Wild Edible Plants Used in Dalmatian Zagora (Croatia).

BACKGROUND: Dalmatian Zagora has experienced significant depopulation trends over recent decades. The area is very interesting because of its rich biodiversity of species as well as its history of the use of wild foods. Since there is a danger of permanent loss of knowledge on the use of wild edibles, we focused our research on recording traditions local to this area. METHODS: We conducted interviews with 180 residents. RESULTS: A record was made of 136 species of wild food plants and 22 species of edible mushrooms gathered in the area. The most frequently collected species are Rubus ulmifolius Schott, Cornus mas L., Portulaca oleracea L., Asparagus acutifolius L., Sonchus spp., Morus spp., Taraxacum spp., Amaranthus retroflexus L., Cichorium intybus L., and Dioscorea communis (L.) Caddick & Wilkin. CONCLUSIONS: The list of taxa used is typical for other (sub-)Mediterranean parts of Croatia; however, more fungi species are used. The most important finding of the paper is probably the recording of Legousia speculum-veneris (L.) Chaix, a wild vegetable used in the area.

Phylogeographic structure of common sage (Salvia officinalis L.) reveals microrefugia throughout the Balkans and colonizations of the Apennines.

Studying the population-genetic and phylogeographic structures of a representative species of a particular geographical region can not only provide us with information regarding its evolutionary history, but also improve our understanding of the evolutionary processes underlying the patterns of species diversity in that area. By analysing eight highly polymorphic microsatellite loci and two chloroplast DNA regions, we have investigated the influence of Pleistocene climate fluctuations on the evolutionary history of Salvia officinalis L. (common sage). The populations with the highest genetic diversity were located in the central parts of the Balkan distribution range. A large group of closely related haplotypes was distributed throughout the Balkans and the central Apennines, while the private lineage occupied the southern Apennines. In addition, two highly differentiated lineages were scattered only over the Balkans. The results suggest that a single refugium of the studied species from the last glacial period was located in the central part of the range in the Balkans. Numerous microrefugia, probably spanning several glaciation cycles, were scattered across the Balkans, while colonisation of the Apennines from the Balkans occurred at least on two occasions.

Population structure and adaptive variation of Helichrysum italicum (Roth) G. Don along eastern Adriatic temperature and precipitation gradient.

Immortelle (Helichrysum italicum (Roth) G. Don; Asteraceae) is a perennial plant species native to the Mediterranean region, known for many properties with wide application mainly in perfume and cosmetic industry. A total of 18 wild H. italicum populations systematically sampled along the eastern Adriatic environmental gradient were studied using AFLP markers to determine genetic diversity and structure and to identify loci potentially responsible for adaptive divergence. Results showed higher levels of intrapopulation diversity than interpopulation diversity. Genetic differentiation among populations was significant but low, indicating extensive gene flow between populations. Bayesian analysis of population structure revealed the existence of two genetic clusters. Combining the results of FST - outlier analysis (Mcheza and BayeScan) and genome-environment association analysis (Samßada, LFMM) four AFLP loci strongly associated with the bioclimatic variables Bio03 Isothermality, Bio08 Mean temperature of the wettest quarter, Bio15 Precipitation seasonality, and Bio17 Precipitation of driest quarter were found to be the main variables driving potential adaptive genetic variation in H. italicum along the eastern Adriatic environmental gradient. Redundancy analysis revealed that the partitioning of genetic variation was mainly associated with the adaptation to temperature oscillations. The results of the research may contribute to a clearer understanding of the importance of local adaptations for the genetic differentiation of Mediterranean plants and allow the planning of appropriate conservation strategies. However, considering that the identified outlier loci may be linked to genes under selection rather than being the target of natural selection, future studies must aim at their additional analysis.

Insular Pharmacopoeias: Ethnobotanical Characteristics of Medicinal Plants Used on the Adriatic Islands.

The Adriatic islands in Croatia, usually divided into two archipelagos - the Kvarner and Dalmatian islands - is one of the largest groups of islands in Europe. Over 40 islands are still inhabited. Unfortunately the traditional use of medicinal plants was never properly documented there. Our data comes from 343 interviews carried out in 36 islands, including the 15 largest islands of the archipelago. The medicinal plants are mainly used to make herbal infusions or decoctions, occasionally medicinal liqueurs, syrups, compresses, or juices squeezed out of raw plants. We recorded the use of 146 taxa, among them 131 with at least one medicinal purpose and 15 only for tea. The frequency curve of use is relatively steep - several plants are used very frequently and most are reported only by one or two informants, which can be explained both by the large geographical spread of the area, and even more so by the devolution of local knowledge and disappearance of gathering practices due to specialization in tourism, modernization and depopulation. Most of the gathered plants already occur in ancient and medieval herbals and are a part of the pan-Mediterranean pharmacopoeia.

Plants in alcoholic beverages on the Croatian islands, with special reference to rakija travarica.

BACKGROUND: This paper aims to record the species used for flavouring and making alcoholic drinks, mainly rakija, on the islands of the Adriatic (Croatia). METHODS: Our data comes from 295 interviews performed on 36 islands, in both the Dalmatian and Kvarner areas of the Adriatic. RESULTS: Altogether, 114 species are used-46% from wild locations only, 15% both wild and cultivated, 38% only cultivated and two species are imported. The most common local alcohol is wine, made without spices, but grape pomace distillate is often flavoured with single or mixed species. The mix is called travarica. The most commonly used species are Foeniculum vulgare Mill., Myrtus communis L., Salvia officinalis L., Ruta graveolens L., Juniperus oxycedrus L., Ceratonia siliqua L., Juglans regia L., Citrus spp., Ficus carica L., Laurus nobilis L., Rosmarinus officinalis L., Artemisia absinthium L., Rosa centifolia L., Mentha × piperita L. and M. spicata L. Unfortunately, the widespread phenomenon of distilling Arbutus unedo L. fruits and fermenting Juniperus 'wine' is now extinct. Apart from grapes, the only commonly distilled fruit now is Ficus carica. CONCLUSIONS: It is striking that nearly all the plants are either wild or cultivated locally, which, in addition to the fact that the alcohol is made locally, shows the incredible local culinary self-sufficiency of the area. The number of species used is also very impressive.

The ethnobotany and biogeography of wild vegetables in the Adriatic islands.

BACKGROUND: Archipelagos of islands have played an important role in shaping some of the paradigms of biology, including the theory of the evolution of species. Later, their importance in biology was further emphasised by the theory of island biogeography, which contributed to a better understanding of the shaping of species richness not only on real islands, but on isolated habitat islands as well. Although ethnobotany is a well-established discipline, patterns of knowledge about plant uses in archipelagos have never been quantitatively analysed, and the whole concept has been only briefly mentioned in the ethnobiological context. The aim of our study was to record which taxa of wild vegetables have been consumed in the Adriatic islands and to establish if such variables as island size, population size, flora or its isolation are correlated with the number of wild vegetables used. METHODS: We interviewed 225 people (15 from each island). RESULTS: Altogether, the use of 89 species of wild vegetables has been recorded. The largest number of wild vegetables is eaten on the islands of Korcula, Vis and Solta, and the lowest on Ugljan, Cres and Dugi Otok. The studied independent variables had a small and statistically not significant effect on the wild vegetable list length. The most visible effect was an increasing trend from north-west to south-east, overrunning the typical biogeographical island patterns. Moreover, one of the large and well-populated islands, Korcula, showed an 'unusually' high level of wild vegetable use. We hypothesise that the current use of so many species on this island has been maintained by the inhabitants' awareness that they are the holders of relic knowledge, an awareness reiterated by ethnographic and popular publications, as well as a strong history of famine. The most interesting edible species used in the Adriatic islands are Bunium alpinum, Cytinus hypocystis (both mainly on Pasman), Lotus edulis (on Vis) and Posidonia oceanica (on Vis and Korcula). CONCLUSIONS: The recorded relationships between the demographic and geographical features of the islands were statistically not significant. We assume that cultural and historical factors diversifying the use of plants in particular islands are stronger than the above-mentioned measurable variables.

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.

Chemical diversity of the natural populations of Dalmatian pyrethrum (Tanacetum cinerariifolium (TREVIR.) SCH.BIP.) in Croatia.

Dalmatian pyrethrum (Tanacetum cinerariifolium (TREVIR.) SCH.BIP.) is a plant species endemic to the east Adriatic coast. The bioactive substance of Dalmatian pyrethrum is a natural insecticide, pyrethrin, a mixture of six active components (pyrethrins I and II, cinerins I and II, and jasmolins I and II). The insecticidal potential of pyrethrin was recognized decades ago, and dried and ground flowers have traditionally been used in Croatian agriculture and households. A total of 25 Dalmatian pyrethrum populations from Croatia were studied to determine the pyrethrin content and composition, and to identify distinct chemotypes. The total pyrethrin content ranged from 0.36 to 1.30% (dry flower weight; DW) and the pyrethrin I/pyrethrin II ratio ranged from 0.64 to 3.33%. The statistical analyses revealed that the correlations between the percentage of pyrethrin I and of all the other components were significant and negative. The total pyrethrin content was positively correlated with the percentage of pyrethrin I and negatively correlated with cinerin II. The multivariate analysis of the chemical variability enabled the identification of five chemotypes among 25 Dalmatian pyrethrum populations. The chemical characterization of indigenous Dalmatian pyrethrum populations may serve as a good background for future breeding and agricultural exploitation.

Development of new microsatellite markers for Salvia officinalis L. and its potential use in conservation-genetic studies of narrow endemic Salvia brachyodon Vandas.

Nine new microsatellite markers (SSR) were isolated from Salvia officinalis L. A total of 125 alleles, with 8 to 21 alleles per locus, were detected in a natural population from the east Adriatic coast. The observed heterozygosity, expected heterozygosity, and polymorphic information content ranged from 0.46 to 0.83, 0.73 to 0.93 and 0.70 to 0.92, respectively. New microsatellite markers, as well as previously published markers, were tested for cross-amplification in Salvia brachyodon Vandas, a narrow endemic species known to be present in only two localities on the Balkan Peninsula. Out of 30 microsatellite markers tested on the natural S. brachyodon population, 15 were successfully amplified. To obtain evidence of recent bottleneck events in the populations of both species, observed genetic diversity (H(E)) was compared to the expected genetic diversity at mutation-drift equilibrium (H(EQ)) and calculated from the observed number of alleles using a two-phased mutation model (TPM). Recent bottleneck events were detected only in the S. brachyodon population. This result suggests the need to reconsider the current threat category of this endemic species.

High diversity of indigenous populations of dalmatian sage (Salvia officinalis L.) in essential-oil composition.

Essential oils of 25 indigenous populations of Dalmatian sage (Salvia officinalis L.) that represent nearly half of native distribution area of the species were analyzed. Plantlets collected from wild populations were grown in the same field under the same environmental conditions and then sampled for essential-oil analysis. The yield of essential oil ranged from 1.93 to 3.70% with average of 2.83%. Among the 62 compounds detected, eight (cis-thujone, camphor, trans-thujone, 1,8-cineole, ß-pinene, camphene, borneol, and bornyl acetate) formed 78.13-87.33% of essential oils of individual populations. Strong positive correlations were observed between camphor and ß-pinene, ß-pinene and borneol, as well as between borneol and bornyl acetate. The strongest negative correlation was detected between camphor and trans-thujone. Principal component analysis (PCA) on the basis of eight main compounds showed that first main component separated populations with high thujone content, from those rich in camphor, while the second component separated populations rich in cis-thujone from those rich in trans-thujone. Cluster analysis (CA) led to the identification of three chemotypes of S. officinalis populations: cis-thujone; trans-tujone, and camphor/ß-pinene/borneol/bornyl acetate. We propose that differences in essential oils of 25 populations are mostly genetically controlled, since potential environmental factors were controlled in this study.