In the present study, we assessed the allelopathic effects of various concentrations (0%, 0.1%, 0.2%, and 0.3%) of horseradish root extract (HRE) on onion root. The average growth of onion root tips during the 0% HRE treatment (deionized water treatment) was 0.9 cm/day, which was the highest among the growth rates obtained with all HRE treatments. Moreover, the average growth during 0.3% HRE treatment was 0.1 cm/day. During cell cycle analysis, the mitotic phase fraction of the control (deionized water treatment) cells was 6.5% of all dividing cells, with this percentage being the highest among the values obtained for all treatment groups. In the control group, all cell cycle phases were identified; however, in the 0.1%, 0.2%, and 0.3% treatment groups, telophase was not identified. The ROS accumulation area of the onion root decreased, as the HRE treatment concentration increased. In the control root, the area of dead tissue was 0%; however, in the 0.1% and 0.2% HRE treatment roots, the ratio was 5% and 50%, respectively. These findings indicate that the allelopathic effect of HRE depends on the concentration of HRE applied to the onion root.