Role of particle size-dependent copper bioaccumulation-mediated oxidative stress on Glycine max (L.) yield parameters with soil-applied copper oxide nanoparticles.

Increased impetus on the application of nano-fertilizers to improve sustainable food production warrants understanding of nanophytotoxicity and its underlying mechanisms before its application could be fully realized. In this study, we evaluated the potential particle size-dependent effects of soil-applied copper oxide nanoparticles (nCuO) on crop yield and quality attributes (photosynthetic pigments, seed yield and nutrient quality, seed protein, and seed oil), including root and seed Cu bioaccumulation and a suite of oxidative stress biomarkers, in soybean (Glycine max L.) grown in field environment. We synthesized three distinct sized (25 nm = S [small], 50 nm = M [medium], and 250 nm = L [large]) nCuO with same surface charge and compared with soluble Cu ions (CuCl) and water-only controls. Results showed particle size-dependent effects of nCuO on the photosynthetic pigments (Chla and Chlb), seed yield, potassium and phosphorus accumulation in seed, and protein and oil yields, with nCuO-S showing higher inhibitory effects. Further, increased root and seed Cu bioaccumulation led to concomitant increase in oxidative stress (HO, MDA), and as a response, several antioxidants (SOD, CAT, POX, and APX) increased proportionally, with nCuO treatments including Cu ion treatment. These results are corroborated with TEM ultrastructure analysis showing altered seed oil bodies and protein storage vacuoles with nCuO-S treatment compared to control. Taken together, we propose particle size-dependent Cu bioaccumulation-mediated oxidative stress as a mechanism of nCuO toxicity. Future research investigating the potential fate of varied size nCuO, with a focus on speciation at the soil-root interface, within the root, and edible parts such as seed, will guide health risk assessment of nCuO.