Abstract: Artemisia annua, which grows in many parts of Chongqing, releases artemisinin, which can inhibit the growth of surrounding plants. To identify pasture grasses that can grow in areas with abundant A. annua, we conducted a pot experiment to study the effects of artemisinin on grass growth. We evaluated the growth, physiology, antioxidant enzyme activity, and nutrient uptake of three pasture grasses (Lolium perenne, Trifolium repens, and Vicia villosa) growing in soil with artemisinin at different concentrations (0, control; 6,12,24,36 mg/kg soil). Artemisinin significantly affected the growth of all three pasture grasses, and the effects differed depending on the artemisinin concentration and the grass species. At 6 mg/kg, artemisinin led to significant decreases in growth indexes and the degree of inhibition increased with increasing artemisinin concentrations. Compared with their respective controls, L. perenne, T. repens, and V. villosa plants grown with artemisinin at 36 mg/kg showed 76.83%, 90.10%, and 65.09% decreases in biomass, respectively, and 47.82%, 38.76% and 44.49% decreases in plant height, respectively. Artemisinin had a negative allelopathic effect on all three pasture grasses, with the strongest effect on T. repens, followed by L. perenne and then V. villosa, indicating that V. villosa was the least sensitive to artemisinin. Therefore, V. villosa may be the most adaptable of the three tested grasses to growth in areas of Chongqing with abundant A. annua. In addition, artemisinin significantly reduced the chlorophyll content, root vigor, nitrate reductase activity, photosynthetic rate, root nutrient uptake, and nitrogen assimilation in all of the tested grasses. As a result, the uptake of nitrogen, phosphorus, and potassium by pasture grasses was decreased by more than 80%. The activity of protective oxidases in pasture grasses varied in response to artemisinin, with superoxide dismutase showing increased activity, peroxidase showing decreased activity, and catalase showing no significant change in activity. However, artemisinin greatly increased the malondialdehyde content in all three pasture grasses, suggesting that more oxyradicals were produced or less oxyradicals were eliminated, leading to cell membrane damage.