Embryological features, pollen and seed viability of Arnica montana (Asteraceae) – a threatened endemic species in Europe

The embryological features, mode of reproduction and reproductive capacity (pollen and seed viability) on two naturalized populations of Arnica montana in Bulgaria were studied. The embryological study shows that A. montana is a facultative apomictic species in which sexual reproduction predominates. In this species, it was established that there is a comparatively high viability of the mature pollen and embryos, which enables the successful realization of its reproductive capacity. The results of the study on A. montana reveal that both sexual and asexual vegetative reproduction with rhizomes undoubtedly play more important roles than the apomixis (namely diplospory) for support and preservation of the populations

Comparative study of in vitro, ex vitro and in vivo grown plants of Arnica montana – polyphenols and free radical scavenging activity

Arnica montana L. is an endangered species rich in sesquiterpene lactones, phenolic acids and flavonoids with high pharmaceutical value. The polyphenolic content and free radical scavenging activity of plants that had passed all stages of cultivation: micropropagation and rooting (in vitro), adaptation in greenhouse (ex vitro) and mountain conditions (in vivo) were evaluated. Four surface flavonoid aglycones [scutellarein 6-methyl ether (hispidulin), scutellarein 6,4’-dimethyl ether (pectolinarigenin), 6-OH luteolin 6-methyl ether and kempferol-6-methyl ether] were detected in the acetone exudates of the studied samples by means of thin layer chromatography.No differences in the accumulation of surface flavonoids were found among the tested leaf extracts of in vitro, ex vitro and in vivo samples. However, the extracts from the flowers were richer in surface flavonoids than extracts from the leaves. The methanol extracts of the samples from ex vitro and in vivo grown A. montana plants had significantly higher radical scavenging activity and polyphenolic content than the extracts of in vitro samples. The observed differences in the contents of these biologically active compounds were related to different growth conditions and stages of plant development. The biotechnological method of A. montana established holds promise for the future production of antioxidants

Multiplication and Conservation of Threatened Medicinal Plant Arnica montana L. by in vitro Techniques

An efficient and reproducible in vitro protocol for mass production of the threatened medicinal plant Arnica montana L. (Asteraceae) was developed. The effectiveness of various combinations of plant growth regulators on A. montana clonal multiplication was assessed, using seedlings stems as initial explants. Among 12 tested nutrient media, the optimum one (MS supplemented with 1.0 mg/l BAP and 0.1 mg/l IAA) increased the organogenesis frequency up to 95% in the best origin, with mean number of shoots per explant 4.25 for 5 weeks. Sub-cultivations on this medium every 4 weeks led to increase of the propagation rate as in the fifth subculture the average number of shoots per explant reached 12.32+-0.82. Rooting of uniform in vitro shoots was 100% successful on half strength MS medium supplemented with 0.5 mg/l IBA. The ex vitro adapted plants showed 90% survival, and were further acclimatized to two mountain ex situ collections. Plants looked healthy and true-to- type and began to bloom in the second or the third year. In addition, a successful protocol for slow-growth storage of in vitro A. montana cultures was elaborated, after testing 8 media with mannitol or sorbitol. The medium 1/2 MS containing 3% sorbitol and 2% sucrose was chosen as the best one, efficiently retarding the growth of the in vitro plantlets, thus allowing 6-month maintenance without sub-cultivation. The developed in vitro protocols could be of great value for commercial propagation and sustainable conservation of this threatened medicinal plant

Razširjenost in ekologija vrst rodu Alchemilla na gorovju Osogovo in Zahodni Balkan v Bolgariji

The present study aims to determine some common species of genus Alchemilla (Rosaceae) in Osogovo Mt. and West Balkan Mt. in Bulgaria. The ecological conditions in seven habitat types according to EUNIS with detected presence of Alchemilla species are compared. According to this classification, the habitats that were found belong to six general types: Transition mires and quaking bogs (D 2.3), Mountain hay meadows (E 2.3), Moist or wet eutrophic and mesotrophic grassland (E 3.4), Acid alpine and subalpine grassland (E 4.3), Subalpine moist or wet tall-herb and fern stands (E 5.5), Evergreen alpine and subalpine heath and scrub (F 2.2). Seven species were reported in the investigated regions: A. crinita Buser, A. erythropoda Juz., A. flabellata Buser, A. glabra Neygenf., A. glaucescens Wallr., A. monticola Opiz and A. viridiflora Rothm. In the paper are submitted some new reports for the presence of A. crinita, A. erythropoda and A. glabra in the investigated areas. Some preliminary results concerning the resources of the examined species are obtained.V raziskavi smo predstavili nekaj vrst rodu Alchemilla (Rosaceae) na gorovjih Osogovo in Zahodni Balkan (Stara Planina) v Bolgariji. Primerjali smo rastiščne razmere v sedmih habitatnih tipih po razvrstitvi EUNIS, v katerih smo ugotovili prisotnost vrst Alchemilla. Po tej razvrstitvi najdene habitate uvrščamo v šest glavnih tipov: prehodna barja (D 2.3), gorski gojeni travniki (E 2.3), mokrotni mezotrofni in eutrofni travniki ali pašniki (E 3.4), alpinska in subalpinska travišča na kisli podlagi (E 4.3), subalpinska in alpinska visoka steblikovja (E 5.5), arktično-alpinske in borealne resave (F 2.2). V raziskovanem območju smo našli sedem vrst: A. crinita Buser, A. erythropoda Juz., A. flabellata Buser, A. glabra Neygenf., A. glaucescens Wallr., A. monticola Opiz and A. viridiflora Rothm. V članku so prikazana nova nahajališča vrst A. crinita, A. erythropoda in A. glabra na raziskovanem območju. Prikazali smo tudi predhodne rezultate koristne uporabe proučevanih vrst rodu Alchemilla

Multiplication and Conservation of Threatened Medicinal Plant Arnica montana L. by in vitro Techniques

An efficient and reproducible in vitro protocol for mass production of the threatened medicinal plant Arnica montana L. (Asteraceae) was developed. The effectiveness of various combinations of plant growth regulators on A. montana clonal multiplication was assessed, using seedlings stems as initial explants. Among 12 tested nutrient media, the optimum one (MS supplemented with 1.0 mg/l BAP and 0.1 mg/l IAA) increased the organogenesis frequency up to 95% in the best origin, with mean number of shoots per explant 4.25 for 5 weeks. Sub-cultivations on this medium every 4 weeks led to increase of the propagation rate as in the fifth subculture the average number of shoots per explant reached 12.32+-0.82. Rooting of uniform in vitro shoots was 100% successful on half strength MS medium supplemented with 0.5 mg/l IBA. The ex vitro adapted plants showed 90% survival, and were further acclimatized to two mountain ex situ collections. Plants looked healthy and true-to- type and began to bloom in the second or the third year. In addition, a successful protocol for slow-growth storage of in vitro A. montana cultures was elaborated, after testing 8 media with mannitol or sorbitol. The medium 1/2 MS containing 3% sorbitol and 2% sucrose was chosen as the best one, efficiently retarding the growth of the in vitro plantlets, thus allowing 6-month maintenance without sub-cultivation. The developed in vitro protocols could be of great value for commercial propagation and sustainable conservation of this threatened medicinal plant

On the distribution of Helichrysum arenarium (l.) Moench (Asreraceae) in Bulgaria

Helichrysum arenarium (L.) Moench is herbaceous perennial plant belonging to Asteraceae family. The plant is well known in phytotherapy for its potential in the treatment of gallbladder disease and is classified as endangered in a number of European countries. In Bulgaria it is a protected species according to the Biodiversity Act (Annex 4) and is in the List of Species of Medicinal Plants under special regimen of conservation and use. The data on the distribution of H. arenarium in the Bulgarian flora have not been updated for more than 20 years. The aim of this study is to determine the current distribution of H. arenarium in Bulgaria on the basis of reviewing the available herbarium specimens in the Bulgarian herbariums, literature data, and personal collections. As a result, the locations of the species on the territory of the country were described and mapped. In Bulgaria H. arenarium is represented with only a few populations located in a limited area in the northeastern part of the country, in a narrow strip between the towns of Shumen and Varna

Plant-soil Water Status-induced Changes in Physiological and Biochemical Properties of Yarrow

WOS: 000392637400021As medicinal plants are generally collected from the wild flora, it is of fundamental importance to identify the optimum growth conditions for increasing the yield and quality potential of ex-situ cultivated plants. Effects of different soil water content [20, 40, 60 and 80 % of water holding capacity (WHC)] on growth, physiology and biochemical profile of Achillea millefolium group-related cv. Proa were investigated in the present study. Alterations in drug herbage, chlorophyll content, quantum efficiency of photosystem II, essential oil rate and composition, proline content and lipid peroxidation, antioxidant activity and flavonoid content of the plants under different plant and soil water status were determined. Decrease in soil water status under 40 % of WHC resulted in degradation of chlorophyll pigments, reduction in photochemical efficiency and lower dry matter production. The essential oil (EO) rate of cv. Proa was higher under limited and excessive water content of soil whereas the main component of EO, chamazulene, was negatively correlated with WHC of soil. Our results suggest a general trend of inverse correlation of antioxidant capacity and flavonoid content with essential oil rate in contrast to the positive correlation of these parameters with chamazulene content. The data obtained may serve as a basis of programming the optimum soil moisture conditions for successful cultivation of cv.Proa.Research Fund of TUBITAK (The Scientific and Technological Research Council of Turkey)Turkiye Bilimsel ve Teknolojik Arastirma Kurumu (TUBITAK); BAS (Bulgarian Academy of Science) [180O290]The authors thank Research Fund of TUBITAK (The Scientific and Technological Research Council of Turkey) and BAS (Bulgarian Academy of Science) for their financial support (Bilateral project No. 180O290)

SOME AGRONOMIC AND QUALITY CHARACTERISTICS OF TWO Achillea asplenifolia Vent. POPULATIONS

WOS: 000392043000017The genus of Achillea L. consists of herbaceous plants are generally distributed in the northern hemisphere. Achillea asplenifolia and Achillea collina are the most important group of A. millefolium due to their highly effective pharmacological properties. Both species, A. asplenifolia and A. collina are distributed in the Europen flora but they don't exist in the Turkish flora. The present study aimed to determine some yield and quality characteristics of two populations, Achillea asplenifolia - 9602 and Achillea asplenifolia - 10403, belonging to Achillea asplenifolia Vent. collected from the flora of Bulgaria at Bornova and to select genotypes with high yield and quality. For this purpose a two years of field experiment was conducted in 2009 and 2010. Plant height (cm), green herbage yield (g/plant), drug herbage yield (g/plant), drug flos yield (g/plant) and essential oil content of populations were determined. Based on the results of two years of experiment minimum and maximum drug flos yield were 2 g/plant and 391.4 g/plant in Achillea asplenifolia - 9602. Essential oil rate ranged between 0.050-3.30%. Minimum and maximum drug flos yield of Achillea asplenifolia (10403) were 0 and 159.8 g/plant respectively and essential oil content ranged between 0-2.5%. A large distribution was found in both population of Achillea asplenifolia and especially population Achillea asplenifolia - 9602 was more remarkable and promising for selection studies.Scientific and Technical Research Council of Turkey (TUBITAK)Turkiye Bilimsel ve Teknolojik Arastirma Kurumu (TUBITAK) [108O290]We thank to the Scientific and Technical Research Council of Turkey (TUBITAK) for its financial support to this study (Project number: 108O290)

Balkan Vegetation Database: historical background, current status and future perspectives

The Balkan Vegetation Database (BVD; GIVD ID: EU-00-019; http://www.givd.info/ID/EU-00- 019) is a regional database that consists of phytosociological relevés from different vegetation types from six countries on the Balkan Peninsula (Albania, Bosnia and Herzegovina, Bulgaria, Kosovo, Montenegro and Serbia). Currently, it contains 9,580 relevés, and most of them (78%) are geo-referenced. The database includes digitized relevés from the literature (79%) and unpublished data (21%). Herein we present descriptive statistics about attributive relevé information. We developed rules that regulate governance of the database, data provision, types of data availability regimes, data requests and terms of use, authorships and relationships with other databases. The database offers an extensive overview about studies on the local, regional and SE European levels including information about flora, vegetation and habitats