Clinopodium L. Taxa from the Balkans-Are There Unique Leaf Micromorphological and Phytochemical Patterns?

The concept of the genus Clinopodium L. has changed considerably since its first description. Most of the currently accepted species of the genus have traditionally been treated as separate genera in the group Satureja sensu lato: Clinopodium L., Calamintha sensu Miller or Moench, and Acinos sensu Miller or Moench. This study aimed to gain a better insight into the species diversity of Clinopodium L. from the Balkans by analyzing the taxa that have traditionally been placed in separate genera. The alkane profile and the micromorphological characteristics of the leaves are analyzed. The leaves are visualized using scanning electron microscopy, and alkanes are isolated using n-hexane as a solvent and analyzed using gas chromatography/mass spectrometry. The alkane profile showed the differentiation of the Acinos-group from the other taxa based on the dominant n-C31, while most of the other taxa contained n-C33 as the dominant alkane. The micromorphological features also showed clear differences between the previously recognized genera, especially in the capitate trichomes. The results showed that micromorphological patterns are highly variable in certain groups of the genus Clinopodium.

Phytochemistry and Micromorphology of Epicuticular Waxes in Four Juniper Species With Fragmented Distribution in the Balkans.

The cuticle is important in the interaction between the plant and its environment, especially in the dry areas. Four species of junipers from the section Sabina wild growing in the Balkans were selected to study leaf wax composition using GC/MS and GC-FID and its surface morphology under SEM to understand the correlation between the distribution and/or habitat of these species and their cuticles. SEM micrographs showed continuous, smooth epicuticular layers with crusts in all species but with a species-specific distribution of different 3D crystalloid types and different cuticle thickness. n-C33 alkane was the most abundant compound, followed by n-C29, n-C31, and n-C35, depending on the species and the site. The average chain length (N) was the lowest in J. phoenicea, but with the greatest dispersion around it. At the same time, the two most continental species (J. foetidissima and J. excelsa) show the N with the lowest dispersion around it. The statistical analyses confirmed the significance of climate on the evolution of the specific epicuticular wax composition in studied junipers.

Interactions between Natural Products-A Review.

Plant-based natural products have been used as a source for therapeutics since the dawn of civilization. According to the World Health Organization (WHO), more than 80% of the world's population relies on traditional medicine for their primary healthcare. Numerous natural extracts, widely known in Traditional Chinese Medicine, Indian Ayurveda medicine and other practices, have led to the modern discovery and development of new drugs. Plants continuously interact with their environment, producing new compounds and ever-changing combinations of existing ones. Interestingly, some of the compounds have shown lower therapeutic activity in comparison to the extract they were isolated from. These findings suggest that the higher therapeutic activity of the source extract was due to the synergistic effect of several compounds. In other words, the total therapeutic potential of the extract cannot be explained only by the sum of its parts alone. In traditional medicine, most herbal remedies are based on a mixture of plants, and it is the interaction between different constituents that amplifies their therapeutic potential. Considering the significant influence traditional medicine has on human healthcare, knowing and studying the synergistic effect of compounds is paramount in designing smart therapeutic agents.

Essential Oil Composition and Micromorphological Traits of Satureja montana L., S. subspicata Bartel ex Vis., and S. kitaibelii Wierzb. Ex Heuff. Plant Organs.

The essential oil (EO) composition of Satureja plants is highly variable. Recent studies suggest that there is an even difference in the EO composition from different plant organs within the same plant. This study aims to examine the chemical profile of EOs and the micromorphological characteristics of different organs of three Satureja species. The relationship between the number of glandular trichomes and EOs profile and relative yield is also investigated. Individuals from five populations were visualized using a scanning electron microscope, while EOs of leaves, calyces, corollas, and whole aerial parts were isolated using simultaneous distillation and extraction and analyzed using gas chromatography/mass spectrometry. Three types of glandular trichomes were detected. Peltate trichomes were present on all plant organs of studied species, while two types of capitate trichomes show different organ and species preferences. The EOs profiles differed across the plant parts, but showed a species specific composition. Univariate and multivariate statistics were used to show a correlation between the peltate trichomes and EO yield, and chemophenetic significance of EO profiles.

Juniperus phoenicea var. turbinata (Guss.) Parl. Leaf Essential Oil Variability in the Balkans.

In the present work, the leaf essential oil from 97 individuals of Juniperus phoenicea var. turbinata (Guss.) Parl. from the Balkan Peninsula was analyzed. The essential oil was dominated by monoterpene hydrocarbons (45.5 - 71.8%), of which α-pinene was the most abundant in almost all of the samples (38.2 - 55.8%). Several other monoterpenes and sesquiterpenes were also present in relatively high abundances in samples such as myrcene, δ-3-carene, ß-phellandrene, α-terpinyl acetate, (E)-caryophyllene and germacrene D. Multivariate statistical analysis suggested the existence of three possible chemotypes based on the abundance of the four components. Even though the intrapopulation variability was high, discriminant analysis (DA) was able to separate populations. DA showed high separation between western and eastern populations but also grouped geographically closer populations along the west Balkan shoreline. The potential influence of the climate on the composition of the essential oil was also studied.

Chemodiversity of Epicuticular n-Alkanes and Morphological Traits of Natural Populations of Satureja subspicata Bartl. ex Vis. along Dinaric Alps - Ecological and Taxonomic Aspects.

Morphological characters and the composition of epicuticular leaf n-alkanes of two Satureja subspicata Bartl. ex Vis. subspecies (subsp. liburnica Silic and subsp. subspicata) from nine natural populations along Dinaric Alps range were studied. Morphological characters were chosen based on Silic's subspecies separation. Seventeen n-alkane homologues (C19 - C35 ) were identified using gas chromatography/mass spectrometry (GC/MS) and GC/flame ionisation detector (FID). The most abundant n-alkane in all populations was n-nonacosane (C29 ), followed by n-hentriacontane (C31 ), with the exception of Divaca population where these two alkanes were co-dominant. Diversity and variability of n-alkane patterns and morphological characters and their relation to different geographic and bioclimatic parameters, including exposure, were analysed by several statistical multivariate methods (PCA, HCA, Discriminant Analysis, Mantel test). These tests showed clear separation of subsp. liburnica from subsp. subspicata, even though population Velebit showed separation from other subsp. liburnica populations based on phytochemical characters. Mantel test showed high correlation with geographical distribution in both investigated data sets. High correlation between morphological and phytochemical characters was also established. However, exposure can influence n-alkane profile, suggesting precaution while taking samples from natural habitats.

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.

Composition of leaf n-alkanes in three Satureja montana L. subspecies from the Balkan peninsula: ecological and taxonomic aspects.

The composition of the epicuticular leaf n-alkanes of eight populations of three Satureja montana subspecies (S. montana L. subsp. pisidica (Wettst.) Silic, S. montana L. subsp. montana, and S. montana L. subsp. variegata (Host) P. W. Ball), from central and western areas of the Balkan Peninsula was characterized by GC-FID and GC/MS analyses. In the leaf waxes, 15 n-alkane homologs with chain-lengths ranging from C21 to C35 were identified. The main n-alkane in almost all samples was n-nonacosane (C29 ), but differences in the contents of three other dominant n-alkanes allowed separating the coastal from the continental populations. The diversity and variability of the epicuticular-leaf-n-alkane patterns and their relation to different geographic and bioclimatic parameters were analyzed by several statistical methods (principal component, discriminant, and cluster analyses as well as the Mantel test). All tests showed a high correlation between the leaf n-alkane pattern and the geographical distribution of the investigated populations, confirming the differentiation between S. montana subsp. pisidica and the other two subspecies. The S. montana subsp. variegata and S. montana subsp. montana populations are geographically closer and their differentiation according to the leaf-n-alkane patterns was not clear, even though there was some indication of discrimination between them. Moreover, most of the bioclimatic parameters related to temperature were highly correlated with the differentiation of the coastal and the continental populations.

Biogeographic variation of foliar n-alkanes of Juniperus communis var. saxatilis Pallas from the Balkans.

The composition of the epicuticular n-alkanes isolated from the leaves of ten populations of Juniperus communis L. var. saxatilis Pallas from central (continental) and western (coastal) areas of the Balkan Peninsula was characterized by GC-FID and GC/MS analyses. In the leaf waxes, 14 n-alkane homologues with chain-lengths ranging from C22 to C35 were identified. All samples were dominated by n-tritriacontane (C33 ), but differences in two other dominant n-alkanes allowed separating the coastal from the continental populations. Several statistical methods (ANOVA, principal component, discriminant, and cluster analyses as well as the Mantel test) were deployed to analyze the diversity and variability of the epicuticular-leaf-n-alkane patterns of the ten natural populations of J. communis var. saxatilis and their relation to different geographic and bioclimatic parameters. Cluster analysis showed a high correlation of the leaf-n-alkane patterns with the geographical distribution of the investigated samples, differentiating the coastal from the continental populations of this taxon. Several bioclimatic parameters related to aridity were highly correlated with this differentiation.

Leaf n-alkanes as characters differentiating coastal and continental Juniperus deltoides populations from the Balkan Peninsula.

The composition of the cuticular n-alkanes isolated from the leaves of nine populations of Juniperus deltoides R.P.Adams from continental and coastal areas of the Balkan Peninsula was characterized by GC-FID and GC/MS analyses. In the leaf waxes, 14 n-alkane homologues with chain-lengths ranging from C22 to C35 were identified. n-Tritriacontane (C33 ) was dominant in the waxes of all populations, but variations between the populations in the contents of all n-alkanes were observed. Several statistical methods (ANOVA, principal component, discriminant, and cluster analyses) were used to investigate the diversity and variability of the cuticular-leaf-n-alkane patterns of the nine J. deltoides populations. This is the first report on the n-alkane composition for this species. The multivariate statistical analyses evidenced a high correlation of the leaf-n-alkane pattern with the geographical distribution of the investigated samples, differentiating the coastal from the continental populations of this taxon.