A novel strategy to study apomixis, automixis, and autogamy in plants.

KEY MESSAGE: The combination of a flow cytometric seed screen and genotyping of each single seed offers a cost-effective approach to detecting complex reproductive pathways in flowering plants. Reproduction may be seen as one of the driving forces of evolution. Flow cytometric seed screen and genotyping of parents and progeny are commonly employed techniques to discern various modes of reproduction in flowering plants. Nevertheless, both methods possess limitations constraining their individual capacity to investigate reproductive modes thoroughly. We implemented both methods in a novel manner to analyse reproduction pathways using a carefully selected material of parental individuals and their seed progeny. The significant advantage of this approach lies in its ability to apply both methods to a single seed. The introduced methodology provides valuable insights into discerning the levels of apomixis, sexuality, and selfing in complex Rubus taxa. The results may be explained by the occurrence of automixis in Rubus, which warrants further investigation. The approach showcased its effectiveness in a different apomictic system, specifically in Taraxacum. Our study presents a comprehensive methodological approach for determining the mode of reproduction where flow cytometry loses its potential. It provides a reliable and cost-effective method with significant potential in biosystematics, population genetics, and crop breeding.

Adapted dandelions trade dispersal for germination upon root herbivore attack.

A plant's offspring may escape unfavourable local conditions through seed dispersal. Whether plants use this strategy to escape insect herbivores is not well understood. Here, we explore how different dandelion (Taraxacum officinale agg.) populations, including diploid outcrossers and triploid apomicts, modify seed dispersal in response to root herbivore attack by their main root-feeding natural enemy, the larvae of the common cockchafer Melolontha melolontha. In a manipulative field experiment, root herbivore attack increased seed dispersal potential through a reduction in seed weight in populations that evolved under high root herbivore pressure, but not in populations that evolved under low pressure. This increase in dispersal potential was independent of plant cytotype, but associated with a reduction in germination rate, suggesting that adapted dandelions trade dispersal for establishment upon attack by root herbivores. Analysis of vegetative growth parameters suggested that the increased dispersal capacity was not the result of stress flowering. In summary, these results suggest that root herbivory selects for an induced increase in dispersal ability in response to herbivore attack. Induced seed dispersal may be a strategy that allows adapted plants to escape from herbivores.

Antidepressive Effects of Taraxacum Officinale in a Mouse Model of Depression Are Due to Inhibition of Corticosterone Levels and Modulation of Mitogen-Activated Protein Kinase Phosphatase-1 (Mkp-1) and Brain-Derived Neurotrophic Factor (Bdnf) Expression.

BACKGROUND Depression is a common disorder linked with high levels of chronicity, psycho-social and physical problems, and suicide. Here, we assessed the antidepressant effects of the hydromethanolic extract of Taraxacum officinale and investigated the underlying mechanism. MATERIAL AND METHODS Antidepressant effects were examined by use of the tail suspension test (TST). Concentrations of corticosterone, dopamine, noradrenaline, and adrenaline were examined by biochemical assays. The mRNA expression was assessed by quantitative RT-PCR. Phytochemical analysis was performed by LC/MS. RESULTS The results showed that the extract at the dosage of 50 and 100 mg/kg significantly (p<0.01) alleviated the TST-induced immobility in the mice, and the effects were comparable to the antidepressant drug Bupropion, which was used as the positive control. Investigation of the underlying mechanism revealed that the T. officinale extract exerts it effects by significantly (p<0.05) decreasing the levels of corticosterone and increasing the concentrations of dopamine, noradrenaline, and adrenaline. Further, the extract also increased the expression of brain-derived neurotrophic factor (Bdnf), which was associated with significant (p<0.05) decrease in the expression of mitogen-activated protein kinase phosphatase-1 (Mkp-1), indicative of the antidepressant potential of T. officinale. Finally, the active constituents of the extract, which include isoetin, hesperidin, naringenin, Kaempferol, sinapinic, and gallic acid, were also identified, which could potentially be responsible for its antidepressant effects. CONCLUSIONS In conclusion, T. officinale exerts significant antidepressant effects in a mouse model of depression by inhibition of corticosterone levels and modulation of Mkp-1 and Bdnf expression.

A separated vortex ring underlies the flight of the dandelion.

Wind-dispersed plants have evolved ingenious ways to lift their seeds1,2. The common dandelion uses a bundle of drag-enhancing bristles (the pappus) that helps to keep their seeds aloft. This passive flight mechanism is highly effective, enabling seed dispersal over formidable distances3,4; however, the physics underpinning pappus-mediated flight remains unresolved. Here we visualized the flow around dandelion seeds, uncovering an extraordinary type of vortex. This vortex is a ring of recirculating fluid, which is detached owing to the flow passing through the pappus. We hypothesized that the circular disk-like geometry and the porosity of the pappus are the key design features that enable the formation of the separated vortex ring. The porosity gradient was surveyed using microfabricated disks, and a disk with a similar porosity was found to be able to recapitulate the flow behaviour of the pappus. The porosity of the dandelion pappus appears to be tuned precisely to stabilize the vortex, while maximizing aerodynamic loading and minimizing material requirements. The discovery of the separated vortex ring provides evidence of the existence of a new class of fluid behaviour around fluid-immersed bodies that may underlie locomotion, weight reduction and particle retention in biological and manmade structures.

Growing cell walls show a gradient of elastic strain across their layers.

The relatively thick primary walls of epidermal and collenchyma cells often form waviness on the surface that faces the protoplast when they are released from the tensile in-plane stress that operates in situ. This waviness is a manifestation of buckling that results from the heterogeneity of the elastic strain across the wall. In this study, this heterogeneity was confirmed by the spontaneous bending of isolated wall fragments that were initially flat. We combined the empirical data on the formation of waviness in growing cell walls with computations of the buckled wall shapes. We chose cylindrical-shaped organs with a high degree of longitudinal tissue stress because in such organs the surface deformation that accompanies the removal of the stress is strongly anisotropic and leads to the formation of waviness in which wrinkles on the inner wall surface are always transverse to the organ axis. The computations showed that the strain heterogeneity results from individual or overlaid gradients of pre-stress and stiffness across the wall. The computed wall shapes depend on the assumed wall thickness and mechanical gradients. Thus, a quantitative analysis of the wall waviness that forms after stress removal can be used to assess the mechanical heterogeneity of the cell wall.

Immunodetection of some pectic, arabinogalactan proteins and hemicellulose epitopes in the micropylar transmitting tissue of apomictic dandelions (Taraxacum, Asteraceae, Lactuceae).

In apomictic Taraxacum species, the development of both the embryo and the endosperm does not require double fertilisation. However, a structural reduction of ovular transmitting tissue was not observed in apomictic dandelions. The aim of this study was to analyse the chemical composition of the cell walls to describe the presence of arabinogalactan proteins (AGPs), hemicellulose and some pectic epitopes in the micropylar transmitting tissue of apomictic Taraxacum. The results point to (1) the similar distribution of AGPs in different developmental stages, (2) the absence of highly methyl-esterified homogalacturonan (HG) in transmitting tissue of ovule containing a mature embryo sac and the appearance of this pectin domain in the young seed containing the embryo and endosperm, (3) the similar pattern of low methyl-esterified pectin occurrence in both an ovule and a young seed with an embryo and endosperm in apomictic Taraxacum and (4) the presence of hemicelluloses recognised by LM25 and LM21 antibodies in the reproductive structure of Taraxacum.

Variation in the strength of reproductive interference from an alien congener to a native species in Taraxacum.

Reproductive interference (RI) may be a contributing factor to the displacement of native species by an alien congener, and RI strength has been shown theoretically to affect distributional relationships between species. Thus, variations in RI strength from alien to native species result in different consequences of invasions and efforts to conserve native species, but the variations have seldom been examined empirically. We therefore investigated RI strength variations from the alien species Taraxacum officinale and its hybrids to eight populations of native dandelions, four T. japonicum populations and two populations each of two subspecies of T. platycarpum. We examined the association between alien relative abundance and native seed set in field surveys, and we also performed hand-pollination experiments to investigate directly the sensitivity of native flowers to alien pollen. We found that the effect of alien relative abundance on native seed set of even the same native species could differ greatly in different regions, and that the sensitivity of native flowers to alien pollen was also dependent on region. Our results, together with those of previous studies, show that RI from the alien to the native species is strong in regions where the alien species outnumbers the native species and marginal where it does not; this result suggests that alien RI can critically affect distributional relationships between native and alien species. Our study highlights the importance of performing additional empirical investigations of RI strength variation and of giving due attention to alien RI in efforts to conserve regional native biodiversity.

Change of plant phenophases explained by survival modeling.

It is known from many studies that plant species show a delay in the timing of flowering events with an increase in latitude and altitude, and an advance with an increase in temperature. Furthermore, in many locations and for many species, flowering dates have advanced over the long-term. New insights using survival modeling are given based on data collected (1970-2010) along a 3000-km long transect from northern to eastern central Europe. We could clearly observe that in the case of dandelion (Taraxacum officinale) the risk of flowering time, in other words the probability that flowering occurs, is higher for an earlier day of year in later decades. Our approach assume that temperature has greater influence than precipitation on the timing of flowering. Evaluation of the predictive power of tested models suggests that Cox models may be used in plant phenological research. The applied Cox model provides improved predictions of flowering dates compared to traditional regression methods and gives further insights into drivers of phenological events.

Temporal variability of the quality of Taraxacum officinale seed progeny from the East-Ural radioactive trace: is there an interaction between low level radiation and weather conditions?

HYPOTHESIS: The multiple stressors, in different combinations, may impact differently upon seed quality, and low-level doses of radiation may enhance synergistic or antagonistic effects. RESULTS: During 1991-2014 we investigated the quality of the dandelion (Taraxacum officinale s.l.) seed progeny growing under low-level radiation exposure at the East-Ural Radioactive Trace (EURT) area (result of the Kyshtym accident, Russia), and in plants from areas exposed to background radiation. The viability of the dandelion seed progeny was assessed according to chronic radiation exposure, accounting for the variability of weather conditions among years. Environmental factors (temperature, precipitation, and their ratio in different months) can modify the radiobiological effects. We found a wide range of possible responses to multiple stressors: inhibition, stimulation, and indifferent effects in different seasons. CONCLUSION: The intraspecific variability of the quality of dandelion seed progeny was greatly increased under conditions of low doses of chronic irradiation. Temperature was the most significant factor for seed progeny formation in the EURT zone, whereas the sums of precipitation and ratios of precipitation to temperature dominantly affected organisms from the background population.

A below-ground herbivore shapes root defensive chemistry in natural plant populations.

Plants display extensive intraspecific variation in secondary metabolites. However, the selective forces shaping this diversity remain often unknown, especially below ground. Using Taraxacum officinale and its major native insect root herbivore Melolontha melolontha, we tested whether below-ground herbivores drive intraspecific variation in root secondary metabolites. We found that high M. melolontha infestation levels over recent decades are associated with high concentrations of major root latex secondary metabolites across 21 central European T. officinale field populations. By cultivating offspring of these populations, we show that both heritable variation and phenotypic plasticity contribute to the observed differences. Furthermore, we demonstrate that the production of the sesquiterpene lactone taraxinic acid ß-D-glucopyranosyl ester (TA-G) is costly in the absence, but beneficial in the presence of M. melolontha, resulting in divergent selection of TA-G. Our results highlight the role of soil-dwelling insects for the evolution of plant defences in nature.

Natural epigenetic variation contributes to heritable flowering divergence in a widespread asexual dandelion lineage.

Epigenetic variation has been proposed to contribute to the success of asexual plants, either as a contributor to phenotypic plasticity or by enabling transient adaptation via selection on transgenerationally stable, but reversible, epialleles. While recent studies in experimental plant populations have shown the potential for epigenetic mechanisms to contribute to adaptive phenotypes, it remains unknown whether heritable variation in ecologically relevant traits is at least partially epigenetically determined in natural populations. Here, we tested the hypothesis that DNA methylation variation contributes to heritable differences in flowering time within a single widespread apomictic clonal lineage of the common dandelion (Taraxacum officinale s. lat.). Apomictic clone members of the same apomictic lineage collected from different field sites showed heritable differences in flowering time, which was correlated with inherited differences in methylation-sensitive AFLP marker profiles. Differences in flowering between apomictic clone members were significantly reduced after in vivo demethylation using the DNA methyltransferase inhibitor zebularine. This synchronization of flowering times suggests that flowering time divergence within an apomictic lineage was mediated by differences in DNA methylation. While the underlying basis of the methylation polymorphism at functional flowering time-affecting loci remains to be demonstrated, our study shows that epigenetic variation contributes to heritable phenotypic divergence in ecologically relevant traits in natural plant populations. This result also suggests that epigenetic mechanisms can facilitate adaptive divergence within genetically uniform asexual lineages.

Temperature and geographic attribution of change in the Taraxacum mongolicum growing season from 1990 to 2009 in eastern China's temperate zone.

Using leaf unfolding and leaf coloration data of a widely distributed herbaceous species, Taraxacum mongolicum, we detected linear trend and temperature response of the growing season at 52 stations from 1990 to 2009. Across the research region, the mean growing season beginning date marginal significantly advanced at a rate of -2.1 days per decade, while the mean growing season end date was significantly delayed at a rate of 3.1 days per decade. The mean growing season length was significantly prolonged at a rate of 5.1 days per decade. Over the 52 stations, linear trends of the beginning date correlate negatively with linear trends of spring temperature, whereas linear trends of the end date and length correlate positively with linear trends of autumn temperature and annual mean temperature. Moreover, the growing season linear trends are also closely related to the growing season responses to temperature and geographic coordinates plus elevation. Regarding growing season responses to temperature, a 1 °C increase in regional mean spring temperature results in an advancement of 2.1 days in regional mean growing season beginning date, and a 1 °C increase in regional mean autumn temperature causes a delay of 2.3 days in regional mean growing season end date. A 1 °C increase in regional annual mean temperature induces an extension of 8.7 days in regional mean growing season length. Over the 52 stations, response of the beginning date to spring temperature depends mainly on local annual mean temperature and geographic coordinates plus elevation. Namely, a 1 °C increase in spring temperature induces a larger advancement of the beginning date at warmer locations with lower latitudes and further west longitudes than at colder locations with higher latitudes and further east longitudes, while a 1 °C increase in spring temperature causes a larger advancement of the beginning date at higher than at lower elevations.

Chromium resistance of dandelion (Taraxacum platypecidum Diels.) and bermudagrass (Cynodon dactylon [Linn.] Pers.) is enhanced by arbuscular mycorrhiza in Cr(VI)-contaminated soils.

In a greenhouse pot experiment, dandelion (Taraxacum platypecidum Diels.) and bermudagrass (Cynodon dactylon[Linn.] Pers.), inoculated with and without arbuscular mycorrhizal fungus (AMF) Rhizophagus irregularis, were grown in chromium (Cr)-amended soils (0 mg/kg, 5 mg/kg, 10 mg/kg, and 20 mg/kg Cr[VI]) to test whether arbuscular mycorrhizal (AM) symbiosis can improve Cr tolerance in different plant species. The experimental results indicated that the dry weights of both plant species were dramatically increased by AM symbiosis. Mycorrhizal colonization increased plant P concentrations and decreased Cr concentrations and Cr translocation from roots to shoots for dandelion; in contrast, mycorrhizal colonization decreased plant Cr concentrations without improvement of P nutrition in bermudagrass. Chromium speciation analysis revealed that AM symbiosis potentially altered Cr species and bioavailability in the rhizosphere. The study confirmed the protective effects of AMF on host plants under Cr contaminations.

Is physiological performance a good predictor for fitness? Insights from an invasive plant species.

Is physiological performance a suitable proxy of fitness in plants? Although, several studies have been conducted to measure some fitness-related traits and physiological performance, direct assessments are seldom found in the literature. Here, we assessed the physiology-fitness relationship using second-generation individuals of the invasive plant species Taraxacum officinale from 17 localities distributed in five continents. Specifically, we tested if i) the maximum quantum yield is a good predictor for seed-output ii) whether this physiology-fitness relationship can be modified by environmental heterogeneity, and iii) if this relationship has an adaptive consequence for T. officinale individuals from different localities. Overall, we found a significant positive relationship between the maximum quantum yield and fitness for all localities evaluated, but this relationship decreased in T. officinale individuals from localities with greater environmental heterogeneity. Finally, we found that those individuals from localities where environmental conditions are highly seasonal performed better under heterogeneous environmental conditions. Contrarily, under homogeneous controlled conditions, those individuals from localities with low environmental seasonality performed much better. In conclusion, our results suggest that the maximum quantum yield seem to be good predictors for plant fitness. We suggest that rapid measurements, such as those obtained from the maximum quantum yield, could provide a straightforward proxy of individual's fitness in changing environments.

Influence of global atmospheric change on the feeding behavior and growth performance of a mammalian herbivore, Microtus ochrogaster.

Global atmospheric change is influencing the quality of plants as a resource for herbivores. We investigated the impacts of elevated carbon dioxide (CO2) and ozone (O3) on the phytochemistry of two forbs, Solidago canadensis and Taraxacum officinale, and the subsequent feeding behavior and growth performance of weanling prairie voles (Microtus ochrogaster) feeding on those plants. Plants for the chemical analyses and feeding trials were harvested from the understory of control (ambient air), elevated CO2 (560 µl CO2 l(-1)), and elevated O3 (ambient × 1.5) rings at the Aspen FACE (Free Air CO2 Enrichment) site near Rhinelander, Wisconsin. We assigned individual voles to receive plants from only one FACE ring and recorded plant consumption and weanling body mass for seven days. Elevated CO2 and O3 altered the foliar chemistry of both forbs, but only female weanling voles on the O3 diet showed negative responses to these changes. Elevated CO2 increased the fiber fractions of both plant species, whereas O3 fumigation elicited strong responses among many phytochemical components, most notably increasing the carbon-to-nitrogen ratio by 40% and decreasing N by 26%. Consumption did not differ between plant species or among fumigation treatments. Male voles were unaffected by the fumigation treatments, whereas female voles grew 36% less than controls when fed O3-grown plants. These results demonstrate that global atmospheric change has the potential to affect the performance of a mammalian herbivore through changes in plant chemistry.

Geographic parthenogenesis and plant-enemy interactions in the common dandelion.

BACKGROUND: Many species with sexual and asexual variants show a pattern of geographic parthenogenesis where asexuals have broader and higher-latitude distribution than sexuals. Because sexual reproduction is often considered a costly evolutionary strategy that is advantageous in the face of selection by coevolving pests and pathogens, one possible explanation for geographic parthenogenesis is that populations at higher latitudes are exposed to fewer pests and pathogens. We tested this hypothesis in the common dandelion (Taraxacum officinale), a species with well-established geographic parthenogenesis, by screening prevalence and effects of several specialized pests and pathogens in natural dandelion populations. RESULTS: We did a population survey of 18 dandelion populations along a geographic transect that ranged from the area where sexual and asexual dandelions co-occur northward into the area where only asexuals occur. In addition we used four southern and four northern populations in a 8x8 cross-inoculation greenhouse experiment in which plants were exposed experimentally to each other's natural field soil microbial communities. The cross-inoculation experiment indicated a higher pathogenicity of soil microbial communities from the southern, mostly sexual, populations compared to soil microbial communities from the northern asexual populations. Northern dandelion populations also showed reduced infestation by a specialized seed-eating weevil. A similar trend of reduced rust fungus infection in northern populations was observed but this trend was not statistically significant. CONCLUSIONS: The prevalence of pests and pathogens decreased along the south-to-north axis of geographic parthenogenesis. This highlights the potential of biotic interactions in shaping patterns of geographic parthenogenesis.

Anatomy of ovary and ovule in dandelions (Taraxacum, Asteraceae).

The genus Taraxacum Wigg. (Asteraceae) forms a polyploid complex within which there are strong links between the ploidy level and the mode of reproduction. Diploids are obligate sexual, whereas polyploids are usually apomictic. The paper reports on a comparative study of the ovary and especially the ovule anatomy in the diploid dandelion T. linearisquameum and the triploid T. gentile. Observations with light and electron microscopy revealed no essential differences in the anatomy of both the ovary and ovule in the examined species. Dandelion ovules are anatropous, unitegmic and tenuinucellate. In both sexual and apomictic species, a zonal differentiation of the integument is characteristic of the ovule. In the integumentary layers situated next to the endothelium, the cell walls are extremely thick and PAS positive. Data obtained from TEM indicate that these special walls have an open spongy structure and their cytoplasm shows evidence of gradual degeneration. Increased deposition of wall material in the integumentary cells surrounding the endothelium takes place especially around the chalazal pole of the embryo sac as well as around the central cell. In contrast, the integumentary cells surrounding the micropylar region have thin walls and exhibit a high metabolic activity. The role of the thick-walled integumentary layers in the dandelion ovule is discussed. We also consider whether this may be a feature of taxonomic importance.

Detecting small-scale genotype-environment interactions in apomictic dandelion (Taraxacum officinale) populations.

Studies of genotype × environment interactions (G × E) and local adaptation provide critical tests of natural selection's ability to counter opposing forces such as gene flow. Such studies may be greatly facilitated in asexual species, given the possibility for experimental replication at the level of true genotypes (rather than populations) and the possibility of using molecular markers to assess genotype-environment associations in the field (neither of which is possible for most sexual species). Here, we tested for G × E in asexual dandelions (Taraxacum officinale) by subjecting six genotypes to experimental drought, mown and benign (control) conditions and subsequently using microsatellites to assess genotype-environment associations in the field. We found strong G × E, with genotypes that performed poorly under benign conditions showing the highest performance under stressful conditions (drought or mown). Our six focal genotypes comprise > 80% of plants in local populations. The most common genotype in the field showed its highest relative performance under mown conditions (the most common habitat in our study area), and almost all plants of this genotype in the field were found growing in mowed lawns. Genotypes performing best under benign experimental conditions were found most frequently in unmown conditions in the field. These results are strongly indicative of local adaptation at a very small scale, with unmown microsites of only a few square metres typically embedded within larger mown lawns. By studying an asexual species, we were able to map genotypes with known ecological characteristics to environments with high spatial precision.

The costs of sex: facing real-world complexities.

Understanding the maintenance of sexual reproduction constitutes a difficult problem for evolutionary biologists because of the immediate costs that sex seems to incur. Typically, general benefits to sex and recombination are investigated that might outweigh these costs. However, several factors can strongly influence the complex balance between costs and benefits of sex; these include constraints on the evolution of asexuality, ecological differentiation, and certain lif-history traits. We review these factors and their empirical support for the first time in a unified framework and find that they can reduce the costs of sex, circumvent them, or make them inapplicable. These factors can even tip the scales to a net benefit for sex. The reviewed factors affect species and species groups differently, and we conclude consequently that understanding the maintenance of sex could turn out to be more species-specific than commonly assumed. Interestingly, our study suggests that, in some species, no general benefits to sex and recombination might be needed to understand the maintenance of sex, as in our case study of dandelions.