Biochar and Nitrogen Synergistically Regulate Soil Carbon Mineralization by Enhancing Aggregate Stability and Altering Microbial Function in Intensive Vegetable Systems

Intensive nitrogen (N) fertilization in greenhouse vegetable systems degrades soil structure and accelerates soil carbon (C) mineralization. Biochar application can alleviate these adverse effects by enhancing aggregate stability and mediating microbially driven nutrient cycling, yet its effects across aggregate fractions remain poorly understood. Here, we investigated how biochar (0, 20, 40 t ha−1) and N interact to affect aggregate stability, C mineralization, nutrient status, and microbial properties in bulk soil and four aggregate classes (large macroaggregates: LMA, > 2000 μm; small macroaggregates: SMA, 250–2000 μm; microaggregates: MA, 53–250 μm; silt + clay: S + C, < 53 μm) in vegetable soil after a 60-day incubation. Results showed that biochar–N co-application increased mean weight diameter by 27.4–30.5% and elevated soil total organic C (TOC) in LMA by 9.11–12.0% and in MA by 8.77–20.2% relative to the N-only treatment. It also reduced β-glucosidase and oxidase activities, as well as fungal and G−-bacterial abundance. Biochar amendment suppressed TOC mineralization by 2.7–24.6% in bulk soil and aggregate fractions, while boosting potentially mineralizable C pools by 12.5–155.7%, and thereby increasing overall mineralization potential. Structural equation modeling revealed the size-dependent regulatory mechanisms underlying these observations. Aggregate stability directly inhibited CO2 emissions in bulk soil and SMA, while the effects in MA and S + C fractions were mediated by shifts in nutrient stoichiometry and hydrolase activities. Our findings clarified the size-dependent mechanisms by which biochar–N co-application promoted soil C sequestration, providing a theoretical basis for the sustainable management of intensive vegetable systems.