A comparative study of the effect of barley stripe mosaic virus (BSMV) and gamma irradiation on mitotic divisions in barley (Hordeum vulgare L.) roots was performed by evaluating the mitotic index (MI), micronucleus (MN) frequency and sister chromatid exchanges (SCE). Results indicate that, similarly to gamma irradiation at doses of 100, 150 and 250 Gy, BSMV reduces the mitotic activity, increases the micronucleus frequency and the rate of SCE and promotes the formation of C-metaphases. In root meristematic cells of the three barley cultivars studied (Galactic, Sonor and Unirea), the mitotic index of infected plants was found to be 52.5, 54.48 and 64.17%, respectively, lower than the uninfected control. An increase in frequency of sister chromatid exchanges was observed in all the experimental variants. In treatments involving viral infection alone or in combination with gamma irradiation chromosomes with three and more chromatid exchanges were observed, while their percentage in the control or in treatments with gamma irradiation alone was reduced. The results of the study indicate that in plants derived from irradiated seeds, BSMV produces an effect that is correlated nonlinearly with the radiation dose applied. Cytological analysis of mitotic divisions in barley roots revealed the genotoxicity of BSMV infection.