An introduction to a novel habitat represents a challenge to plants because they likely would face new interactions and possibly different physical context. When plant populations arrive to a new region free from herbivores, we can expect an evolutionary change in their defense level, although this may be contingent on the type of defense, resistance or tolerance, and cost of defense. Here, we addressed questions on the evolution of tolerance to damage in non-native Spanish populations of Datura stramonium by means of two comparative greenhouse experiments. We found differences in seed production, specific leaf area, and biomass allocation to stems and roots between ranges. Compared to the Mexican native populations of this species, non-native populations produced less seeds despite damage and allocate more biomass to roots and less to stems, and had higher specific leaf area values. Plasticity to leaf damage was similar between populations and no difference in tolerance to damage between native and non-native populations was detected. Costs for tolerance were detected in both regions. Two plasticity traits of leaves were associated with tolerance and were similar between regions. These results suggest that tolerance remains beneficial to plants in the non-native region despite it incurs in fitness costs and that damage by herbivores is low in the non-native region. The study of the underlying traits of tolerance can improve our understanding on the evolution of tolerance in novel environments, free from plants’ specialist herbivores.