Abstract Plants growing outside their native range may be confronted by new regimes of herbivory, but how this affects plant chemical defence profiles has rarely been studied. Using Plantago lanceolata as a model species, we investigated whether introduced populations show significant differences from native populations in several growth and chemical defence traits. Plantago lanceolata (ribwort plantain) is an herbaceous plant species native to Europe and Western Asia that has been introduced to numerous countries worldwide. We sampled seeds from nine native and 10 introduced populations that covered a broad geographic and environmental range and performed a greenhouse experiment, in which we infested half of the plants in each population with caterpillars of the generalist herbivore Spodoptera littoralis. We then measured size‐related and resource‐allocation traits as well as the levels of constitutive and induced chemical defence compounds in roots and shoots of P. lanceolata. When we considered the environmental characteristics of the site of origin, our results revealed that populations from introduced ranges were characterized by an increase in chemical defence compounds without compromising plant biomass. The concentrations of iridoid glycosides and verbascoside, the major anti‐herbivore defence compounds of P. lanceolata, were higher in introduced populations than in native populations. In addition, introduced populations exhibited greater rates of herbivore‐induced volatile organic compound emission and diversity, and similar chemical diversity based on untargeted analyses of leaf methanol extracts. In conclusion, the geographic origin of the populations had a significant influence on morphological and chemical plant traits, suggesting that P. lanceolata populations are not only adapted to different environments in their native range, but also in their introduced range. Read the free Plain Language Summary for this article on the Journal blog.