C input (the relative contributions to the total variance was 20%, hereafter) and mineral protection (47.2%) were dominant factors. In the soil layer of 10–20 cm, both microbial characteristics (41.3%) and mineral protection (32.3%) had substantial effects, whereas the deeper layer (20–40 cm) was predominantly regulated by microbial characteristics (37.9%) and mineral protection (18.8%). Understanding differential drivers of MRC across soil depth, particularly in deeper soil layers, is crucial for accurately predicting the stability and storage of SOC and its responses to chronic N enrichment and/or increased P limitation in (sub)tropical forests. [ABSTRACT FROM AUTHOR]