Anti-breast cancer activity of the essential oil from grapefruit mint (Mentha suaveolens × piperita).

Grapefruit mint (Mentha suaveolens × piperita) is a hybrid, perennial, and aromatic plant widely cultivated all over the world and used in the food, cosmetics, and pharmaceutical industries mostly for its valuable essential oil. Herein, we evaluated the anticancer activity of the grapefruit mint essential oil, cultivated in Iran. For the chemical composition analysis of essential oil, GC-MS was used. MTT assay was utilized for assessing the cytotoxic activity of the essential oil. The type of cell death was determined by annexin V/PI staining. Essential oil effect on the expression of maternally expressed gene 3 (MEG3), a regulatory lncRNA involved in cell growth, proliferation, and metastasis, was studied using qRT-PCR. Linalool (43.9%) and linalool acetate (40.1%) were identified as the dominant compounds of essential oil. Compared with MCF-7, the MDA-MB-231 cells were more sensitive to essential oil (IC50 = 7.6 µg/ml in MCF-7 and 5.9 µg/ml in MDA-MB-231 after 48 h). Essential oil induced cell death by apoptosis. Wound healing scratch assay confirmed the anti-invasive effect of essential oil. In addition, essential oil upregulated the tumor suppressor MEG3 in breast cancer cells. These results provide new insights into grapefruit mint essential oil potential application as an anticancer adjuvant in combination treatments for breast cancer patients.

Phytochemical variations antioxidant, and antibacterial activities among zebaria sumac (Rhus coriaria var. zebaria) populations in Iraq.

Sumac (Rhus coriaria L.) is one of the medicinal plants of Anacardiaceae family and widely used as a spice in Iran and Arab countries. Rhus coriaria var. zebaria is a small tree or large shrub, wildly growing in Iraq and described as a new variety with special characteristics. These increase the importance of studying sumac in these areas. Here, the phytochemical variations and the antibacterial activity of 50 accessions of this variety from five different climatic conditions was evaluated in order to identify the best accession to use and the best area for its cultivation. This is the most comprehensive study on this plant. Essential oil compounds were identified using GC-MS method and according to the results, Z, E-2,13-octadecadien, caryophyllene oxide, 2,4-decadienal, E-caryophyllene and nonanoic acid were among the main compounds. Also, the variety is a rich source of minerals including K, Ca, Mg, Na, P, and N. Sumac fruit extract from Akre Xerds had the highest anthocyanin and the lowest amount was from Kavilca region. The radical scavenging effect of extract from Dostic area in the concentration of 400 µg/mL is closer to the effect of ascorbic acid. The largest inhibition was found in the sumac extracted oil of Xasto Zhere area against S. aureus in compared with penicillin and amoxicillin and enrofloxacin antibiotics.

Intraspecific divergence in essential oil content, composition and genes expression patterns of monoterpene synthesis in Origanum vulgare subsp. vulgare and subsp. gracile under salinity stress.

BACKGROUND: Oregano (Origanum vulgare L.), one of the important medicinal plants in the world, has valuable pharmacological compounds with antimicrobial, antiviral, antioxidant, anti-inflammatory, antispasmodic, antiurolithic, antiproliferative and neuroprotective activities. Phenolic monoterpenes such as thymol and carvacrol with many medical importance are found in Oregano essential oil. The biosynthesis of these compounds is carried out through the methyl erythritol-4 phosphate (MEP) pathway. Environmental stresses such as salinity might improve the secondary metabolites in medicinal plants. The influence of salinity stress (0 (control), 25, 50 and 100 mM NaCl) on the essential oil content, composition and expression of 1-deoxy-D-xylulose-5-phosphate reductoisomerase (DXR), γ-terpinene synthase (Ovtps2) and cytochrome P450 monooxygenases (CYP71D180) genes involved in thymol and carvacrol biosynthesis, was investigated in two oregano subspecies (vulgare and gracile). RESULTS: Essential oil content was increased at low NaCl concentration (25 mM) compared with non-stress conditions, whereas it was decreased as salinity stress intensified (50 and 100 mM). Essential oil content was significantly higher in subsp. gracile than subsp. vulgare. The highest (0.20 mL pot-1) and lowest (0.06 mL pot-1) amount of essential oil yield was obtained in subsp. gracile at 25 and 100 mM NaCl, respectively. The content of carvacrol, as the main component of essential oil, decreased with increasing salinity level in subsp. gracile, but increased in subsp. vulgare. The highest expression of DXR, Ovtps2 and CYP71D180 genes was observed at 50 mM NaCl in subsp. vulgare. While, in subsp. gracile, the expression of the mentioned genes decreased with increasing salinity levels. A positive correlation was obtained between the expression of DXR, Ovtps2 and CYP71D180 genes with carvacrol content in both subspecies. On the other hand, a negative correlation was found between the expression of CYP71D180 and carvacrol content in subsp. gracile. CONCLUSIONS: The findings of this study demonstrated that both oregano subspecies can tolerate NaCl salinity up to 50 mM without significant reduction in essential oil yield. Also, moderate salinity stress (50 mM NaCl) in subsp. vulgare might increase the carvacrol content partly via increment the expression levels of DXR, Ovtps2 and CYP71D180 genes.

Modeling and optimizing concentration of exogenous application of γ-aminobutyric acid on NaCl-stressed pineapple mint (Mentha suaveolens) using response surface methodology: an investigation into secondary metabolites and physiological parameters.

Salinity, a severe worldwide issue, compromises the economic production of medicinal plants including mints and causes drug-yield decline. γ-Aminobutyric acid (GABA) is a tolerance-inducing signaling bio-molecule in various plant physiological processes. Pineapple mint (Mentha suaveolens Ehrh.) is a valuable medicinal herb with an exhilarating scent of citrus fruit. Piperitenone oxide is the major bioactive constituent of its essential oil, having significant demand by pharmaceutical industries. Nonetheless, modeling and optimizing the effective concentration of GABA remain within twin foci of interest. Therefore, a two factor-five level (NaCl 0-150 mM and GABA 0-2.4 mM) central composite design was conducted to model and optimize drug yield and physiological responses of M. suaveolens. Based on the design of experiments (DoE) approach, different linear, quadratic, cubic, and quartic models were assigned to the response variables. Change trends of shoot and root dry weights followed a simple linear model, whereas sophisticated models (i.e., multiple polynomial regression) were fitted to the other traits. NaCl stress inevitably reduced root and shoot dry weight, piperitenone oxide content, relative water content, pigments content, and maximum quantum yield of PSII. However, content of malondialdehyde (MDA) and total flavonoid, and DPPH radical scavenging activity were increased under salinity. Under severe NaCl stress (150 mM), the essential oil content (0.53%) was increased three times in comparison with control (0.18%). Optimization analysis demonstrated that the highest amount of essential oil (0.6%) and piperitenone oxide (81%) as a drug yield-determining component would be achievable by application of 0.1-0.2 mM GABA under 100 mM NaCl. The highest dry weight of root and shoot was predicted to be achieved at 2.4 mM GABA. Overall, extremely severe NaCl stress (i.e., more than 100 mM) in which a sharp drop in yield components value was observed seemed to be out of M. suaveolens salinity tolerance range. Hence, it is rationale to compensate the decrease of drug yield by foliar application of a dilute GABA solution (i.e., 0.1-0.2 mM) under 100 mM NaCl stress or lower levels.

Determining Effective Environmental Factors in the Distribution of Endangered Endemic Medicinal Plant Species Using the BMLR Model: The Example of Wild Celery (Kelussia odoratissima Mozaff., Apiaceae) in Zagros (Iran).

Kelussia odoratissima Mozaff. is a medicinal species native to Iran. The goal of this research was to determine the environmental factors important for the distribution of K. doratissima in Iran using BMLR modeling. Six random transects were established throughout the species' habitat, and 220 quadrats with an area of 4 m2 were plotted. The canopy cover percentages of K. doratissima were estimated in each quadrat. Topographic factors, including elevation, slope, and aspect maps, were generated by creating DEM images. Land use, land evaluation, evaporation, temperature, and precipitation maps of the area were created accordingly. The data collected from the experiments were analyzed using the Minitab and R statistical packages. To determine the effect of the studied factors in the distribution of K. doratissima, we ran a set of backward multiple linear regressions. The results showed that the effects of evaporation, elevation, and slope were significant in the species' distribution, with elevation having a positive effect and evaporation and slope showing negative effects. Further, elevation had the highest effect on distribution (greatest absolute value of beta at 9.660). The next most significant factors in the plant's distribution were evaporation (beta = 8.282) and slope (beta = 0.807), respectively.

Introducing Three New Fruit-Scented Mints to Farmlands: Insights on Drug Yield, Essential-Oil Quality, and Antioxidant Properties.

Mint species are one of the most traded medicinal plants with a wide array of applications in the food, pharmaceutical, and perfumery industries. Here, a field experiment based on completely randomized block design (RCBD) aimed to compare drug yield, antioxidant properties, and essential-oil (EO) quality of three newly introduced mints (i.e., ginger mint, pineapple mint, and grapefruit mint) with a chiefly cultivated one (i.e., peppermint). The results manifested that dry-weight yield and EO yield of grapefruit mint (310 g/m2 and 5.18 g/m2, respectively) was approximately 2 times more than that of others. The highest EO content (i.e., 3.12%, v/w)) was obtained from the ginger mint; however, there were no significant differences among the other three mints. The highest total flavonoids content and 2,2-diphenyl-1-picrylhydrazyl (DPPH) scavenging activity of both methanolic and ethanolic extracts were found in pineapple and grapefruit mint. Methanolic extract of ginger mint yielded the highest total polyphenol content, whereas the ethanolic extract of pineapple mint showed the highest total polyphenol content. According to mean comparisons, the EO of ginger mint exhibited the highest antioxidant activity (EC50 value = 2.23 µL/mL), while EO of peppermint showed the lowest antioxidant activity (EC50 value = 48.23 µL/mL). Gas chromatography analysis identified four EO types among these mints: (i) grapefruit mint EO rich in linalool (51.7%) and linalyl acetate (28.38%); (ii) ginger mint EO rich in linalool (59.16%); (iii) pineapple mint EO rich in piperitone oxide (77.65%); and (iv) peppermint EO rich in menthol (35.65%). The findings of the present study provide new insights into the cultivation of preferable mints possessing desired characteristics for food and drug industries.

Selenium Nanoparticles (Se-NPs) Alleviates Salinity Damages and Improves Phytochemical Characteristics of Pineapple Mint (Mentha suaveolens Ehrh.).

The present study examined the effects of foliar spray of selenium nanoparticles (0, 10 and 20 mg/L) on the yield, phytochemicals and essential oil content and composition of pineapple mint (Mentha suaveolens Ehrh.) under salinity stress (0, 30, 60 and 90 mM NaCl). Obtained results demonstrated that severe salinity stress reduced the fresh weight (FW) and plant height (PH) by 16.40% and 19.10%, respectively compared with normal growth condition. On the other hands, under sever salinity stress relative water content (RWC) and chlorophyll index were reduced by 18.05% and 3.50%, respectively. Interestingly, selenium nanoparticles (Se-NPs; 10 mg/L) application improved the pineapple mint growth. Based on GC-FID and GC-MS analysis, 19 compounds were identified in pineapple mint essential oil. Foliar application of Se-NPs and salinity did not change the essential oil content of pineapple mint, however, the essential oil compounds were significantly affected by salinity and Se-NPs- applications. Foliar application of Se-NPs- had a significant effect on piperitenone oxide, limonene, jasmone, viridiflorol and ß-myrsene under different salinity levels. The highest percentage of piperitenone oxide (79.4%) as the major essential oil component was recorded in the no salinity treatment by applying 10 mg/L of nanoparticle. Interestingly, application of 10 mg L-1 Se-NPs- under 60 mM NaCl increased the piperitenone oxide content by 9.1% compared with non-sprayed plants. Finally, the obtained results demonstrated that foliar application of Se-NPs (10 mg L-1) can improve the pineapple mint growth and secondary metabolites profile under saline conditions.

Natural diversity in phenolic components and antioxidant properties of oregano (Origanum vulgare L.) accessions, grown under the same conditions.

Oregano (Origanum vulgare L.) is a rich source of biologically active components such as phenolic compounds. Here, seven pot grown O. vulgare accessions belonging to three subspecies (subsp. virens, subsp. vulgare and subsp. gracile) were investigated for their content in sixteen bioactive phenolic compounds as well as their antioxidant capacities (DPPH• and FRAP tests), total phenolic content (TPC) and total flavonoid content (TFC) in order to identify the most suitable ones on an industrial level. HPLC analyses showed that rosmarinic acid (659.6-1646.9 mg/100 g DW) was by far the most abundant constituent, followed by luteolin (46.5-345.4 mg/100 g DW), chicoric acid (36.3-212.5 mg/100 g DW), coumarin (65.7-193.9 mg/100 g DW) and quercetin (10.6-106.1 mg/100 g DW), with variability in concentration depending on the accession and subspecies. The highest level of rosmarinic acid and TPC was obtained from Ardabil accession (subsp. virens). There was a significant and positive correlation between rosmarinic acid and antioxidant activity (r = 0.46). TFC significantly correlated to TPC (r = 0.57) as well as to chicoric acid (r = 0.73). Cluster (CA) and principal component (PCA) analyses classified the investigated accessions in three different groups. Such natural variabilities in phenolics provide the possibility of using elite plants for nutraceutical and pharmaceutical industries and domestication of highly antioxidative accessions of oregano.

Improvement of Growth, Yield, Seed Production and Phytochemical Properties of Satureja khuzistanica Jamzad by Foliar Application of Boron and Zinc.

Satureja khuzistanica Jamzad is a valuable and endemic medicinal plant. Boron and zinc are essential elements for the vegetative and reproductive growth of plants and have significant effects on yield, essential oil composition and the seed production of plants. To investigate the effects of the foliar application of zinc and boron on the growth, yield, seed production and phytochemical properties of S. khuzistanica, a study was conducted in a factorial experiment with three replicates in two consecutive years based on a randomized complete block design. The foliar application of boron (B) at three concentrations (control or distilled water, 0.4% and 0.8% as H3BO3) and zinc (Zn) at three concentrations (control or distilled water, 0.3% and 0.6% as ZnSO4) was carried out. Our results showed that the foliar application of B resulted in a significant increase in the fresh and dry weights of plants, the dry weight of stems, drug yield, seed yield, seed germination and 1000-seed weight. At the same time, the application of B resulted in a significant decrease in seed emptiness. The fresh and dry weights of plants, drug yield, seed yield, 1000-seed weight and seed germination were also significantly improved by Zn foliar spraying compared to the control. Application of 0.8% B resulted in a significant decrease in seed emptiness by 14.16% and 22.37%, as compared to the control. The foliar spraying of B and Zn improved the total phenolic content, the essential oil content and the yield and antioxidant activity of S. khuzistanica. Moreover, B application generally concentrated more carvacrol in the essential oil (in the first experimental year). In contrast, no significant differences were observed between Zn treatments in carvacrol content and total flavonoids. The use of several microelements, such as B and Zn, could improve both the quantity and quality of S. khuzistanica. Additionally, improvement of seed set and seed quality by the foliar spraying of Zn and B may be useful for growing plants in arid and semi-arid areas.

Funneliformis mosseae inoculation under water deficit stress improves the yield and phytochemical characteristics of thyme in intercropping with soybean.

Intercropping of medicinal plants/legumes along with bio-fertilizer application is a relatively new sustainable practice for improving the yield and secondary metabolites production. Here, a 2-years field experiment was performed to evaluate the effects of water deficit stress and arbuscular mycorrhizal fungi (AMF) application (as bio-fertilizer) on nutrients concentration, dry matter yield, essential oil quantity and quality of thyme in intercropping with soybean. Three irrigation levels, including (i) irrigation after depletion of 20% (I20) as non-stressed, 50% (I50) as moderate water deficit and 80% (I80) available water as severe water deficit were applied as the main factor. The sub-factor was represented by different cropping patterns including thyme sole culture, replacement intercrop ratio of 50:50 and 66:34 (soybean: thyme) and the third factor was non-usage (control) and usage of AMF. According to our results, the thyme dry yield under moderate and severe water deficit stress decreased by 35 and 44% in the first year, and by 27 and 40% in the second year compared with non-stressed (I20) plants, respectively. Also, the macro- and micro-nutrients of thyme leaves increased significantly in intercropping patterns after application of AMF. The maximum essential oil percentage of thyme was achieved in 50:50 intercropping ratio treated with AMF. Under moderate and severe water deficits, the major constituents of thyme essential oil including thymol, p-cymene and γ-terpinene were increased in intercropping patterns treated with AMF. Generally, AMF application in intercropping ratio of 50:50 may be proposed to farmers as an eco-friendly approach to achieve desirable essential oil quality and quantity in thyme under water deficit stress conditions.

Effects of exogenous application of citrulline on prolonged water stress damages in hyssop (Hyssopus officinalis L.): Antioxidant activity, biochemical indices, and essential oils profile.

This study examined the effects of exogenous citrulline (control, 1 and 2 mM) and water availability (100%, 50% and 25% WA) on antioxidant attributes and essential oil constituents of Hyssopus officinalis L. in two successive harvests. Hyssop tolerantly responded to water deficiency by well-promoted antioxidant enzymes (i.e., superoxide dismutase, and catalase), strong DPPH-scavenging activity, and increasing polyphenols; however, the essential oil content was negatively reduced by water stress. External citrulline further increased the activity of antioxidant enzymes. Citrulline application at 2 mM under severe water stress could also improve essential oil (EO) content in the first and second harvests by about 15 and 30%, respectively. Furthermore, under severe drought, citrulline at 2 mM could obtain the highest yield of isopinocamphone (47%) as the main component of EO. The results showed the high potential of this novel applied metabolite agent to be used in a well-fulfilled production of this medicinal plant.

The volatile oils from the oleo-gum-resins of Ferula assa-foetida and Ferula gummosa: A comprehensive investigation of their insecticidal activity and eco-toxicological effects.

Since time immemorial, the oleo-gum-resins of Ferula assa-foetida and F. gummosa are used in the traditional medical systems as well as in foodstuffs, perfumery and cosmetics. In the present study, we explored the insecticidal efficacy of the essential oils obtained from these oleo-gum-resins to widen their fields of industrial applications. The two essential oils were mainly composed of sulfides [sec-butyl (Z)-propenyl disulfide, sec-butyl (E)-propenyl disulfide, sec-butyl (Z)-propenyl trisulfide and sec-butyl (E)-propenyl trisulfide)] and monoterpenes (α-pinene, ß-pinene and ß-phellandrene), respectively, as determined by GC-MS analysis. The two essential oils were assayed for toxicity on a panel of insects, represented by species of public health relevance (Culex quinquefasciatus and Musca domestica), agricultural (Spodoptera littoralis) and stored-product pests (Prostephanus truncatus and Trogoderma granarium). The ecotoxicological effects of the essential oils were assessed on the aquatic microcrustacean Daphnia magna and the earthworm Eisenia fetida, as well as on human cells. Overall, the two essential oils were effective against important insect pests and vectors. On the other hand, they resulted cytotoxic to fibroblasts and non-target aquatic microcrustaceans. Thus, further insights are needed to determine the full spectrum of their eco-toxicological effects.

Chemical composition, antioxidant activity and cytotoxicity on tumour cells of the essential oil from flowers of Magnolia grandiflora cultivated in Iran.

Magnolia grandiflora (Magnoliaceae) is an evergreen tree with fragrant and showy flowers native to southeastern USA but widely cultivated all over the world and used in cosmetics industry in treatment of skin diseases. Here, we report on the chemical analysis of the essential oil obtained from flowers of plants cultivated in Iran, together with the evaluation of its antioxidant and cytotoxic activities. The essential oil composition was dominated by bioactive sesquiterpenes, namely ß-elemene, bicyclogermacrene, germacrene D and (E)-caryophyllene. The oil exhibited moderate radical scavenging activity towards the [Formula: see text] radical, and mild non-selective inhibitory effects against A375, MDA-MB 231 and T98 G tumour cell lines. The latter were influenced by the presence of the anticancer ß-elemene. These results provided new insights for potential application of M. grandiflora volatile oil in the pharmaceutical and cosmetics industry where only the non-volatile magnolol and honokiol have hitherto been fully exploited.