Chemical and Morphological Inter- and Intrapopulation Variability in Natural Populations of Gentiana pneumonanthe L.

Inter- and intrapopulation variability in six natural populations of the rare species Gentiana pneumonanthe was examined based on morphological and chemical data. Population size and linear morphometric parameters differed significantly among populations, but without a clear connection to habitat conditions, i. e. water supply and light availability. Leaf shape varied from ovate to lanceolate in all populations, and one population was distinctive in having the largest number of leaves of transitional shape. HPLC analyses of six secondary metabolites were performed separately for belowground parts, and aboveground vegetative and reproductive parts of individual plants (6 populations ×7 individuals ×3 plant parts, n=126) in order to examine differences at the population and individual levels. Three secoiridoids (swertiamarin (SWM), sweroside (SWZ), and gentiopicrin (GP)), one xanthone (mangiferin (MGF)), and two flavones (isoorientin (IO) and isovitexin (IV)) were detected and quantified in the analyzed samples: sweroside dominated in the aboveground reproductive part, mangiferin in the aboveground vegetative part, and gentiopicrin in the belowground part. At the population level, differences in contents of the analyzed chemicals among populations were significant only for a few metabolites. At the individual level, a pronounced organ-dependent distribution of secondary metabolites was revealed. The results of this study contribute to a better understanding of natural variability within populations of the rare and threatened G. pneumonanthe, and provide data on the contents and within-plant distribution of secondary metabolites, which are important as pharmacologically active compounds and may be useful for further biotechnological procedures regarding this species.

Essential-Oil Variability in Natural Populations of Pinus mugo Turra from the Julian Alps.

The composition and variability of the terpenes and their derivatives isolated from the needles of a representative pool of 114 adult trees originating from four natural populations of dwarf mountain pine (Pinus mugo Turra) from the Julian Alps were investigated by GC-FID and GC/MS analyses. In total, 54 of the 57 detected essential-oil components were identified. Among the different compound classes present in the essential oils, the chief constituents belonged to the monoterpenes, comprising an average content of 79.67% of the total oil composition (74.80% of monoterpene hydrocarbons and 4.87% of oxygenated monoterpenes). Sesquiterpenes were present in smaller amounts (average content of 19.02%), out of which 16.39% were sesquiterpene hydrocarbons and 2.62% oxygenated sesquiterpenes. The most abundant components in the needle essential oils were the monoterpenes δ-car-3-ene, ß-phellandrene, α-pinene, ß-myrcene, and ß-pinene and the sesquiterpene ß-caryophyllene. From the total data set of 57 detected compounds, 40 were selected for principal-component analysis (PCA), discriminant analysis (DA), and cluster analysis (CA). The overlap tendency of the four populations suggested by PCA, was as well observed by DA. CA also demonstrated similarity among the populations, which was the highest between Populations I and II.

Chemical Composition and Interpopulation Variability of Essential Oils of Taxus baccata L. from Serbia.

The composition of the essential oil of the twigs and needles of Taxus baccata L. from three natural populations in Serbia has been determined by GC/MS analysis. Of the 91 detected compounds, 87 were identified. The most abundant compound classes were aliphatic alcohols, terpenes, aliphatic hydrocarbons, and aliphatic aldehydes, which together comprised ca. 86.92% of the total oil composition. The dominant constituents were oct-1-en-3-ol (23.48%), (3Z)-hex-3-en-1-ol (11.46%; aliphatic alcohols), and myrtenol (11.38%; oxygenated monoterpene). The PCA of 22 selected compounds revealed differentiations of populations based on geographic distribution. The CA showed that Populations I and II from the Dinaric Alps were similar, and that Population III from the Balkan mountain system was distinct. This was the first investigation of interpopulation variability of T. baccata populations based on essential oil composition. The results of this study were compared with those of previous studies concerning volatile compounds produced by Taxus species. The results indicate that the essential oil content of T. baccata populations from this study is unique, mostly resembling the population from southeast Serbia.

A Geometric Morphometrics Approach to the Study of Natural Variations and Hybrid Detection in Populations of Alnus incana (L.) Moench and Alnus rohlenae Vít, Douda and Mandák.

Landmark-based geometric morphometrics (GM) was used to examine, for the first time, spontaneous hybridization between Alnus incana (L.) Moench and Alnus rohlenae Vít, Douda and Mandák, and to assess inter- and intrapopulation variability in leaf shape, leaf size and venation in natural populations in Serbia (Western Balkans). Two geographically distant (30 km) and two close (1.2 km) populations were selected to examine hybridization. The variability in leaf shapes was assessed by canonical variate analysis and linear discriminant analysis performed on the symmetric component of variation. Covariation between the symmetric component of shape variation and the number of pairs of secondary leaf veins was investigated with partial least squares analysis. Static allometry was examined for the first time in the genus Alnus Mill. A higher proportion of A. incana leaves was classified as A. rohlenae in geographically close populations, which is in accordance with the hypothesis about spontaneous hybridization. No single leaf of A. rohlenae was classified as A. incana, indicating that putative hybrids can only be found in grey alder populations. This study demonstrates that GM is a powerful tool for species delimitation and hybrid detection in the genus Alnus and it can be used for preliminary screening in hybrid zones.

Ecologically acceptable usage of derivatives of essential oil of sweet basil, Ocimum basilicum, as antifeedants against larvae of the gypsy moth, Lymantria dispar.

Abstract Ethanol solutions of five fractions obtained from essential oil of sweet basil Ocimum basilicum L. (Lamiales: Lamiaceae) (F1-F5) were tested for their antifeedant properties against 2(nd) instar gypsy moth larvae, Lymantria dispar L. (Lepidoptera: Lymantriidae), in laboratory non-choice and feeding-choice experiments. Prior to bioassays, the chemical composition of each fraction was determined by gas chromatography analyses. Significant larval deterrence from feeding was achieved by application of tested solutions to fresh leaves of the host plant. The most effective were were F1 (0.5%), F4 (0.05, 0.1, and 0.5%), and F5 (0.1 and 0.5%), which provided an antifeedant index > 80% after five days. A low rate of larval mortality was observed in no-choice bioassay. In situ screening of chlorophyll fluorescence as an indicator of plant stress level (assessed by the induced fluorometry) confirmed that the tested compounds did not cause alternations in the photosynthetic efficiency of treated leaves.

Population-Related Variability in Qualitative and Quantitative Secondary Metabolite Profile of Gentianella austriaca (A. & J. Kern.) Holub.

Phytochemical profiling of six natural populations of Gentianella austriaca was performed by HPLC identification and quantification of a number of secondary metabolites, and evaluation of time series of peak areas by chemometric analysis. Phytochemical analysis of G. austriaca revealed the presence of iridoids, flavone-C-glucosides and xanthones. Twelve secondary metabolites were identified in the aerial parts, roots and seeds, including swertiamarin (SWM), gentiopicrin (GP), sweroside (SWZ), isoorientin (ISOOR), swertisin (SWE), demethylbellidifolin-8-O-glucoside (DMB-8-O-glc), bellidifolin-8-O-glucoside (BDF-8-O-glc), mangiferin (MGF), corymbiferin (CBF), corymbiferin-1-O-glucoside (CBF-1-O-glc), bellidifolin (BDF) and campestroside. Multivariate statistical analyses showed relatively low variability among populations according to secondary metabolite content. However, some pharmacologically important compounds were found in higher amounts in a few populations, which could be useful for conservation and future biotechnological procedures.

Ecophysiological and Growth-Related Traits of Two Geophytes Three Years after the Fire Event in Grassland Steppe.

Deliblato Sands is the single largest expanse of sand in Europe. It is the most fire-prone area of Serbia due to the absence of surface water, sandy soils, specific microclimate conditions, and vegetation composition. Post-fire regeneration is a long-term process that includes many aspects of vegetation regrowth and habitat recovery. In the third year following one of the disastrous fires, the growth dynamics of two geophyte species in unburned and burned sites were studied. During the growing season, burned and unburned populations of Crocus reticulatus Steven ex Adam and Iris pumila L. were assessed for growth parameters (biomass production, specific leaf area, leaf area index) and leaf-level ecophysiological traits (photosystem II efficiency, chlorophyll amount, relative water content). Species acclimated differently to changed abiotic and biotic site conditions after the fire event. C. reticulatus burned and unburned populations differed significantly in terms of flowering phenology and ecophysiological traits, whereas I. pumila burned and unburned populations differed significantly in terms of growth parameters. The findings support the assertion that geophytes are generally well adapted to environmental disturbances. Species, however, responded differently to fire-induced changes in the physicochemical and biotic environment, depending on their ecological requirements and adaptive capacity.

Tree species flammability based on plant traits: A synthesis.

The occurrence of large and recurring forest fires has long been associated with fire-prone environments, but this perception has been shifted rapidly in recent decades as Earths' landscapes have become increasingly threatened by severe and unpredictable fires as a result of climate changes. In this regard, the flammability of trees is a topic of great interest for ecology, management, and the development of sustainable restoration and rehabilitation plans. Tree species differ in regard to flammability, and many plant functional traits contribute to flammability at species, community and vegetation level. The relationship between plant traits and flammability at species level is important for a broader understanding of the vegetation-fire dynamic at the local and landscape scales. This review summarizes the current state of knowledge regarding the impact of individual plant traits of tree species on flammability components. By keywords-based searching of academic databases, 85 research papers were collected and analyzed. The literature synthesis shows: i) main issues addressed in studies on plant trait-based tree flammability, ii) general research output and biogeographic regions studied, iii) inventory of tree taxa investigated, iv) relationships between plant traits and flammability components, v) the most relevant plant traits that determine the flammability-related differences between species.

Gentiana asclepiadea L. from Two High Mountainous Habitats: Inter- and Intrapopulation Variability Based on Species' Phytochemistry.

Natural populations of Gentiana asclepiadea L., located at two mountainous sites, were HPLC-analyzed regarding the contents of six representative secondary metabolites. The contents of swertiamarin (SWM), gentiopicrin (GP), sweroside (SWZ), mangiferin (MGF), isoorientin (ISOOR), and isovitexin (ISOV) were determined in six populations (three per study site), and separately for aboveground and belowground plant parts. PCA showed a clear separation of four groups according to the contents of the analyzed secondary metabolites. Out of six analyzed compounds, five were present in all samples and only one (SWZ) was found in Golija populations (belowground parts) but not in Vlasina populations, and its presence can be indicative of the geolocation of populations. Clear separation of groups was mostly affected by the different contents of chemical compounds in plant parts (aboveground versus belowground) and by the differences related to population origin (higher content of SWM and GP in belowground parts of individuals from Vlasina populations and higher content of MGF and ISOOR of individuals from Golija populations). The results of this study contribute to the spatiochemical profiling of G. asclepiadea populations and a better understanding of inter- and intrapopulation variability of pharmacologically important compounds.

The role of plant species on runoff and soil erosion in a Mediterranean shrubland.

Shrubland is a Mediterranean biome characterized by densely growing evergreen shrubs adapted to fire events. To date, scientific research has focused on the impact of vegetation on soil erosion mainly through the control that plant biomass or plant cover exerts on sediment delivery and runoff discharge, being the individual plant species influence on hydrological and erosional processes not achieved in detail. The objective of this research is to determine: i) runoff and soil losses in a shrubland-covered rangeland at Sierra de Enguera, Spain; and ii) how four plant species affect soil and water losses. We measured soil cover, soil properties, runoff discharge and sediment yield under natural rainfall for five years (2010-2014) in a typical shrubland burnt in 1999. Four plant species were selected with 4 plots each: Ulex parviflorus Pourr., Pistacia lentiscus L., Quercus coccifera L. and Rosmarinus officinalis L. Despite that the soil properties and plant cover did not exhibit statistically significant differences among plant species, the runoff discharge was lower on Q. coccifera (4.87%, SE 0.24) and P. lentiscus (6.24%, SE 0.51) than on U. parviflorus (13.41%, SE 0.58) and R. officinalis (13.84%, SE 1.23). Sediment concentrations were, respectively, 3.91, 4.33, 4.31 and 4.88 g l-1, and the differences between R. officinalis and the other species were statistically significant. The runoff discharge determined differences in soil erosion rates among the plant species with lower rates on P. lentiscus (1.36 Mg ha-1 y-1) and Q. coccifera (1.53 Mg ha-1 y-1), than on U. parviflorus (3.17 Mg ha-1 y-1) and R. officinalis (3.85 Mg ha-1 y-1). This long term in situ study indicated that Q. coccifera and P. lentiscus are more efficient in controlling runoff discharge and soil losses than U. parviflorus and R. officinalis one decade after a fire. We discuss these results in light of the recent findings by the scientific community of the role of the canopy cover (rainfall interception), soil macropore and root system, and the water repellency that control the hydrological response of the soil (e.g. runoff generation, infiltration). The information supplied by 5 years of research is relevant for restoration and rehabilitation programs and advise that Q. coccifera and P. lentiscus are the most efficient plant species to control soil and water losses within the Mediterranean shrubland. This is an applied science approach for a better management of rangelands.

Larvicidal and antifeedant activity of some plant-derived compounds to Lymantria dispar L. (Lepidoptera: Limantriidae).

Ethanol solutions of essential oil of Ocimum basilicum and its main component, linalool (both isomer forms), all in three concentrations, as well as botanical standard Bioneem (0.5%), were tested for their toxicity and antifeedant activity against the second instar gypsy moth larvae in the laboratory bioassay. The essential oil of O. basilicum was subjected to gas chromatography analysis, and totally 37 compounds were detected, of which linalool was predominantly present. All tested solutions showed low to moderate larvicidal effect in both residual toxicity test and in chronic larval mortality bioassay. Chronic mortality tests showed that obtained mortality was a consequence of starving rather than ingestion of treated leaves. However, antifeedant index achieved by application of tested solutions in feeding choice assay was remarkable. Foliar application of all tested compounds deterred feeding by L2 in the same percent as Bioneem. Antifeedant index was relatively high at all tested treatments (85-94%); moreover, the larval desensitization to repelling volatiles has not occurred after five days of observation. Low toxic and high antifeedant properties make these plant-derived compounds suitable for incorporation in integrated pest management programs, especially in urban environments.

An ethnobotanical study on the usage of wild medicinal herbs from Kopaonik Mountain (Central Serbia).

An ethnobotanical survey was carried out on the territory of the highest mountain in Central Serbia, Kopaonik, which is characterized by great plant diversity. In total, 83 wild species from 41 families and 96 preparations for use in human therapy were recorded. Among those wild plants which are most commonly used for medicinal purposes, Hypericum perforatum L., Urtica dioica L., Achillea millefolium L., Matricaria chamomilla L., Sambucus nigra L., and Thymus serpyllum L. were particularly highly recommended by the majority of informants as being 'beneficial for all ailments'. The most frequently reported medicinal uses were for treating gastrointestinal ailments (50%), skin injuries and problems (25.6%), followed by respiratory, urinary-genital and cardiovascular problems (20.5%, 20.5%, 19.2%, respectively). Plants with unusual phytotherapeutic uses are Galium verum L. (sedative properties) and Eupatorium cannabinum L. (influenza-like illnesses), while plants with interesting but lesser-known properties include Daphne laureola L. (rheumatism and skin ailments) and Ficaria verna Huds. (tubers for treating haemorrhoids). In addition, 10 wild species used in veterinary medicine, as well as 25 herbs used for human nourishment were noted.

Electroanalytical Sensing of Bromides Using Radiolytically Synthesized Silver Nanoparticle Electrocatalysts.

Monitoring bromides (Br-) is of crucial importance since bromates, potential human carcinogens, are formed during ozonation of water containing bromides in concentrations >100 µg L-1. Within this study, silver (Ag) and four carbon-supported Ag catalysts were synthesized by the γ-radiation method and their morphology and structure examined using transmission electron microscopy, X-ray diffraction, and UV-Vis analysis. The nanocatalysts were tested for Br- sensing in aqueous media using cyclic voltammetry. All five Ag materials exhibited electroactivity for sensing of Br- ions, with pure Ag catalyst giving the best response to Br- ions presence in terms of the lowest limit of detection. Sensing of bromides was also explored in tap water after addition of bromides suggesting that herein prepared catalysts could be used for bromides detection in real samples. Furthermore, sensing of other halogen ions, namely, chlorides and iodides, was examined, and response due to chloride presence was recorded.

Ethnobotany and herbal medicine in modern complementary and alternative medicine: An overview of publications in the field of I&C medicine 2001-2013.

ETHNOPHARMACOLOGICAL RELEVANCE: Ethnobotanical knowledge and traditional medicinal practices from different parts of the world are of global importance and documentation of ethnobotanical, and ethnopharmacological data is a key prerequisite for further research in the area of herbal medicine and its implementation in clinical practice. AIMS: An attempt was made to evaluate the scientific output of research related to ethnobotany and herbal medicine in journals indexed in the subject area "Integrative and Complementary Medicine" in the period 2001-2013, in order to ascertain research trends in both subdisciplines. MATERIALS AND METHODS: All articles related to ethnobotany and herbal medicine, extracted from journals included in the field of I&C Medicine and published in the period 2001-2013, have been analyzed for general bibliometric data, and specific data: ethnobotanical data (geographic, floristic, pharmacological, sociological and other relevant data) and phytotherapeutic data (type of applied herbal medicine, plant species studied, pharmacological activity of studied plant species and disease and disorder type studied on a particular model). RESULTS: In the studied period, the number of articles dealing with ethnobotany and herbal medicine increased 6.3-fold. Articles related to ethnobotanical studies documented medicinal flora from 81 countries, either giving an overview of overall medicinal flora, or presenting the ethnomedicinal aspect of the use of plants for the treatment of ailments typical to the studied area. Additionally, the authors provided significant information on the methods of use and herbal preparations. In herbal medicine studies, plants, traditional plant remedies, herbal medicinal products and active herbal compounds were tested for many of pharmacological activities (146), with the curative activity emerging as most frequently tested. Out of 39 model systems, most of the studies were carried out under controlled in vitro conditions (4589 articles), followed by rat in vivo (2320), human in vivo (1285), mouse in vivo (955), and on agents of pathogenic diseases (887); more than 800 medical disorders were treated. CONCLUSIONS: The study revealed the regions most studied for new records of floristic and ethnomedicinal diversity, the most frequently studied plant species, and the most promising therapeutic indications for the integration of herbal remedies in the curative process, as ascertained from the selected bibliographic databases.

Bioreactor validation and biocompatibility of Ag/poly(N-vinyl-2-pyrrolidone) hydrogel nanocomposites.

Silver/poly(N-vinyl-2-pyrrolidone) (Ag/PVP) nanocomposites containing Ag nanoparticles at different concentrations were synthesized using γ-irradiation. Cytotoxicity of the obtained nanocomposites was determined by MTT assay in monolayer cultures of normal human immunocompetent peripheral blood mononuclear cells (PBMC) that were either non-stimulated or stimulated to proliferate by mitogen phytohemagglutinin (PHA), as well as in human cervix adenocarcinoma cell (HeLa) cultures. Silver release kinetics and mechanical properties of nanocomposites were investigated under bioreactor conditions in the simulated body fluid (SBF) at 37°C. The release of silver was monitored under static conditions, and in two types of bioreactors: perfusion bioreactors and a bioreactor with dynamic compression coupled with SBF perfusion simulating in vivo conditions in articular cartilage. Ag/PVP nanocomposites exhibited slight cytotoxic effects against PBMC at the estimated concentration of 0.4 µmol dm(-3), with negligible variations observed amongst different cell cultures investigated. Studies of the silver release kinetics indicated internal diffusion as the rate limiting step, determined by statistically comparable results obtained at all investigated conditions. However, silver release rate was slightly higher in the bioreactor with dynamic compression coupled with SBF perfusion as compared to the other two systems indicating the influence of dynamic compression. Modelling of silver release kinetics revealed potentials for optimization of Ag/PVP nanocomposites for particular applications as wound dressings or soft tissue implants.