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Photocontrol of flowering and stem extension of the intermediate-day plant Echinacea purpurea.

Intermediate-day plants (IDP) flower most rapidly and completely under intermediate photoperiods (e.g., 12-14 h of light), but few species have been identified and their flowering responses are not well understood. We identified Echinacea purpurea Moench as an IDP and, based on our results, propose a novel mechanism for flowering of IDP. Two genotypes of E. purpurea ('Bravado' and 'Magnus') flowered most completely (>/=79%) and rapidly and at the youngest physiological age under intermediate photoperiods of 13-15 h. Few (</=14%) plants flowered under 10- or 24-h photoperiods, indicating E. purpurea is a strongly quantitative IDP. Plants were also induced to flower when 15-h dark periods were interrupted with as few as 7.5 min of low-intensity lighting (night interruption, NI). Flowering was progressively earlier as the NI increased to 1 h, but was delayed when the NI was extended to 4 h. Stem length increased by >/=230% as the photoperiod or NI duration increased, until plants received a saturating duration (at 14 or 1 h, respectively). Flowering was inhibited when 16-h photoperiods were deficient in red (R, 600-700 nm) light, and was promoted when photoperiods were deficient in far-red (FR, 700-800 nm) light. Because of our results, we propose the flowering behavior of IDP such as E. purpurea is composed of two mechanisms: a light-dependent response operating through light-labile (type I) phytochrome in which flowering is inhibited by an LD, and a light-stable (type II) phytochrome (i.e., phyB, D and E) response in which flowering is promoted by a short-night.

Journal Article↗

In vitro inhibition of cyclooxygenase and 5-lipoxygenase by alkamides from Echinacea and Achillea species.

Polyunsaturated alkamides isolated from Achillea species, Echinacea angustifolia DC., Anacyclus pyrethrum (L.) Link, and Aaronsohnia pubescens (Desf.) Bremer & Humphries, (Compositae) were shown to possess inhibitory activity in in vitro cyclooxygenase (sheep seminal microsomes) and 5-lipoxygenase (porcine leukocytes) assays. Activity appeared to depend on the particular structure of the alkamides.

Amides↗

Echinacea: anatomy, phytochemical pattern, and germination of the achene.

The achenes (fruits) of the therapeutically used Echinacea species E. purpurea, E. angustifolia, and E. pallida can be differentiated structurally (shape, anatomy) as well as phytochemically (essential oil components, alkamides). During germination in all three species dodeca-2E, 4E, 8Z, 10E(10Z)-tetraenoic acid isobutylamide (8/9) is mainly formed. Besides this a number of alkamides typical for the root of E. purpurea are synthesized in moderate amounts. Also, alkene derivatives of isovalerianic acid are produced. It is an interesting fact that neither 2-monoene alkalmides nor polyacetylenes could be detected during achene germination.

Plants, Medicinal↗

Analysis of alkamides and caffeic acid derivatives from Echinacea simulata and E. paradoxa roots.

The constituents of the roots of Echinacea simulata and E. paradoxa were examined by HPLC. The major lipophilic and hydrophilic compounds could be identified. E. paradoxa contained several ketoalkenynes and proved to have almost identical constituents as E. pallida. E. simulata contained alkamides as found in E. angustifolia and in addition ketoalkenynes as in E. pallida. Echinacoside was found in both species.

Amides↗

Macrophage activation by the polysaccharide arabinogalactan isolated from plant cell cultures of Echinacea purpurea.

In this study, acidic arabinogalactan, a highly purified polysaccharide from plant cell cultures of Echinacea purpurea, with a molecular weight of 75,000, was effective in activating macrophages to cytotoxicity against tumor cells and micro-organisms (Leishmania enriettii). Furthermore, this polysaccharide induced macrophages to produce tumor necrosis factor (TNF-alpha), interleukin-1 (IL-1), and interferon-beta 2. Arabinogalactan did not activate B cells and did not induce T cells to produce interleukin-2, interferon-beta 2, or interferon-gamma, but it did induce a slight increase in T-cell proliferation. When injected ip, this agent stimulated macrophages, a finding that may have therapeutic implications in the defense against tumors and infectious diseases.

Animals↗

Dual induction rather than intermediate daylength response of flowering in Echinacea purpurea.

Echinacea purpurea cv. Bravado and Magnus have been reported to be intermediate daylength plants (IDP) which flower in response to photoperiods between 13 and 16 h. The present experiments with E. purpurea cv. Bravado show that E. purpurea is actually a dual induction short-long-day plant which flowers promptly and consistently when grown in short day (SD) followed by long day (LD) conditions, but not with the reverse sequence of photoperiods. The flowering response increased with increasing duration of both the SD and the LD treatments. A minimum of 4 weeks of SD followed by 12 LD was required for complete flowering. No flowering occurred in continuous SD or LD, whereas a high proportion of plants flowered in continuous 14-h daylength. However, flowering was more variable in intermediate daylength than after transition from SD to LD. Furthermore, photoperiods between 13 and 16 h could satisfy both the primary SD induction and the secondary LD induction requirements. As a number of dual induction plants, both short-long-day and long-short-day plants, have such an overlapping window of effective photoperiods that can trigger both the SD and LD responses, the rationale for maintaining IDP as a separate and genuine flowering response group is seriously challenged.

Journal Article↗

Anti-inflammatory activity of a polysaccharidic fraction of Echinacea angustifolia.

The anti-inflammatory activity of a polysaccharidic fraction (EPF) obtained from Echinacea angustifolia roots has been examined using the carrageenan paw oedema and the croton oil ear test. EPF (0.5 mg kg-1 i.v.) almost inhibited the carrageenan-induced oedema over 8 h and furthermore, EPF, topically applied, inhibited mouse ear oedema induced by croton oil. EPF also reduced the leukocytic infiltration of the croton oil dermatitis, evaluated both as peroxidase activity and histologically. After topical application EPF appears to be slightly inferior in potency to indomethacin. The results suggest that the anti-inflammatory activity of E. angustifolia resides in its polysaccharidic content.

Administration, Topical↗

Macrophage activation and induction of macrophage cytotoxicity by purified polysaccharide fractions from the plant Echinacea purpurea.

Purified polysaccharides (EPS) prepared from the plant Echinacea purpurea are shown to strongly activate macrophages. Macrophages activated with these substances develop pronounced extracellular cytotoxicity against tumor targets. The activation is brought about by EPS alone and is independent of any cooperative effect with lymphocytes. Also the production and secretion of oxygen radicals and interleukin 1 by macrophages is increased after activation with EPS. Cells of the macrophages lineage seem to be the main target for the action of these polysaccharides. EPS has no effect on T lymphocytes. B lymphocytes show a comparatively modest proliferation after incubation with E. purpurea EPS. Thus, these compounds, which are at least in tissue culture completely nontoxic, may be suited to activate in vivo cells of the macrophage system to cytotoxicity. They may therefore be of relevance in tumor and infectious systems.

Adjuvants, Immunologic↗

Immunomodulation with Viscum album and Echinacea purpurea extracts.

Extracts of Viscum album (Plenosol) and Echinacea purpurea (Echinacin) are used clinically for their non-specific action on cell-mediated immunity. In vitro we could prove that these two extracts have a stimulating effect on the production of lymphokines by lymphocytes and in the transformation test. A toxic effect on cells was produced only with very high, clinically irrelevant concentrations. Clinical application of these extracts can produce a stimulation of cell-mediated immunity (one therapeutic administration followed by a free interval of one week) or can have a depressive action (daily administrations of higher doses). These observations were confirmed by lymphokine production and assay, 3H-thymidine incorporation and a skin test with recall antigens (Multitest Merieux).

Cell Migration Inhibition↗

Naturally occurring insect growth regulators. III. Echinolone, a highly active juvenile hormone mimic from Echinacea angustifolia roots.

A compound, C14H24O2, which induces strong juvenilizing effects in the yellow mealworm, Tenebrio molitor L., following topical application to the pupal stage, was isolated in pure form from the roots of Echinacea angustifolia DC and tentatively identified as dextrorotatory (E)-10-hydroxy-4,10-dimethyl-4,11-dodecadien-2-one. The compound has been named echinolone.

Biological Assay↗

Patterns of Variation in Alkamides and Cichoric Acid in Roots and Aboveground Parts of Echinacea purpurea (L.) Moench.

We investigated patterns of variation in alkamides and cichoric acid accumulation in the roots and aboveground parts of Echinacea purpurea (L.) Moench. These phytochemicals were extracted from fresh plant parts with 60% ethanol and quantified by high performance liquid chromatography (HPLC) analysis. Concentrations of alkamides and cichoric acid were measured on a dry-weight basis (mg·g(-1)). For total alkamides, concentrations among individual plants varied from 5.02 to 27.67 (mean = 14.4%) in roots, from 0.62 to 3.42 (mean = 1.54) in nearly matured seed heads (NMSH), and 0.22 to 5.25 (mean = 0.77) in young tops (about ½ flower heads, ¼ leaves, and ¼ stems). For cichoric acid, concentrations among individual plants varied from 2.65 to 37.52 (mean = 8.95), from 2.03 to 31.58 (mean = 10.9), and from 4.79 to 38.55 (mean = 18.88) in the roots, the NMSH, and the tops, respectively. Dodeca-2E, 4E, 8Z, 10E-tetraenoic acid isobutylamide and dodeca-2E, 4E, 8Z, 10Z-tetraenoic acid isobutylamide (alkamides 8/9) accounted for only 9.4% of the total alkamides in roots, but comprised 87.9% in the NMSH, and 76.6% in the young tops. Correlations of concentrations of alkamides or cichoric acid between those of roots and those of the NMSH were not statistically significant, and either within the roots, the NMSH, and the young tops. However, a significant negative correlation was observed between the concentration of cichoric acid in the roots and in young tops, and a significant positive correlation was observed between total alkamide concentration in the roots and cichoric acid concentration in the young tops. These results may be useful in the genetic improvement of E. purpurea for medicinal use.

Journal Article↗

Ethephon Promotes Germination of Echinacea angustifolia and E. pallida in Darkness.

Seeds from five lots each of Echinacea angustifolia DC, and E. pallida (Nutt.) Nutt. were germinated in a growth chamber in light (40 μmol·m(-2)· s(-1)) or darkness at 25 °C for 16 to 20 d after soaking in 1 mM ethephon or water for 10 min, or moist stratification at 4 - 6 °C for two weeks. Either light or ethephon promoted seed germination of E. angustifolia and E. pallida, in comparison with darkness in nine of ten lots. Ethephon in the dark had similar or greater germination percentages than water with light. Ethephon with light improved germination in three of ten lots compared with ethephon in the dark. The effect of cold, moist stratification in comparison with darkness varied by seed lot. Five lots of E. purpurea (L.) Moench were tested; however, no treatment differences were measured. The finding that ethethon promoted E. angustifolia and E. pallida seed germination in darkness could be useful in the cultivation of these two species. Chemical name used: 2-chloroethylphosphonic acid (ethephon).

Journal Article↗

Toxicity of Echinacea purpurea. Acute, subacute and genotoxicity studies.

Single oral or intravenous doses of the expressed juice of Echinacea purpurea (EP) proved virtually non-toxic to rats and mice. After 4 weeks of oral administration in doses amounting to many times the human therapeutic dose laboratory tests and necropsy findings gave no evidence of any toxic effects in rats. Tests for mutagenicity carried out in microorganisms and mammalian cells in vitro and in mice all gave negative results. In an in vitro carcinogenicity study EP did not produce malignant transformation in hamster embryo cells.

Animals↗

[Modification of avian humoral immunoreactions by Influex and Echinacea angustifolia extract].

Medicinal complex drugs as well as single ethanolic or aqueous extracts of several plants are commonly used to increase the natural resistance to various infections, though their efficacy and mechanism of action are not yet well elucidated. In the present study, we investigated two problems: firstly, whether the complex drug (Influex) and Echinacea angustifolia extract do stimulate the immunoglobulin and antibody synthesis in chickens immunized with human serum albumin; and secondly, whether a restoration of IgG-synthesis in immunodefective (dysgammaglobulinemic) UM-B 19 chickens is possible with these plant preparations, i.e. if the BG cells which may possibly be present can be polyclonally or antigen specifically stimulated. The preparations were administered orally in two doses, after which the complete immunoglobulin concentration was determined by rocket immunoelectrophoresis and the antibody production by ELISA. The effect of ethanolic solvent was taken into account. The administration of the complex drug to normal Leghorn chickens induced a rise in the serum immunoglobulin concentration, as well as an increase in the three classes of antibody. By the immunodeficient chickens (IgG concentration was below the level of test sensitivity at the start), the administration of the drug led to a slight production in IgG and antibody.

Animals↗

[Immunologic in vivo and in vitro studies on Echinacea extracts].

Ethanolic extracts of Echinacea purpurea, E. pallida and E. angustifolia roots were examined for immunological activity in the carbon clearance test with mice and in the granulocyte test. In the in vivo experiment all extracts, administered orally, were found to enhance phagocytosis significantly. These results correlate with the stimulation of phagocytosis in the in vitro granulocyte test. The lipophilic fractions of the extracts appeared to be more active than the polar fractions. All extracts were analyzed by HPLC in order to correlate the chemical constituents with the immunological activities.

Animals↗

Direct characterization of caffeoyl esters with antihyaluronidase activity in crude extracts from Echinacea angustifolia roots by fast atom bombardment tandem mass spectrometry.

Fast atom bombardment (FAB-MS) and fast atom bombardment tandem mass spectrometry (FAB-MS/MS) techniques (negative ions) have been successfully applied for identification of the constituents responsible for the antihyaluronidase activity of Echinacea angustifolia roots, whose extracts are widely employed for the adjuvant therapy of chronic inflammatory diseases. Crude extracts from different solvents were tested for antihyaluronidase activity, and those with the greatest inhibitory action (the ethylacetate, butylacetate and chloroform fractions, IC50 0.44, 0.50 e 0.62 mg/ml) were directly analyzed by MS. Full scan mass spectra produced intense molecular anions: collisional activation of these resulted in tandem mass spectra rich in significant product ions. Four main caffeoyl conjugates were detected and identified by tandem mass spectrometry (daughter and parent ion mode): 2,3-O-dicaffeoyltartaric acid (chicoric acid) and 5-O-dicaffeoylquinic acid (cynarine) and 2-O-caffeoyltartaric acid (caffaric acid) in the ethylacetate fraction. Among these caffeoyl conjugates, chicoric and caftaric acids had the greatest antihyaluronidase activity: IC50 = 0.42 and 0.61 mM, while the IC50 of cynarine and chlorogenic acid were 1.85 and 2.25 mM.

Acetates↗