Abstract: To investigate the alleviating effects of biochar and Bacillus subtilis-loaded biochar on rape growth under tetracycline stress, a pot experiment was conducted with four treatments: control (CK), tetracycline (K), biochar + tetracycline (TK), and Bacillus subtilis-loaded biochar + tetracycline (WTK). The effects of different treatments on rape growth and physiological characteristics, including photosynthesis, were studied. The results show that compared with CK, the tetracycline stress (K) altered the root architecture of rape with the root tip number, average root diameter, and root volume decreased by 12.5%, 19.1%, and 18.5%, respectively, and the contents of chlorophyll a, chlorophyll b, and carotenoids in rape leaves decreased by 44.1%, 19.0%, and 22.7%, respectively. In contrast, the WTK treatment significantly enhanced the activities of antioxidant enzymes, with catalase (CAT) and peroxidase (POD) activities increasing by 88.7% and 72.7%, respectively, compared to the K treatment. Biochar application effectively promoted chlorophyll synthesis and improved light utilization capacity. Additionally, apparent quantum efficiency and the maximum photochemical efficiency of PSⅡ (F_v/F_m) were improved. Both biochar and Bacillus subtilis-loaded biochar treatments alleviated tetracycline stress, with the Bacillus subtilis-loaded biochar showing the most effective mitigation performance.