The analysis of 19 populations of the grasshopper Eyprepocnemis plorans, collected along four rivers belonging to the Segura basin (Mundo, Benamor, Taibilla and Segura itself), has shown that the presence of B chromosomes ends abruptly in each river, coinciding with the existence of a narrow pass in which this grasshopper cannot live because of the absence of the appropriate habitats. The existence of a broad inland region lacking grasshoppers with B chromosomes suggests that B chromosomes arose after the first colonization of the Iberian Peninsula by E. plorans specimens from North African populations. The B chromosome seems to have spread upstream along each of these four rivers until reaching geographical barriers that have impeded its advance and thus have preserved the non-B chromosome region. The available evidence indicates that the observed geographical distribution of the B chromosome polymorphism in this zone was shaped mainly by historical non-selective events.