Assessment of the mechanism of combined toxicity of imidacloprid and triadimefon to Secale cereale L. seedlings under freeze-thaw cycle conditions

Freeze-thaw (FT) cycles significantly stress crops in Northeast China, exacerbated by pesticide overuse, particularly affecting vulnerable seedlings during these periods. This study investigates the physiological responses of Secale cereale L. seedlings to the insecticide imidacloprid (IMI) and the fungicide triadimefon (T) under simulated FT conditions. Our findings reveal that both pesticides impair photosynthesis in FT environments, resulting in increased malondialdehyde (MDA) and relative conductivity (RC). Furthermore, exposure to IMI and T enhances the activities of superoxide dismutase (SOD), peroxidase (POD), and ascorbate peroxidase (APX), while decreasing reduced glutathione (GSH) and hydrogen peroxide (H₂O₂) levels. Notably, combined stress resulted in significant increases of 80.26%, 16.36%, and 87.7% in RC, SOD, and POD activities, respectively, alongside substantial decreases of 65.87%, 46.34%, 63.74%, and 63.78% in net photosynthesis rate (Pn), transpiration rate (Tr), stomatal conductance (Gs), and water-use efficiency (WUE) in rye seedlings. Molecular docking analyses indicate that IMI and T interact with the active sites of SOD, POD, and APX through hydrogen bonding, compromising membrane integrity and inducing oxidative stress. While Secale cereale L. seedlings exhibit some resistance to IMI and T, FT conditions reduce this resilience. Correlation analysis reveals significant interactions between FT and pesticide stress on seedling physiology, suggesting that the concurrent use of IMI and T should be minimized in FT-prone areas. This study provides new insights into the pathways and mechanisms underlying the combined toxicity of IMI and T, offering a basis for assessing their environmental impacts on crops in regions susceptible to freeze-thaw cycles.