Glomalin-related soil proteins (GRSP) play a crucial role in strengthening soil structure and increasing carbon (C) storage. However, the chemical stability of GRSP and related arbuscular mycorrhizal fungi (AMF) community response to fertilization remains unclear. This study investigated C and nitrogen (N) contents, three-dimensional fluorescence characteristics in GRSP, and AMF properties based on a field experiment that was subjected to 29 years of various fertilizations. The experiment included treatments with no fertilizer (CK), chemical fertilizer (NPK), manure (M), and manure combined with NPK (NPKM) treatments. Results showed that GRSP contained 37–49% C and 6–9% N, respectively. Compared with CK and NPK, the C and N proportions in GRSP significantly increased under M and NPKM. Using the parallel factor model, four fluorescent components of GRSP were identified: one fulvic acid-like component (C2), one tyrosine-like component (C4), and two humic acid-like components (C1, C3). Under M and NPKM, the fluorescent intensity of C2 and C4 decreased, while the humification index (HIX) increased relative to CK and NPK, indicating that organic fertilization could enhance the stability of GRSP. The C and N proportion in GRSP positively associated with soil organic C (SOC), total N (TN), available phosphorus (AP), AMF biomass, and diversity, while C2 and C4 showed negative associations. Structural equation modeling further revealed that manure-induced changes in pH, SOC, TN, and AP increased AMF biomass and diversity, thereby altering GRSP composition and stability. This study provides valuable insights into the compositional traits of GRSP, contributing to sustainable soil management and C sequestration in agroecosystems.
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