Chemical Profiling and Antifungal Activity of Ziziphora clinopodioides Leaf Essential Oil against the Pathogen Verticillium dahliae

Essential oils (EOs) are often used as natural antifungal agents to control the growth of phytopathogenic fungi. The aim of this study was to determine the effect of Ziziphora clinopodioides leaf EO against Verticillium dahliae, a pathogenic fungus of cotton. Gas chromatography-mass spectrometry (GC/MS) analysis revealed the presence of 15 compounds of the total of extracted oil, which was consisted of 98.79 % monoterpenes and 0.61 % sesquiterpenes. The major constituents were pulegone (62.17 %), isomenthone (18.42 %), l-menthone (5.55 %) and piperitenone (3.99 %). The mycelial growth of Verticillium dahliae was completely inhibited at 0.24 μL/mL air under vapor phase condition. Considerable morphological variations were also observed in the fungal sclerotia at the contact phase at 3 μL/mL. This study demonstrated for the first time that Z. clinopodioides EO can effectively inhibit the growth of V. dahliae, implying that it has the potential to be explored as an antifungal agent against Verticillium Wilt of cotton.     

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.     

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.     

Seasonal and Geographical Influences on the Chemical Composition of Juniperus phoenicea L. Essential Oil Leaves from the Northern Tunisia

The essential‐oil composition of 60 individual trees of Juniperus phoenicea L. from four Tunisian populations in three different periods were investigated by GC and GC/MS analyses. 59 Compounds were identified in the oils, and a relatively high variation in their contents was found. All the oils were dominated by the terpenic hydrocarbon fraction, and the main component was α‐pinene (20.28–40.86%). The results of the oil compositions were processed by hierarchical clustering and principal component analysis (PCA) allowing establishing four groups of essential‐oils differentiated by one compound or more. Pattern of geographic variation in essential‐oil composition indicated that individuals from the continental site (Makthar) were clearly distinguished from those from littoral localities (Tabarka, Hawaria, and Rimel).     

Essential‐Oil Composition and Chemical Variability of Senecio vulgaris L. from Corsica

The chemical composition of the essential oils isolated from the aerial parts of Senecio vulgaris plants collected in 30 Corsican localities was characterized using GC‐FID and GC/MS analyses. Altogether, 54 components, which accounted for 95.2% of the total oil composition, were identified in the 30 essential‐oil samples. The main compounds were α‐humulene ( 1 ; 57.3%), (E)‐β‐caryophyllene ( 2 ; 5.6%), terpinolene ( 3 ; 5.3%), ar‐curcumene ( 4 ; 4.3%), and geranyl linalool ( 5 ; 3.4%). The chemical composition of the essential oils obtained from separate organs and during the complete vegetative cycle of the plants were also studied, to gain more knowledge about the plant ecology. The production of monoterpene hydrocarbons, especially terpinolene, seems to be implicated in the plant‐flowering process and, indirectly, in the dispersal of this weed species. Comparison of the present results with the literature highlighted the originality of the Corsican S. vulgaris essential oils and indicated that α‐humulene might be used as taxonomical marker for the future classification of the Senecio genus. A study of the chemical variability of the 30 S. vulgaris essential oils using statistical analysis allowed the discrimination of two main clusters according to the soil nature of the sample locations. These results confirmed that there is a relation between the soil nature, the chemical composition of the essential oils, and morphological plant characteristics. Moreover, they are of interest for commercial producers of essential oil in selecting the most appropriate plants.     

Chemical Variability of the Essential Oil of Juniperus phoenicea var. turbinata from Algeria

The chemical composition of 50 samples of leaf oil isolated from Algerian Juniperus phoenicea var. turbinata L. harvested in eight locations (littoral zone and highlands) was investigated by GC‐FID (in combination with retention indices), GC/MS, and ¹³C‐NMR analyses. The composition of the J. phoenicea var. turbinata leaf oils was dominated by monoterpenes. Hierarchical cluster and principal component analyses confirmed the chemical variability of the leaf oil of this species. Indeed, three clusters were distinguished on the basis of the α‐pinene, α‐terpinyl acetate, β‐phellandrene, and germacrene D contents. In most oil samples, α‐pinene (30.2–76.7%) was the major compound, associated with β‐phellandrene (up to 22.5%) and α‐terpinyl acetate (up to 13.4%). However, five out of the 50 samples exhibited an atypical composition characterized by the predominance of germacrene D (16.7–22.7%), α‐pinene (15.8–20.4%), and α‐terpinyl acetate (6.1–22.6%).     

Chemical Variability and Antioxidant Activity of Limbarda crithmoides L. Essential Oil from Corsica

The chemical compositions of 25 Corsican Limbarda crithmoides ssp. longifolia essential oils were investigated for the first time using GC‐FID, GC/MS, and NMR analyses. Altogether, 65 compounds were identified, accounting for 90.0–99.3% of the total oil compositions. The main components were p‐cymene ( 1 ; 15.1–34.6%), 3‐methoxy‐p‐cymenene ( 4 ; 11.8–28.5%), 2,5‐dimethoxy‐p‐cymenene ( 5 ; 5.9–16.4%), thymol methyl ether ( 6 ; 1.3–14.9%), α‐phellandrene ( 2 ; 0.9–11.9%), and α‐pinene ( 3 ; 0.2–13.4%). The chemical variability of the Corsican oil samples was studied using multivariate statistical analysis, which allowed the discrimination of two main clusters. A direct correlation between the water salinities of the plant locations and the chemical compositions of the L. crithmoides essential oils was evidenced. Indeed, essential oils rich in 1 (30.4–34.6%) were found in samples growing in the wetlands of the southern oriental coast, which exhibit high salinity levels (24.4±0.2–33.9±0.2 ppt), and essential oils with lower contents of 1 (15.1–27.3%) were isolated form samples growing in the wetlands of northern Corsica, which exhibit lower salinity levels (10.90±0.20–15.47±0.15 ppt). The antioxidant potential of L. crithmoides essential oil was also investigated, by assessing the DPPH^.‐ and ABTS^.+‐scavenging activities and the reducing power of ferric ions, and was found to be interesting. Moreover, using bioassay‐guided fractionation of the essential oil, a higher antioxidant activity was obtained for the oxygenated fraction and both ester and alcohol subfractions.     

Chemical Composition Variability of Essential Oils of Daucus gracilis Steinh. from Algeria

The chemical compositions of 20 Algerian Daucus gracilis essential oils were investigated using GC‐FID, GC/MS, and NMR analyses. Altogether, 47 compounds were identified, accounting for 90 – 99% of the total oil compositions. The main components were linalool ( 18 ; 12.5 – 22.6%), 2‐methylbutyl 2‐methylbutyrate ( 20 ; 9.2 – 20.2%), 2‐methylbutyl isobutyrate ( 10 ; 4.2 – 12.2%), ammimajane ( 47 ; 2.6 – 37.1%), (E)‐β‐ocimene ( 15 ; 0.2 – 12.8%) and 3‐methylbutyl isovalerate ( 19 ; 3.3 – 9.6%). The chemical composition of the essential oils obtained from separate organs was also studied. GC and GC/MS analysis of D. gracilis leaves and flowers allowed identifying 47 compounds, amounting to 92.3% and 94.1% of total oil composition, respectively. GC and GC/MS analysis of D. gracilis leaf and flower oils allowed identifying linalool (22.7%), 2‐methylbutyl 2‐methylbutyrate (18.9%), 2‐methylbutyl isovalerate (13.6%), ammimajane (10.4%), 3‐methylbutyl isovalerate (10.3%), (E)‐β‐ocimene (8.4%) and isopentyl 2‐methylbutyrate (8.1%) as main components. The chemical variability of the Algerian oil samples was studied using statistical analysis, which allowed the discrimination of three main Groups. A direct correlation between the altitudes, nature of soils and the chemical compositions of the D. gracilis essential oils was evidenced.     

Composition and Chemical Variability of the Needle Oil from Pinus halepensis growing in Corsica

The composition of oil samples isolated from needles of Pinus halepensis growing in three locations in Corsica (Saleccia, Capo di Feno, and Tre Padule) has been investigated by combination of chromatographic (GC with retention indices) and spectroscopic (MS and ¹³C‐NMR) techniques. In total, 35 compounds that accounted for 77 – 100% of the whole composition have been identified. α‐Pinene, myrcene, and (E)‐β‐caryophyllene were the major component followed by α‐humulene and 2‐phenylethyl isovalerate. Various diterpenes have been identified as minor components. 47 Oil samples isolated from pine needles have been analyzed and were differentiated in two groups. Oil samples of the first group (15 samples) contained myrcene (M = 28.1 g/100 g; SD = 10.6) and (E)‐β‐caryophyllene (M = 19.0 g/100 g; SD = 2.2) as major components and diterpenes were absent. All these oil samples were isolated from pine needles harvested in Saleccia. Oil samples of the second group (32 samples) contained mostly (E)‐β‐caryophyllene (M = 28.7 g/100 g; SD = 7.9), α‐pinene (M = 12.3 g/100 g; SD = 3.6), and myrcene (M = 11.7 g/100 g; SD = 7.3). All these oil samples were isolated from pine needles harvested in Capo di Feno and Tre Padule.     

Essential Oils and Chemical Diversity of Southeast European Populations of Salvia officinalis L.

The essential oils of 25 populations of Dalmatian sage (Salvia officinalis L.) from nine Balkan countries, including 17 indigenous populations (representing almost the entire native distribution area) and eight non‐indigenous (cultivated or naturalized) populations were analyzed. Their essential‐oil yield ranged from 0.25 to 3.48%. Within the total of 80 detected compounds, ten (β‐pinene, 1,8‐cineole, cis‐thujone, trans‐thujone, camphor, borneol, trans‐caryophyllene, α‐humulene, viridiflorol, and manool) represented 42.60 to 85.70% of the components in the analyzed essential oils. Strong positive correlations were observed between the contents of trans‐caryophyllene and α‐humulene, α‐humulene and viridiflorol, and viridiflorol and manool. Principal component analysis (PCA) on the basis of the contents of the ten main compounds showed that four principal components had an eigenvalue greater than 1 and explained 79.87% of the total variation. Performing cluster analysis (CA), the sage populations could be grouped into four distinct chemotypes (A–D). The essential oils of 14 out of the 25 populations of Dalmatian sage belonged to Chemotype A and were rich in cis‐thujone and camphor, with low contents of trans‐thujone. The correlation between the essential‐oil composition and geographic variables of the indigenous populations was not significant; hence, the similarities in the essential‐oil profile among populations could not be explained by the physical proximity of the populations. Additionally, the southeastern populations tended to have higher EO yields than the northwestern ones.     

Chemical Variability and Biological Activities of Essential Oils of Micromeria inodora (Desf.) Benth. from Algeria

The chemical composition of the essential oils isolated from the aerial parts of Micromeria inodora (Desf .) Benth . collected in 24 Algerian localities was investigated from the first time using GC‐FID, GC/MS and ¹³C‐NMR. Altogether, 83 components which accounted for 94.7% of the total oil composition were identified. The main compounds were trans‐sesquisabinene hydrate ( 1 ; 20.9%), α‐terpinyl acetate ( 2 ; 19.8%), globulol ( 3 ; 4.9%), caryophyllene oxide ( 4 ; 4.3%), β‐bisabolol ( 5 ; 2.9%) and trans‐7‐epi‐sesquisabinene hydrate ( 6 ; 2.6%). Comparison with the literature highlighted the originality of the Algerian M. inodora oil and indicated that 1 might be used as taxonomical marker. The study of the chemical variability allowed the discrimination of two main clusters confirming that there is a relation between the essential‐oil compositions and the soil nature of the harvest locations. Biological activity of M. inodora essential oil was assessed against fourteen species of microorganisms involved in nosocomial infections using paper disc diffusion and dilution agar assays. The in vitro study demonstrated a good activity against Gram‐positive strains such as Staphylococcus aureus, Bacillus cereus, Bacillus subtilis, and Enterococcus faecalis, and moderate activity against Candida albicans. These results might be useful for the future commercial valorization of M. inodora essential oil as a promising source of natural products with potential against various nosocomial community and toxinic infections.     

Variability in the Essential‐Oil Composition of Sideritis scardica Griseb. from Native Bulgarian Populations

The essential‐oil composition of six native populations of Sideritis scardica from Bulgaria was studied by GC‐FID and GC/MS analyses. Altogether, 37 components, representing 73.1 to 79.2% of the total oil content were identified. Among them, α‐pinene (4.4–25.1%), β‐pinene (2.8–18.0%), oct‐1‐en‐3‐ol (2.3–8.0%), phenylacetaldehyde (0.5–9.5%), β‐bisabolene (1.3–11.0%), benzyl benzoate (1.1–14.3%), and m‐camphorene ( 1 ; 0.3–12.4%) were the main compounds. All samples were characterized by low contents of oxygenated mono‐ and sesquiterpenes (≤1.6 and 2.3%, resp.). Principal component analysis (PCA) and cluster analysis (CA) showed a significant variability in the chemical composition of the studied samples as well as a correlation between the oil profiles and the ecological conditions of the natural habitats of S. scardica.     

Essential Oil Composition of Phagnalon sordidum (L.) from Corsica,Chemical Variability and Antimicrobial Activity

The chemical composition of Phagnalon sordidum (L.) essential oil was investigated for the first time using gas chromatography and chromatography/mass spectrometry. Seventy‐six compounds, which accounted for 87.9% of the total amount, were identified in a collective essential oil of P. sordidum from Corsica. The main essential oil components were (E)‐β‐caryophyllene (14.4%), β‐pinene (11.0%), thymol (9.0%), and hexadecanoic acid (5.3%). The chemical compositions of essential oils from 19 Corsican locations were investigated. The study of the chemical variability using statistical analysis allowed identifying direct correlation between the three populations of P. sordidum widespread in Corsica and the essential oil compositions they produce. The in vitro antimicrobial activity of P. sordidum essential oil was evaluated and it exhibited a notable activity on a large panel of clinically significant microorganisms.     

Chemical composition and antimicrobial activity of the essential oil of endemic Dalmatian black pine (Pinus nigra ssp. dalmatica)

The chemical composition and the antimicrobial activity of the essential oil isolated from the needles of endemic Dalmatian black pine (Pinus nigra ssp. dalmatica) from Croatia were investigated. The chemical composition of the essential oil was determined by GC and GC/MS analyses, and the main compounds identified were α-pinene, β-pinene, germacrene D, and β-caryophyllene. Disc-diffusion and broth-microdilution assays were used for the in vitro antimicrobial screening. The Dalmatian black pine essential oil exhibited a great potential of antibacterial activity against Gram-positive bacteria (MIC=0.03-0.50% (v/v)) and a less pronounced activity against Gram-negative bacteria (MIC=0.12-3.2% (v/v)). The volatile compounds also inhibited the growth of all fungi tested, including yeast.     

The Chemical Diversity of Lantana camara: Analyses of Essential Oil Samples from Cuba,Nepal, and Yemen

The aerial parts of Lantana camara L. were collected from three different geographical locations: Artemisa (Cuba), Biratnagar (Nepal), and Sana'a (Yemen). The essential oils were obtained by hydrodistillation and analyzed by gas chromatography/mass spectrometry. A cluster analysis of 39 L. camara essential oil compositions revealed eight major chemotypes: β‐caryophyllene, germacrene D, ar‐curcumene/zingiberene, γ‐curcumen‐15‐al/epi‐β‐bisabolol, (E)‐nerolidol, davanone, eugenol/alloaromadendrene, and carvone. The sample from Cuba falls into the group dominated by (E)‐nerolidol, the sample from Nepal is a davanone chemotype, and the sample from Yemen belongs to the β‐caryophyllene chemotype. The chemical composition of L. camara oil plays a role in the biological activity; the β‐caryophyllene and (E)‐nerolidol chemotypes showed antimicrobial and cytotoxic activities.     

The Essential Oils of the Greek Endemic Satureja horvatii ssp. macrophylla in Relation to Bioclimate

The essential oils of the Greek endemic Satureja horvatii subsp. macrophylla collected from 36 areas (among them, 23 are included in twelve sites of the EU network NATURA 2000) belonging to different bioclimatic types were studied. The total essential‐oil content is negatively related to the altitude. The variation of the essential oil composition follows a geographical pattern, which is related to the bioclimatic belts along the taxon's range. Carvacrol dominates in areas with Mediterranean and Submediterranean bioclimate (mainly in the S and C part of the taxon's distribution), linalool or trans‐sabinene hydrate, and/or borneol in the Submediterranean or Temperate Axeric bioclimates (in the N part of distribution), whereas thymol is found as main oil constituent in all three bioclimates.     

Variability in Foliar Essential Oils among Different Morphotypes of Lantana Species Complexes,and Its Taxonomic and Ecological Significance

The genus Lantana has many species complexes, and L. camara is one of the aggressive alien weedy species complexes; species delimitation in these complexes is a nightmare for taxonomists. We examined the diversity in the chemical composition of foliar essential oils among morphotypes of Lantana species complexes inhabiting the same ecological gradient, and its taxonomic and ecological significance. The yields of essential oils varied from 0.1 to 0.79% in foliar hydrodistillates of eleven morphotypes, and a total of 39 chemical constituents were detected by GC/MS. The quantitative and qualitative variability in the composition of essential oils among morphotypes was very high, and hence they represent chemotypes. The diversity observed in the composition of essential oils appears to be of genetic origin and thus of taxonomic value. The formation of distinct clusters and sub‐clusters at high distance cluster combine values also substantiates that the patterns of distribution of chemical constituents among morphotypes can be used in delimiting species and infraspecific taxa within the species complexes. The presence of β‐caryophyllene and other such compounds, which are known to prevent herbivory, in morphotypes of Lantana species complexes suggest that these compounds may provide selective advantage to Lantana over native species in the invasion of new and disturbed habitats.     

Essential Oil Variation among Natural Populations of Lavandula multifida L. (Lamiaceae)

Volatiles from twelve wild Tunisian populations of Lavandula multifida L. growing in different bioclimatic zones were assessed by GC (RI) and GC/MS. Thirty‐six constituents, representing 83.48% of the total oil were identified. The major components at the species level were carvacrol (31.81%), β‐bisabolene (14.89%), and acrylic acid dodecyl ester (11.43%). These volatiles, together with α‐pinene, were also the main compounds discriminating the populations. According to these dominant compounds, one chemotype was revealed, a carvacrol/β‐bisabolene/acrylic acid dodecyl ester chemotype. However, a significant variation among the populations was observed for the majority of the constituents. A high chemical‐population structure, estimated both by principal component analysis (PCA) and unweighted pair group method with averaging (UPGMA) cluster analysis based on Euclidean distances, was observed. Both methods allowed separation of the populations in three groups defined rather by minor than by major compounds. The population groups were not strictly concordant with their bioclimatic or geographic location. Conservation strategies should concern all populations, because of their low size and their high level of destruction. Populations exhibiting particular compounds other than the major ones should be protected first.     

Chemical Composition and Antioxidant Activity of Essential Oils and Methanol Extracts of Different Parts from Juniperus rigida Siebold & Zucc.

The chemical composition and antioxidant activity of essential oils and MeOH extracts of stems, needles, and berries from Juniperus rigida were studied. The results indicated that the yield of essential oil from stems (2.5%) was higher than from needles (0.8%) and berries (1.0%). The gas chromatography/mass spectrometer (GC/MS) analysis indicated that 21, 17, and 14 compounds were identified from stems, needles, and berries essential oils, respectively. Caryophyllene, α‐caryophyllene, and caryophyllene oxide were primary compounds in both stems and needles essential oils. However, α‐pinene and β‐myrcene mainly existed in berries essential oils and α‐ionone only in needles essential oils. The high‐performance liquid chromatography (HPLC) analysis indicated that the phenolic profiles of three parts exhibited significant differences. Needles extracts had the highest content of chlorogenic acid, catechin, podophyllotoxin, and amentoflavone, and for berries extracts, the content of those compounds was the lowest. Meanwhile, three in vitro methods (DPPH, ABTS, and FRAP) were used to evaluate antioxidant activity. Stems essential oil and needles extracts exhibited the powerful antioxidant activity than other parts. This is the first comprehensive study on the different parts of J. rigida. The results suggested that stems and needles of J. rigida are useful supplements for healthy products as new resources.     

Chemical Variability of the Leaf Essential Oil of Xylopia aethiopica (Dunal) A.Rich. from Côte d'Ivoire

The chemical composition of 48 essential‐oil samples isolated from the leaves of Xylopia aethiopica harvested in six Ivoirian forests was investigated by GC‐FID and ¹³C‐NMR analyses. In total, 23 components accounting for 82.5–96.1% of the oil composition were identified. The composition was dominated by the monoterpene hydrocarbons β‐pinene (up to 61.1%) and α‐pinene (up to 18.6%) and the sesquiterpene hydrocarbon germacrene D (up to 28.7%). Hierarchical cluster and principal component analyses allowed the distinction of two groups on the basis of the β‐pinene and germacrene D contents. The chemical composition of the oils of Group I (38 oil samples) was clearly dominated by β‐pinene, while those of Group II (10 samples) were characterized by the association of β‐pinene and germacrene D. The leaves collected in the four inland forests produced β‐pinene‐rich oils (Group I), while the oil samples belonging to Group II were isolated from leaves harvested in forests located near the littoral.