Copper oxide nanoparticles (CuO NPs) are used as a biocide in paints, textiles and plastics. Their application may lead to the contamination of aquatic ecosystems, where potential environmental effects remain to be determined. Toxic effects may be related to interactions of NPs with cellular systems or to particles' solubilisation releasing metal ions. In this report, we evaluated CuO NPs and soluble copper effects on photosynthesis of the aquatic macrophyte Lemna gibba L to determine the role of particle solubility in NPs toxicity. When L. gibba plants were exposed 48 h to CuO NPs or soluble copper, inhibition of photosynthetic activity was found, indicated by the inactivation of Photosystem II reaction centers, a decrease in electron transport and an increase of thermal energy dissipation. Toxicity of CuO NPs was mainly driven by copper ions released from particles. However, the bioaccumulation of CuO NPs in plant was shown, indicating the need to evaluate organisms of higher trophic level.