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Striga hermonthica, a root hemiparasitic Orobanchaceae, severely constrains grain production in sub-Saharan Africa. Host specificity and geography may play important roles in shaping the population structure of S. hermonthica, with the Rift Valley potentially presenting a significant barrier to dispersal. Genetic diversity was assessed in 12 S. hermonthica populations from locations in Ethiopia. Of these, seven populations were parasitic on sorghum, two each on tef and maize, and one on finger millet. Genetic variation was detected using four amplified fragment length polymorphism (AFLP) primer combinations. After correcting for repeatability, 385 fragments were detected across all primer combinations. The percentage of polymorphic loci was relatively high, ranging from 53.2% to 76.4%. Expected heterozygosity ranged from 0.168 to 0.279. Genetic differentiation between populations was relatively high, and all populations were significantly different from each other . FST values ranged from 0.032 to 0.293 and averaged 0.146. Genetic differences between populations could not be attributed to host specificity. Instead, geography was the main determinant of population structure. There was a correlation between geographic and genetic distance. A significant portion of the genetic variance could be apportioned on either side of the Rift Valley (5%; P = 0:001). Also, a significant geographic barrier was identified in the southern portion of the sampled region. [ABSTRACT FROM AUTHOR]