The molecular population structure of Swertia perennis (Gentianaceae) in Central Europe.

Phylogeographic analysis of Swertia perennis, a typical European subalpine springtime species, revealed the existence of at least five major phylogenetic lineages. A large phylogeographic separation exists among these geographical regions, which confirms the existence of glacial refugia in the Pyrenees, but also in the Eastern and Central Alps. The results obtained from the analyzes indicate the existence of three major differences between the populations of the Alps and the Pyrenees, but also between the populations of the Alps and those of other geographical regions (Carpathians, southern Czech Republic, Sudetes and northern Poland). The studied populations from the Black Forest and from north-eastern and southern Poland are a relic of the former wider distribution of these (periglacial) genetic groups within Swertia perennis. Our results also confirm the existence of biogeographical links between the Carpathians and the Hercynian Range and the Alps. Certainly, there was an exchange of genes between populations located in the eastern Alps, the Carpathians and the Hercynian ranges (Czech Republic, Jeseníky, Sudetes, Ore Mountains). This confirms previous results of comparative studies on the genetic diversity of populations of other vascular plant species.

Genetic structure of Trifolium pratense populations in a cityscape.

Urban grasslands provide numerous ecosystem services, and their maintenance should be based on naturally regenerating plant populations. However, the urban environment is challenging for preserving viable populations, mostly because of their high fragmentation and small size, which can lead to genetic drift. We examined red clover (Trifolium pratense) in a medium-size city in Central Europe to test the cityscape effect on within- and among-population genetic diversity. We used eight inter-simple sequence repeat markers to examine the genetic structure of 16 populations, each represented by eight individuals. The isolation by resistance was analysed using a least cost patch approach, focusing on gene flow via pollinators. We found great variation among T. pratense populations, with no discernible geographic pattern in genetic diversity. We linked the diversity to the long history of the city and high stochasticity of land use changes that occurred with city development. In particular, we did not find that the Odra River (ca. 100 m wide) was a strong barrier to gene transfer. However, notable isolation was present due to resistance and distance, indicating that the populations are threatened by genetic drift. Therefore, gene movement between populations should be increased by appropriate management of urban green areas. We also found that small urban grassland (UG) patches with small populations can still hold rare alleles which significantly contribute to the overall genetic variation of T. pratense in the city.

The Role of the Hercynian Mountains of Central Europe in Shaping Plant Migration Patterns in the Pleistocene-A Review.

The climatic changes that took place in Europe during the Quaternary period influenced plant habitats as well as their species and vegetation composition. In this article, biogeographical studies on Hercynian mountain plants that include data for the Alps, Carpathians, and European lowlands are reviewed in order to discuss the phylogeographical structure and divergence of the Hercynian populations from those in other European mountain ranges, Scandinavia, and lowlands. The analyzed studies show specific phylogeographical relations between the Hercynian mountains, Alps, Scandinavia, Carpathians, and European lowlands. The results also indicate that the genetic patterns of plant populations in the Hercynian Mountains may differ significantly in terms of origin. The main migration routes of species to the Hercynian ranges began in the Alps or Carpathians. Some species, such as Rubus chamaemorus L., Salix lapponum L., and Salix herbacea L., are glacial relics that may have arrived and settled in the Hercynian Mountains during the Ice Age and that survived in isolated habitats. The Hercynian Mountains are composed of various smaller mountain ranges and are a crossroads of migration routes from different parts of Europe; thus, intensive hybridization has occurred between the plant populations therein, which is indicated by the presence of several divergent genetic lines.

Pimpinellasaxifragasubsp.rupestris (Apiaceae) - taxonomy and nomenclature of stenoendemic taxon from Karkonosze Mountains (Sudetes, Poland).

Pimpinellasaxifragasubsp.rupestris (Apiaceae) grows in a glacial cirque (Karkonosze Mountains, Sudetes, Poland) on a basalt substrate. Specimens of this species were first collected and described at the end of the 19th century, and their taxonomic distinctiveness and endemic status were determined by Weide in 1962. The typification of the name Pimpinellasaxifragasubsp.rupestris is discussed. The protologue of the name and the diagnostic phrase are evaluated based on herbarium specimen. The lectotype is designated. The paper also presents diagnostic morphological features of this and a closely related species Pimpinellasaxifragasubsp.saxifraga.

Genetic diversity of Salixlapponum populations in Central Europe.

Salixlapponum is a cold-tolerant relict species in Europe that occurs in several sites, probably reflecting previous migration routes of S.lapponum during the Pleistocene. However, only a few data are available on the genetic structures of populations of S.lapponum. In this study, we use PCR-ISSR markers to investigate genetic variation in 19 European populations of S.lapponum L. AMOVA analysis shows that most of the variation (55.8%) occurs within populations; variability among groups accounts for 19.7%. An AMOVA analysis based on four groups determined by STRUCTURE analysis shows similar results: variability of 54.1% within the population and variability of 18.9% between the four population groups, based on geographic regions. Within individual geographic groups, which are characterised by the studied populations, the lowest variability (as well as the highest homogeneity) was found in populations located in Belarus. The obtained results are consistent with our expectations that the European Lowland could be a significant geographic barrier for gene flow over large geographic distances for S.lapponum. Both the Scandinavian and Belarusian populations, as well as those coming from NE Poland, are characterised by significant genetic distinctiveness. However, some populations from NE Poland and the Sudetes show similarities with populations from other geographic regions, indicating existing genetic relationships between them. Moreover, the results suggest a fairly clear division of the population into 4 emerging geographic regions, although separated by a geographical barrier: the Polish lowland, which forms part of the larger geographic unit known as the European Lowland.

Airborne fungi in Longyearbyen area (Svalbard, Norway) - case study.

Studies on the presence of atmospheric fungi in both Arctic and Antarctic polar areas are rare, and many of them were carried out briefly. Currently, when climate change is a fact, polar areas may be subject to various changes and fluctuations, negatively affecting sensitive polar ecosystems. The paper presents the results of tests on presence of fungi in the air over 30 years after the last investigations at the Svalbard Archipelago. A total of fifteen taxa of fungi were isolated in area of Longyearbyen, the majority of which were saprotrophic fungi of the genus Cladosporium that are associated with dead organic matter. Therefore, the presence of this taxon may be a good bioindicator of changes occurring in the Arctic environment, indirectly indicating the melting of glaciers and exposing increasingly larger areas inhabited by microorganisms, including fungi, which increase in number in the air. Additionally, the number of tourists visiting Longyearbyen is increasing, which may significantly affect the number and type of fungi in the air.

Infraspecific variation of some brown Parmeliae (in Poland) - a comparison of ITS rDNA and non-molecular characters.

Infraspecific variation of the ITS rDNA region of some brown Parmeliae occurring in Poland is studied and compared with non-molecular characters. Haplotype networks are used to illustrate the variability within the species. Both newly-produced sequences from Central Europe and from all over the world, downloaded from the GenBank, are used. The number of haplotypes found for each taxon ranged from five in Melaneliastygia to 12 in Melaneliahepatizon and Montaneliadisjuncta; however, their numbers correlate with the number of specimens tested. New haplotypes for Melaneliaagnata, M.hepatizon and Cetrariacommixta are found. Based on our 169-sample dataset, we could not infer any geographical correlation, either locally or world-wide. Many of the analysed haplotypes were widely distributed and the same haplotype was often shared between temperate and polar populations. A comparison of molecular, morphological, anatomical and chemical characters also shows no correlation.

Taxonomic recognition of some species-level lineages circumscribed in nominal Rhizoplaca subdiscrepans s. lat. (Lecanoraceae, Ascomycota).

BACKGROUND: Rhizoplaca subdiscrepans (Nyl.) R. Sant., a saxicolous, placodioid lichen, is considered to have a worldwide distribution in warm-temperate to boreal-arctic areas in Asia, Europe and North America. However, recent studies have revealed that this species includes five unrecognized species-level lineages-'subd A, B, C, D and E'. During research focused on the diversity of saxicolous lichens in mountainous areas of southern Poland, some interesting representatives of the genus Rhizoplaca were found. The main aim of our study was to determine the taxonomic status of the collected specimens by means of molecular tools and a comparative analysis of similar herbarium materials. METHODS: Detailed morphological, anatomical and chemical examinations of reference material from Asia, Europe and North and South America focused primarily on a selected group of lecanoroid taxa with a placodioid thallus. In addition, 21 new generated sequences representing Lecanora pseudomellea, Protoparmeliopsis muralis, Rhizoplaca opiniconensis, R. subdiscrepans s. lat. and R. phaedrophthalma were selected for molecular study using the internal transcribed spacer region (ITS rDNA), together with 95 available GenBank sequences mainly from the genus Rhizoplaca. RESULTS: Polish specimens that clustered with members of a potential species-level lineage 'subd E' of Rhizoplaca subdiscrepans complex were recovered. Comprehensive analyses of the lichen group led us to the conclusion that lineage 'subd E' represents R. subdiscrepans s. str. and that the taxon appears to have a limited geographical distribution and specific habitat preferences. Furthermore, some of the recently defined species candidates within R. subdiscrepans s. lat.-'subd D' and 'subd A'-should be assigned to two previously known species of Rhizoplaca, namely R. opiniconensis (Brodo) Leavitt, Zhao Xin & Lumbsch and R. phaedrophthalma (Poelt) Leavitt, Zhao Xin & Lumbsch, respectively. These two species are characterized by phenotypic features observed as well in analyzed specimens representing lineages 'subd D' and 'subd A'. Moreover, the representatives of these lineages demonstrate some differences in occupied habitat and geographical range that also correspond with the indicated species. Additionally, it was found that Lecanora pseudomellea B.D. Ryan is a strongly supported monophyletic lineage within Rhizoplaca, and therefore an appropriate new combination for the species is proposed.

Taxonomic studies on the Chara section Hartmania in Poland based on morphological and molecular data.

Charophytes are aquatic green macroalgae, which inhabit fresh and brackish water ecosystems. In this study, four species belonging to the genus Chara were examined to determine their taxonomic status. Morphological characteristics of the plant bodies as well as plastid psaB barcoding genes were applied to test the relations among Chara species. Plants were initially classified using morphological features into four species: C. baltica, C. hispida, C. polyacantha and C. rudis, and twelve quantitative characters were used in a principal component analysis and discriminant analysis to determine groupings among the species and to determine the morphological features that best separated the groups. In the component analysis and discriminant analysis, results showed that only C. polyacantha and partly C. baltica formed separate groups. The other species C. hispida and C. rudis were only partially distinguishable. All species from one molecular group, and no differentiation in the psaB variability between them has been found.

Isolation of nucleic acids using silicon dioxide powder as a tool for environmental monitoring.

Isolation and purification of nucleic acids are basic laboratory procedures used in molecular analysis supporting determination of organisms in environmental monitoring. However, many different methods of isolation are commonly used, often being designed for a particular type of DNA extraction. While researchers commonly decide on commercial isolation kits for their ease of use and efficiency, they require large amounts of studied tissue, and the cost of purchasing such kits over a long run can be high. To provide an alternative to using commercial kits, we have developed a simple, rapid, cost-effective, and reliable protocol for DNA isolation from cultured fungi on slants and from dried fungal samples using silica particles (silicon dioxide powder) in chaotropic conditions. With the presented method, it is possible to isolate good-quality DNA from fungi in less than 1.5 h, using easily accessible chemicals. Compared with other methods employing CTAB or commercial kits, it allows fast, easy, and cheap DNA purification from two main sources of fungi routinely used for research. In addition to the method protocol, we also provide advice for further optimization of the isolation process to account for specific conditions, making the procedure more useful.

Neotypification of Protoparmeliopsis garovaglii and molecular evidence of its occurrence in Poland and South America.

Protoparmeliopsis garovaglii is a widely distributed placodioid lichen, which develops a distinctly rosette thallus, composed of elongated and strongly inflated to sinuous-plicate lobes. The taxon is characterised by high morphological plasticity and varied composition of secondary metabolites. However, the epithet was never typified. As such, the identity of P. garovaglii, in its strict sense, was unknown for a long time. Our phylogenetic ITS rDNA analyses, including newly generated sequences, show that European (Austria, Poland), North American (USA) and South American (Bolivia, Peru) specimens of P. garovaglii are placed in a strongly supported monophyletic clade, sister to P. muralis. We provide the first molecular evidence of the occurrence of P. garovaglii in South America (Bolivia and Peru) and the second record in Central Europe (Poland) was also provided. Furthermore, we neotypify P. garovaglii and it is reported here for the first time from Poland.

Phylogeography of Swertia perennis in Europe based on cpDNA markers.

BACKGROUND: Swertia perennis (Gentianaceae) is a perennial diploid and clonal plant species that is discontinuously distributed in peat bogs in the mountains of Europe, Asia and North America as well as in the lowlands of Europe. The current geographical dispersion of S. perennis is probably the result of quaternary climatic changes that have played an important role in determining the distribution of Swertia and other plant and animal species. METHODS: In this study we used molecular techniques and combined data from chloroplast DNA markers (trnLF region and trnH-psbA spacer) to elucidate the phylogeography of S. perennis in Europe. Plants were collected from 28 populations in different locations in the lowlands and mountainous areas of Europe (e.g., the Carpathians, Sudetes, Bohemian Forest and Alps). cDNA was analysed to detect the genetic relationship between specimens from different locations. RESULTS: A total of 20 haplotypes were identified across the dataset. They were characterised by a high level of genetic variability but showed a lack of phylogeographical structure. This pattern may be the result of repeated recolonization and expansion from several areas. Such genetic differentiation may also be attributed to the relatively long-term isolation of S. perennis in Pleistocene refugia in Europe, which resulted in independent separation of different cpDNA phylogenetic lineages and variation in the nucleotide composition of cpDNA. DISCUSSION: The lack of strong phylogeographical structure makes it impossible to indicate the centre of haplotype diversity; however, refugia located in the Carpathians, Sudetes or Alps are the most probable sites where S. perennis existed in Europe. This lack of structure may also indicate a high level of gene flow in times when the landscape and fen systems were not fragmented in numerous geographically-isolated populations. This makes it difficult to speculate about the relationships between Asiatic and European plant populations and the origin and distribution of this species in Europe. Today, it seems to be restricted due to the occurrence of plants which clearly reflects the genetic variability from the ancient period.

Oospore Wall Ornamentation in the Genus Tolypella (Charales, Charophyceae).

The ultrastructural features of oospore wall ornamentation in the genus Tolypella were examined using scanning electron microscopy (SEM). The taxonomic relationships among several species were discussed on the basis of oospore ultrastructure and measurements. In the case of T. glomerata and T. nidifica, our results support the status of separate species. Close relationships and transitional forms may exist between T. nidifica and T. normaniana, and not only in oospore wall ornamentation. Oospores of T. hispanica exhibited the same distinct type of reticulate oospore wall as previously reported, but our results do not support the recognition of T. hispanica as a separate species. Ultrastructure of the oospore walls of T. prolifera and T. intricata was almost identical, suggesting that these species are closely related. We therefore reject previous suggestions that morphological characteristics of oospores as observed in SEM are sufficient for identification of individual species. Although significant differences were found among oospores in individual species of Tolypella, large variation among populations, and among individuals belonging to the same population, caused substantial overlap among species.