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In this work, Caenorhabditis elegans was employed as an in vivo model to determine the toxic effects of atrazine at different concentrations. After the exposure period from the larval stage L1 to adulthood day 1, atrazine (10 mg/L) significantly decreased the body length and lifespan of nematodes. In addition, exposure to ≥0.01 mg/L atrazine remarkably increased the intestinal reactive oxygen species (ROS) levels and reduced locomotion behavior of nematodes, while exposure to ≥ 1 mg/L atrazine decreased the brood size of nematodes. Moreover, atrazine (0.001–0.1 mg/L) upregulated the expression levels of hsp-6::GFP and hsp-6 / 60 in nematodes, indicating the activation of mitochondrial unfolded protein response (mtUPR). On the contrary, atrazine (1–10 mg/L) downregulated the expression levels of hsp-6::GFP and hsp-6 / 60 in nematodes. Furthermore, mtUPR induction governed by the RNAi knockdown of atfs-1 could increase the vulnerability of nematodes against atrazine toxicity. Overall, our findings highlighted the dynamic responses of nematodes toward different concentrations of atrazine, which could be monitored using different sublethal endpoints as bioindicators. [Display omitted] • Dynamic response to atrazine could be monitored by different sublethal endpoints. • mt UPR was very sensitive to atrazine exposure. • mt UPR could provide a protective response to atrazine. [ABSTRACT FROM AUTHOR]