Metabolomic, proteomic and physiological insights into the potential mode of action of thymol, a phytotoxic natural monoterpenoid phenol

Thymol is a natural phenolic monoterpene widely produced by different species belonging to the Labiateae family. Although the thymol phytotoxicity is well known, the knowledge of its potential toxic mechanism is still limited. In this regard, the model species Arabidopsis thaliana was treated for 16 days by sub-irrigation with 300 μM of thymol. The results confirmed the high phytotoxic potential of this phenolic compound, which caused a reduction in plant growth and development. Thymol induced a water status alteration accompanied by an increase in ABA content and stomatal closure. Furthermore, leaves appeared necrotic in the margins and their temperature rinsed. The increase in H2O2 content suggested an oxidative stress experienced by treated plants. Both metabolomic and proteomic analysis confirmed this hypothesis showing a strong increase in osmoprotectants content, such as galactinol and proline, and a significant up-accumulation of proteins involved in ROS detoxification. Furthermore, the down-accumulation of proteins and pigments involved in the photosynthetic machinery, the increase in light sensitivity and the lower PSII efficiency well indicated a reduction in photosynthetic activity. Overall, we can postulate that thymol-induced phytotoxicity could be related to a combined osmotic and oxidative stress that resulted in reduced plant development.