Reducing chemical fertilizers and biochar amendment is essential for achieving carbon neutrality, addressing global warming, and promoting sustainable agricultural development. Biochar amendment, a carbon rich soil additive produced through biomass pyrolysis, enhances soil fertility, increases crop yield, and improves soil carbon storage. However, research on the combined effect of fertilizer reduction and biochar amendment on soil mineral associated organic carbon (MAOC) in jasmine gardens is limited. This study aims to determine if biochar can reduce industrial fertilizer usage without compromising soil quality. This study focuses on jasmine cultivation in southeastern China, employing four treatments: conventional fertilization (CK), biochar amendment without fertilizer (BA), fertilizer reduction (FR), and fertilizer reduction with biochar amendment (FRBA). The effects on MAOC, microbial abundance, and enzyme activity were investigated. The FRBA treatment significantly increased MAOC content by 19.98 % compared to CK (P < 0.05). The BA and FRBA treatments enhanced the diversity of soil bacteria, including Lactobacillus, Azospirillum, and Cutibacterium, which are associated with soil organic carbon sequestration and nutrient decomposition. The RandomForest model identified β-N-acetyl-glucosaminidase (NAG), electric conductivity (EC), β-1, 4-Glucosidase (BG), soil potential of Hydrogen (pH), soil bulk density (BD), and β-D-cellobiosidase (CBH) as key soil traits promoting MAOC accumulation (P < 0.05). The results indicate that BA and FRBA improve soil bacterial community structure, enzyme activity, and MAOC content, promoting soil carbon accumulation through environmental factors and dominant bacteria. This study encourages future fertilization protocols that enhance fertilizer efficiency and carbon storage in crop soils.