Effect of cadmium and polystyrene nanoplastics on the growth, antioxidant content, ionome, and metabolism of dandelion seedlings.

Cadmium is the most prevalent heavy metal pollutant in the environment and can be readily combined with micro/nanoplastics (M/NPs) to change their bioavailability. In the present study, we comprehensively investigated the effect of polystyrene (PS) NPs on dandelion plants grown under Cd stress. Cd exposure significantly inhibited the growth of dandelion seedlings, resulting in a decrease in seedling elongation from 26.47% to 28.83%, a reduction in biomass from 29.76% to 54.14%, and an exacerbation of lipid peroxidation and oxidative stress. The interaction between PS NPs and Cd resulted in the formation of larger aggregates, with the Cd bioavailability reduced by 12.56%. PS NPs affect ion absorption by regulating reactive oxygen production and increasing superoxide dismutase activity, thereby mitigating the adverse effects of Cd. PSCd aggregates induced significant changes in the metabolic profiles of dandelions, affecting various carbohydrates related to alcohols, organic acids, sugar metabolism, and bioactive components related to flavonoids and phenolic acids. Furthermore, based on a structural equation model, exposure to PSCd activated oxidative stress and nutrient absorption, thereby affecting plant growth and Cd accumulation. Overall, our study provides valuable insights into the effects of PS NPs on Cd bioavailability, accumulation, and plant growth, which are crucial for understanding the food safety of medicinal plants in a coexistence environment.

Hysteresis of heavy metals uptake induced in Taraxacum officinale by thiuram.

Dandelion (Taraxacum officinale) yields active substances frequently used in herbal medicinal preparations. Its plantations are exposed to fungal plagues which pose a threat to herbal crops. The aim of this study was to evaluate the long time effects of a fungicide thiuram on dandelion growth and photosynthesis. Additionally, the manganese, iron, copper, zinc, cadmium, and lead uptake and transport were also investigated. Plants were cultivated under greenhouse conditions by the pot method in a universal flowering soil. The elements content in soil and plants were determined by the HR-CS FAAS spectrometer. Thiuram concentrations were established by the HPLC. Those analyses showed that almost 80% of thiuram decomposed within two weeks of its application. The photosynthesis indicators suggested, that plants were in good conditions and the fungicide supplementation facilitated plant growth. The latter could be prompted by thiuram acting as a sulfur rich chemical micro fertilizer. The hypothesis, that thiuram significantly affects heavy metals interactions in dandelion was proved by the one-way analysis of variance. Notable, metals uptake did not completely recover after fungicide decomposition for all investigated elements except iron We suggest to define this chemically induced, time-dependent heavy metals migrations in the soil-plant system as hysteresis of heavy metals uptake.

Immunodetection of Pectic Epitopes, Arabinogalactan Proteins, and Extensins in Mucilage Cells from the Ovules of Pilosella officinarum Vaill. and Taraxacum officinale Agg. (Asteraceae).

The main aim of this study was to compare the cytological difference between ovular mucilage cells in two Asteraceae species-Pilosella officinarum and Taraxacum officinale-in order to determine whether pectic epitopes, arabinogalactan proteins, or extensins are present. The immunocytochemical technique was used. Both the Taracacum and Pilosella genera have been used recently as models for understanding the mechanisms of apomixis. Knowledge of the presence of signal molecules (pectic epitopes, arabinogalactan proteins, and extensins) can help better understand the developmental processes in these plants during seed growth. The results showed that in Pilosella officinarum, there was an accumulation of pectins in the mucilage, including both weakly and highly esterified pectins, which was in contrast to the mucilage of Taraxacum officinale, which had low amounts of these pectins. However, Taraxacum protoplasts of mucilage cells were rich in weakly methyl-esterified pectins. While the mucilage contained arabinogalactan proteins in both of the studied species, the types of arabinogalactan proteins were different. In both of the studied species, extensins were recorded in the transmitting tissues. Arabinogalactan proteins as well as weakly and highly esterified pectins and extensins occurred in close proximity to calcium oxalate crystals in both Taraxacum and Pilosella cells.

Genetic Dissection of Apomixis in Dandelions Identifies a Dominant Parthenogenesis Locus and Highlights the Complexity of Autonomous Endosperm Formation.

Apomixis in the common dandelion (Taraxacum officinale) consists of three developmental components: diplospory (apomeiosis), parthenogenesis, and autonomous endosperm development. The genetic basis of diplospory, which is inherited as a single dominant factor, has been previously elucidated. To uncover the genetic basis of the remaining components, a cross between a diploid sexual seed parent and a triploid apomictic pollen donor was made. The resulting 95 triploid progeny plants were genotyped with co-dominant simple-sequence repeat (SSR) markers and phenotyped for apomixis as a whole and for the individual apomixis components using Nomarski Differential Interference Contrast (DIC) microscopy of cleared ovules and seed flow cytometry. From this, a new SSR marker allele was discovered that was closely linked to parthenogenesis and unlinked to diplospory. The segregation of apomixis as a whole does not differ significantly from a three-locus model, with diplospory and parthenogenesis segregating as unlinked dominant loci. Autonomous endosperm is regularly present without parthenogenesis, suggesting that the parthenogenesis locus does not also control endosperm formation. However, the high recovery of autonomous endosperm is inconsistent with this phenotype segregating as the third dominant locus. These results highlight the genetic complexity underlying apomixis in the dandelion and underline the challenge of introducing autonomous apomixis into sexual crops.

Heterologous activation of the Hevea PEP16 promoter in the rubber-producing laticiferous tissues of Taraxacum kok-saghyz.

Hevea brasiliensis, the most abundant rubber crop, is used widely for the commercial production of natural rubber. To reduce the risk of a shortage in the supply of natural rubber that may arise from a single major rubber crop, rubber dandelion (Taraxacum kok-saghyz) has been developed as an alternative rubber-producing crop by using a transgenic approach. However, it is necessary to identify a suitable promoter for the transfer of rubber biosynthesis-related genes to the species. In this study, the promoter region of H. brasiliensis PEP16, which was isolated as a potentially important component in rubber biosynthesis, was sequenced and a pPEP16::GUS fusion construct was introduced into T. kok-saghyz. Histological and fluorometric studies using transgenic T. kok-saghyz plants indicated that the HbPEP16 promoter was highly activated in a laticiferous tissue-specific manner under normal growth conditions and that promoter activation was tightly regulated by various hormones and external signals. These findings suggested that the HbPEP16 promoter may be a useful molecular tool for the manipulation of gene expression in the laticiferous tissues of T. kok-saghyz.

Loss of function mutation of the Rapid Alkalinization Factor (RALF1)-like peptide in the dandelion Taraxacum koksaghyz entails a high-biomass taproot phenotype.

The Russian dandelion (Taraxacum koksaghyz) is a promising source of inulin and natural rubber because large amounts of both feedstocks can be extracted from its roots. However, the domestication of T. koksaghyz requires the development of stable agronomic traits such as higher yields of inulin and natural rubber, a higher root biomass, and an agronomically preferable root morphology which is more suitable for cultivation and harvesting. Arabidopsis thaliana Rapid Alkalinisation Factor 1 (RALF1) has been shown to suppress root growth. We identified the T. koksaghyz orthologue TkRALF-like 1 and knocked out the corresponding gene (TkRALFL1) using the CRISPR/Cas9 system to determine its impact on root morphology, biomass, and inulin and natural rubber yields. The TkRALFL1 knockout lines more frequently developed a taproot phenotype which is easier to cultivate and harvest, as well as a higher root biomass and greater yields of both inulin and natural rubber. The TkRALFL1 gene could therefore be suitable as a genetic marker to support the breeding of profitable new dandelion varieties with improved agronomic traits. To our knowledge, this is the first study addressing the root system of T. koksaghyz to enhance the agronomic performance.

Kinetics and modeling of cell growth for potential anthocyanin induction in cultures of Taraxacum officinale G.H. Weber ex Wiggers (Dandelion) in vitro

Background: Taraxacum officinale G.H. Weber ex Wiggers is a wild plant used in folk medicine to treat several diseases owing to bioactive secondary metabolites present in its tissue. The accumulation of such molecules in plant cells can occur as a response against abiotic stress, but these metabolites are often deposited in low concentrations. For this reason, the use of a biotechnological approach to improve the yields of technologically interesting bioactive compounds such as anthocyanins is a compelling option. This work focuses on investigating the potential of in vitro T. officinale cultures as an anthocyanin source. Results: To demonstrate the suitability of anthocyanin induction and accumulation in calluses under specific conditions, anthocyanin was induced in the T. officinale callus. A specific medium of 5.5% sucrose supplemented with 6-benzylaminopurine /1-naphthaleneacetic acid in a 10:1 ratio was used to produce an anthocyanin yield of 1.23 mg g-1 fw. An in vitro dandelion callus line was established from this experiment. Five mathematical models were then used to objectively and predictably explain the growth of anthocyanin-induced calluses from T. officinale. Of these models, the Richards model offered the most suitable representation of anthocyanin callus growth in a solid medium and permitted the calculation of the corresponding kinetic parameters. Conclusions: The findings demonstrate the potential of an in vitro anthocyanin-induced callus line from T. officinale as an industrial anthocyanin source.

Host Plant Physiology and Mycorrhizal Functioning Shift across a Glacial through Future [CO2] Gradient.

Rising atmospheric carbon dioxide concentration ([CO2]) may modulate the functioning of mycorrhizal associations by altering the relative degree of nutrient and carbohydrate limitations in plants. To test this, we grew Taraxacum ceratophorum and Taraxacum officinale (native and exotic dandelions) with and without mycorrhizal fungi across a broad [CO2] gradient (180-1,000 µL L-1). Differential plant growth rates and vegetative plasticity were hypothesized to drive species-specific responses to [CO2] and arbuscular mycorrhizal fungi. To evaluate [CO2] effects on mycorrhizal functioning, we calculated response ratios based on the relative biomass of mycorrhizal (MBio) and nonmycorrhizal (NMBio) plants (RBio = [MBio - NMBio]/NMBio). We then assessed linkages between RBio and host physiology, fungal growth, and biomass allocation using structural equation modeling. For T. officinale, RBio increased with rising [CO2], shifting from negative to positive values at 700 µL L-1 [CO2] and mycorrhizal effects on photosynthesis and leaf growth rates drove shifts in RBio in this species. For T. ceratophorum, RBio increased from 180 to 390 µL L-1 and further increases in [CO2] caused RBio to shift from positive to negative values. [CO2] and fungal effects on plant growth and carbon sink strength were correlated with shifts in RBio in this species. Overall, we show that rising [CO2] significantly altered the functioning of mycorrhizal associations. These symbioses became more beneficial with rising [CO2], but nonlinear effects may limit plant responses to mycorrhizal fungi under future [CO2]. The magnitude and mechanisms driving mycorrhizal-CO2 responses reflected species-specific differences in growth rate and vegetative plasticity, indicating that these traits may provide a framework for predicting mycorrhizal responses to global change.

Heterologous expression of chloroplast-localized geranylgeranyl pyrophosphate synthase confers fast plant growth, early flowering and increased seed yield.

Geranylgeranyl pyrophosphate synthase (GGPS) is a key enzyme for a structurally diverse class of isoprenoid biosynthetic metabolites including gibberellins, carotenoids, chlorophylls and rubber. We expressed a chloroplast-targeted GGPS isolated from sunflower (Helianthus annuus) under control of the cauliflower mosaic virus 35S promoter in tobacco (Nicotiana tabacum). The resulting transgenic tobacco plants expressing heterologous GGPS showed remarkably enhanced growth (an increase in shoot and root biomass and height), early flowering, increased number of seed pods and greater seed yield compared with that of GUS-transgenic lines (control) or wild-type plants. The gibberellin levels in HaGGPS-transgenic plants were higher than those in control plants, indicating that the observed phenotype may result from increased gibberellin content. However, in HaGGPS-transformant tobacco plants, we did not observe the phenotypic defects such as reduced chlorophyll content and greater petiole and stalk length, which were previously reported for transgenic plants expressing gibberellin biosynthetic genes. Fast plant growth was also observed in HaGGPS-expressing Arabidopsis and dandelion plants. The results of this study suggest that GGPS expression in crop plants may yield desirable agronomic traits, including enhanced growth of shoots and roots, early flowering, greater numbers of seed pods and/or higher seed yield. This research has potential applications for fast production of plant biomass that provides commercially valuable biomaterials or bioenergy.

Content of Zinc and Copper in Selected Plants Growing Along a Motorway.

In 2011 a study was carried out analyzing the effects of road traffic on bioaccumulation of zinc and copper in selected species of dicotyledonous plants growing on adjacent grasslands. To do the research the plants were sampled from the 9-km-long Siedlce bypass, a part of the international route E-30. They were collected during the flowering stage, at following distances from the road: 1, 5, 10, 15 m. The content of zinc and copper was determined with the AAS method, with dry mineralisation done before. The highest concentration of the elements, regardless of the distance from the road, was found in Taraxacum spec. Among the tested plants, the lowest zinc content was in Vicia cracca, and the lowest copper content in Rumex acetosa. The limit for copper content was exceeded in Taraxacum spec. and, slightly, in Achillea millefolium growing at the roadside, closest to the roadway.

Taraxacum officinale and related species-An ethnopharmacological review and its potential as a commercial medicinal plant.

ETHNOPHARMACOLOGICAL RELEVANCE: Dandelion (Taraxacum spec) is a wild plant that has been used for centuries as a traditional medicine in the relief and treatment of several diseases. This use is due to the presence of sesquiterpenes, saponins, phenolic compounds, flavonoids, and sugars, among others, found in the organs of the plant. AIM OF THE STUDY: The aim of this work is to provide a current review of developments and trends in research on the Taraxacum genus, with a focus on traditional uses and pharmacological properties. This should shed light on the potential of this plant as an attractive commercial herbal medicine. MATERIALS AND METHODS: Documents were collected, analyzed, and classified for information regarding medical, agronomic, genetic, and biological aspects of the Taraxacum species. This process was based on a thorough search of documents indexed by scientific search engines. RESULTS: Two important periods of research on Taraxacum have been identified: the first, between 1930 and 1950; and the second, from 1990 to today. During the former, agricultural and genetics research on this plant were, due to the shortage of natural rubber, the focus. In contrast, the main drive in Taraxacum research is now the recovery of bioactives and/or applications in medicine. Pharmacology is the main area in which these plants have been tested, thanks in part to its widely known traditional uses; however, there is less than enthusiastic interest in further human clinical trials. In other areas, Taraxacum sports an enormous list of compounds of industrial interest; and while it is true that only a small amount of these compounds is immediately available in Taraxacum organs and makes it relatively commercially unattractive, only scarce efforts have been made to improve yields. Compounding this issue, most studies of its growth and cultivation have been focused mainly on controlling it as a weed detrimental to certain industrial crops. To wit, in spite of all the research carried out, less than 1% of all the species identified so far (>2500) have been studied (including Taraxacum officinale, Taraxacum coreanum, Taraxacum mongolicum and Taraxacum platycarpum). This is a indication of the little knowledge that we have about this genus so far. Biotechnology (involving genetics, agriculture, and biology) is the most powerful means by which to take advantage of all the medicinal potential of Taraxacum. Great strides have been made in identifying metabolic pathways for synthesizing terpenes, one of the most important compound families in clinical applications. In order to improve yield and performance of the plant in the field, greenhouse cultivation is another aspect taken into account, deriving an increase in recovery of bioactives from Taraxacum organs. Even while considering that only a few species have been studied, their different biochemical and cultivation profiles indicate huge potential for qualitative improvements in composition through genetic engineering, thus directly impacting pharmacological properties. CONCLUSIONS: Taraxacum is has been traditionally considered a natural remedy, well-inserted into popular knowledge, but with low commercial applicability. Only once the recovery of pure and highly reactive compounds can be pursued at (a qualitatively and quantitatively attractive) economical scale, human clinical trials would be of interest in order to prove their efficacy and safety, positioning Taraxacum as an important commercial source of natural drugs.

Identification, quantification, spatiotemporal distribution and genetic variation of major latex secondary metabolites in the common dandelion (Taraxacum officinale agg.).

The secondary metabolites in the roots, leaves and flowers of the common dandelion (Taraxacum officinale agg.) have been studied in detail. However, little is known about the specific constituents of the plant's highly specialized laticifer cells. Using a combination of liquid and gas chromatography, mass spectrometry and nuclear magnetic resonance spectrometry, we identified and quantified the major secondary metabolites in the latex of different organs across different growth stages in three genotypes, and tested the activity of the metabolites against the generalist root herbivore Diabrotica balteata. We found that common dandelion latex is dominated by three classes of secondary metabolites: phenolic inositol esters (PIEs), triterpene acetates (TritAc) and the sesquiterpene lactone taraxinic acid ß-D-glucopyranosyl ester (TA-G). Purification and absolute quantification revealed concentrations in the upper mgg(-1) range for all compound classes with up to 6% PIEs, 5% TritAc and 7% TA-G per gram latex fresh weight. Contrary to typical secondary metabolite patterns, concentrations of all three classes increased with plant age. The highest concentrations were measured in the main root. PIE profiles differed both quantitatively and qualitatively between plant genotypes, whereas TritAc and TA-G differed only quantitatively. Metabolite concentrations were positively correlated within and between the different compound classes, indicating tight biosynthetic co-regulation. Latex metabolite extracts strongly repelled D. balteata larvae, suggesting that the latex constituents are biologically active.

Deposition of callose in young ovules of two Taraxacum species varying in the mode of reproduction.

Although callose occurs during megasporogenesis in most flowering plants, the knowledge about its general function and the mechanisms by which the callose layer is formed in particular places is still not sufficient. The results of previous studies suggest a total lack of callose in the ovules of diplosporous plants in which meiosis is omitted or disturbed. This report is the first documentation of callose events in dandelions ovules. We demonstrated the pattern of callose deposition during the formation of megaspores through diplospory of Taraxacum type and during normal meiotic megasporogenesis in apomictic triploid Taraxacum atricapillum and amphimictic diploid Taraxacum linearisquameum. We found the presence of callose in the megasporocyte wall of both diplosporous and sexual dandelions. However, in a diplosporous dandelion, callose predominated at the micropylar pole of megaspore mother cell (MMC) which may be correlated with abnormal asynaptic meiosis and may indicate diplospory of the Taraxacum type. After meiotic division, callose is mainly deposited in the walls between megaspores in tetrads and in diplodyads. In subsequent stages, callose gradually disappears around the chalazal functional megaspore. However, some variations in the pattern of callose deposition within tetrad may reflect variable positioning of the functional megaspore (FM) observed in the ovules of T. linearisquameum.

Latitudinal variation in sensitivity of flower bud formation to high temperature in Japanese Taraxacum officinale.

Control of flowering time plays a key role in the successful range expansion of plants. Taraxacum officinale has expanded throughout Japan during the 110 years after it was introduced into a cool temperate region. The present study tested a hypothesis that there is a genetic difference in the bud formation time in relation to temperature along latitudinal gradient of T. officinale populations. In Experiment 1, plants from three populations at different latitudes (26, 36, and 43°N) were grown at three temperatures. Time to flower bud appearance did not significantly differ among the three populations when plants were grown at 14 °C, whereas it increased with increasing latitude when grown at 19 and 24 °C. Rosette diameter was not different among the populations, indicating that the variation in bud formation time reflected a difference in genetic control rather than size variation. The latitudinal variation in bud appearance time was confirmed by Experiment 2 in which plants from 17 population were used. In Experiment 3, the size of plants that exhibited late-flowering was studied to test a hypothesis that the variation in flowering time reflects dormancy of vegetative growth, but the late-flowering plants were found to continue growth, indicating that vegetative dormancy was not the cause of the variation. The results clearly indicate that the degree of suppression of flower bud formation at high temperature decreases with latitude from north to south, which is under genetic control.

Influences on the establishment and dominance of vegetation in stormwater infiltration basins.

Infiltration basins are widely used in urban environments as a technique for managing and reducing the volume of stormwater. These basins can be spontaneously colonized by wild plants, which can be used as bioindicators of edaphic characteristics. As the basins are anthropogenic environments, the description of plant biodiversity allows the determination of which species colonize such environments and identification of the relationships between plants, basin type and operation. Nineteen infiltration basins were selected according to their catchment types (industrial, urban, agricultural). The dominant species were identified and sampled. Rumex sp., Taraxacum sp. and Artemisia sp. are the three types most represented (88, 61 and 55% respectively of the basins studied). Their families and their respective orders are those most commonly found (Caryophyllales, Asterales and Polygonaceae, Asteraceae). Poaceae is the family grouping with the largest number of different species (11). Although each species occupies only 1 or 2 basins, plants of this family occupy 61% of the basins. Although the catchment characteristics of the 19 basins do not play a direct role in the diversity of plant families, they can influence the presence or absence of certain species. Thus, these plants can be used as bio-indicators of basin soil and operating characteristics, such as sediment depths, inundation frequency and duration.

A novel C-S lyase from the latex-producing plant Taraxacum brevicorniculatum displays alanine aminotransferase and l-cystine lyase activity.

We isolated a novel pyridoxal-5-phosphate-dependent l-cystine lyase from the dandelion Taraxacum brevicorniculatum. Real time qPCR analysis showed that C-S lyase from Taraxacum brevicorniculatum (TbCSL) mRNA is expressed in all plant tissues, although at relatively low levels in the latex and pedicel. The 1251 bp TbCSL cDNA encodes a protein with a calculated molecular mass of 46,127 kDa. It is homologous to tyrosine and alanine aminotransferases (AlaATs) as well as to an Arabidopsis thaliana carbon-sulfur lyase (C-S lyase) (SUR1), which has a role in glucosinolate metabolism. TbCSL displayed in vitrol-cystine lyase and AlaAT activities of 4 and 19nkatmg(-1) protein, respectively. However, we detected no in vitro tyrosine aminotransferase (TyrAT) activity and RNAi knockdown of the enzyme had no effect on phenotype, showing that TbCSL substrates might be channeled into redundant pathways. TbCSL is in vivo localized in the cytosol and functions as a C-S lyase or an aminotransferase in planta, but the purified enzyme converts at least two substrates specifically, and can thus be utilized for further in vitro applications.

Transgenerational effects of stress exposure on offspring phenotypes in apomictic dandelion.

Heritable epigenetic modulation of gene expression is a candidate mechanism to explain parental environmental effects on offspring phenotypes, but current evidence for environment-induced epigenetic changes that persist in offspring generations is scarce. In apomictic dandelions, exposure to various stresses was previously shown to heritably alter DNA methylation patterns. In this study we explore whether these induced changes are accompanied by heritable effects on offspring phenotypes. We observed effects of parental jasmonic acid treatment on offspring specific leaf area and on offspring interaction with a generalist herbivore; and of parental nutrient stress on offspring root-shoot biomass ratio, tissue P-content and leaf morphology. Some of the effects appeared to enhance offspring ability to cope with the same stresses that their parents experienced. Effects differed between apomictic genotypes and were not always consistently observed between different experiments, especially in the case of parental nutrient stress. While this context-dependency of the effects remains to be further clarified, the total set of results provides evidence for the existence of transgenerational effects in apomictic dandelions. Zebularine treatment affected the within-generation response to nutrient stress, pointing at a role of DNA methylation in phenotypic plasticity to nutrient environments. This study shows that stress exposure in apomictic dandelions can cause transgenerational phenotypic effects, in addition to previously demonstrated transgenerational DNA methylation effects.

Genotypic diversity effects on the performance of Taraxacum officinale populations increase with time and environmental favorability.

Within-population genetic diversity influences many ecological processes, but few studies have examined how environmental conditions may impact these short-term diversity effects. Over four growing seasons, we followed experimental populations of a clonal, ubiquitous weed, Taraxacum officinale, with different numbers of genotypes in relatively favorable fallow field and unfavorable mowed lawn environmental treatments. Population performance (measured as total leaf area, seed production or biomass) clearly and consistently increased with diversity, and this effect became stronger over the course of the experiment. Diversity effects were stronger, and with different underlying mechanisms, in the fallow field versus the mowed lawn. Large genotypes dominated in the fallow field driving overyielding (via positive selection effects), whereas in the mowed lawn, where performance was limited by regular disturbance, there was evidence for complementarity among genotypes (with one compact genotype in particular performing better in mixture than monoculture). Hence, we predict stronger genotypic diversity effects in environments where intense intraspecific competition enhances genotypic differences. Our four-year field experiment plus seedling establishment trials indicate that genotypic diversity effects have far-reaching and context-dependent consequences across generations.

Density-independent mortality and increasing plant diversity are associated with differentiation of Taraxacum officinale into r- and K-strategists.

BACKGROUND: Differential selection between clones of apomictic species may result in ecological differentiation without mutation and recombination, thus offering a simple system to study adaptation and life-history evolution in plants. METHODOLOGY/PRINCIPAL FINDINGS: We caused density-independent mortality by weeding to colonizer populations of the largely apomictic Taraxacum officinale (Asteraceae) over a 5-year period in a grassland biodiversity experiment (Jena Experiment). We compared the offspring of colonizer populations with resident populations deliberately sown into similar communities. Plants raised from cuttings and seeds of colonizer and resident populations were grown under uniform conditions. Offspring from colonizer populations had higher reproductive output, which was in general agreement with predictions of r-selection theory. Offspring from resident populations had higher root and leaf biomass, fewer flower heads and higher individual seed mass as predicted under K-selection. Plants grown from cuttings and seeds differed to some degree in the strength, but not in the direction, of their response to the r- vs. K-selection regime. More diverse communities appeared to exert stronger K-selection on resident populations in plants grown from cuttings, while we did not find significant effects of increasing species richness on plants grown from seeds. CONCLUSIONS/SIGNIFICANCE: Differentiation into r- and K-strategists suggests that clones with characteristics of r-strategists were selected in regularly weeded plots through rapid colonization, while increasing plant diversity favoured the selection of clones with characteristics of K-strategists in resident populations. Our results show that different selection pressures may result in a rapid genetic differentiation within a largely apomictic species. Even under the assumption that colonizer and resident populations, respectively, happened to be r- vs. K-selected already at the start of the experiment, our results still indicate that the association of these strategies with the corresponding selection regimes was maintained during the 5-year experimental period.

Molecular cloning of mevalonate pathway genes from Taraxacum brevicorniculatum and functional characterisation of the key enzyme 3-hydroxy-3-methylglutaryl-coenzyme A reductase.

Taraxacum brevicorniculatum is known to produce high quality rubber. The biosynthesis of rubber is dependent on isopentenyl pyrophosphate (IPP) precursors derived from the mevalonate (MVA) pathway. The cDNA sequences of seven MVA pathway genes from latex of T. brevicorniculatum were isolated, including three cDNA sequences encoding for 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductases (TbHMGR1-3). Expression analyses indicate an important role of TbHMGR1 as well as for the HMG-CoA synthase (TbHMGS), the diphosphomevalonate decarboxylase and the mevalonate kinase in the provision of precursors for rubber biosynthesis. The amino acid sequences of the TbHMGRs show the typical motifs described for plant HMGRs such as two transmembrane domains and a catalytic domain containing two HMG-CoA and two NADP(H) binding sites. The functionality of the HMGRs was demonstrated by complementation assay using an IPP auxotroph mutant of Escherichia coli. Furthermore, the transient expression of the catalytic domains of TbHMGR1 and TbHMGR2 in Nicotiana benthamiana resulted in a strong accumulation of sterol precursors, one of the major groups of pathway end-products.