Ultrastructural and Proteomic Analyses Revealed the Mechanism by Which Foliar Spraying of Se Nanoparticles Alleviated the Toxicity of Microplastics in Pistia stratiotes L.

The uptake and accumulation of nanoplastics by plants have emerged as a major research focus. Exogenous selenium nanoparticles (SeNPs) are widely used to mitigate the toxicity of abiotic stresses, such as nanoplastics (NPs) and polyethylene (PE-NPs) nanoplastics, and represent a feasible strategy to enhance plant performance. However, the molecular mechanisms by which SeNPs alleviate the phytotoxicity of microplastics and nanoplastics remain poorly defined. To address this gap, we used Pistia stratiotes L. (P. stratiotes) as a model and silicon dioxide nanoparticles (SiO2NPs) as a comparator, integrating physiological assays, ultrastructural observations, and proteomic analyses. We found that NP stress caused ultrastructural damage in root tips, exacerbated oxidative stress, and intensified membrane lipid peroxidation. SeNPs treatment significantly mitigated NP-induced oxidative injury and metabolic suppression. Compared to the NPs group, SeNPs increased T-AOC by 38.2% while reducing MDA and ·OH by 33.3% and 89.6%, respectively. Antioxidant enzymes were also elevated, with CAT and POD rising by 47.1% and 39.2%. SeNPs further enhanced the photosynthetic capacity and osmotic adjustment, reflected by increases in chlorophyll a, chlorophyll b, and soluble sugar by 49.7%, 43.8%, and 27.0%, respectively. In contrast, proline decreased by 17.4%, indicating stress alleviation rather than an osmotic compensation response. Overall, SeNPs outperformed SiO2NPs. These results indicate that SeNPs broadly strengthen anti-oxidative defenses and metabolic regulation in P. stratiotes, effectively alleviating NP-induced oxidative damage. Proteomics further showed that SeNPs specifically activated the MAPK signaling cascade, phenylpropanoid biosynthesis, and energy metabolic pathways, enhancing cell-wall lignification to improve the mechanical barrier and limiting NPs translocation via a phytochelatin-mediated vacuolar sequestration mechanism. SiO2NPs produced similar but weaker alleviative effects. Collectively, these findings elucidate the molecular basis by which SeNPs mitigate NPs' phytotoxicity and provide a theoretical foundation and practical outlook for using nanomaterials to enhance phytoremediation in aquatic systems.