Stable Zn isotopes reveal the uptake and toxicity of zinc oxide engineered nanomaterials inPhragmites australis

The uptake, transport, and toxicity mechanisms of zinc oxide (ZnO) engineered nanomaterials (ZnO-ENMs) in aquatic plants remain obscure. We investigated ZnO-ENM uptake and phytotoxicity in Phragmites australis by combining Zn stable isotopes and microanalysis. Plants were exposed to four ZnO materials: micron-size ZnO, nanoparticles (NPs) of <100 nm or <50 nm, and nanowires of 50 nm diameter at concentrations of 0-1000 mg l −1 . All ZnO materials reduced growth, chlorophyll content, photosynthetic efficiency, and transpiration and led to Zn precipitation outside the plasma membranes of root cells. Nanoparticles <50 nm released more Zn 2+ and were more toxic, thus causing greater Zn precipitation and accumulation in the roots and reducing Zn isotopic fractionation during Zn uptake. However, fractionation by the shoots was similar for all treatments and was consistent with Zn 2+ being the main form transported to the shoots. Stable Zn isotopes are useful to trace ZnO-ENM uptake and toxicity in plants. Environmental Significance Statement Our understanding of zinc oxide nanomaterials interaction with wetland plants is hampered by the lack of scientific consensus about their uptake and toxicity mechanisms in these species. This is a serious concern given the alarming global increase in the discharge of these nanomaterials into the environment and the key ecological roles of wetland plants. The Zn isotopic signature of plant tissue integrates all the Zn metabolic pathway throughout the plant’s life, giving insight about the form of Zn taken up, even if this later transforms into another Zn species. Thus, our findings clarify the exposure routes and the mechanisms of action of zinc oxide engineered nanomaterials in wetland plants while advancing the toolbox for plant physiology and environmental studies. Table of contents The Zn stable isotope composition of plants demonstrates that ZnO engineered nanomaterials dissolve before their uptake and accumulation by the roots (brightest inclusions in root cortex above).