Aquatic plants are generally attributed to have larger ranges than their terrestrial counterparts, but this knowledge is often hindered by insufficient exploration of their diversity. To fill this gap, we investigated the taxonomically extremely challenging aquatic plant group Ranunculus sect. Batrachium in south-western Europe, which is an important glacial refugium, using flow cytometry, chromosome counting and DNA sequencing (ITS nuclear region and two non-coding plastid regions). In a dataset comprising 587 individuals from 117 localities, we detected 36 cytotypes across seven ploidy levels, which included a considerable proportion of previously unrecognized diversity consisting of three high ploidies (7x, 10x, 12x), seven cryptic species, two additional unclassifiable biotypes and nine hybrids. Two thirds of the taxa are polyploid, with many species presumed to be of allopolyploid origin. We discovered a remarkably close relationship between the local cytotype of R. peltatus s.l. and the morphologically distinct, widespread and ecologically specialized species R. fluitans; the latter might have evolved as a result of rapid adaptation to newly colonized river habitats in the early postglacial period. Undeniably, diversity within this group is still incompletely understood and is far more complex than current taxonomic concepts suggest.

• Klíčová slova
• Chromosome number, Cryptic variation, Genome size, Hybridization, Molecular identification, Polyploidy,
• MeSH
• biodiverzita * MeSH
• biologická evoluce * MeSH
• DNA rostlinná genetika   MeSH
• fylogeneze MeSH
• genetická variace MeSH
• molekulární evoluce MeSH
• ploidie MeSH
• polyploidie MeSH
• Ranunculus * genetika   klasifikace   MeSH
• sekvenční analýza DNA MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Evropa MeSH
• Názvy látek
• DNA rostlinná MeSH

Deciduous forests form the dominant natural vegetation of Europe today, but were restricted to small refugia during Pleistocene cold stages, implying an evolutionary past shaped by recurrent range contractions and expansions. Cold-stage forest refugia were probably widespread in southern and central Europe, with the northwestern Balkan Peninsula being of particular importance. However, the actual number and location of deciduous forest refugia, as well as the connections between them, remain disputed. Here, we address the evolutionary dynamics of the deciduous forest understorey species Euphorbia carniolica as a proxy for past forest dynamics. To do so, we obtained genomic and morphometric data from populations representing the species' entire range, investigated phylogenetic position and intraspecific genetic variation, tested explicit demographic scenarios and applied species distribution models. Our data support two disjoint groups linked to separate refugia on the northwestern and central Balkan Peninsula. We find that genetic differentiation between groups started in the early Pleistocene via vicariance, suggesting a larger distribution in the past. Both refugia acted as sources for founder events to the southeastern Alps and the Carpathians; the latter were likely colonised before the last cold stage. In line with traditional views on the pre-Pleistocene origin of many southeastern European deciduous forest species, the origin of E. carniolica was dated to the late Pliocene. The fact that E. carniolica evolved at a time when a period of continuous forestation was ending in much of Eurasia provides an interesting biogeographical perspective on the past links between Eurasian deciduous forests and their biota.

• Klíčová slova
• Alps, angiosperms, demographic modelling, forest understorey, glacial refugia, phylogeography,
• MeSH
• Euphorbia * genetika   MeSH
• fylogeneze MeSH
• fylogeografie MeSH
• genetická variace genetika   MeSH
• haplotypy MeSH
• lesy MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Balkánský poloostrov MeSH
• Evropa MeSH

Odd ploidy-level cytotypes in sexually reproducing species are considered a dead end due to absent or reduced fertility. If sterility is only partial, however, their contribution to the population gene pool can be augmented by longevity and clonal growth. To test this, we investigated the cytotype origin and spatial pattern, and pollen viability in three relict shrub species of the genus Daphne (Thymelaeaceae Juss.) in central Europe. Daphne cneorum subsp. cneorum is a widespread European species that has a broad ecological amplitude, whereas D. cneorum subsp. arbusculoides and D. arbuscula are narrow endemics of the western Pannonian Plain and the Western Carpathians, respectively. Our study confirmed that all three taxa are diploid. However, of more than a thousand analysed individuals of D. cneorum subsp. cneorum, five in four different populations were triploid. Our data indicate that these triploids most likely originate from recurrent autopolyploidization events caused by the fusion of reduced and unreduced gametes. High pollen viability was observed in all three taxa and in both diploid and triploid cytotypes, ranging from 65 to 100 %. Our study highlights the significant role of odd ploidy-level cytotypes in interploidy gene flow, calling for more research into their reproduction, genetic variability, and overall fitness. Interestingly, while the endemic D. arbuscula differs from D. cneorum based on genetic and genome size data, D. cneorum subsp. arbusculoides was indistinguishable from D. cneorum subsp. cneorum. However, our study reveals that the subspecies differ in the number of flowers per inflorescence. This is the first comprehensive cytogeographic study of this intriguing genus at a regional scale, and in spite of its karyological stability, it contributes to our understanding of genomic evolution in plant species with a wide ecological amplitude.

• Klíčová slova
• Carpathians, Daphne, ITS, Pannonian Basin, endemics, genome size stasis, pollen fertility, polyploidy, relicts, triploids,
• Publikační typ
• časopisecké články MeSH

For many arctic species, the spatial (re-)colonization patterns after the last Pleistocene glaciation have been described. However, the temporal aspects of their colonization are largely missing. Did one route prevail early, while another was more important later? The high Arctic archipelago Svalbard represents a good model system to address timeframe of postglacial plant colonization. Svalbard was almost fully glaciated during last glacial maximum and (re-)colonization of vascular plants began in early Holocene. Early Holocene climatic optimum (HCO) supported an expanded establishment of a partly thermophilic vegetation. Today, we find remnants of this vegetation in sheltered regions referred to as "Arctic biodiversity hotspots". The oldest record of postglacial plant colonization to Svalbard is found in Ringhorndalen-Flatøyrdalen. Even though thermophilic species could establish also later in Holocene, only HCO was favorable for vast colonization, and only hotspots offered stable conditions for thermophilic populations throughout Holocene. Thus, these relic populations may reflect colonization patterns of HCO. We investigate whether the colonization direction of thermophilic plants (Arnica angustifolia, Campanula uniflora, Pinguicula alpina, Tofieldia pusilla, and Vaccinium uliginosum ssp. microphyllum) in Ringhorndalen-Flatøyrdalen was uniform and different from later colonization events in other localities and non-thermophilic plants (Arenaria humifusa, Bistorta vivipara, Juncus biglumis, Oxyria digyna, and Silene acaulis). We analyzed plastid haplotypes of the 10 taxa from Ringhorndalen-Flatøyrdalen, from later-colonized localities in Svalbard, and from putative source regions outside Svalbard. Only rare and thermophilic taxa Campanula uniflora and Vaccinium uliginosum ssp. microphyllum provided results suggesting at least two colonization events from different source regions. Tofieldia pusilla and all the non-thermophilic plants showed no clear phylogeographically differentiation within Svalbard. Two of the thermophilic species showed no sequence variation. Based on the results, a uniform colonization direction to Svalbard in early Holocene is not probable; several source areas and dispersal directions were contemporarily involved.

• Klíčová slova
• Arctic, Holocene, Svalbard, diversity hotspot, haplotype, phylogeography,
• Publikační typ
• časopisecké články MeSH

Plastome phylogenomics is used in a broad range of studies where single markers do not bear enough information. Phylogenetic reconstruction in the genus Salix is difficult due to the lack of informative characters and reticulate evolution. Here, we use a genome skimming approach to reconstruct 41 complete plastomes of 32 Eurasian and North American Salix species representing different lineages, different ploidy levels, and separate geographic regions. We combined our plastomes with published data from Genbank to build a comprehensive phylogeny of 61 samples (50 species) using RAxML (Randomized Axelerated Maximum Likelihood). Additionally, haplotype networks for two observed subclades were calculated, and 72 genes were tested to be under selection. The results revealed a highly conserved structure of the observed plastomes. Within the genus, we observed a variation of 1.68%, most of which separated subg. Salix from the subgeneric Chamaetia/Vetrix clade. Our data generally confirm previous plastid phylogenies, however, within Chamaetia/Vetrix phylogenetic results represented neither taxonomical classifications nor geographical regions. Non-coding DNA regions were responsible for most of the observed variation within subclades and 5.6% of the analyzed genes showed signals of diversifying selection. A comparison of nuclear restriction site associated DNA (RAD) sequencing and plastome data on a subset of 10 species showed discrepancies in topology and resolution. We assume that a combination of (i) a very low mutation rate due to efficient mechanisms preventing mutagenesis, (ii) reticulate evolution, including ancient and ongoing hybridization, and (iii) homoplasy has shaped plastome evolution in willows.

• Klíčová slova
• Chamaetia/Vetrix clade, Eurasia, North America, genome skimming, phylogenomics, plastome evolution,
• Publikační typ
• časopisecké články MeSH

Polyploidy has played a crucial role in the evolution of many plant taxa, namely in higher latitudinal zones. Surprisingly, after several decades of an intensive research on polyploids, there are still common polyploid species whose evolutionary history is virtually unknown. Here, we addressed the origin of sweet vernal grass (Anthoxanthum odoratum) using flow cytometry, DNA sequencing, and in situ hybridization-based cytogenetic techniques. An allotetraploid and polytopic origin of the species has been verified. The chromosome study reveals an extensive variation between the European populations. In contrast, an autopolyploid origin of the rarer tetraploid vernal grass species, A. alpinum, has been corroborated. Diploid A. alpinum played an essential role in the polyploidization of both European tetraploids studied.

• Klíčová slova
• FISH, GBSSI, GISH, Poaceae, flow cytometry, genome size, polyploidy,
• MeSH
• biologická evoluce * MeSH
• chromozomy rostlin genetika   MeSH
• diploidie MeSH
• fylogeneze MeSH
• hybridizace genetická * MeSH
• hybridizace in situ MeSH
• lipnicovité cytologie   genetika   MeSH
• molekulární evoluce * MeSH
• polyploidie MeSH
• sekvence nukleotidů MeSH
• tetraploidie MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Glacial refugia of alpine and subnival biota have been intensively studied in the European Alps but the fate of forests and their understory species in that area remains largely unclear. In order to fill this gap, we aimed at disentangling the spatiotemporal diversification of disjunctly distributed black hellebore Helleborus niger (Ranunculaceae). We applied a set of phylogeographic analyses based on restriction-site associated DNA sequencing (RADseq) data and plastid DNA sequences to a range-wide sampling of populations. These analyses were supplemented with species distribution models generated for the present and the Last Glacial Maximum (LGM). We used exploratory analyses to delimit genomically coherent groups and then employed demographic modeling to reconstruct the history of these groups. We uncovered a deep split between two major genetic groups with western and eastern distribution within the Southern Limestone Alps, likely reflecting divergent evolution since the mid-Pleistocene in two glacial refugia situated along the unglaciated southern margin of the Alps. Long-term presence in the Southern Limestone Alps is also supported by high numbers of private alleles, elevated levels of nucleotide diversity and the species' modeled distribution at the LGM. The deep genetic divergence, however, is not reflected in leaf shape variation, suggesting that the morphological discrimination of genetically divergent entities within H. niger is questionable. At a shallower level, populations from the Northern Limestone Alps are differentiated from those in the Southern Limestone Alps in both RADseq and plastid DNA data sets, reflecting the North-South disjunction within the Eastern Alps. The underlying split was dated to ca. 0.1 mya, which is well before the LGM. In the same line, explicit tests of demographic models consistently rejected the hypothesis that the partial distribution area in the Northern Limestone Alps is the result of postglacial colonization. Taken together, our results strongly support that forest understory species such as H. niger have survived the LGM in refugia situated along the southern, but also along the northern or northeastern periphery of the Alps. Being a slow migrator, the species has likely survived repeated glacial-interglacial circles in distributional stasis while the composition of the tree canopy changed in the meanwhile.

• Klíčová slova
• Alps, Helleborus niger, RAD sequencing, demographic modeling, forest understorey, glacial refugia, species distribution modeling,
• Publikační typ
• časopisecké články MeSH

Diploid Alnus glutinosa s. str. and autotetraploid A. rohlenae form a narrow hybrid zone in a study area in southern Serbia, which results in triploid hybrid formation. The vast majority of previous studies have been focused on studies of maternal plants, but the offspring resulting from their crossing have not been much studied. Here, we use the variability of microsatellites and chloroplast DNA between these species and their putative hybrids to create an overall picture of the development of the hybrid zone and its predicted type. To elucidate the gene transfer within both species, the origins of individual ploidies and especially the role of triploid hybrids, a germination experiment was carried out linked with a flow cytometry study of the resulting seedlings. The tension zone model seems to offer the most adequate explanation of our observations, with selection against triploid hybrids and the spatial positioning of the hybrid zone. Despite selection against them, the triploid hybrids play an important role in the exchange of genes between the two species and therefore serve as a bridge for introgression. The presence of fertile triploids is essential for enriching the haplotype diversity between these species and for the development of new genetic lineages.

• Klíčová slova
• chloroplast DNA, germination, introgression, microsatellites, polyploidy, tension zone,
• MeSH
• genetická variace MeSH
• genom chloroplastový * MeSH
• genová introgrese * MeSH
• mikrosatelitní repetice * MeSH
• olše genetika   MeSH
• polyploidie MeSH
• selekce (genetika) MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

BACKGROUND AND AIMS: Archipelagos provide a valuable framework for investigating phenotypic evolution under different levels of geographical isolation. Here, we analysed two co-distributed, widespread plant lineages to examine if incipient island differentiation follows parallel patterns of variation in traits related to dispersal and colonization. METHODS: Twenty-one populations of two anemochorous Canarian endemics, Kleinia neriifolia and Periploca laevigata, were sampled to represent mainland congeners and two contrasting exposures across all the main islands. Leaf size, seed size and dispersability (estimated as diaspore terminal velocity) were characterized in each population. For comparison, dispersability was also measured in four additional anemochorous island species. Plastid DNA data were used to infer genetic structure and to reconstruct the phylogeographical pattern of our focal species. KEY RESULTS: In both lineages, mainland-island phenotypic divergence probably started within a similar time frame (i.e. Plio-Pleistocene). Island colonization implied parallel increases in leaf size and dispersability, but seed size showed opposite patterns of variation between Kleinia and Periploca species pairs. Furthermore, dispersability in our focal species was low when compared with other island plants, mostly due to large diaspore sizes. At the archipelago scale, island exposure explained a significant variation in leaf size across islands, but not in dispersability or seed size. Combined analyses of genetic and phenotypic data revealed two consistent patterns: (1) extensive within-island but very limited among-island dispersal, and (2) recurrent phenotypic differentiation between older (central) and younger (peripheral) island populations. CONCLUSIONS: Leaf size follows a more predictable pattern than dispersability, which is affected by stochastic shifts in seed size. Increased dispersability is associated with high population connectivity at the island scale, but does not preclude allopatric divergence among islands. In sum, phenotypic convergent patterns between species suggest a major role of selection, but deviating traits also indicate the potential contribution of random processes, particularly on peripheral islands.

• Klíčová slova
• Kleinia neriifolia, Periploca laevigata, Allopatric differentiation, Canary Islands, genetic drift, island phylogeography, leaf size, seed size, wind-dispersal traits,
• MeSH
• Asteraceae * MeSH
• biologická evoluce * MeSH
• distribuce rostlin * MeSH
• fylogeografie MeSH
• ostrovy MeSH
• Periploca * MeSH
• zvláštnosti životní historie * MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• ostrovy MeSH
• Španělsko MeSH

The basal position of the small American genus Microtea within the core Caryophyllales was suggested only recently in accordance with molecular phylogeny. However, the specific relationships within the genus were not traced. The results of our phylogenetic analysis based on the matK chloroplast gene suggest the monophyly of Microtea, and Ancistrocarpus and other related genera should be included in it. Microtea is divided into two major sister clades: clade A consisting of M.glochidiata, M.maypurensis and M.tenuifolia, and clade B comprising M.debilis, M.sulcicaulis, M.scabrida, M.celosioides, and M.papillosa. The nrDNA dataset (ITS), although containing only a limited number of accessions, shows the same species number in clade A, and the remaining species studied (M.debilis, M.scabrida and M.celosioides) form clade B. Subgeneric status is assigned to clades A and B corresponding with the names Microteasubgen.Ancistrocarpus subgen. nov. and Microteasubgen.Microtea, respectively. The diagnostic characters at the subgeneric level are as follows: length of pedicels, number of flowers at each node, number of stamens and styles. A multivariate analysis of 13 distinguishing morphological characters supports the results of phylogenetic analysis. All species have similar pericarp and seed ultrasculpture and anatomy, and they share the reticulate pericarp surface (independent of presence or absence of finger-shaped outgrowths on its surface) and rugose or slightly alveolate seed ultrasculpture. On the basis of morphological characters, we accept 10 Microtea species. A checklist includes a new diagnostic key, morphological descriptions and distribution patterns of each species. Galeniacelosioides is the oldest legitimate name available for the plants previously known as Microteapaniculata, for which the combination Microteacelosioides is validated here. The neotypes of Galeniacelosioides and Microteasprengelii were designated from the collections of Prinz Wied at BR. The name M.foliosa is discussed and finally synonymized with M.scabrida. The lectotypes of Ancistrocarpusmaypurensis (≡Microteamaypurensis), Microteadebilisvar.ovata (=M.debilis), M.glochidiata, M.maypurensisvar.angustifolia (=M.tenuifolia), M.glochidiataf.lanceolata (=M.maypurensis), M.longebracteata (=M.celosioides), M.paniculatavar.latifolia (=M.scabrida), M.portoricensis, M.scabrida, M.sulcicaulis, and Potamophilaparviflora (=M.maypurensis) are designated. Microteasulcicaulis is reported for the first time as native to Bolivia, and M.maypurensis is reported from Indonesia (Java), where it is found as an alien plant with an unclear invasion status.

• Klíčová slova
• Caryophyllales, Microtea, molecular phylogeny, reproductive characters, taxonomy,
• Publikační typ
• časopisecké články MeSH