The Triticum aestivum root elongation test can be used as a simple and effective method for the identification of novel potential anti-proliferative agents. The employment of this method is based on the fact that a significant number of oncogenic signal transduction pathways are highly conserved in living organisms. Moreover, the use of phytobiological assays has several advantages, such as rapid outcomes, simple procedure and reduced costs. Previous studies have demonstrated that ultrasound exposure can have lethal effects on small organisms, medium and high frequency exposure, leading to cell destruction. In the present study, the effects of ultrasound exposure on the radicular length of Triticum caryopses were analyzed as a potential tool for further experimentation using ultrasounds as a stress model in phytobiological tests. The influence of the exposure time was determined at different intervals and was proven to exert significant effects. Short intervals of <5 min produced no statistically significant effects on root elongation, and the maximum of inhibition was achieved following 30 min of exposure. The experimental results highlighted that the inhibitory effect of ultrasound irradiation was lower following 15 min of exposure than after a shorter interval of 10 min, emphasizing a complex, non-linear biological response. On the whole, the findings of this study indicated a complex mechanism for the inhibition of the radicular growth by ultrasound exposure. The ultrasound exposure of Triticum seeds may be used as a tool to either assess the effects of toxic agents following an increase in cell permeability, or to examine the synergistic action between ultrasounds and exposure to chemical mixtures that can promote physiological disruptions.