Copper bioaccumulation, photosystem II functioning, and oxidative stress in the seagrass Cymodocea nodosa exposed to copper oxide nanoparticles.

Abstract

Photosynthetic activity, oxidative stress, and Cu bioaccumulation in the seagrass Cymodocea nodosa were assessed 4, 12, 24, 48, and 72h after exposure to two copper oxide nanoparticle (CuO NP) concentrations (5 and 10mgL-1). CuO NPs were characterized by scanning electron microscopy (SEM) and dynamic light scattering measurements (DLS). Chlorophyll fluorescence analysis was applied to detect photosystem II (PSII) functionality, while the Cu accumulation kinetics into the leaf blades was fitted to the Michaelis-Menten equation. The uptake kinetics was rapid during the first 4h of exposure and reached an equilibrium state after 10h exposure to 10mgL-1 and after 27h to 5mgL-1 CuO NPs. As a result, 4-h treatment with 5mgL-1 CuO NPs, decreased the quantum yield of PS II photochemistry (Φ PSΙΙ ) with a parallel increase in the regulated non-photochemical energy loss in PSII (Φ NPQ ). However, the photoprotective dissipation of excess absorbed light energy as heat, through the process of non-photochemical quenching (NPQ), did not maintain the same fraction of open reaction centers (q p ) as in control plants. This reduced number of open reaction centers resulted in a significant increase of H2O2 production in the leaf veins serving possibly as an antioxidant defense signal. Twenty-four-hour treatment had no significant effect on Φ PSΙΙ and q p compared to controls. However, 24h exposure to 5mgL-1 CuO NPs increased the quantum yield of non-regulated energy loss in PSII (Φ NO ), and thus the formation of singlet oxygen (1O2) via the triplet state of chlorophyll, possible because the uptake kinetics had not yet reached the equilibrium state as did 10mgL-1. Longer-duration treatment (48 and 72h) had less effect on the allocation of absorbed light energy at PSII and the fraction of open reaction centers, compared to 4-h treatment, suggesting the function of a stress defense mechanism. The response of C. nodosa leaves to CuO NPs fits the "Threshold for Tolerance Model" with a threshold time (more than 4h) required for induction of a stress defense mechanism, through H2O2 production.