Impact of copper oxide nanoparticles exposure on Arabidopsis thaliana growth, root system development, root lignificaion, and molecular level changes.

The effect of copper oxide nanoparticles (CuONPs) on physiological and molecular level responses were studied in Arabidopsis thaliana. The seedlings were exposed to different concentrations of CuONPs (0, 0.5, 1, 2, 5, 10, 20, 50, and 100 mg/L) for 21 days in half strength Murashige and Skoog medium. The plant biomass significantly reduced under different concentrations (2, 5, 10, 20, 50, and 100 mg/L) of CuONPs stress. Exposure to 2, 5, 10, 20, 50, and 100 mg/L of CuONPs has resulted in significant reduction of total chlorophyll content. The anthocyanin content significantly increased upon exposure to 10, 20, 50, and 100 mg/L of CuONPs. Increased lipid peroxidation was observed upon exposure to 5, 10, and 20 mg/L of CuONPs and amino acid proline content was significantly high in plants exposed to 10 and 20 mg/L of CuONPs. Significant reduction in root elongation was observed upon exposure to 0.5-100 mg/L of CuONPs for 21 days. Exposure to CuONPs has resulted in retardation of primary root growth, enhanced lateral root formation, and also resulted in loss of root gravitropism. Staining with phloroglucionol detected the deposition of lignin in CuONPs-treated roots. Histochemical staining of leaves and roots of CuONPs-exposed plants with nitroblue tetrazolium and 3'3'-diaminobenzidine showed a concentration-dependant increase in superoxide and hydrogen peroxide formation in leaves and roots of CuONPs-exposed plants. Cytotoxicity was observed in root tips of CuONPs-exposed plants as evidenced by increased propidium iodide staining. Real-time PCR analysis showed significant induction of genes related to oxidative stress responses, sulfur assimilation, glutathione, and proline biosynthesis under CuONPs stress.