Characterization of a novel cytosolic sesquiterpene synthase MpTPS4 from Mentha ×piperita as a bioresource for the enrichment of invaluable viridiflorol in mentha essential oil.

Our research addresses the challenge of low concentrations of viridiflorol, a unique and highly valuable sesquiterpene found in various Mentha species. We employed biotechnological strategies to enhance viridiflorol production, which could significantly boost export revenue. Mentha piperita L. sesquiterpene synthase (MpTPS4) was the focus of our study because it is a key enzyme in the biosynthesis of viridiflorol. Through biochemical characterization, we confirmed that MpTPS4 exclusively synthesizes viridiflorol. By overexpressing MpTPS4 in M. ×piperita L. using a glandular trichome-specific promoter, we achieved a notable increase (9-25 %) in viridiflorol content. Additionally, we explored the practical application of viridiflorol as a deterrent against the herbivore Helicoverpa armigera. The RNAi-mediated knockdown of MpTPS4 resulted in a significant reduction in viridiflorol levels in the essential oil. More importantly, these results show how relevant MpTPS4 is for making viridiflorol and how biotechnology could be used to increase biosynthesis. Our research provides valuable insights into enhancing the production of this commercially important sesquiterpene, offering promising opportunities for the mentha industry.

Ethnobotany, phytochemistry, pharmacology and quality control of Peucedanum decursivum (Miq.) Maxim: A critical review.

ETHNOPHARMACOLOGICAL RELEVANCE: Dried roots of Peucedanum decursivum, a traditional Chinese medicine (TCM), has historically respiratory diseases such as cough, thick phlegm, headache, fever, and gynecological diseases, rheumatoid arthritis, and nasopharyngeal carcinoma. AIM OF THE STUDY: Made an endeavor to evaluate the research trajectory of P. decursivum, comprehensively discern its developmental status, and offer a guideline for future investigations. MATERIALS AND METHODS: A meticulous search of literatures and books from 1955 to 2024 via databases like PubMed, Web of Science and CNKI was conducted, including topics and keywords of " P. decursivum" "Angelica decursivum" and "Zihua Qianhu". RESULTS: P. decursivum and its prescriptions have traditionally been used for treating phlegm-heat cough, wind-heat cough, gastrointestinal diseases, pain relief and so on. It contains 234 identified compounds, encompassing coumarins, terpenes, volatile oils, phenolic acids, fatty acids and derivatives. It exhibits diverse pharmacological activities, including anti-asthmatic, anti-inflammatory, antioxidant effects, anti-hypertensive, anti-diabetic, anti-Alzheimer, and anti-cancer properties, primarily attributed to coumarins. Microscopic identification, HPLC fingerprinting, and bioinformatics identification are the primary methods currently used for the quality control. CONCLUSION: P. decursivum demonstrates anti-asthmatic, anti-inflammatory, and antioxidant effects, aligning with its traditional use. However, experimental validation of its efficacy against phlegm and viruses is needed. Additionally, analgesic effects mentioned in historical texts lack modern pharmacological studies. Numerous isolated compounds exhibit highly valuable medicinal properties. Future research can delve into exploring these substances further. Rigorous of heavy metal contamination, particularly Cd and Pb, is necessary. Simultaneously, investigating its pharmacokinetics and toxicity in humans is crucial for the safety.

Novel Study on Chemical Characterization and Antimicrobial, Antioxidant, and Anticholinesterase Activity of Essential Oil from Ecuadorian Bryophyte Syzygiella rubricaulis (Nees) Stephani.

Our research focuses on exploring the chemical composition and some biological properties of the essential oil derived from Syzygiella rubricaulis (Nees) Stephani, a bryophyte species. To conduct a comprehensive analysis, we utilized a DB5MS capillary column along with gas chromatography coupled to mass spectrometry (GC-MS) and flame ionization (GC-FID). The qualitative and quantitative examination revealed the presence of 50 compounds, with hydrocarbon sesquiterpenes (48.35%) and oxygenated sesquiterpenes (46.89%) being the predominant constituents. Noteworthy compounds identified include bicyclogermacrene (12.004%), cedranone <5-> (9.034%), spathulenol (6.835%), viridiflorol (6.334%), silphiperfol-5,7(14)-diene (6.216%), biotol <ß-> (6.075%), guaiol (4.607%), viridiflorene (4.65%), and α-guaienol (3.883%). Furthermore, we assessed the antimicrobial, antioxidant, and anticholinesterase activity of the essential oil, revealing a compelling inhibitory effect against acetylcholinesterase (AChE) with an IC50 value of 26.75 ± 1.03 µg/mL and a moderate antimicrobial (MIC 500 µg/mL, Enterococcus faecium, Lysteria monocytogenes) and antioxidant effect (ABTS: SC50 343.38 and DPPH 2650.23 µg/mL). These findings suggest the potential therapeutic application of the bryophyte essential oil in the treatment of Alzheimer's disease due to its potent anticholinesterase properties.

The chemical composition of the essential oils of two Mediterranean species of Convolvulaceae: Convolvulus althaeoides subsp. tenuissimus collected in Sicily (Italy) and Calystegia silvatica collected in Algeria.

Convolvulus L. and Calystegia R.Br. are two closely related genera of the Convolvulaceae family distributed in Asia, Mediterranean, Macaronesia, East Africa, and Arabia, including about 210 and 30 accepted species, respectively, of flowering plants, present as trees, shrubs, and herbs. The ethnomedical use of Convolvulus species dates to 1730s as they displayed profuse medicinal properties. In the present study, the not previously investigated chemical compositions of the essential oils from aerial parts of Convolvulus althaeoides subsp. tenuissimus (Sm.) Bat., collected in Sicily, and Calystegia sylvatica (Kit.) Griseb., collected in Algeria, were evaluated by GC-MS. The main components of the essential oil of the first one were ß-caryophyllene (28.68%), γ-muurolene (23.75%), and γ-elemene (17.55%), whereas the C. silvatica essential oil was shown to be rich of valeranone (10.77%), viridiflorol (9.45%), and germacrene D (8.61%). Furthermore, a complete literature review on the ethno-pharmacological uses of Convolvulus and Calystegia species was performed.

Viridiflorol induces anti-neoplastic effects on breast, lung, and brain cancer cells through apoptosis.

All active natural molecules are not fully exploited as therapeutic agents, causing delays in the advancement of anticancer drug discovery. Viridiflorol is a natural volatile element that may work as anti-cancer compound. We tested the anticancer properties of viridiflorol at different concentrations ranging from 0.03 to 300 µM in vitro on three cancer cells including breast (MCF-7), lung (A549) and brain (Daoy). The cancer cells responses were documented after treatment using MTT and Annexin V assays. Viridiflorol showed cytotoxic effects against all tested cell lines, reducing cell viability in a concentration-dependent manner with variable IC50 values. Daoy and A549 cell lines were more sensitive to viridiflorol when compared with temozolomide and doxorubicin, respectively. Viridiflorol demonstrated the highest anticancer activity against the Daoy cells with an estimated IC50 of 0.1 µM followed by MCF-7 at 10 µM, and A549 at 30 µM. In addition, upon exposure to concentrations ranging from 30 µM to 300 µM of viridiflorol, early and late apoptotic cell death was induced in a concentration dependent manner in Daoy (55.8%-72.1%), MCF-7 (36.2%-72.7%) and A459 (35%-98.9%) cell lines, respectively. In conclusion, viridiflorol demonstrates cytotoxic and apoptotic ability in three different cancer cell lines (brain, breast and lung).

Comparison of Volatile Organic Compounds of Sideritis romana L. and Sideritis montana L. from Croatia.

A study on the headspace volatile organic compounds (VOCs) profile of native populations of Sideritis romana L. and Sidertis montana L., Lamiaceae, from Croatia is reported herein, to elucidate the phytochemical composition of taxa from this plant genus, well-known for traditional use in countries of the Mediterranean and the Balkan region. Headspace solid-phase microextraction (HS-SPME), using divinylbenzene/carboxene/polydimethylsiloxane (DVB/CAR/PDMS) or polydimethylsiloxane/divinylbenzene (PDMS/DVB) fiber, coupled with gas chromatography-mass spectrometry (GC-MS) was applied to analyze the dried aerial parts of six native populations in total. Furthermore, principal component analysis (PCA) was conducted on the volatile constituents with an average relative percentage ≥1.0% in at least one of the samples. Clear separation between the two species was obtained using both fiber types. The VOCs profile for all investigated populations was characterized by sesquiterpene hydrocarbons, followed by monoterpene hydrocarbons, except for one population of S. romana, in which monoterpene hydrocarbons predominated. To our knowledge, this is the first report on the VOCs composition of natural populations of S. romana and S. montana from Croatia as well as the first reported HS-SPME/GC-MS analysis of S. romana and S. montana worldwide.

A Sesquiterpene Synthase from the Endophytic Fungus Serendipita indica Catalyzes Formation of Viridiflorol.

Interactions between plant-associated fungi and their hosts are characterized by a continuous crosstalk of chemical molecules. Specialized metabolites are often produced during these associations and play important roles in the symbiosis between the plant and the fungus, as well as in the establishment of additional interactions between the symbionts and other organisms present in the niche. Serendipita indica, a root endophytic fungus from the phylum Basidiomycota, is able to colonize a wide range of plant species, conferring many benefits to its hosts. The genome of S. indica possesses only few genes predicted to be involved in specialized metabolite biosynthesis, including a putative terpenoid synthase gene (SiTPS). In our experimental setup, SiTPS expression was upregulated when the fungus colonized tomato roots compared to its expression in fungal biomass growing on synthetic medium. Heterologous expression of SiTPS in Escherichia coli showed that the produced protein catalyzes the synthesis of a few sesquiterpenoids, with the alcohol viridiflorol being the main product. To investigate the role of SiTPS in the plant-endophyte interaction, an SiTPS-over-expressing mutant line was created and assessed for its ability to colonize tomato roots. Although overexpression of SiTPS did not lead to improved fungal colonization ability, an in vitro growth-inhibition assay showed that viridiflorol has antifungal properties. Addition of viridiflorol to the culture medium inhibited the germination of spores from a phytopathogenic fungus, indicating that SiTPS and its products could provide S. indica with a competitive advantage over other plant-associated fungi during root colonization.

New sesquiterpene glycoside ester with antiprotozoal activity from the flowers of Calendula officinalis L.

Continued chromatographic investigation of Calendula officinalis flowers led to the isolation of two sesquiterpenes, including one new, viridiflorol-10-O-ß-quinovopyranoside-2`-O-(3``-methyl-2``-pentenoate) (1), along with a previously reported compound viridiflorol-10-O-ß-fucopyranoside-2`-O-(3``-methyl-2``-pentenoate) (2). The new compound 1 was tested for antiprotozoal activity against Leishmania donovani Amastigote/THP1 and Trypanosoma brucei and it showed IC50 values of 3.57 and 7.84 µg/mL, respectively, while compound 2 exhibited no activity at the highest concentration tested 10 µg/mL.

Natural volatile oils derived from herbal medicines: A promising therapy way for treating depressive disorder.

Depression is a common global mental disorder that seriously harms human physical and mental health. With the development of society, the increase of pressure and the role of various other factors make the incidence of depression increase year by year. However, there is a lack of drugs that have a fast onset, significant effects, and few side effects. Some volatile oils from traditional natural herbal medicines are usually used to relieve depression and calm emotions, such as Lavender essential oil and Acorus tatarinowii essential oil. It was reported that these volatile oils, are easy to enter the brain through the blood-brain barrier and have good antidepressant effects with little toxicity and side effects. In this review, we summarized the classification of depression, and listed the history of using volatile oils to fight depression in some countries. Importantly, we summarized the anti-depressant natural volatile oils and their monomers from herbal medicine, discussed the anti-depressive mechanisms of the volatile oils from natural medicine. The volatile oils of natural medicine and antidepressant drugs were compared and analyzed, and the application of volatile oils was explained from the clinical use and administration routes. This review would be helpful for the development of potential anti-depressant medicine and provide new alternative treatments for depressive disorders.

Melaleuca alternifolia (tea tree) oil and its monoterpene constituents in treating protozoan and helminthic infections.

Australian tea tree (Melaleuca alternifolia) oil (TTO) and its monoterpene constituents such as terpinen-4-ol (T4O), 1,8-cineole, limonene, p-cymene, and α-terpinene have been shown to be effective in controlling a wide range of parasitic infections. The anti-parasitic effects of these compounds are mainly due to their anti-histamine and anti-acetylcholinesterase activities as well as their ability to modulate host inflammatory responses. This review attempts to summarize recent advances in the uses of TTO and its 15 major monoterpene constituents in treating parasitic infections in both humans and animals. Activities against parasitic protozoans (Plasmodium falciparum, Leishmania spp., Trypanosoma spp., Acanthamoeba castellanii, Trichomonas vaginalis, Eimeria, and Ichthyophthirius multifiliis), nematodes (Haemonchus contortus and Anisakis simplex), cestode (Echinococcus ortleppi), and monogeneans (Gasterosteus spp. and Dactylogyrus minutus) have been reported, showing good potentials in treating parasitic infections. Further studies are necessary for developing anti-parasite therapies using TTO or its monoterpenes constituents.

Chemical Composition and Bioactivity of Essential Oil from Blepharocalyx salicifolius.

Natural products represent a source of biologically active molecules that have an important role in drug discovery. The aromatic plant Blepharocalyx salicifolius has a diverse chemical constitution but the biological activities of its essential oils have not been thoroughly investigated. The aims of this paper were to evaluate in vitro cytotoxic, antifungal and antibacterial activities of an essential oil from leaves of B. salicifolius and to identify its main chemical constituents. The essential oil was extracted by steam distillation, chemical composition was determined by gas chromatography/mass spectrometry, and biological activities were performed by a microdilution broth method. The yield of essential oil was 0.86% (w/w), and the main constituents identified were bicyclogermacrene (17.50%), globulol (14.13%), viridiflorol (8.83%), γ-eudesmol (7.89%) and α-eudesmol (6.88%). The essential oil was cytotoxic against the MDA-MB-231 (46.60 µg·mL-1) breast cancer cell line, being more selective for this cell type compared to the normal breast cell line MCF-10A (314.44 µg·mL-1). Flow cytometry and cytotoxicity results showed that this oil does not act by inducing cell death, but rather by impairment of cellular metabolism specifically of the cancer cells. Furthermore, it presented antifungal activity against Paracoccidioides brasiliensis (156.25 µg·mL-1) but was inactive against other fungi and bacteria. Essential oil from B. salicifolius showed promising biological activities and is therefore a source of molecules to be exploited in medicine or by the pharmaceutical industry.

Evaluation of the cytotoxic effects of Cyperus longus extract, fractions and its essential oil on the PC3 and MCF7 cancer cell lines.

Cyperus longus is one of the Iranian endemic species. However, to date, and to the best of our knowledge, there are no availale academic reports on the cytotoxicity of this plant. Thus, this study was carried out to examine the in vitro anti-proliferative and anti-apoptotic effects of Cyperus longus extract, fractions and essential oil (EO) on MCF7 and PC3 cell lines. The chemical constituents of EO were identified using gas chromatography (GC)-mass spectrometry (MS) analysis. The cells were cultured in RPMI-1640 medium and incubated with various concentrations of the plant extract and fractions. Cell viability was quantified by MTT assay following 24, 48 and 72 h of exposure to (12.5-200 µg/ml) of the methanol extract, the dichloromethane (CH2Cl2), ethyl acetate (EtOAc) and water fractions, as well as the EO of the plant. The percentage of apoptotic cells was determined using propidium iodide staining of DNA fragments by flow cytometry (sub-G1 peak). The most effective fraction in the MCF7 cell line was the CH2Cl2 fraction (IC50 after 48 h, 25.34±2.01). The EtOAc fraction (IC50 after 48 h, 35.2±2.69) and the methanol extract (IC50 after 48 h, 64.64±1.64) were also found to be effective. The IC50 values obtained for the PC3 cell line were 37.97±3.87, 51.57±3.87 and 70.33±2.36 for the CH2Cl2 fraction, the EtOAc fraction and the methanol extract, respectively. Based on these data and due to the partial polarity of the most effective fraction (the CH2Cl2 fraction), we also examined the cytotoxicity of the plant EO. The IC50 values after 48 h were 22.25±4.25 and 12.55±3.65 in the PC3 and MCF7 cell lines, respectively. DNA fragmentation assay also confirmed these data. Performing GC-MS analysis for the plant EO revealed that ß-himachalene (10.81%), α-caryophyllene oxide (7.6%), irisone (4.78%), ß-caryophyllene oxide (4.36%), humulene oxide (12%), viridiflorol (4.73%), aristolone (6.39%) and longiverbenone (6.04%) were the main constituents. Our results demonstrated that two of the constituents of Cyperus longus, viridiflorol and longiverbenone, should be investigated further as possible promising chemotherapeutic agents in cancer treatment.

Análisis multivariable de la composición química de aceite esencial dehojas de Melaleuca quinquenervia que crece en Cuba / Multivariate analysis of the chemical composition of the essential oil from leaves of Melaleuca quinquenervia growing in Cuba

El aceite esencial de Melaleuca quinquenervia de la Ciénaga de Zapata, Cuba, obtenido a escala piloto con un rendimiento promedio de 0,8%, presentó una densidad relativa y un índice de refracción promedios de 0,9253g/mL y 1,4827 respectivamente. La determinación de su composición química por cromatografía de gases acoplada a espectrometría de masas permitió identificar 42 compuestos, de los cuales 26 se sometieron al Análisis por Componentes Principales y al Análisis por Conglomerados. Dichos análisis permitieron la separación de las muestras en dos grupos con diferencias significativas (p<0,05) entre los contenidos promedios de sus principales constituyentes. A partir de dichos análisis se estableció la presencia de dos quimiotipos diferentes: uno con elevados contenidos de viridiflorol (33,8 por ciento), seguido por el 1,8-cineol (21,8 por ciento), y otro quimiotipo del tipo 1,8-cineol (30,0 por ciento) con un contenido relativamente elevado de viridiflorol (24,6 por ciento). Ambos quimiotipos presentaron concentraciones similares de los monoterpenos limoneno y αlfa-pineno.

The essential oil of Melaleuca quinquenervia from Ciénaga de Zapata, Cuba, obtained at pilot scale with average yield of 0.8%, showed average relative density and refraction index of 0.9253g/mL and 1.4827 respectively. It ́s chemical composition determination by gas chromatography coupled to Mass Spectrometry allow identifying 42 compounds, 26 of which were submitted to Principal Components Analysis and to Conglomerates Analysis. Such analyses allowed the separation of the samples into two groups with significant differences (p<0.05) between the average contents of its main components. From such analyses, it was established the presence of two different chemotypes: one with high contents of viridiflorol (33.8 percent), followed by 1,8-cineol (21.8 percent), and another chemotype rich in 1,8-cineol (30,0 percent) with a relatively high content in viridiflorol (24.6 percent). Both chemotypes showed similar concentrations of limonene and αlfa-pinene monoterpenes.

Structure and cytotoxic activity of sesquiterpene glycoside esters from Calendula officinalis L.: Studies on the conformation of viridiflorol.

Topic applications of Calendula officinalis L. lipophilic extracts are used in phytotherapy to relieve skin inflammatory conditions whereas infusions are used as a remedy for gastric complaints. Such a different usage might be explained by some cytotoxicity of lipophilic extracts at gastric level but little is known about this. Therefore, we screened the CH2Cl2 extract from the flowers of C. officinalis by MTT and LDH assays in human epithelial gastric cells AGS. This bioassay-oriented approach led to the isolation of several sesquiterpene glycosides which were structurally characterized by spectroscopic measurements, chemical reactions and MM calculations. The conformational preferences of viridiflorol fucoside were established and a previously assigned stereochemistry was revised. The compounds 1a, 2a and 3f showed comparably high cytotoxicity in the MTT assays, whereas the effect on LDH release was lower. Our study provides new insights on the composition of C. officinalis extracts of medium polarity and identifies the main compounds that could be responsible for cytotoxic effects at gastric level.