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PubMed2003

Reversibility of cold- and light-stress tolerance and accompanying changes of metabolite and antioxidant levels in the two high mountain plant species Soldanella alpina and Ranunculus glacialis.

Two high mountain plants Soldanella alpina (L.) and Ranunculus glacialis (L.) were transferred from their natural environment to two different growth conditions (22 degrees C and 6 degrees C) at low elevation in order to investigate the possibility of de-acclimation to light and cold and the importance of antioxidants and metabolite levels. The results were compared with the lowland crop plant Pisum sativum (L.) as a control. Leaves of R. glacialis grown for 3 weeks at 22 degrees C were more sensitive to light-stress (defined as damage to photosynthesis, reduction of catalase activity (EC 1.11.1.6) and bleaching of chlorophyll) than leaves collected in high mountains or grown at 6 degrees C. Light-stress tolerance of S. alpina leaves was not markedly changed. Therefore, acclimation is reversible in R. glacialis leaves, but constitutive or long-lasting in S. alpina leaves. The different growth conditions induced significant changes in non-photochemical fluorescence quenching (qN) and the contents of antioxidants and xanthophyll cycle pigments. These changes did not correlate with light-stress tolerance, questioning their role for light- and cold-acclimation of both alpine species. However, ascorbate contents remained very high in leaves of S. alpina under all growth conditions (12-19% of total soluble carbon). In cold-acclimated leaves of R. glacialis, malate represented one of the most abundant compounds of total soluble carbon (22%). Malate contents declined significantly in de-acclimated leaves, suggesting a possible involvement of malate, or malate metabolism, in light-stress tolerance. Leaves of the lowland plant P. sativum were more sensitive to light-stress than the alpine species, and contained only low amounts of malate and ascorbate.

Acclimatization↗
PubMed2006

Respiratory carbon metabolism in the high mountain plant species Ranunculus glacialis.

Very little is known about the primary carbon metabolism of the high mountain plant Ranunculus glacialis. It is a species with C3 photosynthesis, but with exceptionally high malate content in its leaves, the biological significance of which remains unclear. 13C/12C-isotope ratio mass spectrometry (IRMS) and 13C-nuclear magnetic resonance (NMR) labelling were used to study the carbon metabolism of R. glacialis, paying special attention to respiration. Although leaf dark respiration was high, the temperature response had a Q10 of 2, and the respiratory quotient (CO2 produced divided by O2 consumed) was nearly 1, indicating that the respiratory pool is comprised of carbohydrates. Malate, which may be a large carbon substrate, was not respired. However, when CO2 fixed by photosynthesis was labelled, little labelling of the CO2 subsequently respired in the dark was detected, indicating that: (i) most of the carbon recently assimilated during photosynthesis is not respired in the dark; and (ii) the carbon used for respiration originates from (unlabelled) reserves. This is the first demonstration of such a low metabolic coupling of assimilated and respired carbon in leaves. The biological significance of the uncoupling between assimilation and respiration is discussed.

Carbon↗
PubMed2004

Genetic diversity of Ranunculus acris L. (Ranunculaceae) populations in relation to species diversity and habitat type in grassland communities.

Correlates between genetic diversity at intra- and interpopulation levels and the species diversity in plant communities are rarely investigated. Such correlates may give insights into the effect of local selective forces across different communities on the genetic diversity of local plant populations. This study has employed amplified fragment length polymorphism to assess the genetic diversity within and between 10 populations of Ranunculus acris in relation to the species diversity (richness and evenness) of grassland communities of two different habitat types, 'seminatural' and 'agriculturally improved', located in central Germany. Within-population genetic diversity estimated by Nei's unbiased gene diversity (HE) was high (0.258-0.334), and was not correlated with species richness (Pearson's r = -0.17; P = 0.64) or species evenness (Pearson's r = 0.15; P = 0.68) of the plant communities. However, the genetic differentiation between R. acris populations was significantly correlated with the difference in species evenness (Mantel's r = 0.62, P = 0.02), but not with difference in species richness of plant communities (r = -0.17, P = 0.22). Moreover, we also found that populations of R. acris from the 'seminatural' habitat were genetically different (amova, P < 0.05) from those in 'agriculturally improved' habitats, suggesting that gene flow between these habitat types is limited. The results reported in this study may indicate that habitat characteristics influence the genetic diversity of plant species.

Analysis of Variance↗
PubMed2006

Rare arctic-alpine plants of the European Alps have different immigration histories: the snow bed species Minuartia biflora and Ranunculus pygmaeus.

Minuartia biflora and Ranunculus pygmaeus are circumarctic plants with a few isolated occurrences in the European Alps. We analysed amplified fragment length polymorphism (AFLP) and chloroplast DNA sequence data to unravel the history of their immigration into the Alps and to provide data on their circumpolar phylogeography. In spite of the similar ecological requirements of the two species, they exhibit strikingly different immigration histories into the Alps. In M. biflora, the Alpine populations are most probably derived from source populations located between the Alpine and Scandinavian ice sheets, in accordance with the traditional biogeographic hypothesis. In contrast, the Alpine populations of R. pygmaeus cluster with those from the Tatra Mountains and the Taymyr region in northern Siberia, indicating that the distant Taymyr area served as source for the Alpine populations. Both species showed different levels of genetic diversity in formerly glaciated areas. In contrast to the considerable AFLP diversity observed in M. biflora, R. pygmaeus was virtually nonvariable over vast areas, with a single phenotype dominating all over the Alps and another, distantly related one dominating the North Atlantic area from Greenland over Svalbard to Scandinavia. The same pattern was observed in chloroplast DNA sequence data. Thus, postglacial colonization of R. pygmaeus was accompanied by extreme founder events.

Caryophyllaceae↗
PubMed2005

Seasonal adaptations of the tuberous roots of Ranunculus asiaticus to desiccation and resurrection by changes in cell structure and protein content.

The annual developmental cycle of tuberous roots of Ranunculus asiaticus was studied with respect to structure and content of their cells, to understand how these roots are adapted to desiccation, high temperature and rehydration. Light microscopy, histochemical analysis, and protein analyses by SDS-PAGE were employed at eight stages of annual root development. During growth and maturation of the roots, cortical cells increased in size and their cell walls accumulated pectin materials in a distinct layer to the inside of the primary walls, with pits between adjoining cells. The number of starch granules and protein bodies also increased within the cells. Several discrete proteins accumulated. Following quiescence and rehydration of the roots there was a loss of starch and proteins from the cells, and cell walls decreased in thickness. The resurrection geophyte R. asiaticus possesses desiccation-tolerant annual roots. They store carbon and nitrogen reserves within their cells, and pectin within the walls to support growth of the plant following summer quiescence and rehydration.

Adaptation, Physiological↗
PubMed2006

Connectivity, habitat heterogeneity, and population persistence in Ranunculus nodiflorus, an endangered species in France.

Here, we explore the role of habitat spatial structure in the maintenance of metapopulations of Ranunculus nodiflorus. This rare species grows in puddles that can be connected occasionally by flooded corridors. We monitored five locations in the Fontainebleau forest, France, since 2002 and recorded the presence of corridors among puddles and evaluated their impact on puddle demography and plant fitness. We showed that connections increased population size, by increasing both the number of puddles occupied by the species and the density of individuals within puddles, but seemed to have no direct influence on plant fitness. We found no evidence of a large persistent soil seed bank. Natural corridors are likely to decrease the extinction probability of the populations, most probably by allowing recolonization of empty puddles after extinctions. Therefore, the preservation of corridors appears crucial for the conservation of R. nodiflorus in its natural habitat.

Ecosystem↗
PubMed2006

The role of hybridization, polyploidization and glaciation in the origin and evolution of the apomictic Ranunculus cassubicus complex.

The Ranunculus cassubicus complex, comprising diploids and polyploids, is a good model for studying the role of hybridization and polyploidy in the origin of apomixis. Results from amplified fragment length polymorphism (AFLP) and simple sequence repeat (SSR) analyses performed on 448 individuals were combined with evidence from morphology, isozymes, karyology and distribution. Our results indicated a unique hybrid origin for the apomictic hexaploid R. carpaticola from north-western Slovakia, involving two sexual parents: autotetraploid R. cassubicifolius from the northern pre-Alps, and diploid R. carpaticola from central Slovakia. The hybrids were intermediate to the parents, but unique alleles have resulted from genomic reorganisation in the allopolyploids, which might also have triggered apomixis. Their distribution patterns and estimated ages suggest that hybridization may be correlated with the last glacial period. Hybridization seems to be the major origination for apomicts in the R. cassubicus complex. Polyploidy creates novel sexual genotypes and acts as a springboard for the production of new hybrids, but it only results in a combination with hybridization in apomixis. In turn, asexuality has permitted the perpetuation and establishment of ecologically divergent hybrid genotypes.

Alleles↗
PubMed2006

Evolution of hypervariable microsatellites in apomictic polyploid lineages of Ranunculus carpaticola: directional bias at dinucleotide loci.

Microsatellites are widely used in genetic and evolutionary analyses, but their own evolution is far from simple. The mechanisms maintaining the mutational patterns of simple repeats and the typical stable allele-frequency distributions are still poorly understood. Asexual lineages may provide particularly informative models for the indirect study of microsatellite evolution, because their genomes act as complete linkage groups, with mutations being the only source of genetic variation. Here, we study the direction of accumulated dinucleotide microsatellite mutations in wild asexual lineages of hexaploid Ranunculus carpaticola. Whereas the overall number of contractions is not significantly different from that of expansions, the within-locus frequency of contractions, but not of expansions, significantly increases with allele length. Moreover, within-locus polymorphism is positively correlated with allele length, but this relationship is due solely to the influence of contraction mutations. Such asymmetries may explain length constraints generally observed with microsatellites and are consistent with stable, bell-shaped allele-frequency distributions. Although apomictic and allohexaploid, the R. carpaticola lineages show mutational patterns resembling the trends observed in a broad range of organisms, including sexuals and diploids, suggesting that, even if not of germline origin, the mutations in these apomicts may be the consequence of similar mechanisms.

Cell Lineage↗
PubMed2005

[Studies on the chemical constituents in herb of Ranunculus sceleratus].

OBJECTIVE: To isolate and determine the chemical constituents from Ranunculus sceleratus. METHOD: Six compounds were isolated and purified by extraction and different kinds of column chromatography. The structures were determined on the basis of extensive spectral analysis such as EI-MS, 1H-NMR, 13C-NMR, DEPT and NOEDS. RESULT: The structures were elucidated as stigmasta-4-ene-3,6-dione I, stigmasterol II, isoscopoletin III, scoparone IV, protocatechuic aldehyde V and protocatechuic acid VI, respectively. CONCLUSION: Compound I, III, IV, V and VI were isolated from R. genus for the first time.

Benzaldehydes↗
PubMed2004

[The effect of the extract from Ranunculus japonicus on [Ca2+]i inside rabbit VSMC by serologic pharmacological test].

OBJECTIVE: To observe the influence of serum containing the extract from Ranunculus japonicus (ERj) on the amount of [Ca2+]i inside vascular smooth muscle cells (VSMC). METHODS: Serum containing ERj was prepared by serum pharmacological method. Cultured VSMCs induced by NE were treated with the serum, and the influence of it on the amount [Ca2+]i inside cells indicated with INDO-AM was determined. RESULTS: The serum containing ERj could markedly decrease the increase of the amount of free [Ca2+]i induced by NE in cultured VSMC. CONCLUSION: The serum containing ERj could decrease the intracellular [Ca2+]i.

Animals↗
PubMed2004

[Study on peparation of anemonin from Ranunculus japonicus Thunb].

OBJECTIVE: To study the preparation of anemonin from Ranunculus japonicus Thunb. METHODS: The best conditions of preparation for anemonin were optimized with monosyllabic experiments, such as using the purity of anemonin as a index for best selection of dying tempreture and the extraction ratio as another index for the concentration of NaCl and extracting times. The structures were clucidated on the basis of UV, IR, 1HNMR, 13CNMR, 1H-1H COSY, HMQC and HMQC spectra and chemical reactions and the purity is measured by HPLC. RESULTS: The best conditions for the extraction techniques of anemonin were 15% NaCl, four extracting times, 60 degrees C (0.08 Mpa) for drying. The purity of anemonin was 99% and yield rate was 0. 72 per thousand. Its quality was controlled. CONCLUSION: The preparation techniques are feasible and convenient.

Arabidopsis Proteins↗
PubMed2005

Ecological factors influencing tetraploid speciation in snow buttercups (Ranunculus Adoneus): niche differentiation and tetraploid establishment.

Chromosome doubling plays an important role in generating new species of flowering plants. However, reproductive incompatibilities between newly formed tetraploid plants and their diploid progenitors are expected to create a significant barrier to the persistence and establishment of neopolyploid populations. Ecological differentiation can reduce this barrier via prezygotic isolation arising from spatial separation. Alternatively, superior viability or fecundity of neotetraploid plants might compensate for the reproductive cost of incompatible pollen from diploid neighbors. The performance of plants of both cytotypes can be assessed in their respective habitats through reciprocal transplants, although such experiments have not been used previously in the study of tetraploid speciation. We used a series of seed and seedling transplant experiments to assess ecological differentiation and competitive ability during early establishment phases for tetraploid and diploid forms of the snow buttercup (Ranunculus adoneus). At two sites, seeds from diploids and tetraploids had similar germination probabilities. Tetraploid snow buttercup seedlings had a significant growth advantage in a controlled environment chamber experiment. However, in the field diploid and tetraploid buttercup seedlings did not differ consistently in survival or growth, nor did the two cytotypes show reciprocal advantages in performance, as expected if ecological differentiation has occurred. At the seed and seedling stages, neither niche differentiation nor tetraploid competitive superiority appears sufficient to explain neotetraploid success in the presence of their diploid progenitors.

Adaptation, Biological↗
PubMed1978

Antifungal properties of leaf extract of Ranunculus sceleratus L.

Studies on various antifungal properties of the leaf extraxt of Ranunculus sceleratus L. showed that it was thermostable up to 100 degrees C, retained activity on autoclaving, and remained active up to 15 days at room temperature. It possessed quick fungicidal action, tolerance against heavy fungal inoculum, activity on broad pH range, broad fungicidal spectrum, non-phytotoxicity and non-systemic activity. The extract was lethal at 1:40 dilution and its volatile vapours were also fungitoxic.

Antifungal Agents↗
PubMed1990

New procedure for direct bioautographic TLC assay as applied to a tincture of Ranunculus bulbosus.

A new procedure for TLC bioautographic assay using a flooding method to inoculate the developed plates is described. Details about several microbiological parameters allow good reproducibility for this method which appears rapid, easy to perform and requiring no specialized equipment. Significantly results obtained with a commercial tincture of Ranunculus bulbosus illustrate this method which can be extended to complex mixtures of natural products in order to isolate active fractions.

Biological Assay↗
PubMed2003

Cold- and light-induced changes of metabolite and antioxidant levels in two high mountain plant species Soldanella alpina and Ranunculus glacialis and a lowland species Pisum sativum.

Leaves of the two cold-acclimated alpine plant species Ranunculus glacialis and Soldanella alpina and, for comparison, of the non-acclimated lowland species Pisum sativum were illuminated with high light intensity at low temperature. The light- and cold-induced changes of antioxidants and of the major carbon and phosphate metabolites were analysed to examine which metabolic pathways might be limiting in non-acclimated pea leaves and whether alpine plants are able to circumvent such limitation. During illumination at low temperature pea leaves accumulated high quantities of sucrose, glucose-6-phosphate, fructose-6-phosphate, mannose-6-phosphate and phosphoglycerate (PGA) whereas ATP/ADP-ratios decreased. Although the PGA content also increased in leaves of R. glacialis the other metabolites did not accumulate and ATP/ADP-ratios remained fairly constant in either alpine species. These data indicate a inorganic phosphate (Pi)-limitation in the chloroplasts of pea leaves but not in the alpine species. However, the total phosphate pool and the percentage of free Pi were highest in pea and did not change during illumination in cold. In contrast, free Pi contents declined markedly in R. glacialis leaves, suggesting that Pi is available for metabolism in this species. In S. alpina leaves contents of ascorbate and glutathione doubled in light and cold, while the contents of sugars did not increase. Obviously, S. alpina leaves can use assimilated carbon for ascorbate synthesis, rather than for the synthesis of sugars. A high capacity for ascorbate synthesis might prevent the accumulation of mannose-6-phosphate and Pi-limitation.

Journal Article↗
PubMed2002

Uptake of cadmium by the invasive perennial weeds Ranunculus repens and Geranium robertianum under laboratory conditions.

The objective of this study was to compare the accumulation and partitioning of cadmium (Cd) in a fibrous versus a tap root weed, Ranunculus repens and Geranium robertianum respectively. To meet this objective, we compared the accumulation by and the partitioning of Cd in R repens versus G. robertianum grown in soils spiked with 0.015 grams of Cd for a period of three weeks. The rate of Cd uptake was also compared by following the fate of 109Cd within the root, stem and leaf of the two weeds. Prior to Cd exposure, leaf and stem of control R. repens contained significantly greater amounts of Cd as compared to G. robertianum, whereas Cd concentrations in roots of the control plants for the two species were not significantly different (p > 0.05, student's t-test). Post Cd exposure the two species contained similar amounts of Cd in leaf and stem, however, roots of R. repens contained almost two-fold the amounts of Cd as compared to G. robertianum. Comparison of k (h(-1), rate of 109Cd uptake) for stem, leaf and root of the two species indicated that G. robertianum accumulated 109Cd over the first 24-48 h at a faster rate as compared to R. repens. For both species and all three organs, maximum accumulation of 109Cd occurred within the first 24-48 h. Our findings indicate that the fate of Cd within these two species is quite different with the fibrous root of R. repens serving to accumulate and store Cd whereas in G. robertianum, Cd is rapidly taken up and tends to be accumulated within its leaf.

Cadmium↗
PubMed2003

Out of the Alps: colonization of Northern Europe by East Alpine populations of the Glacier Buttercup Ranunculus glacialis L. (Ranunculaceae).

Ranunculus glacialis ssp. glacialis is an arctic-alpine plant growing in central and southern European and Scandinavian mountain ranges and the European Arctic. In order to elucidate the taxon's migration history, we applied amplified fragment length polymorphism (AFLP) to populations from the Pyrenees, Tatra mountains and Northern Europe and included data from a previous study on Alpine accessions. Populations from the Alps and the Tatra mountains were genetically highly divergent and harboured many private AFLP fragments, indicating old vicariance. Whereas nearly all Alpine populations of R. glacialis were genetically highly variable, the Tatrean population showed only little variation. Our data suggest that the Pyrenees were colonized more recently than the separation of the Tatra from the Alps. Populations in Northern Europe, by contrast, were similar to those of the Eastern Alps but showed only little genetic variation. They harboured no private AFLP fragments and only a subset of East Alpine ones, and they exhibited no phylogeographical structure. It is very likely therefore that R. glacialis colonized Northern Europe in postglacial times from source populations in the Eastern Alps.

DNA Primers↗
PubMed1992

Analgesic and anti-inflammatory effects of Ranunculus japonicus extract.

The analgesic and anti-inflammatory effects of Ranunculus japonicus extract after parenteral administration were determined in several animal models. The extract inhibited the mice writhing responses caused by acetic acid and raised the pain thresholds of mice in the hot-plate test. The extract also inhibited the paw edema of rats induced by carrageenin, ear swelling of mice caused by xylene, mice vascular permeability increase induced by acetic acid, and granuloma formation in rats.

Analgesics↗