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CuO nanoparticles (CuO-NP) were synthesized in a hydrogen diffusion flame. Particle size and morphology were characterized using scanning mobility particle sizing, Brunauer–Emmett–Teller analysis, dynamic light scattering, and transmission electron microscopy. The solubility of CuO-NP varied with both pH and presence of other ions. CuO-NP and comparable doses of soluble Cu were applied to duckweeds, Landoltia punctata. Growth was inhibited 50% by either 0.6 mg L−1 soluble copper or by 1.0 mg L−1 CuO-NP that released only 0.16 mg L−1 soluble Cu into growth medium. A significant decrease of chlorophyll was observed in plants stressed by 1.0 mg L−1 CuO-NP, but not in the comparable 0.2 mg L−1 soluble Cu treatment. The Cu content of fronds exposed to CuO-NP is four times higher than in fronds exposed to an equivalent dose of soluble copper, and this is enough to explain the inhibitory effects on growth and chlorophyll content.