Understanding intraspecific trait variations, particularly for invasive species that occupy large geographic areas with different resource conditions, can enhance our understanding of plant responses to changes in environmental resources. However, most related studies have focused on aboveground traits, while variations in root traits and responses to changes in resources during biological invasion have not been clarified. To fill this knowledge gap, we compared the root traits of Chromolaena odorata from 10 introduced populations in Southeast Asia and 12 native populations in North and Central America under different soil nutrients. The introduced populations of the invader exhibited greater resource-acquisitive root traits, characterized by reduced fine root diameter but increased proportions of absorbing root length and specific root length, compared to the native populations. Although nutrient addition significantly affected root traits, the introduced populations showed greater phenotypic plasticity in four traits (root / shoot ratio, specific root length, absorbing root length proportion, and branching intensity) than the native populations. Different root trait syndromes were observed between the introduced and native populations. These results indicate that after introduction, C. odorata may shift towards a more soil resource-acquisitive strategy and thus respond more positively to increased soils nutrients, thereby showing better performance in high-resource environments. This study provides a better understanding of how species respond to environment changes and reveals the factors underlying exotic plant invasion success.