Although plants are often exposed to atmospheric nanoparticles (NPs), the mechanism of NP deposition and their effects on physiology and metabolism, and particularly in combination with other stressors, are not yet understood. Exploring interactions between stressors is particularly important for understanding plant responses in urban environments where elevated temperatures can be associated with air pollution. Accordingly, 3-year-old spruce seedlings were exposed for 2 weeks to aerial cadmium oxide (CdO) NPs of environmentally relevant size (8-62 nm) and concentration (2 × 105 cm-3). While half the seedlings were initially acclimated to high temperature (35 °C) and vapour pressure deficit (VPD; 2.81 kPa), the second half of the plants were left under non-stressed conditions (20 °C, 0.58 kPa). Atomic absorption spectrometry was used to determine Cd content in needles, while gas and liquid chromatography was used to determine changes in primary and secondary metabolites. Photosynthesis-related processes were explored with gas-exchange and chlorophyll fluorescence systems. Our work supports the hypothesis that atmospheric CdO NPs penetrate into leaves but high temperature and VPD reduce such penetration due to stomatal closure. The hypothesis that atmospheric CdO NPs influences physiological and metabolic processes in plants was also confirmed. This impact strengthens with increasing time of exposure. Finally, we found evidence that plants acclimated to stress conditions have different sensitivity to CdO NPs compared to plants not so acclimated. These findings have important consequences for understanding impacts of global warming on plants and indicates that although the effects of elevated temperatures can be deleterious, this may limit other forms of plant stress associated with air pollution.

Central European Institute of Technology Brno University of Technology Purkyňova 123 CZ 612 00 Brno Czech Republic

Global Change Research Institute of the Czech Academy of Sciences Bělidla 986 4a CZ 603 00 Brno Czech Republic

Institute of Analytical Chemistry of the Czech Academy of Sciences Veveří 967 97 CZ 602 00 Brno Czech Republic

International Clinical Research Center St Anne's University Hospital Brno Pekařská 664 53 CZ 656 91 Brno Czech Republic

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