Euphorbia Characias Research Articles

ECPUB5 Polyubiquitin Gene in Euphorbia characias: Molecular Characterization and Seasonal Expression Analysis.

The spurge Euphorbia characias is known for its latex, which is rich in antioxidant enzymes and anti-phytopathogen molecules. In this study, we identified a novel polyubiquitin protein in the latex and leaves, leading to the first molecular characterization of its coding gene and expressed protein in E. characias. Using consensus-degenerate hybrid oligonucleotide primers (CODEHOP) and rapid amplification of cDNA ends (5'/3'-RACE), we reconstructed the entire open reading frame (ORF) and noncoding regions. Our analysis revealed that the polyubiquitin gene encodes five tandemly repeated sequences, each coding for a ubiquitin monomer with amino acid variations in four of the five repeats. In silico studies have suggested functional differences among monomers. Gene expression peaked during the summer, correlating with high temperatures and suggesting a role in heat stress response. Western blotting confirmed the presence of polyubiquitin in the latex and leaf tissues, indicating active ubiquitination processes. These findings enhance our understanding of polyubiquitin's regulatory mechanisms and functions in E. characias, highlighting its unique structural and functional features.

Effects of Euphorbia characias subsp. characias flower extracts on nociceptive pain and acute inflammatory models in mice

Pain and inflammation are major health issues worldwide, leading to negative consequences. Despite several drugs being available to manage these conditions, their effectiveness can be limited by cost, adverse reactions, and potential tolerance and dependence with long-term use. Euphorbia characias traditionally used in folk medicine for its diverse biological activities - including antiproliferative, antimicrobial, and anti-inflammatory effects - has not been extensively studied in vivo for its analgesic and anti-inflammatory properties. In this study, the antinociceptive and anti-inflammatory properties of the water and ethanolic extracts of E. characias flowers (ECAEFl and ECEEFl) were evaluated using various models. Both extracts significantly reduced paw licking time in a formalin-induced paw licking model, with ECAEFl specifically targeting and ECEEFl affecting both the neurogenic and inflammatory phases. Additionally, in the carrageenan-induced cell migration model, both extracts showed a significant decrease in leukocyte migration, protein extravasation and nitric oxide levels, further demostrating their anti-inflammatory activity. High-Resolution HPLC-ESI-QTOF-MS-MS and HPLC-PDA analysis characterized the chemical composition of the extracts, identifying a significant presence of phenolic compounds, particularly quercetin and its derivatives, which likely contribute to the observed biological activities. These findings highlight the potential of E. characias extracts as natural sources of compounds with antinociceptive and anti-inflammatory properties. Further investigations are needed to elucidate the underlying mechanisms and explore their therapeutic potential in pain and inflammation-related disorders.

Euphorbia characias Extract: Inhibition of Skin Aging-Related Enzymes and Nanoformulation.

Plant extracts have long served as important sources of bioactive compounds, and they are currently the focus of extensive research in the development of novel preventive and therapeutic strategies. However, their health benefits are often limited by low bioavailability. Nanoparticle delivery systems can represent a solution to such limitations. Euphorbia characias is a Mediterranean shrub known to have biological activities, such as inhibiting tyrosinase and showing a potential role as a skin-whitening agent. In this study, an ethanolic extract from E. characias leaves was tested for its inhibitory activity on skin-related enzymes, such as elastase, collagenase, and hyaluronidase, and for sun protection factors. Moreover, the extract was formulated in phospholipid vesicles to improve its local bioavailability and applicability. The vesicles were characterized by size, surface charge, storage stability, and entrapment efficiency. The nanoformulation was also evaluated for antioxidant activity and assayed for cytocompatibility and anti-tyrosinase activity in melanoma cells. Our findings demonstrated that the extract has a photo-protective effect and enzyme-inhibitory properties. E. characias nanoformulation was also cytocompatible and improved the extract’s activity in the cells, suggesting a potential skin application for antimelanogenic treatments and confirming the key role of nanotechnological approaches to maximize plant extract’s potentialities.

Symmetric versus Asymmetric Features of Homologous Homodimeric Amine Oxidases: When Water and Cavities Make the Difference

Symmetry is an intrinsic property of homo-oligomers. Amine oxidases are multidomain homodimeric enzymes that contain one catalytic site per subunit, and that share a high homology degree. In this paper, we investigated, by fluorescence spectroscopy measurements, the conformational dynamics and resiliency in solutions of two amine oxidases, one from lentil seedlings, and one from Euphorbia characias latex, of which the crystallographic structure is still unknown. The data demonstrate that slight but significant differences exist at the level of the local tridimensional structure, which arise from the presence of large internal cavities, which are characterized by different hydration extents. Molecular dynamics and a contact network methodology were also used to further explore, in silico, the structural features of the two proteins. The analysis demonstrates that the two proteins show similar long-range symmetrical connectivities, but that they differ in their local (intra-subunit) contact networks, which appear mostly asymmetric. These features have been interpreted to suggest a new rationale for the functioning of amino oxidases as obligate homodimers.

Euphorbia characias: Phytochemistry and Biological Activities.

The aim of this review is to summarize all the compounds identified and characterized from Euphorbia characias, along with the biological activities reported for this plant. Euphorbia is one of the greatest genera in the spurge family of Euphorbiaceae and includes different kinds of plants characterized by the presence of milky latex. Among them, the species Euphorbia characias L. is an evergreen perennial shrub widely distributed in Mediterranean countries. E. characias latex and extracts from different parts of the plant have been extensively studied, leading to the identification of several chemical components such as terpenoids, sterol hydrocarbons, saturated and unsaturated fatty acids, cerebrosides and phenolic and carboxylic acids. The biological properties range between antioxidant activities, antimicrobial, antiviral and pesticidal activities, wound-healing properties, anti-aging and hypoglycemic properties and inhibitory activities toward target enzymes related to different diseases, such as cholinesterases and xanthine oxidase. The information available in this review allows us to consider the plant E. characias as a potential source of compounds for biomedical research.

Structure of a nucleotide pyrophosphatase/phosphodiesterase (NPP) from Euphorbia characias latex characterized by small-angle X-ray scattering: clues for the general organization of plant NPPs.

Little information is available concerning the structural features of nucleotide pyrophosphatase/phosphodiesterases (NPPs) of plant origin and the crystal structures of these proteins have not yet been reported. The aim of this study was to obtain insight into these aspects by carrying out a comparative analysis of the sequences of two different fragments of an NPP from the latex of the Mediterranean shrub Euphorbia characias (ELNPP) and by studying the low-resolution structure of the purified protein in solution by means of small-angle X-ray scattering. This is the first structure of a plant NPP in solution that has been reported to date. It is shown that the ELNPP sequence is highly conserved in many other plant species. Of note, the catalytic domains of these plant NPPs have the same highly conserved PDE-domain organization as mammalian NPPs. Moreover, ELNPP is a dimer in solution and this oligomerization state is likely to be common to other plant enzymes.

Use of Plant Water Extracts for Weed Control in Durum Wheat (Triticum turgidum L. Subsp. durum Desf.)

The use of plant water extracts to control weeds is gaining attention in environmentally-friendly agriculture, but the study of the effect that such extracts may exert on the yield of durum wheat is still unexplored. In 2014 and 2016, the herbicidal potential of several plant water extracts was field tested on durum wheat (cv Valbelice). In 2014, extracts obtained from Artemisia arborescens, Rhus coriaria, Lantana camara, Thymus vulgaris, and Euphorbia characias were used, whereas in 2016 only A. arborescens and R. coriaria were tested as “donor” plants. In both years, weed incidence was evaluated, together with the major yield parameters of wheat. None of the treatments (including chemicals) could eradicate weeds from the field. In 2014, dicots were in general prevailing in plots treated with extracts of E. characias, while monocots prevailed after treatments with L. camara and R. coriaria. In 2016, lower weed biomass and diversity level were found, and only Avena and Phalaris were detected at harvest time. Treatment with plant water extracts affected grain yields, but it seems likely that those effects are not due to the diverse incidence of weeds in treated and untreated plots, rather to some direct action exerted by allelopathic substances.

Taxonomical and chorological notes 10 (98–110)

The present part of the series provides new records of locally introduced native species in Hungary (Apium repens, Crypsis schoenoides, Limonium gmelinii, Rumex confertus, Sagina apetala). The spread of Crypsis schoenoides and Limonium gmelinii on roadsides is due to winter salting of roads, whereas the appearance of Apium repens and Sagina apetala is likely the consequence of the use of soil or peat material containing their seeds. The reports of Knautia dipsacifolia from the northern part of the Great Hungarian Plain and Spiraea crenata from the Hungarian–Romanian–Ukrainian border region is ascertained here concluding that the report of the first taxon was based on a misidentifi cation of K. drymeia, whereas the record of Spiraea crenata is a consequence of mislocation. New records of two spreading aliens (Eragrostis virescens, Sporobolus vaginifl orus) are discussed here along with a herbarium revision of Eragrostis, which showed that all previous Hungarian records of Eragrostis parvifl ora refer to E. virescens. Further species discussed in the paper escaped from cultivation (Asparagus verticillatus, Euphorbia characias, Nassella tenuissima, the latter two new for the country). The first record of the sedge hybrid Carex ×leutzii in Hungary is also presented.

In vivo wound-healing activity of Euphorbia characias subsp. wulfenii: Isolation and quantification of quercetin glycosides as bioactive compounds

Ethnopharmacological relevanceThe latex and the aerial parts of Euphorbia characias L. (Euphorbiaceae) have been used as medicinal plant to treat wounds and warts in traditional medicine. Aim of the studyThe effect of the plant extract was tested in vivo and in vitro with experimental models to find scientific evidence for traditional use in wound healing. Potentially active wound-healer compounds were isolated from the active fraction using fractionation procedures under the guidance of biological assay and the possible role of the compounds in the wound healing process was also determined. Material and methodsN-hexane, ethyl acetate, and methanol extracts were successively prepared from the aerial parts of E. characias subsp. wulfenii. The extracts were tested with linear incision, circular excision wound models and the hydroxyproline assay method to assess the wound-healing activity. The inhibition of the increase in capillary permeability induced by acetic acid, an acute inflammation model, was used to assay the anti-inflammatory activity. Different chromatographic separation techniques on sephadex and silica gel columns, and bioassay guided assay techniques have been used to isolate the active compounds of the plant. Moreover, hyaluronidase, collagenase and elastase enzymes inhibitory effect of active principle were investigated in vitro to find out the mechanism of action. ResultsThe methanol (MeOH-ex) extract of the aerial parts of E. characias subsp. wulfenii showed significant wound healing activity (linear incision wound model: 43.04%; circular excision wound model 65.24%) and anti-inflammatory activity (34.74%). The methanol extract was separated into its fractions by column chromatography for isolation of efficient compounds. Biological activity of the fractions were assessed and further isolation and purification processes have been carried out in the active fraction. Isolation studies were carried out from the MeOH-ex fraction to obtain active constituents and their structures were elucidated to be quercetin-3-O-rhamnoside (quercitrin), quercetin-3-O-galactoside (hyperoside), and quercetin-3-O-arabinoside (guaijaverin). Further in vitro and in vivo assays showed that quercetin derivatives were responsible for the wound-healing activity of the plant, and also found to be significant anti-elastase and anti-collagenase activities. The amounts of three compounds, isolated from active fraction, were determined by using high performance liquid chromatography. Calibration equation was calculated with dilutions, prepared from pure substances, and assay was performed in total extract, prepared from E. characias subsp. wulfenii. It was detected that the plant had 1.22% quercitrin, 0.35% hyperoside, and 0.11% guaijaverin. The validation of the analytical method was performed by linearity, precision, limit of detection, and limit of quantification parameters. ConclusionPresent study supported the traditional use of the aerial parts E. characias subsp. wulfenii as wound healer and quercetin derivatives were isolated as active components from the active fraction by using bioassay-guided fractionation technique.

Selected Enzyme Inhibitory Effects of Euphorbia characias Extracts.

Extracts of aerial part of Euphorbia characias were examined to check potential inhibitors for three selected enzymes involved in several metabolic disorders. Water and ethanol extracts from leaves and flowers showed in vitro inhibitory activity toward α-amylase, α-glucosidase, and xanthine oxidase. IC50 values were calculated for all the extracts and the ethanolic extracts were found to exert the best effect. In particular, for the α-glucosidase activity, the extracts resulted to be 100-fold more active than the standard inhibitor. The inhibition mode was investigated by Lineweaver-Burk plot analysis. E. characias extracts display different inhibition behaviors toward the three enzymes acting as uncompetitive, noncompetitive, and mixed-type inhibitors. Moreover, ethanolic extracts of E. characias showed no cytotoxic activity and exhibited antioxidant capacity in a cellular model. The LC-DAD metabolic profile was also performed and it showed that leaves and flowers extracts contain high levels of quercetin derivatives. The results suggest that E. characias could be a promising source of natural inhibitors of the enzymes involved in carbohydrate uptake disorders and oxidative stress.

Amine oxidase from Euphorbia characias: Kinetic and structural characterization.

This minireview focuses on a plant copper/2,4,5-trihydroxyphenyl alanine quinone amine oxidase isolated from the latex of the shrub Euphorbia characias (ELAO). This enzyme has been investigated in terms of both molecular structure and kinetic mechanism. The characterization of this enzyme allowed us to identify specific amino acids and domains that play a key role in modulating substrate access into the active site not only for ELAO but also for other plant and mammalian amine oxidases. As mammalian amine oxidases are implicated in several physiological and pathological conditions, the deep structural characterization of their active site accession mechanisms could be the starting point for the development of enzyme modulators with high therapeutic potential. Thus, this paper gives structural/functional insights that open new perspectives in the research about the whole amine oxidase family.

A chitinase from Euphorbia characias latex is a novel and powerful plant‐based pesticide against Drosophila suzukii

ABSTRACTDrosophila suzukii attacks on developing soft fruits have recently caused important economic losses in Europe. This study explores the effectiveness of a new control strategy against this insect pest that is based on a plant chitinase extracted from the latex of the Mediterranean spurge, Euphorbia characias. The ability of the purified Euphorbia latex chitinase (ELC) to degrade the chitin exoskeleton of D. suzukii was assessed using confocal laser scanning microscopy. ELC treatment caused reduced larval growth, higher mortality and notable degradation of external insect structures. Therefore, the chitinase may induce a double effect on the D. suzukii larvae, a direct injury on the larval bodies and an action as antifeedant. The effects of the ELC treatment were also tested on leaves of the insect's host plants, Fragaria × ananassa and Rubus idaeus, using physiological parameters (chlorophyll concentration, chlorophyll fluorescence, leaf gas exchange and water potential) and defence gene expression (FaPGIP, FaChi2_1 and FaChi2_2) as stress indicators. ELC at concentrations effective against D. suzukii did not damage the host plants. Only plant defence gene expression was somewhat enhanced during the early hours after ELC application. In conclusion, ELC, a natural product, proved to be an effective tool for use in the development of an environmentally friendly integrated management strategy against D. suzukii, a pest whose control by conventional chemical insecticides is problematic.

Plant teratologies as a result of phytoplasma infections

The direct correlation between teratological cases and phytoplasma infections was ascertained in spontaneous and cultivated plant species. Plants, belonging to 31 species and 12 families, showing symptoms of growth abnormalities were collected and analysed. Attempted detection of Rhodococcus fascians by isolation, PCR indexing and 16S rRNA sequencing from fasciated tissues allowed to exclude its presence. Nested PCR by universal primers and 16S rRNA sequence analyses indicated the presence of phytoplasmas, belonging to six groups, in the 44% of symptomatic samples. Among the infected species, Austrocylindropuntia exaltata, Opuntia subulata, Euphorbia characias, Euphorbia dendroides, Euphorbia linifolia, Euphorbia myrsinites, Rumex buchephalophorus, Linaria multicaulis and Fedia cornucopiae represent new phytoplasma hosts world-wide. Moreover this is the first report of phytoplasma belonging to subgroup 16SrRNA II-I in Italy. These findings together with the known erratic distribution in plant tissues of these phloem-restricted prokaryotes indicate a close correlation between fasciation and similar growth disorders and phytoplasma infections.

Two different in vitro tests confirm the blocking activity of d-galactose lectins on the adhesion of Escherichia coli F4 to pig brush border receptors

The pathogenicity of Enterotoxigenic Escherichia coli (ETEC) expressing F4ac (K88) fimbrial adhesin and its adhesion to the pig jejunum surface is genetically controlled by the presence of a specific receptor glycoprotein on brush borders of the epithelium. We firstly screened the degree of inhibition obtained with ten galactose-recognising lectins – respectively from Abrus precatorius (APA-I); Coregonus maraena; Euphorbia characias; Crotalaria juncea; Sambucus ebulus (SEL1d); Sambucus nigra (SNAV); Coregonus peled; Momordica charantia (MCL); Trychosanthes kirilowii; Adenia racemosa –, by an ELISA test based on intestinal brush border sensitive for E. coli F4ac adhesion and ETEC F4ac fimbrial antigen. Three lectins significantly inhibited the fimbrial adhesion and the highest value was obtained by Crotalaria juncea lectin (p < .01). The results were then confirmed by an in vitro inhibition test of ETEC adhesion to villi isolated from ETEC susceptible pigs. The favourable results obtained with the supplementation of two d-galactose lectins against the ETEC adhesion to intestinal villi stimulate in vivo testing of the addition of these lectins in the piglet feeding

Quality Characteristics of Wholemeal Flour and Bread from Durum Wheat (Triticum turgidum L subsp. durum Desf.) after Field Treatment with Plant Water Extracts.

The use of selected plant water extracts to control pests and weeds is gaining growing attention in organic and sustainable agriculture, but the effects that such extracts may exert on the quality aspects of durum wheat are still unexplored. In 2014, 5 plant water extracts (Artemisia arborescens, Euphorbia characias, Rhus coriaria, Thymus vulgaris, Lantana camara) were prepared and distributed on durum wheat cv Valbelice to evaluate their potential herbicidal effects. After crop harvesting, the major physicochemical and technological parameters of wholemeal flours obtained from each treatment were measured and compared with those from chemical weeding and untreated controls. A baking test was also performed to evaluate the breadmaking quality. In wholemeal flours obtained after the treatment with plant extracts protein and dry gluten content were higher than in control and chemical weeding. Wholemeal flours obtained after chemical weeding reached the highest Mixograph parameters, and that from durum wheat treated with R. coriaria extract demonstrated a very high α-amylase activity. We concluded that the treatments with plant water extracts may influence many quality traits of durum wheat. This occurrence must be taken into account in overall decisions concerning the use of plant extracts in pest and weed management practice.

Anti-inflammatory and wound healing activity of Euphorbia characias and bioassay-guided isolation of some flavonoids

Anti-inflammatory and wound healing activity of Euphorbia characias and bioassay-guided isolation of some flavonoids

Chitinase III in Euphorbia characias latex: Purification and characterization

This paper deals with the purification of a class III endochitinase from Euphorbia characias latex. Described purification method includes an effective novel separation step using magnetic chitin particles. Application of magnetic affinity adsorbent noticeably simplifies and shortens the purification procedure. This step and the subsequently DEAE-cellulose chromatography enable to obtain the chitinase in homogeneous form. One protein band is present on PAGE in non-denaturing conditions and SDS–PAGE profile reveals a unique protein band of 36.5±2kDa. The optimal chitinase activity is observed at 50°C, pH 5.0. E. characias latex chitinase is able to hydrolyze colloidal chitin giving, as reaction products, N-acetyl-d-glucosamine, chitobiose and chitotriose. Moreover, we observed that calcium and magnesium ions enhance chitinase activity. Finally, we cloned the cDNA encoding the E. characias latex chitinase. The partial cDNA nucleotide sequence contains 762bp, and the deduced amino acid sequence (254 amino acids) is homologous to the sequence of several plant class III endochitinases.

Euphorbia characias as Bioenergy Crop: a Study of Variations in Energy Value Components According to Phenology and Water Status

Euphorbia characias has drawn much attention as a potential bioenergy crop given its considerable amount of latex, rich in hydrocarbon-like compounds, and its ability to grow in large areas of semiarid lands. Compositions of major constituents with an energy value have been determined for the three phenological stages of this plant (preflowering, flowering, and postflowering) and different irrigation treatments. Metabolites from both nonpolar and polar extracts have been identified and quantified by GC-MS, GC-FID, HPLC-ELSD, and UPLC-PDA-MS. The results highlight that the end of the flowering period is the optimal harvesting time to maximize the yields of E. characias as a potential energy crop. The total water requirements to obtain the maximum yields of hexane- and methanol-extractables were determined for its annual development cycle.

Molecular Analysis of a Copper- and Zinc-Containing Superoxide Dismutase Gene Isolated From the Latex of Euphorbia Characias: Another Piece in the Molecular Puzzle of Euphorbiaceae Latex Proteins

A copper- and zinc-containing superoxide dismutase (Cu/Zn-SOD) cDNA was isolated from the Euphorbia characias latex (Elx) using consensus degenerate hybrid oligonucleotide primer (CODEHOP) design and RT-PCR strategy. Both 3'and 5'untraslated regions (UTR) were isolated by rapid amplification of cDNA ends (RACE) method. Analysis of the nucleotide sequence revealed that the Cu/Zn-SOD cDNA contains an open reading frame encoding a protein of 152 amino acids. Bioinformatic analyses of Elx SOD gene revealed a high identity rate with a large number of plant Cu/Zn-SODs in the deduced amino acid sequence. Since isoenzymes may be generated through the multiplicity of SOD genes or result from post-trascriptional events, genomic Southern blot in conjunction with northern blot experiments were also performed. The genomic analysis showed that the E. characias genome contains a single SOD gene. Northern blot analyses confirmed the presence of a single SOD mRNA demonstrating that alternative splicing does not occur. Quantita- tive real time-PCR (qRT-PCR) experiments showed that SOD gene expression in latex reaches maximum levels during the summer. These results suggest that the Cu/Zn-SOD of Euphorbia characias latex probably may be involved in the antioxid- ative process triggered by oxidative stress induced by the conditions of environmental change in which the plant lives.

Spatial variation in the fatty acid composition of elaiosomes in an ant-dispersed plant: Differences within and between individuals and populations

Euphorbia characias is a common myrmecochorous plant of the western Mediterranean Basin whose seeds are dispersed by ants following fruit explosion. The variation in elaiosomes’ fatty acid composition of this species was studied at three hierarchical levels (sub-individual, individual and population) in four populations from the Iberian Peninsula. We found that differences in fruit location on the inflorescence do not seem to influence the fatty acid composition of elaiosomes, providing to each propagule an equal chance of being dispersed. However, significant differences in elaiosome fatty acid composition between individuals and populations were found for most of the compounds identified. The content of oleic acid, a key mediator in the ant–seed interaction, differed widely between populations, probably reflecting geographic variations in co-adaptation between plants and their dispersers. The finding that the fatty acid composition of E. characias elaiosomes is distinct from that of the seed itself, but very similar to that of elaiosomes from unrelated species, reinforces the idea of convergent evolution in the chemical composition of these structures.