Using "Suizhu" strawberries as test materials, two cultivation modes (soil-based and substrate-based) were established. Zinc sulfate solutions at concentrations of 0% (CK), 0.1% (Zn 0.1), 0.2% (Zn 0.2), and 0.3% (Zn 0.3) were applied via foliar spray during the initial flowering stage. Fruits were harvested in two separate batches. The contents of zinc, iron manganese, copper, and molybdenum in the strawberry fruits were determined using Inductively Coupled Plasma Mass Spectrometry. Combined with Random Forest modeling, Principal Component Analysis, and Three-way ANOVA, this study investigated the effects of foliar zinc application on trace element accumulation and mineral nutritional quality under different cultivation modes. The results indicated that foliar zinc application significantly increased the Zn content in strawberry fruits, with the biofortification efficiency significantly influenced by the ZnSO4 concentration, cultivation mode, and harvest batch. In the soil-based mode, the effectiveness of zinc fortification showed significant batch variations: the highest Zn content in the first batch was observed under the 0.2% treatment, increasing by 181% compared to the control (P < 0.05), while only the 0.3% treatment was effective for the second batch, increasing by 122% (P < 0.05). In the substrate-based mode, both batches exhibited a consistent dose-response effect, with the 0.3% treatment reaching the highest Zn content, significantly increasing by 151% and 125% respectively compared to the control (P < 0.05). Zinc fortification triggered changes in multi-element concentrations: a synergistic effect was observed between Zn and Mn accumulation, while an antagonistic effect was found between Zn and Fe. The accumulation of Cu and Mo was significantly affected by the interaction between cultivation mode and harvest batch. Evaluation using the General Index of Nutritional Quality (GINQ) showed that the second batch of fruits under substrate cultivation with 0.3% foliar Zn achieved the optimal mineral nutritional quality, with a GINQ value of 5.94. These findings suggest that substrate cultivation combined with appropriate foliar Zn concentrations can achieve efficient and sustainable Zn biofortification while maintaining high mineral nutritional quality. For soil-cultivated strawberries, management should prioritize the fortification of the first batch and address the Zn-Fe antagonism. This study provides a theoretical basis and technical reference for zinc biofortification and nutrition-oriented production in strawberries.