Hydrochorous Seed Transport in the Lower Traisen River before and after Riverbed Restoration.

Hydrological restoration was carried out in a Lower Traisen, a small river within the floodplain of the Danube. The main goal was the reestablishment of typical riparian plant communities by using the potential of natural dispersal processes. We studied the transport of plant diaspores in the river water before and after the reconstruction of the riverbed. Aquatic seed traps were placed upstream and downstream of the restoration site. We identified the transported species and tested the viability of propagules. Functional species traits were analyzed to predict the probability of successful hydrochorous dispersal and changes in the transport pool due to the restoration. One-third of the local species pool was detected as being diaspores in the river. We observed a significant increase of ruderal species and neophytes, while the competitors and stress-tolerant competitors declined. Hydrochory is an important dispersal pathway for numerous plant species in the study area, including those without specific adaptations to this vector. Hydrochorous transport appears to be a sink for large-seeded species, primarily adapted to endozoochory. Follow-up management should be recommended to control the invasive species and to improve the structural and biological diversity of the Traisen Valley by supporting target species, which are also represented in the transport pool.

Genotyping-by-sequencing reveals range expansion of Adonis vernalis (Ranunculaceae) from Southeastern Europe into the zonal Euro-Siberian steppe.

The Euro-Siberian steppe flora consists of warm- and cold-adapted species, which may have responded differently to Pleistocene glacials and interglacials. Genotyping-by-sequencing individuals from across the distribution range of the pheasant's eye (Adonis vernalis), we aimed to gain insight into steppe florogenesis based on the species' evolutionary history. Although the primary area of origin of the species group comprising A. vernalis, A. villosa and A. volgensis is in Asia, our results indicate that recent populations of A. vernalis are not of Asian origin but evolved in the southern part of Europe during the Pleistocene, with Spanish populations clearly genetically distinct from the Southeastern European populations. We inferred that A. vernalis migrated eastwards from the sub-Mediterranean forest-steppes of Southeastern Europe into the continental forest-steppe zone. Eastern European populations had the highest private allelic richness, indicating long-term large population sizes in this region. As a thermophilic species, A. vernalis seems unlikely to have survived in the cold deserts of the Last Glacial Maximum in Western Siberia, so this region was likely (re)colonized postglacially. Overall, our results reinforce the importance of identifying the area of origin and the corresponding ecological requirements of steppe plants in order to understand the composition of today's steppe flora.

Genetic Diversity of Stratiotes aloides L. (Hydrocharitaceae) Stands across Europe.

Intense land use and river regulations have led to the destruction of wetland habitats in the past 150 years. One plant that is affected by the reduction in appropriate habitats is the macrophyte Stratiotes aloides which has become rare in several areas. The preservation of genetic diversity within a species is a prerequisite for survival under changing environmental conditions. To evaluate the level of genetic diversity within and among populations of Stratiotes aloides, we investigated samples from waterbodies across Europe using AFLP. Low genetic diversity among samples from the same population was found, proving that stands consist of few clones which propagate clonally. Nevertheless, most populations showed differences compared to other populations indicating that there is genetic diversity within the species. The analyzed samples formed two groups in STRUCTURE analyses. The two groups can be further subdivided and mainly follow the major river systems. For conserving the genetic diversity of Stratiotes aloides, it would thus be preferable to focus on conserving individuals from many different populations rather than conserving selected populations with a higher number of individuals per population. For reintroductions, samples from the same river system could serve as founder individuals.

The phylogeographic history of Krascheninnikovia reflects the development of dry steppes and semi-deserts in Eurasia.

Constituting one of Earth's major biomes, steppes are characterised by naturally treeless extra-tropical vegetation. The formation of the Eurasian steppe belt, the largest steppe region in the world, began in Central Asia during the Neogene. In the glacial stages of the Pleistocene, steppe displaced forest vegetation, which in turn recolonised the area during the warmer interglacial periods, thus affecting the distribution of plants adapted to these habitats. Krascheninnikovia ceratoides (Chenopodiaceae) is a plant characteristic of dry steppe and semi-desert formations. Earlier studies showed that the ancestor of this autochthonous steppe element originated in Central Asia during the Miocene/Pliocene, i.e., in the same region and at the same time as the first appearance of steppe vegetation. However, as the extant lineages of Krascheninnikovia ceratoides diversified only 2.2 ± 0.9 Mya, it may represent a modern element of current dry steppe and semi-desert formations, rather than a component of the first steppe precursors of the Miocene. As such, it may have capitalised on the climatic conditions of the cold stages of the Quaternary to expand its range and colonise suitable habitats outside of its area of origin. To test this hypothesis, phylogeographic methods were applied to high-resolution genotyping-by-sequencing data. Our results indicate that Krascheninnikovia originated in western Central Asia and the Russian Altai, then spread to Europe in the West, and reached North America in the East. The populations of eastern Central Asia and North America belong to the same clade and are genetically clearly distinct from the Euro-Siberian populations. Among the populations west of the Altai Mountains, the European populations are genetically distinct from all others, which could be the result of the separation of populations east and west of the Urals caused by the Pleistocene transgressions of the Caspian Sea.

Genetic variation in an ephemeral mudflat species: The role of the soil seed bank and dispersal in river and secondary anthropogenic habitats.

Many ephemeral mudflat species, which rely on a soil seed bank to build up the next generation, are endangered in their natural habitat due to the widespread regulation of rivers. The aim of the present study was to elucidate the role of the soil seed bank and dispersal for the maintenance of genetic diversity in populations of near-natural river habitats and anthropogenic habitats created by traditional fish farming practices using Cyperus fuscus as a model. Using microsatellite markers, we found no difference in genetic diversity levels between soil seed bank and above-ground population and only moderate differentiation between the two fractions. One possible interpretation is the difference in short-term selection during germination under specific conditions (glasshouse versus field) resulting in an ecological filtering of genotypes out of the reservoir in the soil. River populations harbored significantly more genetic diversity than populations from the anthropogenic pond types. We suggest that altered levels and patterns of dispersal together with stronger selection pressures and historical bottlenecks in anthropogenic habitats are responsible for the observed reduction in genetic diversity. Dispersal is also supposed to largely prohibit genetic structure across Europe, although there is a gradient in private allelic richness from southern Europe (high values) to northern, especially north-western, Europe (low values), which probably relates to postglacial expansion out of southern and/or eastern refugia.

Isolation and characterization of microsatellite markers for Hypochaeris incana (Asteraceae) and close relatives.

PREMISE OF THE STUDY: We developed microsatellite markers to study clonal growth and interspecific hybridization in the Patagonian and subantarctic plant Hypochaeris incana (Asteraceae) and its closest relatives. METHODS AND RESULTS: We developed primers for microsatellite loci from 454 sequence reads of genomic DNA of H. incana. We tested them on individuals of H. acaulis, H. hookeri, H. incana, H. palustris, and H. tenuifolia. We selected 15 polymorphic microsatellite loci, which delivered clearly scorable fragments in most or all species. With mean values between 0.7 and 0.8, the expected heterozygosity in populations of H. incana is high. CONCLUSIONS: Due to high levels of polymorphism, the developed markers make it possible to distinguish between genets and ramets in H. incana. In some markers, null alleles complicate the scoring of genotypes in tetraploids. All of the developed markers are suitable to study interspecific hybridization among this group of closely related species.

Fitness and growth of the ephemeral mudflat species Cyperus fuscus in river and anthropogenic habitats in response to fluctuating water-levels.

Cyperus fuscus is a representative of threatened ephemeral wetland plant communities in summer-dry shoreline habitats. We compared variation and plasticity in traits related to fitness and growth of plants germinating from the soil seed bank and established plants from river and secondary anthropogenic habitats. Plants from sites at rivers, fishponds and fish storage ponds were cultivated and selfed to get homogenous seed material for a germination and an environmental manipulation experiment involving three different water regimes. Differences in traits and their plasticities were evaluated by means of linear mixed models. Cyperus fuscus followed a low-oxygen escape strategy when flooded. Seeds of plants derived from the soil seed bank germinated faster than seeds of plants derived from established plants suggesting that short-term selection of genotypes is mediated by the particular conditions on the site during germination. The experiment revealed significant differences between river and secondary habitats as well as between the soil seed bank and established plants. For example, plants from river habitats produced the highest number of culms with inflorescences. The difference was most evident under partial submergence. Plants from fish storage ponds rapidly reached the reproductive phase, but produced less culms with inflorescences. This seemingly allows them to cope with numerous and irregular disturbances and intensive substrate moisture changes. Our results suggest that populations have adapted to conditions at secondary habitats provided by fish farming during the last centuries.

Isolation of nuclear microsatellite markers for Cyperus fuscus (Cyperaceae).

PREMISE OF THE STUDY: Microsatellite markers were characterized in the extremely specialized ephemeral wetland plant species Cyperus fuscus (Cyperaceae). The markers will be used for studying population genetics in natural vs. anthropogenic habitats, on a European scale, and the role of the soil seed bank in the life cycle of this ephemeral species. METHODS AND RESULTS: Twenty-one microsatellite loci were established and scored in two populations, with mean number of alleles of 2.6 and 2.9 and mean expected heterozygosity of 0.405 and 0.470, respectively. Forty-four additional loci with the number of alleles ranging from one to four (mean = 2.1) were successfully amplified in seven individuals. CONCLUSIONS: The novel microsatellite markers will be useful for studying the genetic structure of populations of this ephemeral plant as well as their seed bank.

Comparative biogeography of the cytotypes of annual Microthlaspi perfoliatum (Brassicaceae) in Europe using isozymes and cpDNA data: refugia, diversity centers, and postglacial colonization.

In the last few years, the biogeography of many European plant species has been analyzed using molecular markers, and some consistent patterns of Pleistocenic differentiation and range fluctuations have been established. These studies mostly focused on perennial herbs or woody species, rarely considering annual taxa. This study focused on the annual Microthlaspi perfoliatum, which is distributed all over Europe and comprises three cytotypes. Morphologically, these cytotypes are hard to distinguish, although, based on molecular markers, they should be treated as two different species. Diploid and polyploid cytotypes had a different biogeographical history, with distinct glacial refugia. For the polyploids, a well-known distribution pattern of relict areas was confirmed, with Iberia, Italy, and the Balkans serving as primary Pleistocene refugia. Secondary refuge areas have been detected in southeastern France and neighboring Switzerland, with closer affinity to the Iberian refugium than to any other region based on allozyme and cpDNA haplotype data. For the diploids, two refugia have been characterized, one of which is congruent to the secondary refugium of the polyploids in France and Switzerland. The second refuge of diploid populations is located in unglaciated lowland areas of East Austria and Croatia. Isozyme and cpDNA haplotype data favor a postglacial colonization of diploid populations into Germany from Austrian lowland areas along the Danube River as well as from Switzerland. This scenario is also true for polyploids in Germany, Belgium, and Sweden.