Abstract: Maternal habitat and resource availability can jointly influence how invasive plants respond to environmental stress. Seeds produced in contrasting habitats may carry maternal-environmental legacies that affect subsequent growth, physiology, and stress tolerance, whereas nutrient inputs can modify the capacity of plants to withstand toxic substrates. However, how maternal habitat mediates the response of invasive plants to heavy metal stress under different nutrient conditions remains poorly understood. This study examined the highly invasive Ambrosia artemisiifolia L. using seeds collected from heavy metal-contaminated and non-contaminated habitats. A greenhouse experiment was conducted with mine tailings as the growth substrate and three nutrient treatments: no nutrient addition, nitrogen and phosphorus addition, and powdered A. artemisiifolia biomass addition. Growth traits, leaf physicochemical properties, and substrate physicochemical properties were measured to determine how nutrient availability and maternal habitat interact under heavy metal stress. Results showed that nutrient addition and maternal habitat significantly affected total biomass and plant height, with a significant interactive effect. Nutrient inputs strongly promoted the growth of plants derived from non-contaminated habitats, whereas plants derived from contaminated habitats showed a more stress-tolerant resource-utilization strategy. Addition of powdered A. artemisiifolia biomass significantly increased substrate carbon and nitrogen contents and reduced the C/N ratio. Correlation and redundancy analyses further indicated that substrate nutrient status was the dominant factor regulating trait variation in A. artemisiifolia. Overall, the response of A. artemisiifolia to heavy metal stress depended strongly on maternal habitat of the seeds, and nutrient addition amplified phenotypic divergence among populations from different maternal habitats. These findings reveal that maternal habitat and nutrient availability interact to regulate invasive plant performance in contaminated substrates and provide insight into invasion mechanisms and management strategies in polluted ecosystems.