In Vitro Propagation of Rhododendron tomentosum – an Endangered Essential Oil Bearing Plant from Peatland

Rhododendron tomentosum Harmaja (formerly Ledum palustre L.) is a medicinal peat bog plant native to northern Europe, Asia and North America. This plant has a distinctive aroma thanks to the presence of essential oil, to which it also owes its anti-inflammatory, analgesic, antimicrobial and insecticidal properties. However, in Europe R. tomentosum is classified as an endangered species, mainly due to degradation of peatlands. In the present work, the micropropagation protocol for R. tomentosum was established for the first time, providing both an ex situ conservation tool and a means of continuous production of in vivo and in vitro plant material for further studies. R. tomentosum microshoots were initiated from leaf explants and further multiplied using Schenk- Hildebrandt (SH) medium supplemented with 9.84 μM 2iP and 1.00 μM TDZ. The shoots were elongated on the SH medium supplemented with 24.6 μM 2iP and subsequently rooted using the perlite substrate saturated with half-strength Woody Plant medium supplemented with 1.0% sucrose and 4.92 μM IBA. The regenerated plants were hardened on the phytohormone-free SH medium and acclimatized using 3:1:1 deacidified peat:perlite:gravel substrate. The identity of the mother plant was confirmed at morphological and molecular levels and Random Amplified Polymorphic DNA (RAPD) method was implemented to assess the genetic fidelity of the regenerants. The essential oil content of the maternal plant, in vitro shoots and the regenerants was determined by steamdistillation, and the obtained volatile fractions were analyzed by GC/MS

Morphological, Karyological and Molecular Characteristics of Festuca arietina Klok. – a Neglected Psammophilous Species of the Festuca valesiaca agg. from Eastern Europe

Until recently, Festuca arietina was practically an unknown species in the flora of Eastern Europe. Such a situation can be treated as a consequence of insufficient studying of Festuca valesiaca group species in Eastern Europe and misinterpretation of the volume of some taxa. As a result of a complex study of F. arietina populations from the territory of Ukraine (including the material from locus classicus), Belarus and Lithuania, original anatomy, morphology and molecular data were obtained. These data confirmed the taxonomical status of F. arietina as a separate species. Eleven morphological and 12 anatomical characters, ITS1-5.8S-ITS2 cluster of nuclear ribosomal genes, as well as the models of secondary structure of ITS1 and ITS2 transcripts were studied in this approach. It was found for the first time that F. arietina is hexaploid (6x = 42), which is distinguished from all the other narrow-leaved fescues by specific leaf anatomy as well as in ITS1-5.8S-ITS2 sequences. Molecular data indicating possible hybridogenous origin of F. arietina, fall in line with the anatomical-morphological data and explain the tendency toward sclerenchyma strands fusion with formation of a continuous ring in F. arietina, as well as F. arietina ecological confinement to psammophyte biotopes

Adenophora liliifolia: Condition of its Populations in Central Europe

This study deals with populations of the European-South-Siberian geoelement Adenophora liliifolia (L.) A. DC. in the Czech Republic, Slovakia, Hungary, Romania, and Poland, where this species has its European periphery distribution. We studied the population size, genetic variability, site conditions, and vegetation units in which A. liliifolia grows. Recent and historical localities of A. liliifolia were ranked into six vegetation units of both forest and non-forest character. A phytosociological survey showed differences in the species composition among localities. Only a weak pattern of population structure was observed (only 22% of total genetic variation present at the interpopulation level, AMOVA analysis), with moderate values for gene diversity (Hj = 0.141) and polymorphism (P = 27.6%). Neighborjoining and Bayesian clusterings suggest a similar genetic background for most of the populations from Slovakia, the Czech Republic, and Poland, contrary to the populations from Hungary, Romania, as well as two populations from Central and South Slovakia. This might be explained by a relatively recent fragmentation of the A. liliifolia populations in Central Europe. Nevertheless, it seems that several populations in Romania, South Hungary, and Slovakia were isolated for a longer period of time and their genetic differentiation is more evident

Spatial Genetic Structure within Populations of Sorbus torminalis (L.) Crantz: Comparative Analysis of the Self-incompatibility Locus and Nuclear Microsatellites

Distribution of genetic diversity among and within plant populations may depend on the mating system and the mechanisms underlying the efficiency of pollen and seed dispersal. In self-incompatible species, negative frequency- dependent selection acting on the self-incompatibility locus is expected to decrease intensity of spatial genetic structure (SGS) and to reduce population differentiation. We investigated two populations (peripheral and more central) of wild service tree (Sorbus torminalis (L.) Crantz), a self-incompatible, scattered tree species to test the differences in population differentiation and spatial genetic structure assessed at the self-incompatibility locus and neutral nuclear microsatellites. Although, both populations exhibited similar levels of genetic diversity regardless of the marker type, significant differentiation was noticed. Differences between FST and RST suggested that in the case of microsatellites both mutations and drift were responsible for the observed differentiation level, but in the case of the S-RNase locus drift played a major role. Microsatellites indicated a similar and significant level of spatial genetic structure in both populations; however, at the S-RNase locus significant spatial genetic structure was found only in the fragmented population located at the north-eastern species range limits. Differences in SGS between the populations detected at the self-incompatibility locus were attributed mainly to the differences in fragmentation and population history