Chemical profile and antioxidant activity of the species Veronica saturejoides Vis. ssp. saturejoides (Plantaginaceae)—which is endemic to Croatia, Bosnia and Herzegovina and Montenegro —were investigated. Volatile compounds produced by glandular trichomes (composed of one stalk cell and two elliptically formed head cells according to scanning electron microscope investigation) were isolated from the plants collected in two locations. Additionally, as a part of specialized metabolites, total polyphenols, total tannins, total flavonoids and total phenolic acids were determined spectrophotometrically. In the lipophilic volatile fractions-essential oils, the most abundant compounds identified were hexahydrofarnesyl acetone, caryophyllene oxide and hexadecanoic acid. In total, the class of oxygenated sesquiterpenes and the group of fatty aldehydes, acids and alcoholic compounds dominated in the essential oils. In the hydrophilic volatile fractions-hydrosols, the most abundant compounds identified were trans-p-mentha-1(7),8-dien-2-ol, allo-aromadendrene and (E)-caryophyllene. A group of oxygenated monoterpenes and the sesquiterpene hydrocarbons dominated in the hydrosols. Antioxidant activity of essential oils and hydrosols was tested with two methods: 2,2′-diphenyl-1-picrylhydrazyl (DPPH) and oxygen radical absorbance capacity (ORAC). Essential oils showed higher antioxidant activity than hydrosols and showed similar antioxidant activity to Rosmarinus officinalis essential oil. Obtained results demonstrate that this genus is a potential source of volatiles with antioxidant activity.
Pinus heldreichii (Bosnian pine) is a Balkan-Apennine endemic and relict pine species that inhabits high mountains in the Mediterranean and sub-Mediterranean regions. Nineteen populations of P. heldreichii from the Western Balkans encompassing 187 individual trees were examined to evaluate morphological variation, a rarely studied aspect of the species. Univariate and multivariate statistics were applied in order to assess the variation of morphological traits of cones and seeds, evaluate the relationships among the sampled populations and verify geographic differentiation in the Dinaric Alps versus Scardo-Pindic mountains. The observations of P. heldreichii covering the populations from the western margins and the centre of the species distribution range indicated a morphological variation among populations and their geographic structure. In general, the southern populations (Scardo-Pindic group) had lower values for the most of morphological traits than the northern ones (Dinaric group). The observed geographic differences between these populations exhibit a north-western to south-eastern gradient, with a few inconsistencies. The southernmost sampled population, Tomorr in Albania, showed remarkable morphological divergence from the other studied populations and appeared to be a distinct morphological group. The pattern of morphological variation in Bosnian pine most likely resulted from multiple effects of long-term isolation and fragmentation in high mountain systems, adaptation to extreme environments and human disturbances.
Leaf and shoot characteristics of the following four European barberry taxa from the Balkan Peninsula and Sicily were investigated in the present study: Berberis croatica, B. vulgaris, B. aetnensis and B. cretica. Analyses were based on 10 populations of B. croatica, five of B. vulgaris and two populations of both B. aetnensis and B. cretica. Populations were randomly selected within the natural distribution area of these species. Eight leaf traits, three shoot traits and the blade length/width ratio were analysed. Multivariate analysis (principal component analysis, canonical discriminant analysis and cluster analysis) distinguished B. cretica and B. aetnensis populations and, to a lesser extent, the populations of B. croatica and B. vulgaris. ANOVA showed that the analysed populations of both B. aetnensis and B. cretica were homogeneous within the species. All populations of both B. croatica and B. vulgaris showed different degrees of intraspecies variability. Lack of complete separation, the observed grouping of populations and high intraspecies variability in B. vulgaris and B. croatica may reflect the fact that the sampled B vulgaris and B.croatica populations were located at environmentally variable sites (unlike B. aetnensis and B. cretica), resulting in high phenotypic plasticity in those populations. Even though the observed patterns of morphological variation support the idea of four barberry taxa on the Balkan Peninsula and in Sicily, because of suspected adaptive phenotypic plasticity of the analysed Berberis taxa, the true taxonomic status of these taxa needs to be additionally confirmed by molecular methods.
This paper presents new records and noteworthy data on the following taxa in SE Europe and adjacent regions: the diatom alga Eunotia boreoalpina; the saprotrophic fungus Clitocybe truncicola; the liverwort Haplomitrium hookeri; the moss Leptodon smithii: the monocots Epipactis purpurata, Stipa tirsa, Typha laxmannii and T. shuttleworthii; and the dicots Krascheninnikovia ceratoides, Polygonum albanicum and Sorbus latifolia.
Of the four species of the genus Satureja (Lamiaceae) that are recognized in Bosnia and Herzegovina, S. subspicata has the the widest distribution. It is taxonomically challenging species of geographically limited distribution and little data on its genetic diversity throughout its range is available. We sampled six geographically distinct populations from Bosnia and Herzegovina and applied nrDNA (ITS1, ITS2), chloroplast markers (matK and trnL) and AFLP to examine genetic diversity of S. subspicata in the center of its distribution range and to explore the possibility of establishing the species DNA barcode. AFLP analysis showed large genetic differentiation among populations as well as moderate correlation between genetic distance among populations and geographic distance among locations. MatK has not proven useful in distinguishing S. subspicata from sympatric species. However, nrDNA sequences provided necessary resolution power, with ITS2 being more informative. Estimates of evolutionary divergence between nrDNA sequences obtained in our research and homologous sequences of sympatric Satureja deposited in the GenBank reveal closer relationship between geographically proximate populations of different species and slight divergence within S. subspicata sequences pool. This outcome highlights the importance of considering overall genetic diversity across the distribution range of a species when assigning DNA barcode.
Abstract Epigenetic variation in natural populations with contrasting habitats might be an important element, in addition to the genetic variation, in plant adaptation to environmental stress. Here, we assessed genetic, epigenetic, and cytogenetic structure of the three Lilium bosniacum populations growing on distinct habitats. One population was growing under habitual ecological conditions for this species and the other two were growing under stress associated with high altitude and serpentine soil. Amplified fragment length polymorphism and methylation-sensitive amplification polymorphism analyses revealed that the three populations did not differentiate genetically, but were clearly separated in three distinct clusters according to DNA methylation profiles. Principal coordinate analysis showed that overall epigenetic variation was closely related to habitat conditions. A new methylation-sensitive amplification polymorphism scoring approach allowed identification of mainly unmethylated (φST = 0.190) and fully CpG methylated (φST = 0.118) subepiloci playing a role in overall population differentiation, in comparison with hemimethylated sites (φST = 0.073). In addition, unusual rDNA repatterning and the presence of B chromosomes bearing 5S rDNA loci were recorded in the population growing on serpentine soil, suggesting dynamic chromosome rearrangements probably linked to global genome demethylation, which might have reactivated some mobile elements. We discuss our results considering our earlier data on morphology and leaf anatomy of several L. bosniacum populations, and suggest a possible role of epigenetics as a key element in population differentiation associated with environmental stress in these particular lily populations.
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