Levels of inter- and intrapopulation genetic variation were determined in five North American populations of leafy spurge using chloroplast DNA (cpDNA) RFLPs and RAPD markers. Thirteen plastome types were identified among 123 individuals collected from five geographically separated populations. Number of plastomes within a population ranged from one to seven, with four of the populations having a predominate type plus one or more rarer types. Some plastome types were shared by populations, but plastome distribution among populations was nonrandom. RAPD markers indicated greatest relatedness among individuals within a population. Relatedness among populations as established through RAPDs was greater for geographically closer populations; this relationship was not observed for cpDNA markers. Differences in the range of movement for pollen and seed may account for differences between results of the cpDNA and RAPD analyses. The high degree of genetic variability among North American leafy spurge suggests possible multiple introductions or a high degree of variability within leafy spurge populations in its native range.