Time Matters: the Toxicity of Zinc Oxide Nanoparticles to Lemna minor L. Increases with Exposure Time

The use of zinc oxide nanoparticles (nano-ZnO) has rapidly increased in recent years, and this has triggered the need for versatile toxicity tests that can be used to test a range of different exposure scenarios. Acute exposure studies, using a variety of plant species, have overwhelmingly demonstrated nano-ZnO-induced toxicity, but substantial differences in the degree of phytotoxicity are reported in different studies. Here, we analysed the role of exposure time in determining the variation in phytotoxic effects. Using the model species Lemna minor, the effects of short-term (24 h), standardised (1 week) and chronic (up to 6 weeks) nano-ZnO exposure were compared. Nano-ZnO effects on Lemna minor growth indicators (biomass growth rate, root length), chlorophyll content and photosynthetic efficiency were measured. Rapid inhibitory effects of nano-ZnO on the maximal quantum yield of photosystem II could be measured after just 24-h exposure. Standardised (1 week) experiments revealed phytotoxic effects on Lemna minor biomass growth. More severe inhibitory effects on growth developed gradually over 4 to 6 weeks exposure to nano-ZnO, and these were qualitatively associated with increased zinc content in the plant. Such dynamics of nano-ZnO toxicity have not been elucidated before, and this study emphasises the importance of exposure time in studies of nanoparticle toxicity. We conclude that short-term, standardised experiments can potentially underestimate the environmental phytotoxicity, which may result from chronic exposure to nano-ZnO.