Search

Biostimulants improve yield, quality, and stress acclimation in crops. In this work, we tested the possibility of using phenolics-rich extracts from spelt (Triticum dicoccum L.) husks to attenuate the effects of salt stress (100–200 mM NaCl) in maize. Two methanolic extracts were prepared from the soluble-conjugated (SC), and the insoluble-bound (IB) phenolic acid fractions of the spelt husk, and their effects were investigated on several stress-associated biochemical parameters, such as proline, lipid peroxidation, H2O2, GSH levels, and ion content. Results show that SC and IB fractions of husk extracts behaved very differently, no doubt due to their greatly divergent chemical composition, as revealed by both GC-MS and HPLC analyses. The efficacy of treatments in mitigating salt stress was also dose- and timing-dependent. IB, even at the lower concentration tested, was able to recover the performance of stressed plants in terms of growth, photosynthetic pigments content, and levels of salt stress markers. Recovery of shoot growth to control levels and reduction of stress-induced proline accumulation occurred regardless of whether plants were pre-treated or post-treated with IB, whereas only pre-treatment with the higher dose of IB was effective in mitigating oxidative stress. Although in some cases SC and even methanol alone exerted some positive effects, they could also be deleterious whereas IB never was. Overall, results indicate that a polyphenol-containing extract obtained from spelt by-products can behave as biostimulant in maize plants and can mitigate their response to salt stress, by acting on different biochemical targets.

The importance of polyamines (PAs) in plant growth and development was recognised several decades ago and, since then, their role in cell proliferation, embryogenesis, organogenesis, flowering, fruit development and ripening, etc. has been investigated to a great extent. In more recent years, most of the attention on PAs has been focussed on their functions in biotic and, especially, abiotic stress responses. Exogenous application, transgenic plants over- or under-expressing PA biosynthetic genes, and mutants have been used to unveil their essential contribution to plant tolerance to salinity, drought, chilling, and heavy metal stresses, among others. In parallel, knowledge on their mechanisms of action has increased greatly and it is today evident that PA functions depend upon their ability to (a) bind electrostatically or covalently to numerous compounds thereby modulating membrane, cell wall, nucleic acid, and protein structure and functions, (b) produce hydrogen peroxide via their catabolic pathways, (c) interact with the biosynthetic and signaling pathways of practically all known phyto hormones, and (d) interact with nitric oxide. In this review, a state-of-the-art overview of PA functions in plants and their possible applications in mitigating stress in crop plants is provided. The potential of seed priming with PAs as an economically and environmentally valid approach for enhancing plant tolerance to adverse environmental conditions is discussed and some results from our study on quinoa and salt stress are presented.

In indoor vertical farms, energy consumption represents a bottleneck for both a system’s affordability and environmental footprint. Although switching frequency (sf) represents a crucial factor in determining the efficacy of light emitting diodes (LED) lighting systems in converting electricity into light, the impact of sf is still underexplored. The aim of this work was to investigate the effect of LEDs sf on the productive and qualitative responses of lettuce (Lactuca sativa L.), also considering the resource use efficiency. Plants were grown for 14 days under red and blue LEDs (215 μmol m−2 s−1 and 16/8 h light/dark, with a red:blue ratio of 3) characterized by two different sf for the blue diode, namely high sf (850 kHz) and low sf (293 kHz). A fluorescent light (same light intensity and photoperiod) was included. LED sf did not alter plant morphological parameters, including fresh or dry biomass, leaf number, leaf area, or water use efficiency. A low sf increased the energy use efficiency (EUE) by 40% as compared to high sf. The latter enhanced the leaf antioxidant capacity, as a consequence of increased concentrations of caftaric and chicoric acids, isoquercetin, and luteolin, consistent with the upregulation of a few genes related to the biosynthetic pathway of phenolic compounds (4C3H and DFR). The study highlights that different sf may significantly affect the EUE as well as crop nutritional properties.

Activity-guided fractionations of

Sweet cherries (Prunus avium L.) are highly appreciated fruits for their taste, color, nutritional value, and beneficial health effects. In this work, seven new cultivars of sweet cherry were investigated for their main quality traits and nutraceutical value. The phytochemical profile of three classes of phenolic compounds and the antioxidant activity of the new cultivars were investigated through high-performance liquid chromatography with diode array detection (HPLC-DAD) and spectrophotometric assays, respectively, and compared with those of commonly commercialized cultivars. Cyanidine-3-O-rutinoside was the main anthocyanin in all genotypes, and its levels in some new cultivars were about three-fold higher than in commercial ones. The ORAC-assayed antioxidant capacity was positively correlated with the total anthocyanin index. The nutraceutical value of the new cultivars was investigated in terms of antioxidant/neuroprotective capacity in neuron-like SH-SY5Y cells. Results demonstrated that the new cultivars were more effective in counteracting oxidative stress and were also able to upregulate brain-derived neurotrophic factor (BDNF), a pro-survival neurotrophin, suggesting their potential pleiotropic role in counteracting neurodegenerations.

Extensive documentation is available on plant essential oils as a potential source of antimicrobials, including natural drugs against Candida spp. Yeasts of the genus Candida are responsible for various clinical manifestations, from mucocutaneous overgrowth to bloodstream infections, whose incidence and mortality rates are increasing because of the expanding population of immunocompromised patients. In the last decade, although C. albicans is still regarded as the most common species, epidemiological data reveal that the global distribution of Candida spp. has changed, and non-albicans species of Candida are being increasingly isolated worldwide. The present study aimed to review the anti-Candida activity of essential oils collected from 100 species of the Lamiaceae family growing in the Mediterranean area and the Middle East. An overview is given on the most promising essential oils and constituents inhibiting Candida spp. growth, with a particular focus for those natural products able to reduce the expression of virulence factors, such as yeast-hyphal transition and biofilm formation. Based on current knowledge on members of the Lamiaceae family, future recommendations to strengthen the value of these essential oils as antimicrobial agents include pathogen selection, with an extension towards the new emerging Candida spp. and toxicological screening, as it cannot be taken for granted that plant-derived products are void of potential toxic and/or carcinogenic properties.

This paper describes the use of 1H NMR profiling and chemometrics in order to facilitate the selection of medicinal plants as potential sources of collagenase inhibitors. A total of 49 plants with reported ethnobotanical uses, such as the healing of wounds and burns, treatment of skin-related diseases, rheumatism, arthritis, and bone diseases, were initially chosen as potential candidates. The in vitro collagenase inhibitory activity of hydroalcoholic extracts of these plants was tested. Moreover, their phytochemical profiles were analyzed by 1H NMR and combined with the inhibitory activity data by an orthogonal partial least squares model. The results showed a correlation between the bioactivity and the concentration of phenolics, including flavonoids, phenylpropanoids, and tannins, in the extracts. Considering the eventual false-positive effect on the bioactivity given by tannins, a tannin removal procedure was performed on the most active extracts. After this procedure, Alchemilla vulgaris was the most persistently active, proving to owe its activity to compounds other than tannins. Thus, this plant was selected as the most promising and further investigated through bioassay-guided fractionation, which resulted in the isolation of a flavonoid, quercetin-3-O-β-glucuronide, as confirmed by NMR and HRMS spectra. This compound showed not only a higher activity than other flavonoids with the same aglycone moiety, but was also higher than doxycycline (positive control), the only Federal Drug Administration-approved collagenase inhibitor. The approach employed in this study, namely the integration of metabolomics and bioactivity-guided fractionation, showed great potential as a tool for plant selection and identification of bioactive compounds in natural product research.

Neurodegenerative diseases, such as Alzheimer’s Disease (AD), are characterized by a multifactorial etiology and one of the most impacting factors is represented by oxidative stress, a biochemical process that is given by an imbalance between reactive oxygen species (ROS) and endogenous antioxidant defenses. Cherries, and in particular sweet cherries, are a nutritionally dense food rich in anthocyanins, quercetin, hydroxycinnamates, potassium, fiber, vitamin C, carotenoids, and melatonin. In the present study, we investigated the antioxidant activity of different Sweet Unibo cherries (Prunus avium L.), obtained through a natural breeding program at the University of Bologna, in neuron-like SH-SY5Y cells differentiated with Retinoic Acid (RA). Five different cherry extracts were characterized by HPLC-DAD/FL analysis. Cells were treated with the different cherry extracts for 24 h before H2O2 exposure. All the extracts, except one, were able to significantly increase cell viability, measured by MTT assay, in respect to cells exposed to H2O2. Moreover, the same four extracts reduced ROS production as measured by the dichlorofluorescein diacetate assay meanwhile only three of five extracts significantly increased GSH levels in respect to H2O2 exposed cells as measured by monochloro bimane assay. The same extracts that significantly reduced ROS production were also able to up-regulate phase II antioxidant enzymes glutathione reductase and NQO1 as measured by RT-PCR. Our findings support the idea that cherries can have a beneficial health effect in counteracting oxidative stress in neurons and these effects seem to be related to the specific phenolic pattern of the different cherry extracts. This work was supported by MIUR-PRIN 2015 (No. 20152HKF3Z).

Neurodegenerative diseases, such as Alzheimer’s and Parkinson’s diseases, are caused by the progressive death of neurons in different regions of the nervous system. Their etiology is multifactorial with oxidative stress as one of the most impacting factors. The brain, due to its high oxygen consumption and high lipid content, is highly vulnerable to the effects of reactive oxygen species. The increase in the generation of free radicals damages the cellular biomolecules and induces necrosis or apoptosis. Sweet cherry fruits are a nutritionally dense food rich in anthocyanins, quercetin, hydroxycinnamates, potassium, fiber, vitamin C, and carotenoids. In the present study, we investigated the antioxidant activity of different sweet cherries (Prunus avium L.), obtained through a natural breeding program at the University of Bologna, in neuron-like SH-SY5Y cells differentiated with retinoic acid. The phytochemical pattern of five different cherry extracts has been characterized by HPLC-DAD/FL analysis. To study the protective effects of the extracts, SH-SY5Y cells were treated with different concentrations of the extracts for 24 h before H2O2 exposure. All the extracts, except one, were able to significantly increase cell viability and decrease ROS levels in respect to cells exposed to H2O2 as measured by MTT and DCFH-DA assays, respectively. Three of the five extracts significantly increased GSH levels in respect to H2O2 exposed cells as measured by monochlorobimane assay. The same extracts that significantly reduced ROS production were also able to up-regulate some important phase II antioxidant enzymes, such as GR and NQO1, as measured by RT-PCR. Our findings support the idea that these new sweet cherries can be considered a new functional food with a high antioxidant and neuroprotective effect and these effects seem to be related to the specific phenolic pattern of the different cherry extracts. This work was supported by MIUR-PRIN 2015 (No. 20152HKF3Z).

“This is an Accepted Manuscript version of the following article, accepted for publication in Natural Product Research. Natural Product Research 35.24 (2021): 5843-5847. It is deposited under the terms of the Creative Commons Attribution-NonCommercial-NoDerivatives License (http://creativecommons.org/licenses/by-nc-nd/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited, and is not altered, transformed, or built upon in any way”, Activity-guided fractionations of Jatropha macrantha Müll. Arg. led to the isolation of pomolic acid (1) and euscaphic acid (2). The potential for inhibition against NF-κB and HIF-1α production of these two compounds was tested in different tumour cell lines. Compounds 1 and 2 showed an inhibitory activity of HIF-1α in the SK-MEL-28 (IC50=3.01 ± 0.02 μM and 3.78 ± 0.02 μM), A549 (IC50=9.97 ± 0.01 μM and 10.25 ± 0.01 μM) and U-373 MG (IC50=6.34 ± 0.02 μM and 8.85 ± 0.02 μM) cell lines. In addition, compounds 1 and 2 showed an inhibitory activity on NF-κB in SK-MEL-28 (IC50=1.05 ± 0.02 μM and 2.71 ± 0.01 μM), A549 (IC50=3.63 ± 0.01 μM and 3.73 ± 0.02 μM) and U-373 MG (IC50=2.55 ± 0.02 μM and 3.39 ± 0.01 μM) cell lines. This is the first report that isolates these compounds from J. macrantha and tests their antitumor potential

Nowadays the high nutritional value of whole grains is recognized, and there is an increasing interest in the ancient varieties for producing wholegrain food products with enhanced nutritional characteristics. Among ancient crops, einkorn could represent a valid alternative. In this work, einkorn flours were analyzed for their content in carotenoids and in free and bound phenolic acids, and compared to wheat flours. The most promising flours were used to produce conventional and sourdough fermented breads. Breads were in vitro digested, and characterized before and after digestion. The four breads having the best characteristics were selected, and the product of their digestion was used to evaluate their anti-inflammatory effect using Caco-2 cells. Our results confirm the higher carotenoid levels in einkorn than in modern wheats, and the effectiveness of sourdough fermentation in maintaining these levels, despite the longer exposure to atmospheric oxygen. Moreover, in cultured cells einkorn bread evidenced an anti-inflammatory effect, although masked by the effect of digestive fluid. This study represents the first integrated evaluation of the potential health benefit of einkorn-based bakery products compared to wheat-based ones, and contributes to our knowledge of ancient grains.

The aim of this study was to determine the in vitro antimicrobial activity of crude extracts of three plants from Ayurveda tradition (Tinospora cordifolia, Alstonia scholaris, Crataeva nurvala) against reference microbial strains and clinical isolates of methicillin-resistant Staphylococcus aureus (MRSA) and carbapenemase-producing Klebsiella pneumoniae. IC50 values were obtained by micro-dilution methods meeting the requirements of the NCCLS standard. The cytotoxicity of the extracts was also investigated on a mammalian cell line. Extracts displayed a variable degree of antimicrobial activity and did not interfere with mammalian cell proliferation. T. cordifolia and A. scholaris exhibited a higher inhibitory activity against clinical isolates of MRSA and carbapenemase-producing K. pneumoniae compared with reference strains, while C. nurvala exhibited a different behaviour. An antifungal activity towards Candida albicans was observed for A. scholaris extract. Results indicate that constituents from T. cordifolia and A. scholaris may be a potential source of new therapeutic strategies for infectious diseases.

Amongst the processing technologies able to improve the functional features of cereal-based foods, sourdough fermentation using Lactic Acid Bacteria (LAB) has been recently rediscovered for its beneficial effects. Wheat (Triticum aestivum L.) bread doughs were prepared using LAB strains belonging to different Lactobacillus species and changes in phenolic acid, carotenoid content and antioxidant capacity were evaluated. Two L. plantarum strains out of six were able to significantly increase carotenoid content in the dough, suggesting that a higher mobilization/solubilisation of these antioxidant compounds occurs. Within different fractions (free, soluble-conjugated, insoluble-bound), the relative distribution of ferulic acid and antioxidant activity changes depending on the specific strain. Overall, results indicate that some LAB strains cause in situ changes, significantly increasing the content of functional compounds in doughs during fermentation. This, in turn, could improve the functional features of bakery foods characterised by a high content in carotenoids and other bioactive compounds.

The aim of the present research was to study the effects of an ethanolic extract of Salvia sagittata Ruiz & Pav (SSEE), an endemic Ecuadorian plant traditionally used to treat inflammation and different intestinal affections, on primary cultures of porcine aortic endothelial cells (pAECs). pAECs were cultured in the presence of different concentrations (1-200 μg/mL) of SSEE for 24 h, and cytotoxicity was evaluated by the MTT assay. SSEE did not negatively affect cellular viability at any concentration tested. Cell cycle was analyzed and no significant change was observed. Then, the anti-inflammatory effects of SSEE on pAECs were analyzed using a lipopolysaccharide (LPS) as the inflammatory stimulus. Different markers involved in the inflammatory process, such as cytokines and protective molecules, were evaluated by real-time quantitative PCR and Western blot. SSEE showed the ability to restore pAEC physiological conditions reducing interleukin-6 and increasing Heme Oxygenase-1 protein levels. The phytochemical composition of SSEE was also evaluated via HPLC-DAD and spectrophotometric assays. The presence of different phenolic acids and flavonoids was revealed, with rosmarinic acid as the most abundant component. SSEE possesses an interesting antioxidant activity, as assessed through both the Oxygen Radical Absorbance Capacity (ORAC) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) assays. In conclusion, results suggest that SSEE is endowed with an in vitro anti-inflammatory effect. This represents the initial step in finding a possible scientific support for the traditional therapeutic use of this plant.

Plants belonging to the Amaryllidaceae contain an exclusive group of alkaloids, known as sources of important biological activities. In the present work, Pancratium illyricumL., a species belonging to this family and endemic of Sardinia (Italy), was investigated for its alkaloid content. Fresh bulbs and leaves were processed separately. Standard extraction and purification procedures were applied to obtain fractions and compounds for GC–MS and NMR analysis. In addition to eight already known alkaloids (1–8), 11α-hydroxy-O-methylleucotamine (9) was isolated for the first time and its structure completely determined by one and two-dimensional 1H and 13C NMR spectroscopy. This new galanthamine-type compound exhibited a pronounced in vitro acetylcholinesterase (AChE) inhibitory activity (IC50 = 3.5 ± 1.1 μM) in comparison to the reference standard galanthamine hydrobromide (IC50 = 1.5±0.2 μM).

Introduction The argan tree (Argania spinosa) is an endemic species from south-western Morocco. Argan-based preparations have been widely used in Moroccan traditional medicine for their biological properties, as well as for several cosmetic purposes. Whereas kernel, pulp of fruit and trunk have been extensively studied for their nutritional and pharmacological effects, relatively little is known about argan tree leaves. Objective The main purpose of the present study is to investigate and characterise the bioactive phenolic fractions in both crude and aqueous extracts derived from argan tree leaves. Methodology A qualitative profile of the antioxidant phenolic compounds in argan leaves was obtained by means of structural hypothesis based on UV spectra and mass spectrometric fragmentation patterns. Moreover, selected phenolics were quantified in argan leaves by using a fully validated method based on liquid chromatography coupled to diode array detection and tandem mass spectrometry (LC-DAD-MS/MS). All the extracts were purified by a fast and reliable microextraction by packed sorbent (MEPS) procedure, before analysing them by LC-MS/MS. Results Based on retention times, mass spectrometric fragmentation and UV spectra, 13 phenolic compounds were identified or tentatively elucidated from crude and aqueous extracts derived from Argania spinosa leaves, while seven compounds were quantified in both extracts. Conclusion The obtained results could represent a first step towards a complete characterisation of the argan plant, its bioactive profiling and the valorisation of its by-products as a source of potentially beneficial bioactive molecules. Copyright © 2015 John Wiley & Sons, Ltd.

The phytochemical profile, antioxidant activity and α-glucosidase inhibitory activity of Hypericum scruglii Bacchetta, Brullo et Salmeri and Hypericum hircinum L. were analyzed and compared to those of the best-known Hypericum perforatum L. Samples were prepared using three different extraction methods (maceration in 70% ethanol, extraction in MeOH/phosphate buffer and decoction), the influence of which on the metabolic profile and bioactivities are discussed. H. scruglii and H. hircinum extracted in 70% ethanol strongly inhibited α-glucosidase (IC50 7.25 and 14.05 μg/ml, respectively), which is a valuable enzymatic target for treating metabolic disorders, while H. perforatum was found to be less powerful than the other two species in all the performed biological tests. The phytochemical profile was analyzed by NMR, HPLC-DAD and HPLC-FLD, revealing remarkable differences among the species. In particular, H. scruglii, which is a species endemic to Sardinia Island (Italy), was particularly enriched in two phloroglucinol-derivatives identified by means of 2D NMR and LC/MS/MS experiments as 3-geranyl-1-(2′-methylbutanoyl)-phloroglucinol and 3-geranyl-1-(2′-methylpropanoyl)-phloroglucinol.

Herpes simplex virus types 1 (HSV-1) and 2 (HSV-2) cause several clinically relevant syndromes in both adults and neonates. Despite the availability of efficient anti-HSV agents, the search for new therapeutic approaches is highly encouraged due to the increasing drug resistance of virus strains. Medicinal plants represent a source of potential bioactive compounds. In this context we evaluated the anti-herpetic activity of Hemidesmus indicus (L.) R. Br., a plant widely used in traditional Indian medicine. The hydroalcoholic extract prepared from roots was characterized by NMR and HPLC analysis and assayed in vitro by CPE reduction and virus infectivity assays to define its anti-viral effect. The extract's mechanism of action was investigated by virucidal and time-of-addition assays and by in vitro α-glucosidase inhibitory assay. The extract exhibited a remarkable anti-herpetic activity at 100 mg/mL, at non-cytotoxic concentration, through multiple mechanisms: it reduced the infectivity of viral particles released from infected cells possibly through its anti-ER α-glucosidase inhibitory activity and it inhibited the beginning stage of HSV infection acting as a virucide agent and/or preventing virus attachment to the host cell surface.

Recently, increasing interest has been paid to the biostimulant properties of plant extracts as they have been shown to improve crop yield and quality. Biolchim SpA is a leading company in the production and commercialization of biostimulants. The working approach to the development of innovative products is focused on the research of plant extracts with biostimulant properties. To this aim, Biolchim SpA has created a collection of botanical extracts whose potential use as biostimulants has to be screened according to their biological activity. Bioassays are small-scale tests that assess the biological activity of substances by evaluating their effect on living model organisms. In this study, an array of in vitro (root-tip elongation test) and in vivo (greenhouse growth tests) bioassays was set up and used to test the biostimulant properties of two botanical extracts. Overall results indicate that the array of bioassays is able to screen and differentiate the extracts according to their biological activity. The root-tip elongation bioassay is a first and rapid screening method of their hormone-like properties; the greenhouse growth bioassays provide information on their biostimulant performances in growth conditions similar to the open field and on their possible mode of application. The array of bioassays will be improved by including further in vitro and in vivo tests to fully unravel the biological properties of the extracts. The final goal is to screen the whole collection and classify the extracts according to their potential use as biostimulants. After the screening, prototypes will be formulated by combining different extracts and their efficacy will be tested in open field conditions.

Argania spinosa (L.) Skeels is an endemic Moroccan species belonging to Sapotaceae family. In this work, lipophilic and aqueous extracts were obtained from leaves and subjected to a chemical profiling by MS and LC-MS/MS. Pentacyclic terpenoids were identified and quantified in the lipophilic fraction, while phenolic compounds (mainly belonging to flavonols and flavan-3-ols) were identified in the aqueous fraction. The antibacterial activities of fractions were evaluated in vitro against both reference Gram-positive and -negative bacterial strains and clinical isolates of methicillin-sensitive and methicillin-resistant Staphylococcus aureus (MSSA and MRSA); in addition, the compounds quantified as main components in each extract were assayed against reference strains. A relevant antibacterial activity was observed against reference MSSA and MRSA strains of S. aureus: for the lipophilic fraction, MIC50 values obtained were 177.8 and 170.6 μg/mL for the former and the latter, respectively, while for the aqueous fraction were 215.5 and 233.3 μg/mL. These inhibitory activities could be mainly ascribed to ursolic and oleanolic acids, among pentacyclic terpenoids, and to quercetin concerning phenolic compounds. A remarkable antibacterial activity was also observed against clinical isolates, thus argan leaves can be considered of interest in the chemotherapy of human infections.

Ethnopharmacological relevance The top flowering aerial parts of the Hypericum species are traditionally used to prepare ointments to heal cuts and burns. Sometimes even the fruits are used for these purposes. Hypericum androsaemum L., commonly known as tutsan or shrubby St. John's Wort, is a Mediterranean medicinal plant which has been traditionally used to prepare an ointment for treating cuts and wounds. Aim of the study To evaluate the extracts obtained from H. androsaemum red berries as functional ingredients for skin care formulations. Materials and methods The methanolic extract was obtained by Soxhlet extraction while the aqueous extract was prepared by decoction; their composition was determined by HPLC analysis. Their biological activities were measured in terms of proliferation and migration of human fibroblasts, inhibition of collagenase activity, and immunomodulatory effects on human peripheral blood mononuclear cells (PBMCs). In addition, we evaluated their photostability by UV spectroscopy and their protective effects against APPH-induced hemolysis in red blood cells (RBC). Results The polar extracts contained significant amounts of shikimic (108,143.7–115,901.3 mg/kg) and chlorogenic acids (45,781.1–57,002.7 mg/kg). The main components of these extracts made an important contribution to a significant increase in human fibroblast migration. Both extracts were also active as collagenase inhibitors, with the aqueous one showing a greater inhibitory capacity (IC 50 value of 88.1 µg/mL), similar to that of chlorogenic acid. The kinetic parameters determined for the enzymatic reaction revealed for both aqueous extract and chlorogenic acid an uncompetitive mechanism of inhibition. The methanolic extract showed important effects on PBMCs by modulating IL-6. Both extracts proved to be photostable in the UVA/B range and protected RBC against peroxidation at low concentrations. Conclusions H. androsaemum red berries were proven to contain phytochemicals that improve skin regeneration, hence potentially employable in skin care formulations.

Teucrium chamaedrys, one of the most common and investigated species of the genus Teucrium, has been used for centuries in traditional medicine for many purposes. Its phytochemical components comprise, among others, phenylethanoid glycosides (PGs) and neo-clerodane diterpenoids. Several reports have demonstrated a wide range of beneficial biological and pharmacological activities of the phenylethanoid components, while the diterpenes were shown to be strongly hepatotoxic. In this work, in vitro cultures were established from leaf explants of T. chamaedrys. Both solid (callus) and liquid (cell suspension) cultures maintained the capacity to produce PGs, with teucrioside (TS) representing the most abundant one. Cell suspensions had a lower TS content than that found in leaf extracts, but higher than that of calli. An NMR-based metabolomics approach was used to compare the product profile of intact plants vs. cell suspension cultures, and results showed that neo-clerodane diterpenes, present in the intact plant, were not detected in cell cultures. Several elicitors were supplied to cell cultures with the aim of increasing TS production, and elicitation was tested at different growth phases and by exposing cells for different periods. Methyl jasmonate and fungal mycelia from Trichoderma viridae and Fusarium moniliforme were able to significantly increase TS production if supplied at the early-exponential growth phase for 24 h. Based on the proposed link between proline and the phenylpropanoid pathways, proline accumulation in cell cultures was followed throughout a 14-day culture period, showing that it strictly reflected that of TS production. Moreover, exogenously supplied proline, and its analogue hydroxyproline, turned out to be very effective in increasing teucrioside production. © 2012 Elsevier Ltd. All rights reserved.

3,5-O-dicaffeoyl-4-O-malonilquinic acid (1) (irbic acid) has been isolated for the first time from cell cultures of Centella asiatica and till now it has never been reported to be present in the intact plant. Evidence of its structure was obtained by spectroscopic analyses (MS/NMR). Besides 1, cell cultures produce also the known 3,5-O-dicaffeoylquinic acid, chlorogenic acid, and the triferulic acid 2 (4-O-8′/4′-O-8″-didehydrotriferulic acid). Biological activities were evaluated for compound 1, which showed to have a strong radical scavenging capacity, together with a high inhibitory activity on collagenase. This suggests a possible utilization of this substance as a topical agent to reduce the skin ageing process.

Pollination of pummelo (Citrus grandis L. Osbeck) pistils has been studied in planta by adding compatible and self-incompatible (SI) pollen to the stigma surface. The pollen germination has been monitored inside the pistil by fluorescent microscopy showing SI altered morphologies with irregular depositions of callose in the tube walls, and heavy callose depositions in enlarged tips. The polyamine (PA) content as free, perchloric acid (PCA)-soluble and -insoluble fractions and transglutaminase (TGase) activity have been analyzed in order to deepen their possible involvement in the progamic phase of plant reproduction. The conjugated PAs in PCA-soluble fraction were definitely higher than the free and the PCA-insoluble forms, in both compatible and SI pollinated pistils. In pistils, pollination caused an early decrease of free PAs and increase of the bound forms. The SI pollination, showed highest values of PCA-soluble and -insoluble PAs with a maximum in concomitance with the pollen tube arrest. As TGase mediates some of the effects of PAs by covalently binding them to proteins, its activity, never checked before in Citrus, was examined with two different assays. In addition, the presence of glutamyl-PAs confirmed the enzyme assay data and excluded the possibility of a misinterpretation. The SI pollination caused an increase in TGase activity, whereas the compatible pollination caused its decrease. Similarly to bound PAs, the glutamyl-PAs and the enzyme activity peaked in the SI pollinated pistils in concomitance with the observed block of the pollen tube growth, suggesting an involvement of TGase in SI response.

Chemical fingerprinting of commercial Pelargonium capitatum (Geraniaceae) essential oil samples of south African origin was performed by GC, GC/MS, and (13) C- and (1) H-NMR. Thirty-seven compounds were identified, among which citronellol (32.71%) and geraniol (19.58%) were the most abundant. NMR Spectra of characteristic chemicals were provided. Broad-spectrum bioactivity properties of the oil were evaluated and compared with those of commercial Thymus vulgaris essential oil with the aim to obtain a functional profile in terms of efficacy and safety. P. capitatum essential oil provides a good performance as antimicrobial, with particular efficacy against Candida albicans strains. Antifungal activity performed against dermatophyte and phytopathogen strains revealed the latter as more sensitive, while antibacterial activity was not remarkable against both Gram-positive and Gram-negative bacteria. P. capitatum oil provided a lower antioxidant activity (IC(50) ) than that expressed by thyme essential oil, both in the 1,1-diphenyl-2-picrylhydrazyl (DPPH) and β-carotene bleaching tests. Results in photochemiluminescence (PCL) assay were negligible. To test the safety aspects of P. capitatum essential oil, mutagenic and toxicity properties were assayed by Ames test, with and without metabolic activation. Possible efficacy of P. capitatum essential oil as mutagenic protective agent against NaN(3) , 2-nitrofluorene, and 2-aminoanthracene was also assayed, providing interesting and significant antigenotoxic properties.

In this work, we have analyzed the changes in the protein expression profile elicited by chromium (Cr) exposure in the freshwater green alga Pseudokirchneriella subcapitata, a well known bio-indicator of water pollution. We tested two experimental conditions, namely 0.2 and 1 ppm of potassium dichromate; this concentration range includes the environmentally-relevant concentrations. Results show that neither concentration of potassium dichromate tested inhibited algal growth. However, the proteomic approach allowed the identification of relevant modifications in protein expression. In fact, among 800 protein spots detected by two-dimensional electrophoresis, 16 Cr-regulated proteins, including predicted and novel ones, were identified using tandem mass spectromic protein analysis. The results demonstrate a Cr-specific action in altering several photosynthetic proteins, such as ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO), RuBisCO activase, Light Harvesting Chla/b protein complex, and stress related Chla/b binding protein1. Although Cr toxicity with respect to photosynthesis has been already documented, here we have identified, for the first time, the target proteins of this toxicity. Cr also induced a modulation of some proteins involved in the metabolism of the amino acids glutamine, arginine and methionine. These data are supported by changes in cellular polyamine (PA) accumulation. Present findings provide new insight into the molecular mechanisms underlying Cr toxicity in P. subcapitata.

In the search for novel antibacterial agents against multidrug-resistant bacteria, an alkaloid extract obtained from whole plants of Crinum angustum Steud., containing six different groups of alkaloids, was analysed by using gas chromatography/mass spectrometry, and its in vitro activity against American Type Culture Collection bacterial strains and clinical isolates was evaluated. The antimicrobial activity of the extract was tested against nine standard strains of microorganisms and two drug-resistant clinical isolates, methicillin-resistant Staphylococcus aureus and carbapenemase-producing Klebsiella pneumoniae. The extract exhibited a significant activity against six of the examined strains of microorganisms. Antibacterial activity was more pronounced on Gram-positive bacteria than on Gram-negative bacteria, and IC50 values ranged from 156 to 625 μg/mL. Mycostatic activity against Candida albicans was also found, with an IC50 of 78 μg/mL after 48 h of incubation . Promising results were also obtained from tests carried out on both clinical isolates investigated.

Micellar electrokinetic chromatography was applied to the determination of the major phytomarkers, namely 2-hydroxy-4-methoxybenzaldehyde, 2-hydroxy-4-methoxybenzoic acid, and 3-hydroxy-4-methoxybenzaldehyde, of Hemidesmus indicus root, an Indian medicinal plant. H. indicus bioactive preparations were analyzed by reverse flow micellar electrokinetic chromatography (MEKC) using sodiumtaurodeoxycholate as the surfactant. A pH 2.5 phosphate buffer (50mM) was supplemented with 65mM of sodium taurodeoxycholate to produce the MEKC pseudostationary phase; because of the suppression of the electroosmotic flow, the migration of the partitioned analytes was toward the capillary anodic end. The use of a short fused-silica capillary (8.5 cm effective length; 50 µm i.d.) allowed the separation of phytomarkers, including vanillin and salicylaldehyde (reported as additional metabolites of H. indicus roots), in less than 8 min. The method showed good validation parameters and was applied to the analysis of methanol extracts and a root decoction of H. indicus, a promising botanical drug. The obtained results were compared to those from an independent high-performance liquid chromatography–mass spectrometry method. The 2-Hydroxy-4-methoxybenzaldehyde, 2-hydroxy-4-methoxybenzoic acid, and 3-hydroxy 4-methoxybenzaldehyde were found in all samples confirming their roles as phytomarkers. The absence of vanillin and salicylaldehyde suggested that these latter compounds should not be regarded as characteristic components of the bioactive preparations from the plant roots.

The argan tree (Argania spinosa) is an endemic species from south-western Morocco. Argan-based preparations have been widely used in Moroccan traditional medicine for their biological properties, as well as for several cosmetic purposes. Whereas kernel, pulp of fruit and trunk have been extensively studied for their nutritional and pharmacological effects, relatively little is known about argan tree leaves.The main purpose of the present study is to investigate and characterise the bioactive phenolic fractions in both crude and aqueous extracts derived from argan tree leaves.A qualitative profile of the antioxidant phenolic compounds in argan leaves was obtained by means of structural hypothesis based on UV spectra and mass spectrometric fragmentation patterns. Moreover, selected phenolics were quantified in argan leaves by using a fully validated method based on liquid chromatography coupled to diode array detection and tandem mass spectrometry (LC-DAD-MS/MS). All the extracts were purified by a fast and reliable microextraction by packed sorbent (MEPS) procedure, before analysing them by LC-MS/MS.Based on retention times, mass spectrometric fragmentation and UV spectra, 13 phenolic compounds were identified or tentatively elucidated from crude and aqueous extracts derived from Argania spinosa leaves, while seven compounds were quantified in both extracts.The obtained results could represent a first step towards a complete characterisation of the argan plant, its bioactive profiling and the valorisation of its by-products as a source of potentially beneficial bioactive molecules.

We investigated the mechanisms underlying severe bleeding occurring upon consumption of Ferula communis. The prenylated coumarin ferulenol extracted from this plant did not directly affect blood coagulation but showed hepatocyte cytotoxicity and, at non-cytotoxic concentrations (less than 100 nM), impaired factor X biosynthesis (40%reduction). Studies with ferulenol derivatives indicated the prenyl residue as major determinant of ferulenol activity.

Dihydroasparagusic acid (DHAA) is the reduced form of asparagusic acid, a sulfur-containing flavor component produced by Asparagus plants. In this work, DHAA was synthetically produced by modifying some published protocols, and the synthesized molecule was tested in several in vitro assays (DPPH, ABTS, FRAP-ferrozine, BCB, deoxyribose assays) to evaluate its radical scavenging activity. Results show that DHAA is endowed with a significant in vitro antioxidant activity, comparable to that of Trolox. DHAA was also evaluated for its inhibitory activity toward tyrosinase, an enzyme involved, among others, in melanogenesis and in browning processes of plant-derived foods. DHAA was shown to exert an inhibitory effect on tyrosinase activity, and the inhibitor kinetics, analyzed by a Lineweaver-Burk plot, exhibited a competitive mechanism. Taken together, these results suggest that DHAA may be considered as a potentially active molecule for use in various fields of application, such as pharmaceutical, cosmetics, agronomic and food.

Both trivalent and hexavalent chromium, i.e., Cr(III) and Cr(VI), respectively, were previously demonstrated to affect in vitro germination and ultrastructure of kiwifruit (Actinidia deliciosa) pollen. In the present work, the response to chromium in germinating pollen was evaluated in terms of changes in the polyamine profile. Slight, though significant, increases in free spermidine and spermine occurred after exposure to Cr(III), while the levels remained almost unchanged after Cr(VI) treatment. The spermidine synthase inhibitor cyclohexylamine (CHA) caused a dramatic increase in free putrescine in both chromium-treated and untreated samples, while spermidine content was not affected. Interestingly, CHA positively affected the performance of chromium-treated pollen by partially, though significantly, restoring pollen tube growth. The major growth recovery was registered with 1 mM CHA in the presence of Cr(VI), concomitant with a considerable reduction in uptake of the metal. Conversely, endogenous calcium levels were more heavily affected in Cr(III)-treated pollen. The effect of CHA on production of reactive oxygen species also varied depending on the chromium species. The response of pollen to the CHA-induced putrescine excess was compared with that exerted by an exogenous supply of the same diamine. Results show that in Cr(III)-treated pollen, putrescine over-accumulation induced by CHA exerted similar effects as exogenous putrescine, while this was not true in the Cr(VI) treatment. It appears that the diamine was able to improve pollen tolerance to metal stress through different mechanisms, mostly depending upon the chromium species, namely via reduced metal uptake or by substituting for calcium.

In the present study callus cultures of Zanthoxylum stenophyllum were initiated in vitro and the effect of different growth regulators and elicitors was tested both upon callus growth and secondary metabolite production. On a medium containing naphthalenacetic acid (NAA), kinetin and 2,4 dichlorophenoxyacetic acid (2,4D), a yellowish and friable callus was obtained from 90% of cotyledon explants, while no callus appeared from other portions of the seedlings. The time course of callus growth and secondary metabolite accumulation was followed after subculturing the established callus culture on different media containing various hormonal combinations. Results indicate that medium containing NAA and a higher concentration of 2,4-D compared to maintenance medium gave the highest stimulation of callus growth, suggesting that this auxin has a critical role in influencing growth. Addition of an organic nitrogen source such as casein hydrolysate also had a positive effect on growth. HPLC analysis of methanol extractable secondary metabolites from Z. stenophyllum calli grown on different media showed that phytohormones and nutrients were able to modify the chromatographic pattern of compounds. Calli grown on the medium giving the highest stimulation of growth show a reduced accumulation of some secondary products. Under the same hormonal conditions, the presence or absence of casein also altered the metabolites produced, both qualitatively and quantitatively. In response to elicitation by methyl jasmonate (MJ), metabolite production was different for the different classes of compounds, and hormonal composition of the culture medium turned out to be very important in determining the response to MJ. Thus, results confirm the importance of the reciprocal interactions between hormones, nutrients and elicitors when attemps are made to enhance secondary metabolite accumulation in in vitro cultures.

Flavonols are phytochemicals widely found in commonly consumed foods. In spite of their beneficial effects on human health, however, cytotoxicity and even suspected genotoxicity have also been reported for the flavonol, quercetin. This points to the need for preventive studies to identify any cytotoxic effects associated with pure flavonol intake. This work was performed with the aim of verifying whether a plant-based in vitro system, the pollen tube, could be used to evaluate the cytotoxic potential of exogenous flavonols. Increasing concentrations of the aglycone, quercetin, and its glycoside, rutin, were assayed with regard to tube growth of kiwifruit pollen, determined by applying the pollen tube growth test protocol. This test, based on the photometric quantification of pollen tube mass production in suspension cultures, has already been applied in the sensitive and reliable toxicological evaluation of a wide range of chemicals. Whereas 60–800μM rutin promoted kiwifruit pollen tube elongation, 10–50μM quercetin strongly inhibited growth, and also produced irreversible malformations, such as screw-like tube growth, abnormal vacuolation, alteration of organelle streaming, and nuclear positioning. Thus, the cytotoxic potentials of the two flavonols have been confirmed to differ. Pollen tubes seem to afford a promising test system for a preventive, rapid in vitro biosafety assessment of antioxidant nutritional supplements, without using laboratory animals.

Hypericum hircinum L. is a shrub traditionally used in some Mediterranean areas as a remedy for several diseases, mainly linked to the airway apparatus. In this work, an ethanolic extract prepared from aerial parts of plants collected in Sardinia (Italy), and six fractions isolated by a chromatographic separation of the extract, were evaluated for their antioxidant activity using three in vitro assays. Based on the ethnobotanical use of the plant and in view of the emerging role that enzymes belonging to matrix metalloproteinases (MMPs) play in the pathogenesis of some respiratory diseases, the effect of the crude extract and of several fractions on in-vitro collagenase activity was also evaluated. Results show that the ethanolic extract and fractions containing quercetin, chlorogenic acid, and 5,7,3′,5′-tetrahydroxyflavanone, a component recently identified in this species, are endowed with the highest free radical scavenging activity. Conversely, fractions containing shikimic acid as the main component did not show this property. The crude extract was able to inhibit in-vitro collagenase activity with an IC50 value of 156 μg/mL. A Lineaweaver–Burk plot, built to obtain the kinetic parameters of the enzymatic reaction, revealed that the inhibitory mechanism is non-competitive. Single fractions were also evaluated for their inhibitory activity on collagenase; fractions mainly containing flavonols and the substituted flavanone showed the highest inhibitory effect. Thus, H. hircinum can be considered as a new natural source of molecules able to inhibit enzymes of the MMP family, which could enter as active ingredients in wrinkle-care cosmetics.

In this work [14C]spermidine binding to total proteins solubilized from plasma membrane purified from zucchini (Cucurbita pepo L.) hypocotyls was investigated. Proteins were solubilized using octyl glucoside as a detergent. Specific polyamine binding was thermolabile, reversible, pH dependent with an optimum at pH 8.0, and had a Kd value of 5 μm, as determined by glass-fiber-filter assays. Sephadex G-25 M gel-filtration assays confirmed the presence of a spermidine-protein(s) complex with a specific binding activity. By sodium dodecyl sulfate-polyacrylamide gel electrophoresis and native polyacrylamide gel electrophoresis of collected fractions having the highest specific spermidine-binding activity, several protein bands (113, 75, 66, and 44 kD) were identified. The specificity of spermidine binding was examined by gel-filtration competition experiments performed using other polyamines and compounds structurally related to spermidine. Partial purification on Sephadex G-200 led to the identification of 66- and 44-kD protein bands, which may represent the putative spermidine-binding protein(s) on the plasmalemma.

Polyamine content and enzyme activities in the biosynthetic and degradative pathways of polyamine metabolism were investigated in sieve-tube sap, xylem sap and tissues of seedlings and adult plants of Ricinus communis L. Polyamines were present in tissues and translocation fluids of both seedlings and adult plants in relatively high amounts. Only free polyamines were translocated through the plant, as indicated by the finding that only the free form was detected in the phloem and the xylem sap. Removal of the endosperm increased the polyamine content in the sieve-tube exudate of seedlings. The level and pattern of polyamines in tissue of adult leaves changed during leaf age, but not, however, in the sieve-tube sap. Xylem sap was relatively poor in polyamines. Polyamine loading in the phloem was demonstrated by incubating cotyledons with [14O]putrescine and several unlabelled polyamines. Feeding cotyledons with cadaverine and spermidine led to a decrease in the level of putrescine in sieve-tube sap, indicating a competitive effect. Comparison of polyamine content in the tissue and export rate showed that the export would deplete the leaves of polyamines within 1–3 d, if they were not replenished by biosynthesis. Polyamine biosynthesis in Ricinus proceeds mostly via arginine decarboxylase, which in vitro is 100-fold more active than ornithine decarboxylase. The highest arginine decarboxylase, ornithine decarboxylase and diamine oxidase activities were detected in cotyledons, while in sieve-tube sap only a slight arginine decarboxylase activity was found.

International audience; Climate change is rapidly degrading the conditions of crop production. For instance, increasing salinization and aridity is forecasted to increase in most parts of the world. As a consequence, new stress-tolerant species and genotypes must be identified and used for future agriculture. Stress-tolerant species exist but are actually underutilized and neglected. Many stress-tolerant species are indeed traditional crops that are only cultivated by farmers at a local scale. Those species have a high biodiversity value. Besides, the human population will probably reach nine billion within coming decades. To keep pace with population growth, food production must increase dramatically despite the limited availability of cultivable land and water. Here, we review the benefits of quinoa, Chenopodium quinoa Willd., a seed crop that has endured the harsh bioclimatic conditions of the Andes since ancient times. Although the crop is still mainly produced in Bolivia and Peru, agronomic trials and cultivation are spreading to many other countries. Quinoa maintains productivity on rather poor soils and under conditions of water shortage and high salinity. Moreover, quinoa seeds are an exceptionally nutritious food source, owing to their high protein content with all essential amino acids, lack of gluten, and high content of several minerals, e.g., Ca, Mg, Fe, and health-promoting compounds such as flavonoids. Quinoa has a vast genetic diversity resulting from its fragmented and localized production over the centuries in the Andean region, from Ecuador to southern Chile, and from sea level to the altiplano. Quinoa can be adapted to diverse agroecological conditions worldwide. Year 2013 has therefore been declared the International Year of Quinoa by the United Nations Food and Agriculture Organization. Here, we review the main characteristics of quinoa, its origin and genetic diversity, its exceptional tolerance to drought and salinity, its nutritional properties, the reasons why this crop can offer several ecosystem services, and the role of Andean farmers in preserving its agrobiodiversity. Finally, we propose a schematic model integrating the fundamental factors that should determine the future utilization of quinoa, in terms of food security, biodiversity conservation, and cultural identity.

Chenopodium quinoa Willd. (quinoa) is a halophyte for which some parameters linked to salt tolerance have been investigated separately in different genotypes and under different growth conditions. In this study, several morphological and metabolic responses were analysed in parallel after exposure to salinity. In vitro seed germination was initially delayed by a 150 mM NaCl treatment but eventually reached the same level as the control (0 mM NaCl), whereas seedling root growth was enhanced; both parameters were moderately inhibited (~35–50%) by 300 mM NaCl. In pot grown plants, plant size was reduced by increasing salinity (0–750 mM NaCl). Transpiration and stomatal conductance were decreased at the highest salinity levels tested, consistent with reduced stomatal density and size. The density of epidermal bladder cells (EBCs) on the leaf surface remained unaffected up to 600 mM NaCl. Tissue contents of Na+ and Cl– increased dramatically with salt treatment, but resulted in only a 50% increase in Na+ from 150 to 750 mM NaCl. Internal K+ was unaffected up to 450 mM NaCl but increased at the highest salinity levels tested. Excretion through sequestration into EBCs was limited (generally ≤20%) for all ions. A modest dose-dependent proline accumulation, and concomitant reduction in total polyamines and putrescine efflux occurred in NaCl-treated plants. Results confirm the importance of inorganic ions for osmotic adjustment, the plant’s ability to maintain K+ levels and the involvement of putrescine efflux in maintaining ionic balance under high salinity conditions. Conversely, ion excretion and proline appear to play a minor role. Taken together these results indicate which parameters could be used for future comparison among different genotypes.

Chenopodium quinoa (Willd.) is an Andean plant showing a remarkable tolerance to abiotic stresses. In Chile, quinoa populations display a high degree of genetic distancing, and variable tolerance to salinity. To investigate which tolerance mechanisms might account for these differences, four genotypes from coastal central and southern regions were compared for their growth, physiological, and molecular responses to NaCl at seedling stage. Seeds were sown on agar plates supplemented with 0, 150 or 300mM NaCl. Germination was significantly reduced by NaCl only in accession BO78. Shoot length was reduced by 150mM NaCl in three out of four genotypes, and by over 60% at 300mM (except BO78 which remained more similar to controls). Root length was hardly affected or even enhanced at 150mM in all four genotypes, but inhibited, especially in BO78, by 300mM NaCl. Thus, the root/shoot ratio was differentially affected by salt, with the highest values in PRJ, and the lowest in BO78. Biomass was also less affected in PRJ than in the other accessions, the genotype with the highest increment in proline concentration upon salt treatment. Free putrescine declined dramatically in all genotypes under 300mM NaCl; however (spermidine+spermine)/putrescine ratios were higher in PRJ than BO78. Quantitative RT-PCR analyses of two sodium transporter genes, CqSOS1 and CqNHX, revealed that their expression was differentially induced at the shoot and root level, and between genotypes, by 300mM NaCl. Expression data are discussed in relation to the degree of salt tolerance in the different accessions.