Metabolomics Reveals the "Invisible" Responses of Spinach Plants Exposed to CeO₂ Nanoparticles

Engineered nanoparticles (NPs) with activities that mimic antioxidant enzymes have good prospects in agriculture because they can increase photosynthesis and improve stress tolerance. Here, the interaction between cerium oxide NPs with spinach plants ( Spinacia oleracea) was investigated by integrating phenotypic and metabolomic analyses. Soil-grown, four-week-old spinach plants were foliar exposed for 3 weeks to CeO₂ NPs at 0, 0.3, and 3 mg per plant. Phenotypic parameters (chlorophyll fluorescence, photosynthetic pigment contents, plant biomass, lipid peroxidation, and membrane permeability) were not affected. However, metabolomics analysis revealed that both doses of CeO₂ NPs induced metabolic reprogramming in leaves and roots in a non-dose-dependent manner. The low dose of CeO₂ NPs (0.3 mg per plant) induced stronger metabolic reprogramming in spinach leaves than high dose of CeO₂ NPs. However, the high dose of CeO₂ NPs triggered more metabolic changes in roots, compared to the low dose. Foliar spray of CeO₂ NPs at 3 mg/plant induced marked down-regulation of a number of amino acids (threonine, tryptophan, l-cysteine, methionine, cycloleucine, aspartic acid, asparagine, tyrosine, and glutamic acid). In addition, Zn decreased by 44% and 54% in leaves and Ca decreased by 38% and 32% in roots under exposure to CeO₂ NPs at 0.3 and 3 mg/plant, respectively. These results provide better understanding of the intrinsic phenotypic and metabolic changes imposed by CeO₂ NPs in spinach plants.