Dea Baričevič

Evaluation of the antioxidants activities of four Slovene medicinal plant species by traditional and novel biosensory assays

We investigated the antioxidant activity of methanolic and water extracts of Slovene accessions of four medicinal plant species (Salvia officinalis, Achillea millefolium, Origanum vulgare subsp. vulgare and Gentiana lutea). Their free radical-scavenging activity against the DPPH. free radical was studied with a spectrophotometric assay, while their biological activity with the help of a laboratory-made biosensor based on immobilized fibroblast cells (assay duration: 3 min). The observed antioxidant activity of the extracts from the four investigated medicinal plant species was dependent on both the solvent used for extraction and the assay method (conventional or biosensor-based). Independently from the assay method and the solvent used for extraction, the lowest scavenging activity was observed in root extracts of G. lutea. Treatment of the immobilized cells with the plant extracts resulted in an increase of the cell membrane potential (membrane hyperpolarization), possibly due to the reduction of membrane damage due to oxidation. The novel cell biosensor could be utilized as a rapid, high throughput tool for screening the antioxidant properties of plant-derived compounds.

Effect of water stress and nitrogen fertilization on the content of hyoscyamine and scopolamine in the roots of deadly nightshade (Atropa belladonna)

Abstract The study intended to elaborate the optimal environmental conditions of water supply and nitrogen fertilization for maximum content of hyoscyamine (% dw) and scopolamine (% dw). Plants grown from seeds of Slovene autochthonous population of deadly nightshade (Atropa belladonna), were treated with different water regimes (35–95% depletion of available soil water) together with enhanced nitrogen supply (0.37–1.60 g/pot N) in a greenhouse experiment. Dry plant extracts from 32-week old roots were analysed with capillary electrophoresis (CE) for the presence of tropane alkaloids (hyosciamyne, scopolamine). The results of the plant treatment responses showed that the maximal yield of tropane alkaloids (hyoscyamine: 54 mg/plant; scopolamine: 7 mg/plant) was achieved in plants grown under an optimal irrigation regime (35% depletion of available soil water) accompanied with total nitrogen supply of 0.37 g/pot. By contrast, the maximal content of alkaloids was achieved with 95% depletion of available soil water and a nitrogen supply of 1.60 g/pot.

Free radical scavenging activities of yellow gentian (Gentiana lutea L.) measured by electron spin resonance

Yellow gentian (Gentiana lutea L.) is a herbal species with a long-term use in traditional medicine due to its digestive and stomachic properties. This paper presents an investigation of the free radical scavenging activity of methanolic extracts of yellow gentian leaves and roots in two different systems using electron spin resonance (ESR) spectrometry. Assays were based on the stable free radical 1,1-diphenyl-2-picrylhydrazyl (DPPH) and the superoxide radicals (O2 +) generated by the xanthine/ xanthine oxidase (X/XO) system. The results of gentian methanolic extracts were compared with the antioxidant capacity of synthetic antioxidant butylated hydroxyanisole (BHA). This study proves that yellow gentian leaves and roots exhibit considerable antioxidant properties, expressed either by their capability to scavenge DPPH or superoxide radicals.

The Impact of Drought Stress and/or Nitrogen Fertilization in Some Medicinal Plants

SUMMARY Several pot experiments were set up to follow the impact of drought stress and/or nitrogen fertilization on the yield and secondary metabolites content of medicinal and aromatic plants (MAP) accessions held within Genebank for MAP in Slovenia. The results of pot trials with fenugreek and with deadly nightshade are presented. In fenugreek (Trigonella foenumgraecum L., Fabaceae) the maximal yield of dios-genin was achieved in plants grown under an optimal irrigation regime (35% depletion of available soil water). Drought stressed seeds of cv. Margaret contained less diosgenin (p = 0.0021) than irrigated plants (0.123%, dw). In deadly nightshade (Atropa belladonna L., Solanaceae), the content of tropane alkaloids was determined. The results of the plant treatment responses showed that the maximal yield of tropane alkaloids (hyoscyamine: 54 mg/plant; scopolamine: 7 mg/plant) was achieved in plants grown under an optimal irrigation regime (35% depletion of available soil water) accompanied with a total nitrogen supply of 0.37 g/pot. By contrast, the maximal content of alkaloids was achieved with 95% depletion of available soil water and a nitrogen supply of 1.60 g/pot.

Determination of seco-iridoid and 4-pyrone compounds in hydro-alcoholic extracts of Gentiana lutea L. subsp. symphyandra Murb. leaves and roots by using high performance liquid chromatography.

In view of the increasing interest in herbal remedies and their important role in health maintenance and well-being, a study was carried out to clarify the role and importance of the solvent in the extraction process. Hydro-alcoholic (methanol or ethanol) or water extraction procedures for secondary metabolites from Gentiana lutea subsp. symphyandra leaves and roots were optimized for the yield of compounds that have proved to act antioxidatively. Using high performance liquid chromatography (HPLC), the contents of seco-iridoid glucosides (gentiopicroside, amarogentin) and 4-pyrones (mangiferin (xanthone), homoorientin (flavone C-glycoside), isovitexin (flavone C-glycoside)) in Gentiana lutea subsp. symphyandra extracts were determined. The analysis revealed that the contents of observed secondary metabolites in Gentiana lutea subsp. symphyandra extracts varied significantly based on the solvent (methanol, ethanol, and water) used in the extraction procedure. Seco-iridoids and 4-pyrone contents in extract...

Genetic Diversity and Demographic History of Wild and Cultivated/Naturalised Plant Populations: Evidence from Dalmatian Sage (Salvia officinalis L., Lamiaceae)

Dalmatian sage (Salvia officinalis L., Lamiaceae) is a well-known aromatic and medicinal Mediterranean plant that is native in coastal regions of the western Balkan and southern Apennine Peninsulas and is commonly cultivated worldwide. It is widely used in the food, pharmaceutical and cosmetic industries. Knowledge of its genetic diversity and spatiotemporal patterns is important for plant breeding programmes and conservation. We used eight microsatellite markers to investigate evolutionary history of indigenous populations as well as genetic diversity and structure within and among indigenous and cultivated/naturalised populations distributed across the Balkan Peninsula. The results showed a clear separation between the indigenous and cultivated/naturalised groups, with the cultivated material originating from one restricted geographical area. Most of the genetic diversity in both groups was attributable to differences among individuals within populations, although spatial genetic analysis of indigenous populations indicated the existence of isolation by distance. Geographical structuring of indigenous populations was found using clustering analysis, with three sub-clusters of indigenous populations. The highest level of gene diversity and the greatest number of private alleles were found in the central part of the eastern Adriatic coast, while decreases in gene diversity and number of private alleles were evident towards the northwestern Adriatic coast and southern and eastern regions of the Balkan Peninsula. The results of Ecological Niche Modelling during Last Glacial Maximum and Approximate Bayesian Computation suggested two plausible evolutionary trajectories: 1) the species survived in the glacial refugium in southern Adriatic coastal region with subsequent colonization events towards northern, eastern and southern Balkan Peninsula; 2) species survived in several refugia exhibiting concurrent divergence into three genetic groups. The insight into genetic diversity and structure also provide the baseline data for conservation of S. officinalis genetic resources valuable for future breeding programmes.

Medicinal and Aromatic Plants in Scientific Databases

This chapter provides assessment of uses and retrieval of terms related to medicinal and aromatic plants in databases available on different platforms/interfaces: ABI/Inform Global (ProQuest; business, management, finance, trade, markets), Agricola (NAL, American National Agricultural Library), Agris-Agrovoc (FAO, Food and Agriculture Organization of the United Nations), CAB Abstracts/OVID (CABI, CAB International; agriculture), Compendex (Ei/Engineering Index, Elsevier; engineering, technical sciences), FSTA (IFIS, International Food Information System; food/drink sciences, technology, human nutrition), Medline/Ebsco-MeSH (NLM, American National Library of Medicine; health, medical sciences (biomedicine), veterinary medicine), Scopus/SciVerse (Elsevier; citation database), Sociological Abstracts (ProQuest; social sciences, e.g. human-environment interactions, rural sociology, ethnology), Web of Science/Web of Knowledge (Thomson Reuters; citation database). Database features and functionalities are reviewed. The applicable terms (used for indexing, controlled glossaries, descriptors, subjects headings, keywords) are located in respective thesauri, e.g.: drug crops, drug plants, essential oil crops, essential oil plants, herbal drugs, herbal medicine, medicinal plants, phytotherapy, “plants, medicinal”. Several additional terms and non-descriptors are identified, e.g. aromatic plants, herbaceous agents, herbal remedies, ethnobotany, herbal products, herbal preparations, plant drugs, herbal therapies. Search utilities, principles and rules are tested, e.g. search syntax (query), operators (Boolean, proximity, context), field codes, truncation (wildcard), phrase search, stemming (lemmatization). Bibliometric (scientometric) analysis is conducted in selective databases in order to tentatively assess the numbers and growth of potentially relevant records.

Conservation of Wild Crafted Medicinal and Aromatic Plants and Their Habitats

The use of Medicinal and aromatic plants (MAPs) in the form of traditional medicine, is still the preferred method of medical treatment in developing countries, owing to the unavailability and/or high prices of modern medicines. In the developed economies, as a contract, the market sectors for plant products seem to show new potentials, like in the case of different processing industries. The market for herbal medicines and supplements is expected to reach 107 billion USD by 2017. Most of the MAPs supply required for meeting the global demand, however, still originates from natural sources. Overharvesting, in addition to the destruction of natural habitats, lead to serious losses in biodiversity. Many countries have introduced measures to protect species under the guidance of the CBD and GSPC targets. Nevertheless, the numbers of rare or threatened wild species have been on an increase, especially in recent years. Numerous in situ and ex situ conservation strategies to preserve rare or threatened MAP species have been proposed. Some complementary conservation measures include restrictions on wildcrafting, protection in botanical, ethnobotanical gardens or other areas (national parks, nature reserves). Involvement of local communities in development programs of conservation, for example on-farm cultivation instead of wildcrafting, is essential for preservation of valuable genetic resources for the future.

Densitometric DNA analysis in agarose electrophoretic gels

Agarose gel electrophoresis is a basic separation tool used in molecular biology, mostly for qualitative DNA analysis. There are constraints limiting its use in quantitative analysis, namely low repeatability and a narrow linear range. However, by using an internal standard or internal normalization, repeatability and linear range could be significantly improved. In the work discussed in this paper it was shown that an approximately fivefold improvement in repeatability and an over threefold wider linear range could be achieved by applying internal normalization. Using the proposed approach, genetic markers, for example RAPD and PCR—RFLP, or even microsatellite markers, could be conveniently quantitatively assessed using agarose gel electrophoresis.