GC/MS Based Metabolite Profiling and Antioxidant Activity of Balkan and Bulgarian Endemic Plants

Balkan and Bulgarian endemic plants Viola rhodopaea Becker (Violaceae), Veronica rhodopaea (Velen.) Degen ex Stoj. & Stefanov (Plantaginaceae), Silene roemeri Friv. (Caryophyllaceae), Jasione bulgarica Stoj. & Stef and Campanula lanata Friv. (Campanulaceae) were examined. Metabolite profiles of methanolic extracts of studied species were analyzed by GC/MS and HPTLC. Total flavonoid and phenol contents were determined by aluminum chloride (AlCl3) and Folin-Ciocalteu’s reagent, respectively. Antioxidant potential of the extracts was assayed by DPPH test. Fatty acids and alcohols, phenolic and organic acids, flavonoid aglycones and glycosides, sterols and carbohydrates were identified. Jasione bulgarica and Viola rhodopaea extracts were determined to have the highest flavonoid and phenol content. Significant radical scavenging activity was estimated for the extracts of J. bulgarica, Veronica rhodopaea and Viola rhodopaea. The present information of chemical composition and antiradical potential of studied species are reported for the first  time

The status and role of genetic diversity of trees for the conservation and management of riparian ecosystems: a European experts' perspective

Riparian vegetation supports high biodiversity providing many services and is, therefore, an important landscape element. Riparian ecosystems are subject to numerous pressures leading to population decline and genetic erosion of riparian plants. This may have cascading effects at various ecosystem levels, including decreasing ecosystem services, so identifying the current status of genetic diversity of riparian tree species is vital to improve the effectiveness of restoration efforts. We aimed to elicit expert views on the status and importance of genetic diversity of tree species, and conservation needs across European riparian ecosystems. Sharing of such information among researchers, managers and policymakers has the potential to enhance ecological restoration and management of riparian ecosystems. We identified experts in riparian genetic resources conservation and management across Europe. These included stakeholders with different perspectives, ranging from researchers to practitioners. We designed a set of questionnaires where our identified experts were asked to answer questions related to the status and conservation of genetic diversity of riparian tree species in their respective countries. Specifically, we asked about societal awareness, legislative tools, good practices and conservation or restoration projects accounting for intraspecific genetic diversity and differentiation of tree species in riparian ecosystems. Questionnaire responses were analysed and discussed in light of the scientific literature to define needs and priorities related to the management and conservation of genetic diversity of riparian tree species. The experts recognized that a combination of in situ and ex situ measures and/or integrative conservation of riparian ecosystems is the most appropriate option for conserving the genetic diversity of riparian tree species. Simultaneous application of conservation measures at the level of priority species, identified by experts, and protection of riparian areas are required. Synthesis and applications. This study revealed the importance of recognizing the ecological processes that shape the genetic diversity of riparian tree species in hydrographic networks (dendritic spatial configuration, specific patterns of gene flow among riparian populations, fragmentation of river by dams) but also the need to overcome socio-economic barriers, such as lack of policy priority, deficiency in funding and weak legislation framewor

Tracing the origin of Oriental beech stands across Western Europe and reporting hybridization with European beech : implications for assisted gene flow

The benefits and risks of human-aided translocation of individuals within the species range, assisted gene flow (AGF), depend on the genetic divergence, on the rate and direction of hybridization, and on the climate transfer distance between the host and donor populations. In this study, we explored the use of Oriental beech (Fagus sylvatica subsp. orientalis), growing from Iran to the Balkans, for AGF into European beech populations (F. sylvatica subsp. sylvatica) that increasingly suffer from climate warming. Using samples from natural populations of Oriental and European beech and microsatellite loci, we identified 5 distinct genetic clusters in Oriental beech with a divergence (FST) of 0.15 to 0.25 from European beech. Using this knowledge, we traced the origin of 11 Oriental beech stands in Western Europe established during the 20th century. In two stands of Greater Caucasus origin, we found evidence for extensive hybridization, with 18% and 41% of the offspring having hybrid status. Climate data revealed higher seasonality with warmer and drier summers across the native Oriental beech sites in comparison to the planting sites in Western Europe. Accordingly, we found that bud burst of Oriental beech occurred four days earlier than in European beech. Overall, our results suggest that AGF of Oriental beech could increase the genetic diversity of European beech stands and may foster introgression of variants adapted to expected future climatic conditions. Our study showcases the evaluation of the benefits and risks of AGF and call for similar studies on other native tree species

Genetic Differentiation of the Western Capercaillie Highlights the Importance of South-Eastern Europe for Understanding the Species Phylogeography

The Western Capercaillie (Tetrao urogallus L.) is a grouse species of open boreal or high altitude forests of Eurasia. It is endangered throughout most mountain range habitat areas in Europe. Two major genetically identifiable lineages of Western Capercaillie have been described to date: the southern lineage at the species' southernmost range of distribution in Europe, and the boreal lineage. We address the question of genetic differentiation of capercaillie populations from the Rhodope and Rila Mountains in Bulgaria, across the Dinaric Mountains to the Slovenian Alps. The two lineages' contact zone and resulting conservation strategies in this so-far understudied area of distribution have not been previously determined. The results of analysis of mitochondrial DNA control region sequences of 319 samples from the studied populations show that Alpine populations were composed exclusively of boreal lineage; Dinaric populations of both, but predominantly (96%) of boreal lineage; and Rhodope-Rila populations predominantly (>90%) of southern lineage individuals. The Bulgarian mountains were identified as the core area of the southern lineage, and the Dinaric Mountains as the western contact zone between both lineages in the Balkans. Bulgarian populations appeared genetically distinct from Alpine and Dinaric populations and exhibited characteristics of a long-term stationary population, suggesting that they should be considered as a glacial relict and probably a distinct subspecies. Although all of the studied populations suffered a decline in the past, the significantly lower level of genetic diversity when compared with the neighbouring Alpine and Bulgarian populations suggests that the isolated Dinaric capercaillie is particularly vulnerable to continuing population decline. The results are discussed in the context of conservation of the species in the Balkans, its principal threats and legal protection status. Potential conservation strategies should consider the existence of the two lineages and their vulnerable Dinaric contact zone and support the specificities of the populations

Diameter growth and survival of local half-sib families of Scots pine (Pinus sylvestris L.) in Yundola, Bulgaria

The paper presents results of a study on a progeny test in Yundola established in 1966 and comprising 26 local plus trees of Scots Pine. Diameters at breast height (DBH) were measured in 1996 and 2012. There were substantial differences among the performance of half-sib families regarding the diameter growth. The best performing families 30 and 46 years after planting were the same, but there were statistically significant changes in the rank of the families as a whole. Mean survival rate was 91% at the age of 30 and 79% at the age of 46, which is relatively high. Individual heritability increased from 0.2 to 0.5-0.6 from the age 30 to 46 and additive genetic coefficient of variation was 5.2% at the age of 30 and 10.1% at the age of 46. Heritabilities and estimated response to selection were within the range of the results reported in other studies on Scots Pine. The main inferences of the study are that individual selection could be a promising tool for genetic improvement of Scots Pine in the region, and genetic parameters could change over time even after 30 years of age

Outcrossing rates in two seed orchards of Pinus nigra Arn. in Bulgaria

Seed orchards are an important source of seeds with improved genetic quality. Here we present results of a study on the genetic composition of seed orchards crop of Austrian black pine (Pinus nigra Arn.). Two seed orchards were included in the study – a clonal seed orchard established near Sliven, and a seedling seed orchard, established in the region of Simitli. The outcrossing rates were higher in the clonal seed orchard (Sliven), 0.873 and 0.806 multi-locus (tm) and single-locus (ts) estimates, respectively. The same values were 0.623 and 0.530, respectively, in the seedling seed orchard (Simitli). The inbreeding coefficient was positive in both cases, but was significantly different from zero in the clonal seed orchard only (0.101 vs. 0.032). Thus, the two studied seed orchards demonstrate different genetic efficiency. The results are discussed in relation to seed orchard management and efficiency.

Sideritis elica, a New Species of Lamiaceae from Bulgaria, Revealed by Morphology and Molecular Phylogeny

Sideritis elica, from the Rhodope Mountains, is described as a species new to science. Results of a detailed morphological analysis were combined with the data of molecular analyses using DNA barcoding as an efficient tool for the genetic, taxonomic identification of plants. The combination of morphological features distinguishes the new species well: Its first three uppermost leaf pairs are significantly shorter and wider, the branchiness of the stems is much more frequent, the whole plant is much more lanate, and it looks almost white, as opposed to the other closed species of section Empedoclia, which look grayish green. The molecular analysis, based on the rbcL and trnH-psbA regions, supports the morphological data about the divergence of Sideritis scardica and Sideritis elica. The studied populations of the two taxa were found to be genetically distant (up to 6.8% polymorphism for trnH-psbA) with distinct population-specific nucleotide patterns, while no polymorphism in the DNA barcodes was detected within the Sideritis elica population. The results confirm the existence of a new species called Sideritis elica, which occurs in the nature reserve Chervenata Stena, located in the northern part of the Central Rhodope Mountains. There were only 12 individuals found in the locality, which underlines the necessity of conservation measures

Halofitska vegetacija v jugovzhodni Bolgariji in ob obali Črnega morja

The paper presents results of a syntaxonomic analysis of the herbaceous phytocoenoses on the salt steppes, meadows and marshes in Southeastern Bulgaria, and along the Southern Black Sea Coast. The halophytic vegetation is distributed mostly in the Tundzha Lowland and the large salt lakes along the Black Sea Coast, where the saline soils occupy large territories. Most syntaxa identified in the paper are well-known and widespread in Central and Eastern Europe (Salicornietum prostratae, Suaedetum maritimae, Juncetum maritimae etc.), but there are also some specific vegetation types, which are presented by endemic associations as Petrosimonio brachiatae-Puccinellietum convolutae, Bupleuro tenuissimae-Camphorosmetum monspeliacae. The most widespread one is Diantho pallidiflori-Puccinellietum convolutae. It demonstrates a big variation in the appearance and the dominant structure of the described phytocoenoses. The comparision of the Bulgarian halophytic vegetation with other places in Europe shows its similarities with these vegetation types distributed in the Eastern Mediterranean and Central and Eastern Europe.V članku so predstavljeni rezultati sintaksonomske analize zeliščnih fitocenoz slanih step, travišč in močvirij v jugovzhodni Bolgariji in ob južni črnomorski obali. Halofitska vegetacija je najbolj pogosta v nižini Tundzha in v velikih slanih jezerih ob obali Črnega morja, kjer slana tla zavzemajo velika območja. Večina obravnavanih sintaksonov je dobro znanih in splošno razširjenih v srednji in vzhodni Evropi (Salicornietum prostratae, Suaedetum maritimae, Juncetum maritimae itd.), nekaj pa je posebnih vegetacijskih tipov, ki jih predstavljajo endemične asociacije Petrosimonio brachiatae-Puccinellietum convolutae, Bupleuro tenuissimae-Camphorosmetum monspeliacae. Najbolj razširjena je združba, ki jo uvrščamo v asociacijo Diantho pallidiflori-Puccinellietum convolutae, za katero je značilna velika variabilnost v videzu in dominantni strukturi. Primerjava halofitske vegetacije iz Bolgarije je pokazala podobnosti z vegetacijskimi tipi, razširjenimi v vzhodnem Sredozemlju ter srednji in vzhodni Evropi

On the Reproductive Potential in <i>Primula veris</i> L. (Primulaceae): Embryological Features, Pollen and Seed Viability, Genetic Diversity

Primula veris (Primulaceae) is a valuable medicinal plant. The main characteristics for assessing the reproductive potential (embryological features; mode of reproduction; pollen and seed viability) and the genetic diversity of populations of the species from Bulgaria were studied. The anthers are tetrasporangiate. Their wall development follows the Dicotyledonous-type and consists of: epidermis, a fibrous endothecium, an ephemeral middle layer and a secretory (glandular) tapetum. After meiosis in pollen mother cells and simultaneous microsporogenesis tetrahedral tetrads are formed predominantly in the anthers. Many ovules (approximately 20) develop in the unilocular ovary and are anatropous, tenuinucellate and bitegmic. The embryo sac (ES) develops after Polygonum (monosporic)-type from the chalazal cell of linear megaspore tetrad in the ovule. After double fertilization, a Caryophyllad-type embryo and initially nuclear endosperm form. In the studied populations, high pollen viability of more than 95% was established. Extremely low viability (about 4%) of the seeds obtained from natural populations was established. The results reveal P. veris to be a predominantly amphimictic (sexually reproducing) species, although rare vegetative propagation is also observed. As a result of this study, essential data were obtained about the reproductive structures and processes and for assessing the reproductive potential of P. veris