Particle size and concentration dependent toxicity of copper oxide nanoparticles (CuONPs) on seed yield and antioxidant defense system in soil grown soybean (Glycinemax cv. Kowsar).

Increasing applications of engineered nanomaterials (ENMs) warrant lifecycle assessment of their potential toxicity. Herein, we investigated potential phytotoxicity of copper oxide nanoparticles (CuONPs) on seed yield, focusing on particle size- and concentration-dependent responses of multiple antioxidant defense biomarkers, in soil-grown Glycinemax (cv. Kowsar) during its lifecycle. To this end, we synthesized three distinct sizes CuONPs (25, 50 and 250 nm): all with high purity, monoclinic crystal structure, and same surface charge. Each pot received two seeds, placed in soil inoculated with N-fixing bacteria (Rhizobium japonicum) and grown outdoor for 120 days. Our results show lipid peroxidation (MDA) and several antioxidant biomarkers (SOD, CAT, POX, APX) were differentially altered by the copper compound type, concentrations, and their interactions (p < 0.01). We show particle size- and concentration-dependent influence of CuONPs on lipid peroxidation, and such antioxidant biomarkers including SOD, CAT, POX, and APX, in soybean leaf at 120-day post-plantation. Particularly, the effects of CuONP-25 were consistently higher for most antioxidant biomarkers tested compared to the two larger size CuONPs (CuONP-50, CuONP-250) or Cu2+ ions treatments. We show that the concentration-response curves for CuONP-25 and Cu2+ ions were linear (R2 > 0.65), unlike for the larger size CuONPs (CuONP-50, CuONP-250) the relationships were nonlinear (R2 < 0.45), for most antioxidant biomarkers. The concentration-response curves for seed yield for all types of Cu compounds were linear (R2 > 0.65). Soybean seed yield also mirrored particle size- and concentration-dependent inhibition with CuONPs, and inhibition of CuONP-25 was significantly higher than the two larger size CuONPs or Cu2+ ions at all concentrations tested. All in all, our findings indicate differential nano-specific toxicity compared to ionic Cu2+ toxicity in soybean. These results may guide researchers and regulators on how best to tailor ENMs with specific particle characteristics rendering them more or less toxic, and better inform risk assessment of CuONPs in soil grown food crops such as soybean.