Leontopodium Alpinum Research Articles

Exploring Actinobacteria Associated With Rhizosphere and Endosphere of the Native Alpine Medicinal Plant Leontopodium nivale Subspecies alpinum.

The rhizosphere of plants is enriched in nutrients facilitating growth of microorganisms, some of which are recruited as endophytes. Endophytes, especially Actinobacteria, are known to produce a plethora of bioactive compounds. We hypothesized that Leontopodium nivale subsp. alpinum (Edelweiss), a rare alpine medicinal plant, may serve as yet untapped source for uncommon Actinobacteria associated with this plant. Rhizosphere soil of native Alpine plants was used, after physical and chemical pre-treatments, for isolating Actinobacteria. Isolates were selected based on morphology and identified by 16S rRNA gene-based barcoding. Resulting 77 Actinobacteria isolates represented the genera Actinokineospora, Kitasatospora, Asanoa, Microbacterium, Micromonospora, Micrococcus, Mycobacterium, Nocardia, and Streptomyces. In parallel, Edelweiss plants from the same location were surface-sterilized, separated into leaves, roots, rhizomes, and inflorescence and pooled within tissues before genomic DNA extraction. Metagenomic 16S rRNA gene amplicons confirmed large numbers of actinobacterial operational taxonomic units (OTUs) descending in diversity from roots to rhizomes, leaves and inflorescences. These metagenomic data, when queried with isolate sequences, revealed an overlap between the two datasets, suggesting recruitment of soil bacteria by the plant. Moreover, this study uncovered a profound diversity of uncultured Actinobacteria from Rubrobacteridae, Thermoleophilales, Acidimicrobiales and unclassified Actinobacteria specifically in belowground tissues, which may be exploited by a targeted isolation approach in the future.

Ultrastructural changes in in vitro callus plant cells producing secondary metabolites

The objective of our research was to analyze the ultrastructure of the callus cells of Leontopodium alpinum (Edelweiss), Fragaria x ananassa (strawberry) and Cotinus coggygria (smokebush) producing large amounts of secondary metabolites. Unlike plants, the in vitro callus has the peculiarity of proliferating (meristem-like) and at the same time synthesizing tissue-specific compounds with physiological significance in different stages of development. The cell population of the callus consists of two types of cells: those which divide and those which are producing the secondary metabolites, in different ratios, depending on the signals from the culture environment. Our TEM observations showed that in the metabolic active callus cells a complex network of membrane-bound organelles are involved in the synthesis, sorting, transport, degradation, storing or exocytosis of secondary metabolites such as anthocyanin pigments, flavonoids, polyphenols, tannins, oils, polysaccharides, alkaloids, many of them with antioxidant activity and with potential biotechnological value. Our results indicate that in the experimental conditions of the in vitro culture, callus cells have developed a complex endomembrane system that includes endoplasmic reticulum, Golgi apparatus, trans-Golgi network, multivesicular bodies, multilamellar bodies, membranous vesicles and vacuoles (vacuom) as an end point of the catabolic pathway. In the biosynthetic active callus cells, the entire cellular machinery is subordinated to the production of secondary metabolites, evidencing phenomena of autophagy, through which different useful cellular components are recycled. Finally, the secondary metabolites are irreversibly stored in large amounts in the vacuoles, at the tonoplast level, some of them into the cytoplasm, into the modified plastids or on the external face of the cell wall. In the intercellular space we noticed excreted vesicles, or most often, a reticulated extracellular matrix, probably involved in cell protection, recognition and integration.

The chemistry and pharmacology of Edelweiss: a review

Leontopodium nivale ssp. alpinum (syn. Leontopodium alpinum) is a perennial herb commonly known as Edelweiss, which has a long tradition in Alpine countries and adjacent regions as a medicinal plant. This review discusses current knowledge on the traditional uses, chemistry, biological activities and toxicology of this species. Several different classes of compounds such as terpenoids (analogues of sesquiterpenes, bisabolenes), phenylpropanoids (phenolic acids, flavonoids, coumarins, lignans), fatty acids and polyacetylenes were previously isolated from various parts of Edelweiss. Different types of extracts and compounds derived from this plant have been found to possess a broad spectrum of pharmacological activities on the cardiovascular and nervous systems. Furthermore, the plant have known anti-inflammatory, antimicrobial, antioxidant and chemo-protective effects. The observed pharmacological activities as well as toxicological profile of preparations and isolated compounds of Edelweiss support the view that these might be used in the development of agents with therapeutic benefit in various human diseases. Some suggestions for further research on chemical characterization and pharmacological properties are also given in this review.

Leoligin, the Major Lignan from Edelweiss (Leontopodium nivale subsp. alpinum), Promotes Cholesterol Efflux from THP-1 Macrophages.

Leoligin is a natural lignan found in Edelweiss (Leontopodium nivale ssp. alpinum). The aim of this study was to examine its influence on cholesterol efflux and to address the underlying mechanism of action. Leoligin increases apo A1- as well as 1% human plasma-mediated cholesterol efflux in THP-1 macrophages without affecting cell viability as determined by resazurin conversion. Western blot analysis revealed that the protein levels of the cholesterol efflux transporters ABCA1 and ABCG1 were upregulated, whereas the SR-B1 protein level remained unchanged upon treatment with leoligin (10 μM, 24 h). Quantitative reverse transcription PCR further uncovered that leoligin also increased ABCA1 and ABCG1 mRNA levels without affecting the half-life of the two mRNAs in the presence of actinomycin D, a transcription inhibitor. Proteome analysis revealed the modulation of protein expression fingerprint in the presence of leoligin. Taken together, these results suggest that leoligin induces cholesterol efflux in THP-1-derived macrophages by upregulating ABCA1 and ABCG1 expression. This novel activity suggests leoligin as a promising candidate for further studies addressing a possible preventive or therapeutic application in the context of atherosclerosis.

Subpicosecond surface dynamics in genomic DNA from in vitro-grown plant species: a SERS assessment.

In this work the surface-enhanced Raman total half band widths of seven genomic DNAs from leaves of chrysanthemum (Dendranthema grandiflora Ramat.), common sundew (Drosera rotundifolia L.), edelweiss (Leontopodium alpinum Cass), Epilobium hirsutum L., Hypericum richeri ssp. transsilvanicum (Čelak) Ciocârlan, rose (Rosa x hybrida L.) and redwood (Sequoia sempervirens D. Don. Endl.) have been measured. We have shown that surface-enhanced Raman spectroscopy (SERS) can be used to study the fast subpicosecond dynamics of DNA in the proximity of a metallic surface. The dependencies of the total half band widths and the global relaxation times, on the DNA molecular subgroup structure and on the type of genomic DNA, are reported. In our study, the full widths at half-maximum (FWHMs) for the SERS bands of genomic DNAs from different leaf tissues are typically in the wavenumber range from 15 to 55 cm(-1). Besides, it can be observed that molecular relaxation processes studied in this work have a global relaxation time smaller than 0.71 ps and larger than 0.19 ps. A comparison between different ranges of FT-Raman and SERS band parameters, respectively, corresponding to DNA extracted from leaf tissues is given. It is shown that the interaction between DNA and a metallic surface has the potential to lead to a shortening of the global relaxation times, as compared with molecular dynamics in solution. We have found that the surface dynamics of molecular subgroups in plant DNA is, in some cases, about two times faster than the solution dynamics of nucleic acids. This can be rationalized in a qualitative manner by invoking the complex landscape of the interaction energy between the molecule and the silver surface.

Clonal growth and demography of a hemicryptophyte alpine plant: Leontopodium alpinum Cassini

Clonally reproducing hemicryptophytic rosette plants are common in the alpine belt. However, their demography and, indirectly, their growth and reproductive strategy in these harsh conditions were rarely studied. We analysed the morphology, clonal reproduction and demography of one such species, Leontopodium alpinum, in two populations of the Swiss Alps. The species forms small colonies of 1–5 (maximum 30) sterile rosettes with a few flowering stalks. After flowering, the apical meristem dies and one or two new axillary buds grow below the previous rosette in the following year, developing into short rhizomes (<2 cm), which decay after 4 years. The new stalk produces sterile rosettes before flowering after 2–4 years, depending on climatic conditions. The apical meristem often dies during the sterile stage, and is replaced by a new axillary bud. Levkovitch matrices on two stages (sterile and flowering rosettes) showed that rosette survival and clonal reproduction maintain long-lived populations (λ = 0.96). Elasticities indicated that a change in the survival of sterile rosettes had the strongest effect on population dynamics, and this stage lasts, on average, 6.8 years at 2,480 m. Altogether, L. alpinum is following Tomlinson’s architectural model. This growth form appears perfectly adapted to harsh alpine conditions: the clonal ramification ensures longevity to genets and the semelparous behaviour of the rosettes allows an efficient flowering, whatever the climatic conditions. L. alpinum appears to follow a common growth model among rosette possessing hemicryptophytes in the alpine belt.

The role of GA3 in the germination process of high-mountain endemic and threatened species: Leontopodium nivalePinguicula fiorii and Soldanella minima subsp. samnitica (central Apennines, Italy)

The ex situ conservation of biodiversity is an essential tool for environmental protection interventions. Germination studies of seeds that belong to endangered species are essential for ex situ conservation strategies. In this study, we investigate the germination responses of three high-altitude endemic and vulnerable species (Leontopodium nivale, Pinguicula fiorii and Soldanella minima subsp. samnitica). Specifically we identified potential dormancy mechanisms by investigating the responses of germination percentage and rate to different concentrations of gibberellic acid (GA3), by performing a general linear model. L. nivale reached a germination percentage of 98.0 ± 2.0% (mean ± SE) under control conditions (20°C; 12/12 photoperiod; no GA3 addition). P. fiorii showed the highest germination percentage (78.0 ± 2.0%) in the treatment with GA3 500 ppm. S. minima subsp. samnitica did not show sensitivity to GA3 but responded positively to cold stratification (6 month at 5°C) with a germination percentage of 90 ± 6%. This study made it possible to acquire important information on the germination process of threatened and rare endemic taxa.

Lignan formation in hairy root cultures of Edelweiss (Leontopodium nivale ssp. alpinum (Cass.) Greuter)

A hairy root line of Edelweiss (Leontopodium nivale ssp. alpinum (Cass.) Greuter) was obtained upon transformation with Agrobacterium rhizogenes strain ATCC15834. Elicitation of this line with silver nitrate, sucrose, methyl jasmonate and yeast extract at various concentrations in most cases resulted in a stimulation of lignan biosynthesis. Through elicitation with 6% sucrose the roots accumulated the pharmacologically active lignans leoligin and 5-methoxy-leoligin at levels of 0.0678% and 0.0372%, respectively, without significant growth inhibition. These lignan levels were comparable to those found in intact roots of cultivated Edelweiss. The biotechnological production of leoligin could be an attractive option for the continuous, field culture-independent production of the valuable secondary metabolites leoligin and 5-methoxy-leoligin.

A Better Understanding of the Ecological Conditions for Leontopodium alpinum Cassini in the Swiss Alps

Although Leontopodium alpinum is considered to be threatened in many countries, only limited scientific information about its autecology is available. In this study, we aim to define the most important ecological factors which influence the distribution of L. alpinum in the Swiss Alps. These were assessed at the national scale using species distribution models based on topoclimatic predictors and at the community scale using exhaustive plant inventories. The latter were analysed using hierarchical clustering and principal component analysis, and the results were interpreted using ecological indicator values. Leontopodium alpinum was found almost exclusively on base-rich bedrocks (limestone and ultramafic rocks). The species distribution models showed that available moisture (dry regions, mostly in the Inner Alps), elevation (mostly above 2,000 m a.s.l.) and slope (mostly >30°) were the most important predictors. The releves showed that L. alpinum is present in a wide range of plant communities, all subalpine-alpine open grasslands, with a low grass cover. As a light-demanding and short species, L. alpinum requires light at ground level; hence, it can only grow in open, nutrient-poor grasslands. These conditions are met in dry conditions (dry, summer-warm climate, rocky and draining soil, south-facing aspect and/or steep slope), at high elevations, on oligotrophic soils and/or on windy ridges. Base-rich soils appear to also be essential, although it is still unclear whether this corresponds to physiological or ecological (lower competition) requirements.

Generation of Leontopodium alpinum differentiated biomass as a new active biotechnological ingredient

Generation of Leontopodium alpinum differentiated biomass as a new active biotechnological ingredient

Surface‐enhanced Raman scattering assessment of DNA from leaf tissues adsorbed on silver colloidal nanoparticles

In this work, the surface‐enhanced Raman scattering (SERS) spectra of seven genomic DNAs from leaves of chrysanthemum (Dendranthema grandiflora Ramat.), common sundew (Drosera rotundifolia L.), edelweiss (Leontopodium alpinum Cass), Epilobium hirsutum L., Hypericum richeri ssp. transsilvanicum (Čelak) Ciocârlan, rose (Rosa x hybrida L.) and redwood (Sequoia sempervirens D. Don. Endl.), respectively, have been analyzed in the wavenumber range 200–1800 cm−1. The surface‐enhanced Raman vibrational modes for each of these cases, spectroscopic band assignments and structural interpretations of genomic DNAs are reported. A high molecular structural information content can be found in the SERS spectra of these DNAs from leaf tissues. Based on this work, specific plant DNA–ligand interactions or accurate local structure of DNA might be further investigated using surface‐enhanced Raman spectroscopy. Besides, this study will generate information which is valuable in the development of label‐free DNA detection for chemical probing in living cell. Copyright © 2013 John Wiley &amp; Sons, Ltd.

Chemical profiling of Edelweiss (Leontopodium alpinum Cass.) extracts by micellar electrokinetic capillary chromatography

This manuscript reports on the first CE-method suitable for analyzing twelve pharmacologically relevant compounds (flavonoids, caffeic acids and leontopodic acid) in the aerial parts of Leontopodium alpinum. By using a 60mM borax buffer containing 25mM SDS and 35% acetonitrile the analytes could be well resolved in 25min and accurately determined in different Edelweiss specimens. Method validation assured the compliance of the assay to ICH standards (correlation coefficients ≥0.999, LOD ≤7.2μg/ml, LOQ ≤23.0μg/ml, recovery rates from 96.6 to 101.1%, intermediate precision ≤7.01%, and repeatability ≤5.52%). The quantitative results revealed a strong dependence of the content of phenolics on the growth status of the plants. In early stages no flavonoids were found, while during blooming all metabolites reached higher levels; commercially cultivated plant material showed the overall highest values (flavonoids: 0.25–0.34%, caffeic acids: 4.99–6.21%, leontopodic acid: 4.21–5.05%).

LAW PROTECTED PLANTS OF NATIONAL NATURE RESERVE DREVENÍK (HORNÁD BASIN) IN PROJECT LEARNING

The National nature reserve in Drevenik has an area of 101.82 ha with the highest degree of plant protection. Travertine formation is located at an altitude of 609 m a.s.l. in the eastern part of Hornad basin in East Slovakia. In the Drevenik area there are around 270,470 species and subspecies of vascular plants. Approximately 60 species are classified as endangered species of Slovakia and are classified as mountain, lowland, forest and steppe species. The plants are of phytogeographical significance, some reaching their northern limit of range in this area. Protected species of the area include (Pulsatilla slavica G. Reuss.), Alpine aster (Aster alpinus L.), Carpathian harebell (Campanula carpatica Jacq.), Pontic dragonhead (Dracocephalum austriacum L.), Hungarian iris (Iris aphylla subsp. hungarica), European columbine (Aquilegia vulgaris L.), Snowdrop windflower (Anemone sylvestris L.), Manchurian monk’s-hood (Aconitum variegatum L.), Turk’s cap lily (Lilium martagon L.), St. Lucie cherry (Cerasus mahaleb (L.) Mill.), Yellow Monkshood (Aconitum anthora L.), Bladdernut (Staphylea pinnata L.) and Edelweiss (Leontopodium alpinum Cass.). The aim of project learning is the interconnection between school theory and practice, and the upbringing towards independence and responsibility. The pupils in the second primary education stage developed these skills by elaborating a card file of plants and a phytogeographical map of the range of the protected species.

Leoligin, the major lignan from Edelweiss, activates cholesteryl ester transfer protein

ObjectiveCholesteryl ester transfer protein (CETP) plays a central role in the metabolism of high-density lipoprotein particles. Therefore, we searched for new drugs that bind to CETP and modulate its activity. MethodsA preliminary pharmacophore-based parallel screening approach indicated that leoligin, a major lignan of Edelweiss (Leontopodium alpinum Cass.), might bind to CETP. Therefore we incubated leoligin ex vivo at different concentrations with human (n=20) and rabbit plasma (n=3), and quantified the CETP activity by fluorimeter. Probucol served as positive control. Furthermore, we dosed CETP transgenic mice with leoligin and vehicle control by oral gavage for 7 days and measured subsequently the in vivo modulation of CETP activity (n=5 for each treatment group). ResultsIn vitro, leoligin significantly activated CETP in human plasma at 100pM (p=0.023) and 1nM (p=0.042), respectively, whereas leoligin concentrations of 1mM inhibited CETP activity (p=0.012). The observed CETP activation was not species specific, as it was similar in magnitude for rabbit CETP. In vivo, there was also a higher CETP activity after oral dosage of CETP transgenic mice with leoligin (p=0.015). There was no short-term toxicity apparent in mice treated with leoligin. ConclusionCETP agonism by leoligin appears to be safe and effective, and may prove to be a useful modality to alter high-density lipoprotein metabolism.

Apomixis is not prevalent in subnival to nival plants of the European Alps.

Background and AimsHigh alpine environments are characterized by short growing seasons, stochastic climatic conditions and fluctuating pollinator visits. These conditions are rather unfavourable for sexual reproduction of flowering plants. Apomixis, asexual reproduction via seed, provides reproductive assurance without the need of pollinators and potentially accelerates seed development. Therefore, apomixis is expected to provide selective advantages in high-alpine biota. Indeed, apomictic species occur frequently in the subalpine to alpine grassland zone of the European Alps, but the mode of reproduction of the subnival to nival flora was largely unknown.MethodsThe mode of reproduction in 14 species belonging to seven families was investigated via flow cytometric seed screen. The sampling comprised 12 species typical for nival to subnival plant communities of the European Alps without any previous information on apomixis (Achillea atrata, Androsace alpina, Arabis caerulea, Erigeron uniflorus, Gnaphalium hoppeanum, Leucanthemopsis alpina, Oxyria digyna, Potentilla frigida, Ranunculus alpestris, R. glacialis, R. pygmaeus and Saxifraga bryoides), and two high-alpine species with apomixis reported from other geographical areas (Leontopodium alpinum and Potentilla crantzii).Key ResultsFlow cytometric data were clearly interpretable for all 46 population samples, confirming the utility of the method for broad screenings on non-model organisms. Formation of endosperm in all species of Asteraceae was documented. Ratios of endosperm : embryo showed pseudogamous apomixis for Potentilla crantzii (ratio approx. 3), but sexual reproduction for all other species (ratios approx. 1·5).ConclusionsThe occurrence of apomixis is not correlated to high altitudes, and cannot be readily explained by selective forces due to environmental conditions. The investigated species have probably other adaptations to high altitudes to maintain reproductive assurance via sexuality. We hypothesize that shifts to apomixis are rather connected to frequencies of polyploidization than to ecological conditions.

Metabolic fingerprinting of Leontopodium species (Asteraceae) by means of 1H NMR and HPLC–ESI-MS

The genus Leontopodium, mainly distributed in Central and Eastern Asia, consists of ca. 34–58 different species. The European Leontopodium alpinum, commonly known as Edelweiss, has a long tradition in folk medicine. Recent research has resulted in the identification of prior unknown secondary metabolites, some of them with interesting biological activities. Despite this, nearly nothing is known about the Asian species of the genus. In this study, we applied proton nuclear magnetic resonance (1H NMR) spectroscopy and liquid chromatography–mass spectrometry (LC–MS) metabolic fingerprinting to reveal insights into the metabolic patterns of 11 different Leontopodium species, and to conclude on their taxonomic relationship. Principal component analysis (PCA) of 1H NMR fingerprints revealed two species groups. Discriminators for these groups were identified as fatty acids and sucrose for group A, and ent-kaurenoic acid and derivatives thereof for group B. Five diterpenes together with one sesquiterpene were isolated from Leontopodium franchetii roots; the compounds were described for the first time for L. franchetii: ent-kaur-16-en-19-oic acid, methyl-15α-angeloyloxy-ent-kaur-16-en-19-oate, methyl-ent-kaur-16-en-19-oate, 8-acetoxymodhephene, 19-acetoxy-ent-kaur-16-ene, methyl-15β–angeloyloxy-16,17-epoxy-ent-kauran-19-oate. In addition, differences in the metabolic profile between collected and cultivated species could be observed using a partial least squares-discriminant analysis (PLS-DA). PCA of the LC–MS fingerprints revealed three groups. Discriminating signals were compared to literature data and identified as two bisabolane derivatives responsible for discrimination of group A and C, and one ent-kaurenoic acid derivative, discriminating group B. A taxonomic relationship between a previously unidentified species and L. franchetii and Leontopodium sinense could be determined by comparing NMR fingerprints. This finding supports recent molecular data. Furthermore, Leontopodium dedekensii and L. sinense, two closely related species in terms of morphology and DNA-fingerprints, could be distinguished clearly using 1H NMR and LC–MS metabolic fingerprinting.

Free radicals and antioxidants in two oxidative-stress cell models exposed to ochratoxin A and amyloid β: unexpected results

Oxidative stress is reported to be closely related to pathogenetic mechanisms and plays a central role in several molecular toxicities. It seems reasonable to hypothesize that antioxidant molecules and/or free radical scavengers can protect from oxidative stress-induced damage by quenching free-radical reactions. Leontopodic acid (LA) is a fully substituted, hexaric acid derivative isolated from Leontopodium alpinum Cass., commonly known as ‘Edelweiss’ (Asteraceae family), and exhibits significant chemical antioxidant power. The present study was designed to evaluate LA’s capacity to counteract induced oxidative stress in two in vitro cell models: one exposed the LLC-PK1 cell line to ochratoxin A and the other exposed the differentiated SH-SY5Y cells to amyloid β. The effects of a 24 h pretreatment with LA on induced ROS (reactive oxygen species) production and cytotoxicity were evaluated in the two models. Surprisingly, the results showed that while LA decreased ROS production, it did not increase cell survival.

MOLECULAR PHYLOGENY OF THE EDELWEISS (LEONTOPODIUM, ASTERACEAE – GNAPHALIEAE)

Leontopodium is a genus of approximately 30 species with a conspicuous Asian–European disjunct distribution. In this study samples from the Himalayan/Tibetan centre of diversity of the genus, as well as from Europe, were analysed to infer a phylogeny of the genus using sequences of nuclear ribosomal (ITS and ETS) and plastid (matK and trnL/F) DNA. The Southeast Tibetan monotypic Sinoleontopodium [Leontopodium lingianum (Y.L.Chen) Dickoré, comb. nov.] falls into Leontopodium. Monophyly of Leontopodium, including Sinoleontopodium, is supported. Due to low rates of sequence divergence, intrageneric relationships in general are weakly supported, a pattern frequently observed in plant groups centred in the Tibetan Plateau. Three phylogenetic groups can be identified, however, and these are also supported by morphology. The low levels of nucleotide divergence suggest a young age for the group, which has been influenced by the turbulent geological history of the Tibetan Plateau. Leontopodium is a characteristic Sino-Himalayan element that appears to have found its way into the mountains of Europe in geologically recent times. The two European taxa, Leontopodium alpinum and L. nivale, form a genetically distinct group, which, considering the wide geographic disjunction, shows surprisingly little divergence from its Asian relatives.

New Ex Situ Collection of Rare and Threatened Medicinal Plants in the Pirin Mts. (Bulgaria)

This study reports the results of our efforts to create a stable and vigorous collection of rare and endemic plants from the Pirin Mountains on a plot located at Dobrinishte village (the North-East foothills of the Pirin) from 2005 to 2008. Some threatened plants from the Pirin Mountains well known for their medicinal properties were included in the ex situ collection. Here we share our observations on the pre-adaptation of adult plants and our experience in growing seedlings. Adult plants were transplanted from their native populations in the summer of 2005 and they developed amazingly well. A total of 88.2% of the transplants survived until the next season and 31.7% of the survivals bloomed the next year. Many of these set seed for the following seasons. Good germination and early stage development was observed for Leontopodium alpinum but later many of the young seedlings did not survive. Linum capitatum demonstrated poor germination without treatment other than stratification. Good germination occurred in Sideritis scardica, Stachys recta, after treatment with gibberellic acid. Germination of Hypericum linarioides was poor. In total 462 seedlings of the studied plant species were prepared for planting in the garden. The seedlings at the 2-6 true leaf stage, were transported to Dobrinishte village and planted singly into the flower beds with fine marble gravel top-dressing (up to 1 cm size). Most transplanted seedling survived the first summer. Although the number of survivals gradually decreased each following year and the critical points were mainly winters seed grown plants bloomed and set seed next seasons. Thus seed was collected for the next year from the flower beds, rather than from nature.

Biodiversity in the high ranges of the Alps: Ethnobotanical and climate change perspectives

Alpine plants, those predominantly occurring above a treeline, contribute significantly to biodiversity in mountain systems. Thus, one might expect a considerable part of the Alpine flora to be ethnobotanically relevant as has been documented for the Eastern Himalaya. As climate change, especially global warming, may lead to the extinction of Alpine plants, at least locally, loss of alpine species might be important to local people who rely on traditional medicine. How this holds for the Alps will be discussed in this contribution. The Alps are different from the Himalayas in that land use has changed dramatically during the past century. Rural life and culture has changed and much land has been abandoned or is under modern land use systems (industrial farming, tourism, urbanisation). Though much traditional knowledge has disappeared, traditional medicinal practices are still popular as supplemental to modern medicine. In a representative region in the Central Alps, traditional healers use 268 plants species of which 158 can be considered native to that area. Of the 25 predominantly Alpine species three are restricted to the highest Alps where warming might lead to their extinction. In the Alps some ornamental plants are of particular interest from an ethnobotanical point of view. People are apparently much more interested in what happens to Edelweiss ( Leontopodium alpinum) than to medicinal plants. Other famous ornamentals ( Leontopodium, Rhododendron, Gentiana, Anemone species) do not grow at critical high elevations, but could disappear locally from mountains which are too low in elevation.