Modulations in protein phosphorylation explain the physiological responses of barley (Hordeum vulgare) to nanoplastics and ZnO nanoparticles.

To address the knowledge gap on the effects of the co-existence of nanomaterials on plant growth, barley (Hordeum vulgare L.) plants were irrigated with zinc oxide nanoparticles (0.5 g L-1), nanoplastics (1 g L-1), and the combination of these two nanomaterials for 10 days. The co-existence of nanoplastics and ZnO nanoparticles increased H2O2 concentration by 12.76% and 38.30%, compared with the ZnO nanoparticles and nanoplastics exposure. The concentration of abscisic acid (ABA) in plants under the co-existence of nanoplastics and ZnO nanoparticles was 29.53% and 10.42% higher than that in ZnO nanoparticles treated plants and nanoplastics treated plants. The global analysis of phosphoproteomics identified 132 phosphorylated proteins and 173 phosphorylation sites in barley leaves exposed to the nanomaterial combination, which were related to photosynthesis, carbon fixation, nitrogen metabolism, and arginine and proline metabolisms. Further physiological analysis indicated that the combination of ZnO nanoparticles and nanoplastics caused larger damage to the systems of antioxidant and carbohydrate metabolisms as exemplified by decreased activities of apoplastic peroxidases (25.10%-48.60%), glutathione reductase (91.07%-94.94%), and sucrose synthase (53.59%-61.19%) in roots and increased cell wall invertase activity (12.97%-17.61%) in leaves, compared with the single nanomaterial treatments. These results indicate that the modulations in protein phosphorylation were closely related to the physiological responses to nanomaterial exposure, suggesting that the co-existence of nanomaterials may lead to greater impacts than single ones.