Striga gesnerioides is a root hemiparasite that primarily parasitizes dicotyledonous species, including cowpea (Vigna unguiculata L.) and other legumes. Based on the differential resistance response of various cultivars, landraces, and breeding lines, it has been proposed that several distinct races of cowpea-parasitic S. gesnerioides exist in West Africa. In this study, we used amplified fragment length polymorphism profile analysis to examine the genetic variability within and among populations of cowpea-parasitic S. gesnerioides within the suspected distribution range of a particular race, and statistical clustering methods to define the phenetic relationships of the various races in West Africa. Our data indicate that genetic variability within and among populations of each of the previously recognized races of cowpea-parasitic S. gesnerioides is extremely low. On the basis of genotypic profile and host differential resistance responses, 2 previously unknown races were identified. Of the 7 races now identifiable, races SG1 (from Burkina Faso) and SG5 (from Cameroon) are the most closely related, and SG4 (from Benin) and SG3 (from Niger/Nigeria) are the most divergent. SG6, a new race of the parasite identified in Senegal, was found to be the most genetically similar to SG4 from Benin. We also demonstrate that a hypervirulent isolate of the S. gesnerioides from Zakpota, in the Republic of Benin, is genotypically distinct from other populations of SG4, thereby warranting designation as a separate race, which we called SG4z. To further support our race classification scheme, we identified a group of molecular markers that effectively discriminate each of the various races. Finally, we show that an isolate (designated SG4i) of the wild legume Indigofera hirsuta--parasitic S. gesnerioides is genetically distinct and significantly diverged from the various races of cowpea-parasitic S. gesnerioides. Our data suggest that both geographic isolation and host-driven selection are critical factors defining race formation in S. gesnerioides in West Africa.