Widespread lead (Pb) contamination of agricultural soils is a global issue stemming from human activities. The remediation of Pb-contaminated soils used for agricultural purposes is critically important to safeguard food crop safety. Despite the modulating effects of sulfur (S) on plant responses to toxic heavy metals, the ecological, physiological, and molecular mechanisms driving such modulation in the Pb hyperaccumulator Arabis alpina L. remain unclear. Here, we investigated the effects of five S concentrations (0, 50, 100, 150, and 200 mg kg−1) on A. alpina grown in Pb-contaminated soil from a lead-zinc mining area. Under S50 (i.e., 50 mg kg−1) and S100 treatments, the Pb concentration in both shoots and roots of A. alpina significantly decreased compared to the control (S0). Specifically, the S50 treatment significantly enhanced Pb accumulation, plant biomass, and plant height, indicating that low S applications facilitate Pb accumulation from the soil and alleviate Pb toxicity. Additionally, S50, S100, and S150 treatments significantly improved photosynthetic rate, stomatal conductance, and intercellular CO2 concentration in A. alpina. Transcriptomic analysis showed that S50 and S100 treatments increased the expression of the LHCA, LHCB, psa, and psb genes, which had a significant impact on photosynthetic efficiency. S50 and S100 boosted glutathione (GSH) levels in A. alpina roots, and the increased expression of GST gene enhanced tolerance to environmental stress. In summary, these results suggest that an appropriate supply of S (S50 and S100) not only alleviates Pb toxicity by enhancing plant biomass, height, photosynthetic features, and sulfur metabolites but also stimulates Pb accumulation in the hyperaccumulator A. alpina. Our study elucidated the specific concentrations of sulfur that optimally enhance both Pb accumulation and stress tolerance in the hyperaccumulator A. alpina, providing novel insights into the practical application of sulfur in phytoremediation strategies and advancing our understanding of the underlying molecular mechanisms. Display omitted •Sulfur supply enhanced plant growth and lead accumulation simultaneously.•S supply increased photosynthetic efficiency and expression of related genes.•S supply boosted GSH level, enhanced GST expression, improved plant tolerance.