Improved tillage management in fragrant rice cropping systems can enhance soil organic carbon (SOC) and the content of 2-Acetyl-1-Pyrroline (2-AP), a crucial volatile compound contributing to the aroma of fragrant rice. Despite this, the interplay between 2-AP content in fragrant rice and SOC metabolism, alongside the influences exerted by soil microbial functions, remains poorly understood. This study introduces a comprehensive 6-year field experiment which aims to correlate SOC with rice grain 2-AP content by analyzing soil microbial KEGG functions, such as carbon and amino acid metabolism, using metagenomic sequencing. The experiment assessed three tillage practices, conventional tillage (CT), reduced tillage (RT), and no tillage (NT), with soil samples collected on three dates in 2022. The results indicated that NT significantly (p < 0.05) enhanced SOC content and modified carbon metabolism by upregulating the Calvin cycle (K01601) and reducing hemicellulose degradation (K01710). Additionally, NT notably increased the soil levels of alkaline amino acids, such as histidine and ornithine, which were 165.17% and 1218.42% higher, respectively, than those in CT, possibly linked to an increase in soil pH. Furthermore, the 2-AP content in fragrant rice under NT was significantly higher by 52.02% and 13.90% compared to under RT and CT, respectively. NT also upregulated K00250 (alanine, aspartate, and glutamate metabolism) and K00290 (valine, leucine, and isoleucine biosynthesis), leading to significantly higher levels of 2-AP biosynthesis-related amino acids proline and glutamate in fragrant rice grain. This study links SOC and 2-AP biosynthesis via soil microbial functions, presenting a novel strategy for improving the quality of fragrant rice through soil management practices.