The Red Queen hypothesis for the advantage of sex predicts that pathogens will evolve by increasing fitness with frequent encounters with specific host genotypes. In this study, BMV population size, measured as an indicator of fitness, was investigated during repeated passages through the same, or different host genotypes of the crop host, Hordeum vulgare (barley). Overall, mean BMV concentration within individual hosts was significantly higher in genetically homogeneous compared to heterogeneous host passage lines. In addition, BMV populations, passaged through a specific host variety, showed higher growth in that host variety compared to BMV passaged through varying varieties. These results supports the Red Queen hypothesis. However, the decrease in viral populations during passages contradicts the Red Queen. Nevertheless, the results found here show that even under simplified conditions, pathogens do not evolve in simple, predictable ways. Constraints on pathogen evolution may lead to counterintuitive results.