Essential Oils of Micromeria dalmatica Benth., a Balkan Endemic Species of Section Pseudomelissa

The essential oils of 13 Greek populations of Micromeria dalmatica, a Balkan endemic species and member of the section Pseudomelissa, were examined for the first time. Among the studied populations, two main oil types could be distinguished. Type I was found to be rich in β‐pinene, limonene, and germacrene D (accounting for 55.6–70.2% of the total oil), and Type II was characterized by the preponderance of p‐menthane compounds (accounting for 64.2–89.9% of the oil). The latter oil type could be further divided into two subtypes, one comprising oils with predominance of piperitenone and piperitenone oxide and another composed of oils containing high proportions of pulegone, menthone, and isomenthone. The abundance of p‐menthane compounds is a common feature of the oils of all members of the section Pseudomelissa studied to date. However, the existence of oils of Type I has not been previously reported for M. dalmatica, neither for other members of the section Pseudomelissa.     

Biocidal Compounds from Mentha sp. Essential Oils and Their Structure–Activity Relationships

Essential oils from Greek Mentha species showed different chemical compositions for two populations of M. pulegium, characterized by piperitone and pulegone. Mentha spicata essential oil was characterized by endocyclic piperitenone epoxide, piperitone epoxide, and carvone. The bioactivities of these essential oils and their components have been tested against insect pests (Leptinotarsa decemlineata, Spodoptera littoralis and Myzus persicae), root‐knot nematodes (Meloydogine javanica) and plants (Lactuca sativa, Lolium perenne, Solanum lycopersicum). The structure–activity relationships of these compounds have been studied including semi‐synthetic endocyclic trans‐carvone epoxide, exocyclic carvone epoxide, a new exocyclic piperitenone epoxide and trans‐pulegone epoxide. Leptinotarsa decemlineata feeding was affected by piperitenone and piperitone epoxide. Spodoptera littoralis was affected by piperitone epoxide and pulegone. The strongest nematicidal agent was piperitenone epoxide, followed by piperitone epoxide, piperitenone and carvone. Germination of S. lycopersicum and L. perenne was significantly affected by piperitenone epoxide. This compound and carvone epoxide inhibited L. perenne root and leaf growth. Piperitenone epoxide also inhibited the root growth of S. lycopersicum. The presence of a C(1) epoxide resulted in strong antifeedant, nematicidal and phytotoxic compounds regardless of the C(4) substituent. New natural crop protectants could be developed through appropriate structural modifications in the p‐menthane skeleton.     

Chemical Characterization and Insecticidal Properties of Essential Oils from Different Wild Populations of Mentha suaveolens subsp. timija (Briq.) Harley from Morocco

The present study is the first investigation of the volatile‐oil variability and insecticidal properties of the endemic Moroccan mint Mentha suaveolens subsp. timija (mint timija). The yield of essential oils (EOs) obtained from different wild mint timija populations ranged from 0.20±0.02 to 1.17±0.25% (v/w). GC/MS Analysis revealed the presence of 44 oil constituents, comprising 97.3–99.9% of the total oil compositions. The main constituents were found to be menthone (1.2–62.6%), pulegone (0.8–26.6%), cis‐piperitone epoxide (2.9–25.5%), piperitone (0.3–35.5%), trans‐piperitone epoxide (8.1–15.7%), piperitenone (0.2–9.6%), piperitenone oxide (0.5–28.6%), (E)‐caryophyllene (1.5–11.0%), germacrene D (1.0–15.7%), isomenthone (0.3–7.7%), and borneol (0.2–7.3%). Hierarchical‐cluster analysis allowed the classification of the EOs of the different mint timija populations into four main groups according to the contents of their major components. This variability within the species showed to be linked to the altitude variation of the mint timija growing sites. The results of the insecticidal tests showed that all samples exhibited interesting activity against adults of Tribolium castaneum, but with different degrees. The highest toxicity was observed for the EOs belonging to Group IV, which were rich in menthone and pulegone, with LC₅₀ and LC₉₀ values of 19.0–23.4 and 54.9–58.0 μl/l air in the fumigation assay and LC₅₀ and LC₉₀ values of 0.17–0.18 and 0.40–0.52 μl/cm² in the contact assay.     

Mentha longifolia in vitro cultures as safe source of flavouring ingredients

In vitro plantlets and callus of M. longifolia were established and their volatile constituents characterized by GC-MS analysis of their headspaces (HSs) and essential oils (EOs). Significant quali-quantitative differences were found in the aromatic fingerprints in comparison with the M. longifolia parent plants. In fact, limonene and carvone were the main constituents in the EOs of the mother plants, while the aroma of the in vitro plant material were especially enriched in oxygenated terpenes. In particular, huge amounts of piperitenone and piperitenone oxide (75 %) were found for in vitro plantlets, while trans-carvone oxide (19 %) and trans-piperitone epoxide (9 %) were found in callus EO. However, the established in vitro plant material showed lack of pulegone and menthofurane, thus preserving an important feature observed in the volatile fingerprint of the parent plants. In fact, because of their well-known toxicity significant amounts of pulegone and menthofurane may compromise the safety using of mint essential oil. Therefore the in vitro M. longifolia plantlets and callus may be regarded as a potential source of a safe flavouring agent.     

Essential‐Oil Variability of Juniperus deltoides R.P.Adams along the East Adriatic Coast – How Many Chemotypes Are There?

The composition of the essential oils isolated from twigs of ten Juniperus deltoides R.P . Adams populations from the east Adriatic coast was determined by GC‐FID and GC/MS analyses. Altogether, 169 compounds were identified, representing 95.6–98.4% of the total oil composition. The oils were dominated by monoterpenes (average content of 61.6%), which are characteristic oil components of species of the Juniperus section. Two monoterpenes, α‐pinene and limonene, were the dominant constituents, comprising on average 46.78% of the essential oils. Statistical methods were deployed to determine the diversity of the terpene classes and the common terpenes between the investigated populations. These statistical analyses revealed the existence of three chemotypes within all populations, i.e., a α‐pinene, limonene, and limonene/α‐pinene type.     

Biological activities of the essential oils and methanol extract of tow cultivated mint species (<Emphasis Type="Italic">Mentha longifolia</Emphasis> and <Emphasis Type="Italic">Mentha pulegium</Emphasis>) used in the Tunisian folkloric medicine

The composition of the essential oils and methanolic extracts of two cultivated mint species (M. longifolia and M. pulegium), as well as the in vitro antimicrobial and antioxidant activities of the essential oil and methanol extract of Mentha longifolia and Mentha pulegium were compared. GC-MS analysis of the essential oil identified 41 compounds constituting 96.66 and 96.13% of the total oil from M. longifolia and M. pulegium, respectively. The later oils were rich on pulegone (47.15 and 61.11%, respectively). Moreover, 1,8 cineole (11.54%), menthone (10.7%), α-pinene (3.57%), α-terpineol (3.17%) and d-cadinene (3.53%) were only present in M. longifolia oil, while isomenthone (17.02%), and piperitone (2.63%), were characteristic of M. pulegium oil. Shoot extract of the two species showed significantly different contents in total polyphenols (89.1 and 37.41 mg GAE/g DW), flavonoids (63.93 and 33.83 mg CE/g DW) and tannins (1.47 and 3.07 mg CE/g DW), respectively in M. longifolia and M. pulegium. The essential oils showed strong antimicrobial activity against all 16 microorganisms tested, whereas the methanol extracts were inactive. Moreover, the essential oils showed higher antioxidant activity than the methanolic extracts against the DPPH and superoxide radical scavenging. In fact, antioxidant activities of the oils were the same for both M. longifolia and M. pulegium against DPPH (IC₅₀ = 9 and 10 μg/ml, respectively) and 2-fold and 4-fold higher than shoot extracts (IC₅₀ = 20 and 48 μg/ml, respectively). Moreover, both oils showed the same antioxidative abilities as compared to the positive control (butylated hydroxytoluene). In the same way, the capacity to inhibit superoxide anion was very significant for the two oils (0.1 μg/ml for M. longifolia and 0.11 μg/ml for M. pulegium).     

Glandular trichomes and essential oil characteristics of in vitro propagated <Emphasis Type="Italic">Micromeria pulegium</Emphasis> (Rochel) Benth. (Lamiaceae)

Main conclusion

In vitro conditions and benzyladenine influenced both content and composition of micropropagated Micromeria pulegium essential oils, with pulegone and menthone being the main essential oil components. The content and chemical composition of Micromeria pulegium (Rochel) Benth. essential oils were studied in native plant material at vegetative stage and in micropropagated plants, obtained from nodal segments cultured on solid MS medium supplemented with N⁶–benzyladenine (BA) or kinetin at different concentrations, alone or in combination with indole-3-acetic acid. Shoot proliferation was achieved in all treatments, but the highest biomass production was obtained after treatment with 10 μM BA. Phytochemical analysis identified up to 21 compounds in the essential oils of wild-growing and in vitro cultivated plants, both showing very high percentages of total monoterpenoids dominated by oxygenated monoterpenes of the menthane type. Pulegone and menthone were the main essential oil components detected in both wild-growing plants (60.07 and 26.85 %, respectively) and micropropagated plants grown on either plant growth regulator-free medium (44.57 and 29.14 %, respectively) or BA-supplemented medium (50.77 and 14.45 %, respectively). The percentage of total sesquiterpenoids increased in vitro, particularly owing to sesquiterpene hydrocarbons that were not found in wild-growing plants. Differences in both content and the composition of the essential oils obtained from different samples indicated that in vitro culture conditions and plant growth regulators significantly influence the essential oils properties. In addition, the morphology and structure of M. pulegium glandular trichomes in relation to the secretory process were characterized for the first time using SEM and light microscopy, and their secretion was histochemically analyzed.

     

Chemical Diversity among the Essential Oils of Wild Populations of Stachys lavandulifolia Vahl (Lamiaceae) from Iran

The variation of the essential‐oil composition among ten wild populations of Stachys lavandulifolia Vahl (Lamiaceae), collected from different geographical regions of Iran, was assessed by GC‐FID and GC/MS analyses, and their intraspecific chemical variability was determined. Altogether, 49 compounds were identified in the oils, and a relatively high variation in their contents was found. The major compounds of the essential oils were myrcene (0.0–26.2%), limonene (0.0–24.5%), germacrene D (4.2–19.3%), bicyclogermacrene (1.6–18.0%), δ‐cadinene (6.5–16.0%), pulegone (0.0–15.1%), (Z)‐hex‐3‐enyl tiglate (0.0–15.1%), (E)‐caryophyllene (0.0–12.9), α‐zingiberene (0.2–12.2%), and spathulenol (1.6–11.1%). For the determination of the chemotypes and the chemical variability, the essential‐oil components were subjected to cluster analysis (CA). The five different chemotypes characterized were Chemotype I (germacrene D/bicyclogermacrene), Chemotype II (germacrene D/spathulenol), Chemotype III (limonene/δ‐cadinene), Chemotype IV (pulegone), and Chemotype V (α‐zingiberene). The high chemical variation among the populations according to their geographical and bioclimatic distribution imposes that conservation strategies of populations should be made appropriately, taking into account these factors. The in situ and ex situ conservation strategies should concern all populations representing the different chemotypes.     

Variations in composition of peppermint oil in relation to production areas

Compositional data obtained for peppermint oil fromMentha piperita L. produced in Tasmania as well as oils produced in other major production areas, were analysed by principal coordinate analysis. The 6 variates included in the analysis were the oil compounds limonene, cineole, menthone, menthofuran, menthyl acetate and menthol. In general, Tasmanian oils were characterised by low menthofuran, low limonene, low menthyl acetate and to a lesser extent high menthone, low cineole and high menthol concentrations relative to most major production areas. When the variation in composition of oil samples from within Tasmania was displayed in 3 dimensions (using the first 3 principal coordinates) it was not possible to achieve any pronounced separation of oils produced at different locations within southern Tasmania. Principal coordinates which were based mainly on cineole and menthol concentrations, respectively, did allow a degree of separation between oils produced in southern and northern Tasmania. Generally, oils produced in northern Tasmania had lower cineole and, in some cases, higher menthol than southern Tasmanian oils. Oil extracted from regrowth herb after commercial harvest was distinguished by having very high menthyl acetate, low menthone, high menthol, high menthofuran, low cineole and low limonene concentrations. The effect of some cultural and environmental factors on oil composition is also discussed.     

Intraspecific Variability of the Essential Oil of Ziziphora clinopodioides ssp. rigida from Iran

Hydrodistillated essential oils of Ziziphora clinopodioides ssp. rigida from nine populations of the Lashgardar protected region (Hamedan Province, Iran) were analyzed by using GC and GC/MS techniques to determine the intraspecific chemical variability. Altogether, 39 compounds were identified in the oils, and a relatively high variation in their contents was found. The main constituents of the essential oils were pulegone (0.7–44.5%), 1,8‐cineole (2.1–26.0%), neomenthol (2.5–22.5%), 4‐terpineol (0.0–9.9%), 1‐terpineol (0.0–13.2%), neomenthyl acetate (0.0–7.1%), and piperitenone (0.0–5.4%). For the determination of the chemotypes and the intraspecific chemical variability, the essential oil components were subjected to cluster analysis (CA). The five different chemotypes characterized were Chemotype I (pulegone/neomenthol), Chemotype II (pulegone), Chemotype III (pulegone/1,8‐cineole), Chemotype IV (neomenthol), and Chemotype V (1,8‐cineole/4‐terpineol).     

Variability in Chemical Composition and Antimicrobial Activity of Mentha pulegium L. Essential Oil,Cultivated under Different Plant Growth Promoting Rhizobacteria

The aim of present work was to study the essential oil chemical composition and antimicrobial activity of cultivated Mentha pulegium L. under different plant growth promoting rhizobacteria (Pseudomonas fluorescens, Bradyrhizobium sp. and Sinorhizobium meliloti) individually and in consortium. Yield, in plants inoculated with Bradyrhizobium sp. and S. meliloti in consortium, increase significantly relative to control plants. GC and GC/MS analyses pointed to a qualitative and quantitative variability of components. The investigated essential oils were clustered into three chemotypes: piperitenone/1,8-cineol (40.9/29.4 %) chemotype in plants inoculated with Bradyrhizobium sp. individually, S. meliloti individually, and Bradyrhizobium sp. and S. meliloti in consortium, piperitone/menthone (41.8/33.8 %) chemotype in plants inoculated with P. fluorescens individually, P. fluorescens and Bradyrhizobium sp. in consortium, and P. fluorescens and S. meliloti in consortium and pulegone/menthol (47.9/31.5 %) chemotype in control plants. The antimicrobial activity, carried out by the disc diffusion method and the determination of the Minimum Inhibitory Concentration (MIC) against ten microorganisms, varied significantly according to the tested microorganism and the rhizobacterial species used individually or in consortium (inhibition zone: 8.5–33.5 mm; MIC: 0.25–2.5 μL/mL). Our findings provided useful indications to select interesting chemotype within M. pulegium, especially in perspective of its cultivation.     

Composition and biovariability of Deverra scoparia volatile oil and its potential use as a source of bioactive phthalide components

Chemical biodiversity of essential oils of natural populations of Deverra scoparia growing in different regions in Algeria, and spreading from of semiarid to Saharan territories, was assessed. More than 60 compounds were identified by GC and GC-MS. Both phenylpropanoids and phathalids classes of bioactive components in leaves part were exhibiting drastic variations: (0.77–59.46%) and (0.04–28.55%), respectively. More specifically, the following bioactive components reached a very high level for this plant species: myristicin “up to 58.19%”, dill apiole “up to 25.52%” and (Z)-ligustilide “up to 27.5%”. Seven major compounds identified for the flowers part were varying from very low to very high percentages: α-pinene (2.49–42.32%), sabinene + β-pinene (1.11–30.31%), α-phellandrene (0.99–20.63%), p-cymene (0.0–44.22%), limonene (0.0–42.99%), myristicin (0.03–58.19%) and dill apiole (0.4–25.52%). Similarly, for the stems part, larges variations of both major and minor components were also registered: α-pinene (17.97–53.25%), sabinene + β-pinene (3.44–24.99%), α-phellandrene (0.75–10.32%), limonene (1.35–59.68%), terpinen-4-ol (0.29–15.85%) and spathulenol (0.46–11.29%). Agglomerative Hierarchical Cluster analysis was conducted, showing the existence of two main groups. Obtained data for volatile components in each plant part gave an insight of the existence of simultaneous clear and large biodiversity of the chemical profiles of the studied plant parts.     

Screening the essential oil composition of wild Sicilian fennel

Fifty-six samples of wild fennel (Foeniculum vulgare Mill.) have been collected in different localities of Sicily and analysed for their content in seed essential oils. The GC-FID-MS analyses allowed identifying 78 compounds, representing more than 98% of the oils. Phenylpropanoids are the most highly represented components – 55 samples show estragole as the main compound ranging between 34 and 89%, while (E)-anethole is the other phenylpropanoid ranging between 0.1 and 36%, although it only reaches appreciable values in a few samples, largely being present below 1%. The oxygenated monoterpene, fenchone, a typical fennel oil component, has been found in all samples, in the 2–27% range; α-pinene (1–21%), limonene (1–17%) and γ-terpinene (<1–4%) were the monoterpene hydrocarbons found in all samples. Only one sample showed piperitenone oxide and limonene as main components, with 42 and 34%, respectively, and with the lack of any phenylpropanoid. Sesquiterpenes and others compounds class had only negligible values or were not detected in all samples. Statistical analyses allowed to single out five groups, reflecting the compositional differences of the essential oil profiles of the wild Sicilian fennel.     

Effect of Growth Stage on Essential‐Oil Yield and Composition of Daucus sahariensis

The essential oils obtained by hydrodistillation from Daucus sahariensis Murb . harvested at three different growth stages were characterized by GC/MS analysis. In total, 88 compounds were identified, with myristicin (29.8–51.7%), myrcene (6.7–31.1%), α‐pinene (11.6–14.8%), and limonene (5.3–11.5%) as main constituents. Monoterpene hydrocarbons were the most represented compounds in the oils of the plant samples collected during the flower‐budding and full‐flowering periods. On the contrary, during the fruiting stage, the oils were dominated by phenylpropanoids. The essential oils were subject of considerable variation in their composition during the various developmental stages, particularly concerning the content of myrcene that decreased significantly passing from the vegetative to the fruiting stage. Conversely, for myristicin, the opposite trend was observed. Furthermore, the essential‐oil yields were quite low during the flower‐budding phase (0.27%), but rapidly increased during plant development (0.63 and 0.68% for the flowering and fruiting phases, resp.).     

Chemical Variability of the Needle Oil of Juniperus communis ssp. alpina from Corsica

The composition of 109 samples of essential oil isolated from the needles of Juniperus communis ssp. alpina growing wild in Corsica was investigated by GC (in combination with retention indices), GC/MS, and ¹³C‐NMR. Forty‐four compounds accounting for 86.7–96.7% of the oil were identified. The oils consisted mainly of monoterpene hydrocarbons, in particular, limonene (9.2–53.9%), β‐phellandrene (3.7–25.2%), α‐pinene (1.4–33.7%), and sabinene (0.1–33.6%). The 109 oil compositions were submitted to k‐means partitioning and principal component analysis, which allowed the distinction of two groups within the oil samples. The composition of the major group (92% of the samples) was dominated by limonene and β‐phellandrene, while the second group contained mainly sabinene beside limonene and β‐phellandrene.     

Preliminary studies of monoterpenoid variability in Satureja douglasii

Monoterpenoid composition of samples from 10 populations of Satureja douglasii in a 250-ha area were analyzed in Spring 1974. Two distinct compositional types were found: Type A contained large amounts of limonene and carvone but very small quantites of piperitenone, piperitone, pulegone and isomenthone (3-oxy compounds), whereas Type B was essentially the reverse. This variation is attributed to a genotypic difference. The amounts of 3-oxy compounds varied greatly among the Type B populations. Increasing shade conditions were correlated with increased amounts of the more reduced compounds isomenthone and piperitone. Clonal lines were established from two Type B populations whose habitats represented extremes of sun and shade conditions. The two clonal lines, which had very different compositions in the field, produced similar oils under controlled conditions. Growth under different temperature regimes had little effect on composition. Developmental changes in composition implied the precursor-product relationships piperitenone→piperitone and pulegone→isomenthone. The data suggest that S douglasii monoterpenoid compositional patterns are significantly influenced by genetic and environmental factors (probably light intensity) as well as developmental processes.     

Volatile secondary metabolites of Micromeria dalmatica Benth. (Lamiaceae): biosynthetical and chemotaxonomical aspects

Analysis by GC and GC/MS of the essential oil obtained from above-ground parts of Micromeria dalmatica Benth. allowed the identification of 116 components, comprising 93.6% of the total oil composition. The major compounds are 3-oxygenated p-menthane monoterpenes and were identified as pulegone (29.6%), menthone (11.7%), and piperitenone (10.8%). The chemical composition of this and additional 30 oils obtained from selected Micromeria Benth. taxa were compared by using multivariate statistical analysis (agglomerative hierarchical cluster analysis and principal component analysis (PCA)). The results of statistical analyses, as well as the domination of different concurrent p-menthane-skeleton-type monoterpene biosynthetical sub-branches in the compared M. dalmatica samples, implied the occurrence of at least two different chemotypes of the mentioned species.     

Comparative Study of Chemical Composition,Cholinesterase Inhibition and Antioxidant Potential of Mentha pulegium L. Essential Oil

Perennial plant Mentha pulegium L. (pennyroyal, Lamiaceae) can be found in Europe and Mediterranean. In areas where it thrives, M. pulegium is used in nutrition and as medicinal plant. Essential oil of M. pulegium is also a frequent constituent of foods and fragrances, because of mint-like odor. Regarding the use of M. pulegium in traditional medicine and nutrition, as well as fact that essential oils are potential sources of bioactive components, this study was conducted to examine the chemical composition of essential oil of M. pulegium wild growing in Bosnia and Herzegovina and its biological activity. The chemical profile testing was made using GC/MS and GC/FID technique. Potential of cholinesterase inhibition was tested by Ellman's assay. The antioxidant activity was tested by DPPH and FRAP assay. The dominant components in analyzed oil were pulegone 54.4 %, p-menthone 14.0 % and piperitenone 12.8 %. Good antioxidant activity and moderate cholinesterase inhibition potential of tested essential oil indicates to possibility of its use in treatment of diseases related to free radicals, Alzheimer disease and as lipid protecting antioxidant.     

Phytochemical Analysis and in vitro Free‐Radical‐Scavenging Activities of the Essential Oils from Leaf and Fruit of Melaleuca leucadendra L.

The phytochemical profile of Melaleuca leucadendra L. leaf and fruit oils from Cuba was investigated by GC and GC/MS. Forty‐one and sixty‐four volatile compounds were identified and quantified, accounting for 99.2 and 99.5% of the leaf‐oil and fruit‐oil total composition, respectively. The main components were 1,8‐cineol (43.0%), viridiflorol (24.2%), α‐terpineol (7.0%), α‐pinene (5.3%), and limonene (4.8%) in the leaf oil, and viridiflorol (47.6%), globulol (5.8%), guaiol (5.3%), and α‐pinene (4.5%) in the fruit oil. The antioxidant capacity of these essential oils was determined by three different in vitro assays (2,2‐diphenyl‐1‐picrylhydrazyl (DPPH) radical, thiobarbituric acid reactive species (TBARS), and 2,2′‐Azinobis(3‐ethylbenzothiazoline‐6‐sulfonic acid) (ABTS) radical cation), and significant activities were evidenced for all of them.     

Chemical Composition of Juniperus communis L. Cone Essential Oil and Its Variability among Wild Populations in Kosovo

Ripe cones of Juniperus communis L. (Cupressaceae) were collected from five wild populations in Kosovo, with the aim of investigating the chemical composition and natural variation of essential oils between and within wild populations. Ripe cones were collected, air dried, crushed, and the essential oils obtained by hydrodistillation. The essential‐oil constituents were identified by GC‐FID and GC/MS analyses. The yield of essential oil differed depending on the population origins and ranged from 0.4 to 3.8% (v/w, based on the dry weight). In total, 42 compounds were identified in the essential oils of all populations. The principal components of the cone‐essential oils were α‐pinene, followed by β‐myrcene, sabinene, and D ‐limonene. Taking into consideration the yield and chemical composition, the essential oil originating from various collection sites in Kosovo fulfilled the minimum requirements for J. communis essential oils of the European Pharmacopoeia. Hierarchical cluster analysis (HCA) and principal component analysis (PCA) were used to determine the influence of the geographical variations on the essential‐oil composition. These statistical analyses suggested that the clustering of populations was not related to their geographic location, but rather appeared to be linked to local selective forces acting on the chemotype diversity.