Wheat stripe mosaic virus (WhSMV) is the causal agent of soilborne wheat mosaic disease (SBWMD) in Brazil, which is a serious threat to wheat production in the southern part of the country. WhSMV also occurs in Paraguay and South Africa. The virus is soilborne, transmitted by Polymyxa graminis, and management strategies are mainly based on genetic resistance. Variation in the reaction (type and severity of symptoms) of cultivars has been reported depending on the region and/or year of cultivation, leading wheat breeding programmes to test their materials in several locations, which increases costs and is time‐consuming. One hypothesis is that this variation in symptoms is a result of the genetic variability of WhSMV. We assessed the genetic variability and population structure of WhSMV infecting wheat in Brazil and South Africa. In field experiments conducted in different locations in southern Brazil, a consistent reproducibility of symptoms was observed in a set of cultivars, and molecular analysis showed a low degree of genetic variability of the Brazilian viral population. The hypothesis that the variation in SBWMD symptoms in Brazil is due to genetic variability of the virus was thus rejected. Comparison of the Brazilian and South African WhSMV isolates indicated that they comprise distinct subpopulations and that the Brazilian subpopulation is more variable than the South African subpopulation. The lower genetic variability of the South African subpopulation suggests genetic stability or a recent emergence of WhSMV in that region.