Danijela Stešević

Ethnobotanical study on traditional uses of wild medicinal plants in Prokletije Mountains (Montenegro)

AIM OF THE STUDY The main objectives were to collect information on the use of wild growing medicinal plants by local people living in high mountain region of Montenegro and conduct local botanical and ecological surveys. Active ingredients of plant species officinal in European Pharmacopoeia 6.0 (Ph. Eur. 6.0) were studied and we assessed possibilities for commercial exploitation for local economic development. MATERIALS AND METHODS The 75 people that were interviewed (40-82 years old) identified 94 species for treatment of various human ailments. For each named species, the following elements are provided: botanical name, family, part(s) used, medicinal use and perceived property, listing in published pharmacopoeias, the relative abundance of each species and locality where the plant was collected. Chemical analyses were done according to prescriptions of Ph. Eur. 6.0 in order to estimate potential commercial use of native plants. RESULTS AND CONCLUSION The most common in traditional usage were Rosaceae (11 species) making 11.7%, Asteraceae (10 species) 10.6% and Lamiaceae (7 species) 7.4%. From 94 species reported, 35 (37.2%) are officinal in Ph. Eur. 6.0 and 12 in national pharmacopoeias (12.8%). Aerial parts were mostly used (43.6%). The most frequently reported medicinal uses were for treating gastrointestinal (57.4%) and respiratory diseases (41.5%).

Chemotype Diversity of Indigenous Dalmatian Sage (Salvia officinalis L.) Populations in Montenegro

To identify how many chemotypes of Salvia officinalis exist in Montenegro, the chemical composition of the essential oils of 12 wild‐growing populations was determined by GC‐FID and GC/MS analyses. Among the 40 identified constituents, the most abundant were cis‐thujone (16.98–40.35%), camphor (12.75–35.37%), 1,8‐cineol (6.40–12.06%), trans‐thujone (1.5–10.35%), camphene (2.26–9.97%), borneol (0.97–8.81%), viridiflorol (3.46–7.8%), limonene (1.8–6.47%), α‐pinene (1.59–5.46%), and α‐humulene (1.77–5.02%). The composition of the essential oils under study did not meet the ISO 9909 requirements, while the oils of populations P02–P04, P09, and P10 complied with the German Drug Codex. A few of the main essential‐oil constituents appeared to be highly intercorrelated. Strong positive correlations were observed between α‐pinene and camphene, camphene and camphor, as well as between cis‐thujone and trans‐thujone. Strong negative correlations were evidenced between cis‐thujone and α‐pinene, cis‐thujone and champhene, cis‐thujone and camphor, as well as between trans‐thujone and camphene. Multivariate analyses allowed the grouping of the populations into three distinct chemotypes, i.e., Chemotype A, rich in total thujones, Chemotype B, with intermediate contents of thujones, α‐pinene, camphene, and camphor and high borneol contents, and Chemotype C, rich in camphor, camphene, and α‐pinene. The chemotypes did not significantly differ in the total essential‐oil content and the cis/trans‐thujone ratio.

Cytotoxic Guaianolide from Anthemis segetalis (Asteraceae)

Two epimeric guaianolides were isolated from the aerial parts of Anthemis segetalis. These compounds have not been detected in any Anthemis species so far. One of them was tested for cytotoxicity against human cervical cancer (HeLa) and murine melanoma (B16) cell lines showing appreciable activity. Copyright

How many taxa? Spatiotemporal evolution and taxonomy of Amphoricarpos (Asteraceae, Carduoideae) on the Balkan Peninsula

Amphoricarpos Vis. is an early diverging genus within tribe Cardueae (Carduoideae, Asteraceae), which is disjunctly distributed in the Balkan Peninsula, Anatolia and the Caucasus; the Anatolian and Caucasian taxa are sometimes treated as separate genus Alboviodoxa. We focus on the monophyletic Balkan populations, which have been treated very inconsistently in previous taxonomic accounts (one polymorphic species with or without varying sets of intraspecific taxa vs. two species, one of them with two subspecies). In order to disentangle relationships among populations across the entire distribution area of Amphoricarpos on the Balkan Peninsula, we employed amplified fragment length polymorphisms (AFLPs) as well as nuclear and plastid DNA sequences (ITS and rps16–trnK) to a dense sampling of populations. ITS was also used to reconstruct the genus’ spatiotemporal evolution. In addition, we contrasted the genetic results with morphological data to provide a sound taxonomic revision of Amphoricarpos on the Balkan Peninsula. The split between the Balkan populations and the Anatolian A. exsul took place in the late Miocene or early Pliocene, whereas diversification within the Balkan lineage is much younger and likely started in the Pleistocene. The deepest splits seen in AFLPs and/or ITS separate the geographically disjunct northern- and southern-most populations. Divergence within the continuous distribution area in the centre is shallower, but allowed recognition of three largely allopatric clusters. Morphometric data, however, were neither in line with previous multi-taxon treatments nor with patterns of genetic divergence. We therefore refrain from recognising any of the genetic groups as a distinct taxonomic entity and rather suggest treating all Balkan populations as a single, genetically, morphologically and ecologically variable species, Amphoricarpos neumayerianus (Vis.) Greuter, without intraspecific taxa.

Morphological, genetic and phytochemical variation of the endemic Teucrium arduini L. (Lamiaceae)

Analysis of the morphological traits of leaves, genetic variability (analyzed by AFLP) and chemical composition of essential oils (analyzed by GC-MS) was conducted on eleven populations of the endemic Illyric-Balcanic species Teucrium arduini L. in Croatia, Bosnia and Herzegovina, and Montenegro. Average blade length and width ranged from 20.00 to 31.47mm and from 11.58 to 15.66mm, respectively. Multivariate analysis (PCA, UPGMA) of morphological traits distinguished two continental Bosnian populations from the remaining populations. AFLP analysis separated the investigated populations into two groups based primarily on geographical distance. Essential oil analysis showed a total of 52 compounds, with two chemotypes distinguished based on the essential oil profile. The first was a sesquiterpene chemotype, with β-caryophyllene, germacrene D or caryophyllene oxide as the major compounds, while the second was an oxygenated monoterpene chemotype, with pulegone and piperitone oxide as the main components. The Mantel test showed a stronger correlation between the morphological traits and AFLP than between the essential oil profile and AFLP. The test also showed a stronger association between the essential oil profile and geographical position than between the morphological traits and geographical position.

Epiphytic lichens and lichenicolous fungi from the northern part of Montenegro

Abstract: Bilovitz, R O., Stešević, D. & Mayrhofer, H. 2010. Epiphytic lichens and lichenicolous fungi from the northern part of Montenegro. - Herzogia 23: 249–256. A list of 120 taxa of lichens (117 species) and three species of lichenicolous fungi from the northern part of Montenegro is presented. The lichens Biatora chrysantha, Caloplaca monacensis, Candelariella efflorescens, Loxospora elatina, Micarea adnata, Ochrolechia arborea, O. microstictoides, Phaeophyscia nigricans, Physconia enteroxantha, Ph. grisea, Rinodina capensis, R. polysporoides, R. pyrina, Scoliciosporum umbrinum var. corticolum, Xanthoria candelaria, X. ulophyllodes and the lichenicolous fungi Lichenodiplis lecanorae, Telogalla olivieri, and Xanthoriicola physciae are new to Montenegro.

Trampling impact on vegetation of embryonic and stabilised sand dunes in Montenegro

Trampling is one of the human activities that are harmful for plant species and communities of sand dune ecosystems. The aim of this study was to compare the vegetation of embryonic and shifting Ammophila sand dunes with and without fencing to limit trampling. Fenced sand dunes appeared to be richer in species but differences were more prominent in embryonic sand dunes. Some species (Cakile maritima, Pancratium maritimum) were missing on trampled embryonic dunes. The positive impact of trampling exclusion on embryonic sand dunes was indicated by a lowered slope in a Whittaker graph as well as by rarefaction curves that showed higher species richness on the lower slope. Changes in the vegetation of more stabilised shifting Ammophila sand dunes due to trampling are not evident, although species composition is also impoverished. Fencing of parts of sand dunes proved to be an effective measure for vegetation conservation. In addition to physical exclusion of visitors, fences can also have symbolic value for raising public awareness.

Micromorphological traits and essential oil contents of Micromeria kerneri Murb. and M. juliana (L.) Benth. (Lamiaceae)

The chemical composition of the essential oil (analysed by GC and GC-MS), the types and distribution of trichomes and pollen morphology (analysed by scanning electron microscopy) were investigated in two closely related species, Micromeria kerneri Murb. and Micromeria juliana (L.) Benth. (Lamiaceae) from Southeast Europe as a contribution to their taxonomy. The essential oil of M. kerneri was characterized by a high concentration of oxygenated sesquiterpenes, with caryophyllene-oxide as the major compound. Caryophyllene-oxide was also the major component of the essential oil of M. juliana from all localities, except from Mt Krivošije (Montenegro), where piperitone oxide was the major constituent. Non-glandular trichomes, peltate trichomes, and two types of capitate trichomes (type 1 composed of one basal epidermal cell, and one head cell with subcuticular space; type 2 composed of one basal epidermal cell, two stalk cells, and one head cell with subcuticular space) were observed on leaves, the calyx and on the stem. Pollen of both species had six apertures (hexacolpate) set in the equatorial pollen belt (zonocolpate) and showed medium reticulate ornamentation. Multivariate analysis (PCA and UPGMA) of essential oil components clearly separated the investigated M. kerneri and M. juliana populations, and confirmed the opinion that they are different taxa. On the other hand, micromorphological traits between these species were the same. Nevertheless, definitive conclusions about the taxonomic relationships among these species will require genetic analysis.

Human impact on sandy beach vegetation along the southeastern Adriatic coast

Abstract We compared the zonation of vegetation and connectivity of coastal plant communities between two distinct areas, one in Montenegro and another in Albania, that differ in terms of human impact, mainly through tourism activities. Transect plots were used to gather data about plant cover and communities, their zonation and connectivity. For description of communities multivariate methods were used and for distribution zonation we used gamma connectivity and richness of boundaries. We found that the transects of vegetation zonation from Albania, with better preserved sites, were richer in the number of boundaries, with more varied combinations of boundaries and the pattern of zonation was also more diverse. On the other hand, there were two plant communities found only in Montenegro. The more impacted transects on the disturbed beaches from Montenegro were also more unidirectional from sea to hinterland but with less ideal zonation. Plant communities from Albania were distributed more straightforwardly but contacts between them were in both directions. The less disturbed beach had zonation very similar to potential vegetation, while plant communities of the touristic beach were fragmented or even substituted by replacement communities. Coastal dune systems in Albania are still well preserved, therefore monitoring and protection measures are recommended.

Lichenized fungi of a chestnut grove in Livari (Rumija, Montenegro)

Abstract - Sixty taxa (59 species and 1 variety) of lichenized fungi are reported from a chestnut grove in Livari. The majority of them (55 species and 1 variety) occurred on Castanea sativa. The recently described Xylographa soralifera is new to the Balkan Peninsula. The lichenicolous fungus Monodictys epilepraria growing on Lepraria rigidula is new to Montenegro. The lichen mycota is compared with similar localities in Italy and Switzerland. The species composition in Livari is most similar to the Montieri site in Tuscany.