Thymus is one of the best known genera within the Labiatae (Lamiaceae) family, with more than 200 species and many medicinal and culinary uses. The effects of prolonged drought on lipid profile were investigated in tolerant and sensitive thyme plants (Thymus serpyllum L. and Thymus vulgaris L., respectively). Non-targeted non-polar metabolite profiling was carried out using Fourier transform ion cyclotron resonance (FT-ICR) mass spectrometry with one-month-old plants exposed to drought stress, and their morpho-physiological parameters were also evaluated. Tolerant and sensitive plants exhibited clearly different responses at a physiological level. In addition, different trends for a number of non-polar metabolites were observed when comparing stressed and control samples, for both sensitive and tolerant plants. Sensitive plants showed the highest decrease (55%) in main lipid components such as galactolipids and phospholipids. In tolerant plants, the level of lipids involved in signaling increased, while intensities of those induced by stress (e.g., oxylipins) dramatically decreased (50–60%), in particular with respect to metabolites with m/z values of 519.3331, 521.3488, and 581.3709. Partial least square discriminant analysis separated all the samples into four groups: tolerant watered, tolerant stressed, sensitive watered and sensitive stressed. The combination of lipid profiling and physiological parameters represented a promising tool for investigating the mechanisms of plant response to drought stress at non-polar metabolome level.

Thyme (Thymus spp.) volatiles predominantly consisting of monoterpenes and sesquiterpenes, serve as antimicrobial, antiseptic and antioxidant in phytomedicine. They also play a key role in plants as secondary metabolites via their potential role against herbivores, attracting pollinators and abiotic stress tolerance. Plant volatiles are affected by different environmental factors including drought. Here, the effect of prolonged water deficit stress on volatile composition was studied on the sensitive and tolerant thyme plant cultivars (T. vulgaris Var. Wagner and T. vulgaris Var. Varico3, respectively). Volatile sampling along with morpho–physiological parameters such as soil moisture, water potential, shoot dry weight, photosynthetic rate and water content measurements were performed on one-month-old plants subsequent to water withholding at 4-day intervals until the plants wilted. The tolerant and sensitive plants had clearly different responses at physiological and volatile levels. The most stress-induced changes on the plants’ physiological traits occurred in the photosynthetic rates, where the tolerant plants maintained their photosynthesis similar to the control ones until the 8th day of the drought stress period. While the analysis of the volatile compounds (VOCs) of the sensitive thyme plants displayed the same pattern for almost all of them, in the tolerant plants, the comparison of the pattern of changes in the tolerant plants revealed that the changes could be classified into three separate groups. Our experimental and theoretical studies totally revealed that the most determinant compounds involved in drought stress adaptation included α-phellandrene, O-cymene, γ-terpinene and β-caryophyelene. Overall, it can be concluded that in the sensitive plants trade-off between growth and defense, the tolerant ones simultaneously activate their stress response mechanism and continue their growth.

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