The pattern of genetic variation within and among natural populations of broomrape (Orobanche crenata Forsk.) from southern Spain was analysed by RAPD markers. Hierarchical analysis of phenotypic diversity using AMOVA was performed to analyse the partitioning of the variation among populations and among individuals. Although most of the genetic diversity was attributable to differences among individuals within a population (94.29%), significant st values among populations suggested the existence of phenotypic differentiation. Moreover, corresponding HOMOVA analysis revealed that molecular variances were significantly heterogeneous among populations although no clear grouping pattern could be established. These results are to be expected considering the predominant outcrossing behaviour of O. crenata.