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The potential of Thymus pulegioides L. as an alternative and valuable source of functional ingredients has been assessed. For this purpose, the phenolic constituent profiles and the antioxidant, anti-proliferative, neuroprotective, anti-aging and anti-diabetic activities of both T. pulegioides aqueous decoctions (AD) and hydro-ethanolic extracts (HE) were studied and compared for the first time. Rosmarinic acid was the main phenolic compound, accounting for 35.2% or 47.8% of total identified phenols in AD or HE, respectively (58.5 and 151.9 mg g-1 extract). Furthermore, large amounts of luteolin-O-hexuronide (AD: 39.9 ± 2.5 mg g-1, HE: 60.8 ± 7.0 mg g-1), eriodictyol-O-hexuronide (AD: 19.9 ± 2.5 mg g-1, HE: 26.8 ± 7.0 mg g-1) and chrysoeriol hexoside (AD: 23.5 ± 0.7 mg g-1, HE: 16.0 ± 0.7 mg g-1) were found. Both extracts showed significant in vitro antioxidant activity and anti-proliferative activity against Caco-2 cells (IC50 82.25 μg mL-1 and 105.44 μg mL-1, AD and HE, respectively), and reduced hepatotoxicity (HepG2 cells). In general, both T. pulegioides extracts showed poor anti-diabetic activity, moderate anti-aging effects and high neuroprotective activity with both AD and HE extracts, at 0.5 mg mL-1, showing 80% inhibition of the acetylcholinesterase activity and 94% inhibition of the tyrosinase activity. The present study highlights the important potential of this herb as a functional food ingredient.

Thymus mastichina, also called mastic thyme or Spanish marjoram, is endemic to the Iberian Peninsula, where it is widely used in folk medicine especially for treating digestive and respiratory systems disorders, and as a condiment to season olives. This work describes for the first time the detailed phenolic composition of exhaustive hydroethanolic extracts and aqueous decoctions of Thymus mastichina. Unlike other species of the Thymus genera, Thymus mastichina extracts contain high amounts of salvianolic acid derivatives, with salvianolic acid A isomer being the main derivative. This isomer was identified in extracts from Thymus mastichina for the first time. Also, an undescribed salvianolic acid derivative in Thymus mastichina was identified and its structure was tentatively described. Extracts from Thymus mastichina showed significant scavenging activity of 2,2-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) radical cation, hydroxyl, and nitric oxide radicals. The anti-proliferative effect of both T. mastichina extracts were tested against Caco-2 and HepG2 cells; the hydroethanolic extract showed a high anti-proliferative activity against Caco-2 cells compared to HepG2 cells (at 24 h exposure, the concentration that inhibits 50% of proliferation, IC50, was 71.18 ± 1.05 µg/mL and 264.60 ± 11.78 µg/mL for Caco-2 and HepG2, respectively). Thus, these results make this species a promising candidate for further investigation of its anti-tumoral potential. Therefore, Thymus mastichina can be potentially used as a functional food (used as a decoction or herbal tea) or as a source of bioactive ingredients with antioxidant and anti-proliferative properties.

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Medicinal and aromatic plants are important sources for plant secondary metabolites. The genetic manipulation of plants associated with in vitro plant regeneration systems facilitates efforts to engineer secondary product metabolic pathways. The fungal infections have been increasing in recent years due to several factors, namely, the increased incidence of high-risk patients, particularly immunocompromised hosts. Aromatic plants have been empirically used as antimicrobial agents, but the mechanisms of action are still unknown. Thyme has a great interest due to the possibility of its use in different applications, in medicine, in the cosmetic industry, or as food additives. Several studies have shown that thyme oils possess antimicrobial activity. Increasingly, plant breeding has taken advantage of molecular biology developments in order to genotype the species of interest to accelerate their selection. These approaches consist in choosing desired genotypes based on molecular markers or the knowledge of the genes involved in a particular trait. The in vitro culture techniques can be used to multiply plants selected after molecular and antifungal studies. The course of the investigation and the current state in relation to micropropagation, molecular studies, and antifungal action of the Thymus genus plants will be presented.