AbstractBACKGROUNDDigitaria insularis is a weed species that has gained considerable importance in Brazil's soybean production areas that rely on glyphosate‐resistant cultivars. Herbicide‐resistant weed populations of this species have been reported in many regions in Brazil, first in the south, followed by later reports in the north. We hypothesized that the spread of herbicide‐resistant D. insularis is facilitated by movement of agricultural machinery from the southern regions of Brazil.RESULTSPopulation genomics revealed a weak or no genetic structure (FST = [0; 0.16]), moderate expected heterozygosity (HE = 0.15; 0.44) and low inbreeding (FIS = [−0.1; 0.1]) in D. insularis populations. Our data supported the hypothesis that herbicide resistance gene flow predominantly occurred in a south‐to‐north direction based on a migration analysis. We also found evidence of local adaptation of resistant populations in the northern soybean‐growing regions of Brazil.CONCLUSIONEvidence in our work suggests that gene flow of glyphosate‐resistant D. insularis is associated with movement of agricultural machinery, although local selection pressure seems to play an important role in the evolution of herbicide resistance throughout the country. Our results suggest preventive practices such as equipment sanitation should be implemented to limit the spread of herbicide resistant D. insularis. © 2021 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.