Abstract: To optimize nutrient assimilation and the accuracy of fertilization regimes in Prunus persica, the‘Jinlinghuanglu’ cultivar was used as experimental material. A field investigation incorporating a two-factor split-plot design, comparing conventional versus stratified fertilization and straw mulching versus clean tillage, was conducted in the Yellow River oldchannel region. This research elucidated the modulatory influences of these agronomic techniques on soil fertility parameters, enzymatic kinetics, shoot elongation, and fruit physicochemical properties within soil profile of the peach orchard. The results showed that: (1) Straw mulching can significantly improve the soil fertility of the 0~20 cm surface layer. The increase in organic matter content and EC value is relatively high, followed by the contents of total nitrogen, available phosphorus and available potassium. Straw mulching facilitated the optimization of the soil microhabitat, stimulated soil enzymatic activity, increased the proliferation of fruiting spurs, and promoted the ontogenetic shift from vegetative to reproductive development in peach trees. (2) Stratified fertilization proved efficacious in alleviating localized nutrient sequestration prevalent with conventional single-layer applications, thereby diminishing the spatial heterogeneity of nutrient concentrations, pH, and EC throughout the soil profile. This strategy promoted nutrient translocation into deeper soil horizons, enhanced nutrient bioavailability and biological utilization in the lower strata, improved nutrient uptake by peach trees, and ultimately augmented fruit quality. (3) Correlation analysis among organic matter, nutrient elements, and enzymatic activity demonstrated discernible layer-specific differentials. Surface organic matter exerted a beneficial influence on nutrient enrichment, whereas deep-stratum organic matter augmented the activity of soil functional enzymes. In conclusion, straw mulching promoted the fertility and functional activation of the topsoil, while stratified fertilization achieved a homogenized nutrient distribution within the tilled layer. The synergistic application of these two methodologies effectively ameliorated the orchard soil environment, thereby underpinning the high-quality and highefficiency cultivation of peach trees.