Abstract

Long-term fertilization regulates soil microbial communities and is essential for black soil health and sustainable productivity, yet its key drivers remain unclear. Using a 39-year field experiment, we evaluated the effects of four fertilization regimes: no fertilizer (CK), chemical fertilizer (NPK), organic fertilizer (M), and combined organic-inorganic fertilizer (MNPK). Soil properties and bacterial communities were analyzed using Illumina MiSeq sequencing, quantitative real-time PCR (qRT-PCR), and multivariate analyses. Proteobacteria, Actinobacteriota, Acidobacteriota, Chloroflexi, and Gemmatimonadota dominated (>80% of the community), and all treatments significantly altered their relative abundances. Compared with CK, NPK reduced soil pH by 8.3% and bacterial abundance by 29.7%, increased soil organic matter (SOM) by 22.9%, and decreased community evenness. MNPK reduced pH by only 2.0%, increased SOM by 53.8% and bacterial abundance by 38.9%, and improved community evenness, mitigating acidification while maintaining high diversity. M increased pH by 2.3%, SOM by 73.3%, and bacterial abundance by 71.8%. Soil pH, available phosphorus, and SOM were the main drivers of community structure. Overall, MNPK showed the strongest synergistic effects on soil fertility and microbial stability, making it an optimal strategy for sustainable black soil management.

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