Optimization of phenolics and antioxidants extraction from Centaurium erythraea using response surface methodology

In recent years, Centaurium erythraea extracts have attracted much research attention in the context of prevention or treatment of many diseases due to its bioactive compounds content and antioxidant activity. The antioxidants of C. erythraea are very effective as they possess excellent antioxidant activity. Thus, it can be used as a safe and natural food preservative. The aim of this study is to make extracts more effective by optimizing the extraction conditions of the phenolics and antioxidants from C. erythraea using response surface methodology (RSM) based on a central composite design (CCD). Two process variables (Methanol volume fraction and solid - solvent ratio) were evaluated at five levels (13 experimental designs). Multiple regression analyses were performed to obtain quadratic polynomial equations using RSM; each response was fitted by a quadratic model. The adequacy of the models was proven using the analysis of variance (ANOVA). The significant effects of the factors and their interactions on the extraction efficiency were investigated at 95% confidence interval. RSM indicated that the optimal extraction conditions were 71% methanol volume fraction and 2.2:10 solid:solvent ratio. Predicted values thus obtained were close to the experimental values indicating suitability of the model.

Phytohormone profiles in non-transformed and AtCKX transgenic centaury (Centaurium erythraea Rafn) shoots and roots in response to salinity stress in vitro

Plant hormones regulate numerous developmental and physiological processes. Abiotic stresses considerably affect production and distribution of phytohormones as the stress signal triggers. The homeostasis of plant hormones is controlled by their de novo synthesis and catabolism. The aim of this work was to analyse the contents of total and individual groups of endogenous cytokinins (CKs) as well as indole-3-acetic acid (IAA) in AtCKX overexpressing centaury plants grown in vitro on graded NaCl concentrations (0, 50, 100, 150, 200 mM). The levels of endogenous stress hormones including abscisic acid (ABA), salicylic acid (SA) and jasmonic acid (JA) were also detected. The elevated contents of total CKs were found in all analysed centaury shoots. Furthermore, increased amounts of all five CK groups, as well as enhanced total CKs were revealed on graded NaCl concentrations in non-transformed and AtCKX roots. All analysed AtCKX centaury lines exhibited decreased amounts of endogenous IAA in shoots and roots. Consequently, the IAA/bioactive CK forms ratios showed a significant variation in the shoots and roots of all AtCKX lines. In shoots and roots of both non-transformed and AtCKX transgenic centaury plants, salinity was associated with an increase of ABA and JA and a decrease of SA content.

Changes in Photosynthetic Pigments Content in Non-Transformed and AtCKX Transgenic Centaury (Centaurium erythraea Rafn) Shoots Grown under Salt Stress In Vitro

The effects of graded sodium chloride (NaCl) concentrations (0-, 50-, 100-, 150-, and 200-mM) on photosynthetic pigment contents in in vitro grown shoots of important medicinal plant species (Centaurium erythraea Rafn) were investigated. Non-transformed, one AtCKX1 and two AtCKX2 transgenic centaury lines, with altered cytokinin profiles, were used in this study. The chlorophyll (Chl) and carotenoid contents differed in the non-transformed and transgenic lines. In general, salinity significantly reduced the Chl a and Chl b contents in comparison to the NaCl-free medium. The lowest Chl content was observed in AtCKX2 transgenic shoots grown on all the culture media. The total carotenoid content was increased in shoots of non-transformed and both AtCKX2 transgenic lines grown in 50-mM NaCl. On the other hand, in concentrations ˃50-mM NaCl, the total carotenoid content was decreased in all analysed centaury shoots. The Chl a/Chl b ratio in all the shoots increased progressively in the graded NaCl concentrations. Contrarily, the addition of NaCl in the culture medium reduced the Chl/carotenoid ratio in centaury shoots. Taken together, the results of this study partly explained the mode of centaury plant adaptations to salt stress in vitro. Thus, the results on centaury shoots confirmed that the determination of the photosynthetic pigment contents can be a very useful non-destructive screening method in order to discriminate susceptible and resistant plant species/lines to salt stress conditions.

The Arabinogalactan Protein Family of Centaurium erythraea Rafn

Centaurium erythraea (centaury) is a medicinal plant with exceptional developmental plasticity in vitro and vigorous, often spontaneous, regeneration via shoot organogenesis and somatic embryogenesis, during which arabinogalactan proteins (AGPs) play an important role. AGPs are highly glycosylated proteins belonging to the super family of O-glycosylated plant cell surface hydroxyproline-rich glycoproteins (HRGPs). HRGPs/AGPs are intrinsically disordered and not well conserved, making their homology-based mining ineffective. We have applied a recently developed pipeline for HRGP/AGP mining, ragp, which is based on machine learning prediction of proline hydroxylation, to identify HRGP sequences in centaury transcriptome and to classify them into motif and amino acid bias (MAAB) classes. AGP sequences with low AG glycomotif representation were also identified. Six members of each of the three AGP subclasses, fasciclin-like AGPs, receptor kinase-like AGPs and AG peptides, were selected for phylogenetic and expression analyses. The expression of these 18 genes was recorded over 48 h following leaf mechanical wounding, as well as in 16 tissue samples representing plants from nature, plants cultivated in vitro, and developmental stages during shoot organogenesis and somatic embryogenesis. None of the selected genes were upregulated during both wounding recovery and regeneration. Possible functions of AGPs with the most interesting expression profiles are discussed.

Observations on Antagonism in the Arbuscular Mycorrhizal Systems

The majority of land plants are colonised by arbuscular mycorrhizal fungi, however ,members of few families notably Cruciferae, Chenopodiaceae, Caryophyllaceae and Polygonaceae, so called non-host are not colonized by these fungi. Previous studies have shown that the growth and development of non-host species were severely inhibited when grown in the presence of active AMF mycelia. There is therefore a need to understand the mechanistic bases of adverse effects of AMF mycelia upon seedlings of non-host species. In this experiment young roots of non-hosts Spergula arvensis and Arabis hirsuta, and host Centaurium erythraea were exposed to mycorrhizal and non-mycorrhizal extracts. The results of this experiment showed that mycorrhizal extracts significantly reduced the growth of radicles and root hairs development of non-host species whereas had no effect on radicles and root hairs development of host species. These results indicate that toxicity effects rather the nutritional factors are the drivers of the negative responses of non-host species to the presence of the AMF. Keywords: mycorrhizal fungi, host and non-host plants, Plantago lanceolata , Glomus mossae , mycelium

A novel comparative chromatographic research of secoiridoid glycosides in two species of centaury herb

The aim: A novel comparative analysis of the secoiridoid glycosides composition in Centaurium erythraea Rafn. and Cеntaurium pulchellum (Sw.) Druce has being described. Swertiamarin has been chosen as an active marker and its comparative quantitative estimation in two species of the raw material has been carried out by HPLC method. Materials and methods: The quantitative HPLC analysis of swertiamarin was conducted using a chromatographic column ACE 5 C18. Methanol and 0,5 % acetic acid aqueous solution were used as mobile phases; the chromatographic procedure was carried out in a gradient mode. Results: The content of swertiamarin in the C. pulchellum herb ranges from 2.51 to 3.07 mg/100 mg. In that time swertiamarin content in C. erythraea herb varies and depends from 3.83 to 8.94 mg/100 mg. The content of swertiamarin in C. pulchellum herb is much lower than in C. erythraea herb. Taking into account this fact the possibility of using of C. pulchellum herb for medicine instead of C. erythraea herb must be proven additionally by biological experiments. Also it could be preliminary concluded that the most appropriate climate for swertiamarin accumulation in C. erythraea herb is located in Central and Eastern parts of Ukraine. Conclusions: It was discovered that the main representative of secoiridoid glycosides in C. erythraea is swertiamarin when in C. pulchellum Druce that are represented by both sweroside and swertiamarin. A more perspective raw material – C. erythraea herb, according to the content of swertiamarin, was chosen

Antioxidant and Antibacterial Effects of Pollen Extracts on Human Multidrug-Resistant Pathogenic Bacteria

The present work was designed to search the possible antibacterial effect of the ethanolic extract of pollens from six botanical origins, Punica granatum, Quercus ilex, Centaurium erythraea, Coriandrum sativum, Ruta graveolens, and Citrus aurantium, against multidrug pathogenic bacteria, Escherichia coli, Pseudomonas aeruginosa, Enterobacter cloacae, Acinetobacter baumannii, Klebsiella pneumoniae, and Staphylococcus aureus. The content of phenolic compounds, flavones, and flavonols was measured. The antioxidant activities were evaluated using four assays: total antioxidant capacity, DPPH, ABTS, and reducing power. Antibacterial activity was studied using the agar disk diffusion method, and the MIC and MBC were determined. Results obtained showed a positive correlation between the antioxidant content of pollen extracts and the antibacterial capacity, Punica granatum and Quercus ilex pollen extracts were the most efficient against Gram-positive and Gram-negative bacteria, and Centaurium erythraea, Coriandrum sativum, and Ruta graveolens had a moderate effect, while Citrus aurantium had no antibacterial effect. It is concluded that pollens can be a good source of bioactive molecules that exhibit potent antioxidant effects and strong antibacterial activities.

Determination of amino acids of some plants from Gentianaceae family

Medicines from plants are widely used in the complex treatment of different diseases every day. Therefore, theoretical and practical interest is the in-depth study of the herb of perspective plants like Centaurium erythraea Rafn. and Gentiana cruciata L. These plants have a long history of usage and interest of people. The aim of the study was to determine the content of amino acids in these plants. The amino acids composition and content in the herb of study species of the family Gentianaceae determined by the HPLC method. The results of the research revealed that the raw material of Centaurium erythraea Rafn. and Gentiana cruciata L. contains free and bound amino acids. Sixteen free and seventeen bound amino acids were identified in the herb of Centaurium erythraea Rafn. The herb of Gentiana cruciata L. contained thirteen free and fifteen bound amino acids. L-glutamic acid, L-arginine, L-aspartic acid, and L-cystine were predominant of Centaurium erythraea Rafn. herb. Amino acids L-lysine, L-serine, L-aspartic acid, and L-phenylalanine were present in the herb of Gentiana cruciata L. in the greatest amount. The metabolic processes in which these amino acids are involved connected to the medicinal properties of the study plants according to their use in official or nontraditional medicine.

Chemical Composition, Antioxidant, and α-Glucosidase-Inhibiting Activity of Aqueous and Hydroethanolic Extracts of Traditional Antidiabetics from Croatian Ethnomedicine

Type 2 diabetes (T2D) is a chronic disease with a growing prevalence worldwide. In addition to the conventional therapy, many T2D patients use phytotherapeutic preparations. In the present study, chemical composition, antioxidant, and α-glucosidase inhibiting activity of traditional antidiabetics from Croatian ethnomedicine (Achillea millefolium, Artemisia absinthium, Centaurium erythraea, Morusalba, Phaseolus vulgaris, Sambucus nigra, and Salvia officinalis) were assessed. The efficacy of water and 80% ethanol as extraction solvents for bioactive constituents was compared. HPLC analysis revealed that the prepared extracts were rich in phenols, especially rutin, ferulic, and chlorogenic acid. Antiradical (against DPPH and ABTS radicals), reducing (towards Mo6+ and Fe3+ ions), and enzyme inhibiting properties were in linear correlation with the content of phenolic constituents. Ethanolic extracts, richer in phenolic substances, showed dominant efficacy in those assays. Aqueous extracts, on the other hand, were better Fe2+ ion chelators and more active in the β-carotene linoleic acid assay. Extracts from S. officinalis and A. millefolium were particularly active antioxidants and α-glucosidase inhibitors. A. absinthium, another potent α-glucosidase inhibitor, contained chromium, a mineral that promotes insulin action. The investigated plants contained significant amounts of minerals useful in management of T2D, with negligible amounts of heavy metals deeming them safe for human use.