settingsOrder Article ReprintsOpen AccessArticleBiochemical Responses to the Long-Term Impact of Copper Sulfate (CuSO4) in Tobacco Plantsby Anastasia S. Tugbaeva *ORCID,Alexander A. ErmoshinORCID andIrina S. KiselevaORCIDDepartment of Experimental Biology and Biotechnology, Institute of Natural Sciences and Mathematics, Ural Federal University, Ekaterinburg 620002, Russia*Author to whom correspondence should be addressed.Int. J. Mol. Sci. 2023, 24(20), 15129; https://doi.org/10.3390/ijms242015129Received: 8 September 2023 / Revised: 28 September 2023 / Accepted: 1 October 2023 / Published: 13 October 2023(This article belongs to the Special Issue Advances in the Physiology of Primary and Secondary Plant Metabolism under Abiotic and Biotic Stress)Downloadkeyboard_arrow_down Browse Figures Versions NotesAbstractMetabolic changes under stress are often studied in short-term experiments, revealing rapid responses in gene expression, enzyme activity, and the amount of antioxidants. In a long-term experiment, it is possible to identify adaptive changes in both primary and secondary metabolism. In this study, we characterized the physiological state of tobacco plants and assessed the amount and spectrum of phenolic compounds and the lignification of axial organs under excess copper stress in a long-term experiment (40 days). Plants were treated with 100 and 300 μM CuSO4, as well as a control (Knop solution). Copper accumulation, the size and anatomical structure of organs, stress markers, and the activity of antioxidant enzymes were studied. Lignin content was determined with the cysteine-assisted sulfuric method (CASA), and the metabolite profile and phenolic spectrum were determined with UHPLC-MS and thin-layer chromatography (TLC). Cu2+ mainly accumulated in the roots and, to a lesser extent, in the shoots. Copper sulfate (100 μM) slightly stimulated stem and leaf growth. A higher concentration (300 μM) caused oxidative stress; H2O2 content, superoxide dismutase (SOD), and guaiacol peroxidase (GPOX) activity increased in roots, and malondialdehyde (MDA) increased in all organs. The deposition of lignin increased in the roots and stems compared with the control. The content of free phenolics, which could be used as substrates for lignification, declined. The proportions of ferulic, cinnamic, and p-coumaric acids in the hydrolysate of bound phenolics were higher, and they tended toward additional lignification. The metabolic profile changed in both roots and stems at both concentrations, and changed in leaves only at a concentration of 300 μM. Thus, changes in the phenolic spectrum and the enhanced lignification of cell walls in the metaxylem of axial (root and stem) organs in tobacco can be considered important metabolic responses to stress caused by excess CuSO4.
