Sickleweed (Falcaria vulgaris), an introduced species native to Europe and Asia, grows as an aggressive weed in some areas of the upper Midwest in the United States. We are reporting genetic diversity and population structure of sickleweed populations using microsatellite markers and nuclear and chloroplast DNA sequences. Populations showed high genetic differentiation but did not show significant geographic structure, suggesting random establishment of different genotypes at different sites was likely due to human mediated multiple introductions. Three genetic clusters revealed by microsatellite data and presence of six chlorotypes supported our hypothesis of multiple introductions. Chloroplast DNA sequence data revealed six chlorotypes nested into two main lineages suggesting at least two introductions of sickleweed in the upper Midwest. Some individuals exhibited more than two alleles at several microsatellite loci suggesting occurrence of polyploidy, which could be a post-introduction development to mitigate the inbreeding effects. High genetic variation in the introduced range attributable to multiple introductions and polyploidy may be inducing the evolution of invasiveness in sickleweed. Results of this study provide valuable insights into the evolution of sickleweed and baseline data for designing proper management practices for controlling sickleweed in the United States.