Abstract: This study investigates the physicochemical properties, microbial community diversity, species composition, and environmental drivers of pyrite tailings from 3 locations: Yunfu City in Guangdong (GD), Hefei City in Anhui (AH), and Ankang City in Shaanxi (SX) in China. Results indicate significant differences (P < 0.05) in pH, NAG-pH, and total and available heavy metal contents among regions. GD pyrite tailings exhibit a significantly higher microbial Shannon index compared to AH and SX (P < 0.05), and distinct microbial community compositions (P < 0.05). Dominant phyla include Proteobacteria and Actinomycetota, while key genera such as Ralstonia, Sulfobacillus, and Leptospirillum demonstrate acid tolerance, heavy metal resistance, and iron-sulfur oxidation. The main environmental drivers influencing microbial community structure and composition include average annual temperature, average annual precipitation, pH, γ, DTPA-Zn, DTPA-Cr, and Cu. These findings provide theoretical support for pyrite tailings management and environmental restoration.