stilbenes and methyl-jasmonate

Tools to address the mismatch between technological and phenolic maturity of grapes are needed. Application of elicitors could be an effective alternative. This work compares the effect of the application of methyl jasmonate (MeJ) in conventional form and, as a novelty, in the form of MeJ-doped nanoparticles (ACP-MeJ) on the phenolic composition of Tempranillo grapes. Results showed that, regardless of season, both treatments increased the grape total phenols content. In 2019, most of the anthocyanins, and to a lesser extent the flavanols, increased with the application of MeJ, and several hydroxycinnamic acids increased in the grapes treated with ACP-MeJ, with dose 10 times lower than those of the MeJ conventional. In 2020, anthocyanins were not affected by the treatments, but total flavanols, flavonols, hydroxybenzoic acid, and stilbenes increased after ACP-MeJ application. Thus, foliar application of ACP-MeJ could serve to increase grape phenolic composition, reducing maturity decoupling and the environmental impact.

Topics: Anthocyanins; Coumaric Acids; Flavonols; Fruit; Hydroxybenzoates; Incidence; Nanoparticles; Phenols; Polyphenols; Seasons; Stilbenes; Vitis; Wine

Peanut cultivars are known to produce stilbene compounds. Transcriptional control plays a key role in the early stages of the stress response mechanism, involving both PR-proteins and stilbene compounds. In this study, the production of stilbenoid compounds, especially prenylated, was investigated in two cultivars of peanut hairy root lines, designated as K2-K599 and T9-K599 elicited with a combination of chitosan (CHT), methyl jasmonate (MeJA), and cyclodextrin (CD): CHT + MeJA + CD. The antioxidant activities and stilbenoid content of both K2-K599 and T9-K599 hairy root lines increased significantly during the elicitation period. The T9-K599 hairy root line expressed higher ABTS and FRAP antioxidant activities than the K2-K599 line while the latter exhibited greater total phenolic content than the former at all-time points. Additionally, the K2-K599 line exhibited more stilbene compounds, including trans-resveratrol, trans-arachidin-1, and trans-arachidin-3 than the T9-K599 line, which showed statistically significant differences at all-time points. Gene expression of the enzyme involved in the stilbene biosynthesis pathway (PAL, RS, RS3) was observed, responding early to elicitor treatment and the metabolic production of a high level of stilbenoid compounds at a later stage. The antioxidant enzyme (CuZn-SOD, APX, GPX) and pathogenesis-related protein (PR; PR4A, PR5, PR10, chitinase) genes were strongly expressed after elicitor treatment at 24 h and decreased with an increasing elicitation time. Investigation of the response mechanism illustrates that the elicitor treatment can affect various plant responses, including plant cell wall structure and integrity, antioxidant system, PR-proteins, and secondary plant metabolites at different time points after facing external environmental stimuli.

Topics: Acetates; Antioxidants; Arachis; Chitosan; Cyclodextrins; Cyclopentanes; Fabaceae; Oxylipins; Plant Roots; Stilbenes

The present study is aimed at determining whether leaf volatile organic compounds (VOCs) are good markers of the grapevine response to defence elicitors in the field. It was carried out in two distinct French vineyards (Burgundy and Bordeaux) over 3 years. The commercial elicitor Bastid

Topics: Acetates; Copper Sulfate; Cyclopentanes; Farms; Oxylipins; Plant Leaves; Stilbenes; Terpenes; Volatile Organic Compounds

Prenylated stilbenoids such as arachidin-1 and arachidin-3 are stilbene derivatives that exhibit multiple pharmacological activities. We report an elicitation strategy using different combinations of cyclodextrin, hydrogen peroxide, methyl jasmonate and magnesium chloride to increase arachidin-1 and arachidin-3 production in peanut hairy root cultures. The treatment of hairy root cultures with cyclodextrin with hydrogen peroxide selectively enhanced arachidin-1 yield (132.6 ± 20.4 mg/L), which was 1.8-fold higher than arachidin-3. Similarly, cyclodextrin combined with methyl jasmonate selectively enhanced arachidin-3 yield (178.2 ± 6.8 mg/L), which was 5.5-fold higher than arachidin-1. Re-elicitation of the hairy root cultures further increased the levels of arachidin-1 and arachidin-3 by 24% and 42%, respectively. The ethyl acetate extract of the culture medium was consecutively fractionated by normal- and reversed-phase column chromatography, followed by semi-preparative HPLC purification on a C18 column to yield arachidin-1 with a recovery rate of 32% and arachidin-3 with a recovery rate of 39%, both at higher than 95% purity. This study provided a sustainable strategy to produce high-purity arachidin-1 and arachidin-3 using hairy root cultures of peanuts combined with column chromatography and semi-preparative HPLC.

Topics: Acetates; Arachis; Cyclodextrins; Cyclopentanes; Hemiterpenes; Hydrogen Peroxide; Magnesium Chloride; Oxylipins; Plant Roots; Stilbenes

Plants counteract Cd toxicity by activating cellular stress responses. The simultaneous exogenous application of methyl jasmonate (MeJA) and methyl-β-cyclodextrin (CD) before Cd exposure improved the response of Arachis hypogaea hairy root culture to the unfavourable effects of Cd toxicity. At 24 h after elicitation, genes that encode key enzymes in the phenylpropanoid biosynthesis pathway (i.e., PAL and RS3) were up-regulated to 3.2- and 5.4-fold changes respectively, thereby inducing stilbene production. The up-regulation of genes that encode transcription factors (i.e., ERF1 and ERF6) significantly increased the expression of several genes (PR4A, PR5, PR10, and chitinase) that encode the pathogenesis-related (PR) proteins to 25.8-, 45-, 5- and 12.6-fold changes, respectively. The more dramatic up-regulation of PR protein-encoding genes demonstrated the significant role of defence proteins in plant protective mechanisms. The prolonged (i.e., 72-h) treatment with MeJA + CD_Cd triggered adaptive responses by substantially increasing the levels of antioxidants, stilbenes, and other phenolic substances. These findings suggest that the interaction between signalling elicitors (MeJA and CD) and Cd modulates a complex signalling network for plant defence system.

Topics: Acetates; Adaptation, Physiological; Arachis; Cadmium; Chromatography, Liquid; Cyclopentanes; Oxylipins; Plant Roots; Real-Time Polymerase Chain Reaction; Signal Transduction; Stilbenes; Stress, Physiological; Tandem Mass Spectrometry; Transcriptome

Plants encounter diverse stressors simultaneously with changing environmental factors. The combined effect of different types of stresses can have a wide range of effects on plants. The present study demonstrated that various stress factors such as the combination of chemical elicitors, namely paraquat (PQ), methyl jasmonate (MeJA) and methyl-β-cyclodextrin (CD), light exposure versus darkness, and mechanical shearing stress affected the defence response in peanut hairy root culture. The antioxidant activities were dramatically increased at all time points after hairy roots were subjected to elicitation with PQ + MeJA + CD under root cutting in both light and dark conditions. The stilbene compounds were highly increased in the culture medium after elicitor treatment of uncut hairy roots under dark conditions. In contrast to the high stilbene contents detected in culture medium under dark conditions, the transcription of the stilbene biosynthesis genes PAL, RS and RS3 was enhanced by the effect of light in uncut hairy root tissues. The antioxidant enzyme genes APX, GPX and CuZn-SOD of uncut and cut hairy roots were more highly expressed in light conditions than in dark conditions. The pathogenesis-related protein (PR)-encoding genes chitinase, PR4A, PR5 and PR10 of uncut hairy roots were highly expressed in response to light conditions compared to dark conditions at all time points. Recent evidence of the production of antioxidant stilbene compounds and defence response genes has implicated plant protective functions through defence responses under different stress challenges. Plant responses might therefore be regulated by the coordination of different signal responses through dynamic pathways.

Topics: Acetates; Arachis; beta-Cyclodextrins; Cyclopentanes; Darkness; Gene Expression Regulation, Plant; Light; Oxylipins; Paraquat; Plant Roots; Stilbenes; Stress, Mechanical; Tissue Culture Techniques

Topics: Acetates; Biosynthetic Pathways; Catechin; Cell Culture Techniques; Cell Line; Cyclopentanes; Electric Conductivity; Flavonoids; Glucosides; Hydrogen-Ion Concentration; Light; Oxylipins; Phenols; Phenylalanine; Resveratrol; Secondary Metabolism; Stilbenes; Suspensions; Vitis

The objective of the present work was to synthesize biopolymeric nanoparticles (NPs) entrapping the resistance-inductor methyl jasmonate (MeJA) to be employed as a novel and alternative strategy in integrated pest management. NPs were prepared by using a continuous flow microfluidic reactor that allows to precisely control some features that are crucial for applications such as size, polydispersion, morphology and reproducibility. Poly(lactic-co-glycolic acid) (PLGA), a biopolymer largely studied for its use in biological applications, was chosen for the production of NPs entrapping MeJA, a biotic endogenous elicitor able to trigger plant's defense responses. The effect of different fluid-dynamic conditions, PLGA molecular weight and concentration on NP properties (dimensions, polydispersion, morphology, stability) was evaluated. DLS and SEM were employed to characterize the obtained NPs. MeJA-loaded PLGA NPs ranging from 40 to 70 nm were administered to Vitis vinifera cell cultures, in order to evaluate the biological response in terms of stilbene biosynthesis. HPLC investigations showed a faster response when the elicitor was administered by PLGA NPs in comparison with free MeJA. This result demonstrates that the encapsulation in PLGA NPs significantly promotes MeJA cell uptake and the activation of MeJA-induced responses.

Topics: Acetates; Cyclopentanes; Microfluidic Analytical Techniques; Nanoparticles; Oxylipins; Pest Control; Plant Cells; Polyglycolic Acid; Stilbenes; Vitis

Vitis rupestris is used as rootstock or to obtain hybrids with Vitis vinifera, due to its resistance to certain pathogens. Its resistance mechanisms are poorly understood, while it is known that stilbene neo-synthesis is a central defense strategy in V. vinifera. In the present study, the response to methyl jasmonate (MeJa) and light treatment in terms of stilbene biosynthesis and the expression of genes involved in polyphenol biosynthesis was investigated in V. vinifera and V. rupestris cells. The two species exhibited a similar constitutive stilbene content [2.50-2.80 mg g

Topics: Acetates; Cyclopentanes; Gene Expression Regulation, Plant; Light; Oxylipins; Stilbenes; Vitis

Morus alba L. (Moraceae) has been used in traditional medicine for the treatment of several illnesses. Recent research also revealed several pharmacological activities from many groups of secondary metabolites, including the stilbenoids mulberroside A, oxyresveratrol, and resveratrol, which are promising compounds for cosmetic and herbal supplement products. In our previous study, cell cultures of M. alba showed high productivity of these compounds. In this study, we attempted to develop immobilized cell cultures of M. alba and to test the effect of elicitors and precursors on the production of stilbenoids. The immobilization of the M. alba cells significantly promoted the secretion of mulberroside A into the extracellular matrix and culture media to 60%, while enhancing the level of oxyresveratrol and resveratrol by 12- and 27-fold, respectively. The elicitation of immobilized cells with a combination of 50 µM methyl jasmonate and 0.5 mg/mL yeast extract for 24 h promoted a twofold increase in the production of all three stilbenoids. Furthermore, the addition of 0.05 mM L-phenylalanine, 0.03 mM L-tyrosine, or a combination resulted in the enhancement of mulberroside A production for up to twofold. The addition of L-tyrosine significantly enhanced the production of oxyresveratrol and resveratrol. This is the first report of stilbenoid production using immobilized cell cultures of M. alba. The cultures have benefits over normal cell suspension cultures by promoting the secretion of mulberroside A and enhancing the levels of oxyresveratrol and resveratrol. Thus, it could be a candidate method for the production of these stilbenoids.

Topics: Acetates; Alginates; Cell Culture Techniques; Culture Media; Cyclopentanes; Disaccharides; Morus; Oxylipins; Phenylalanine Ammonia-Lyase; Plant Extracts; Plant Leaves; Resveratrol; Stilbenes; Tyrosine

Grape stilbenes are a well-known family of plant polyphenolics that have been confirmed to have many biological activities in relation to health benefits. In the present study, we investigated the effect of methyl jasmonate (MeJA) elicitor at four different concentrations (25, 50, 100 and 200 μM) in combination or not with high-level light irradiation (10,000 LUX) on a cell line obtained from the pulp of Vitis vinifera cv. Shahani. Our results showed that the stilbene synthesis pathway is inhibited by high-light conditions. A concentration of 50 μM MeJA was optimum for efficient production and high accumulation of total phenolics and total flavonoids as well as total stilbenoids. Furthermore, we showed that there is a significant negative correlation between the production of these metabolites and cell growth. These data provide valuable information for the future scale-up of cell cultures for the production of these very high value compounds in bioreactor system.

Topics: Acetates; Biosynthetic Pathways; Cell Culture Techniques; Cell Line; Cyclopentanes; Darkness; Flavonoids; Light; Oxylipins; Phenols; Plant Cells; Stilbenes; Vitis

Stilbenes have a significant biological activity and are one of the most important non-flavonoid contributors to grape and wine health-related properties. The accumulation of this class of compounds could be favored by viticultural practices such as the application of biostimulants. However, stilbene concentration also depends on several factors, including, for example, grape variety. Therefore, the aim of this work was to study the influence of foliar treatments carried out with elicitors (methyl jasmonate (MeJ) and a commercial foliar spray (YD)) and nitrogen compounds (phenylalanine and urea) on the grape stilbene composition of three varieties: Grenache, Graciano and Tempranillo. An ultra-high-pressure liquid chromatographic methodology was validated for stilbene determination.. Results showed that, despite the huge influence of the grape variety, YD significantly improved stilbene composition in Grenache and Graciano, while MeJ increased the stilbene content in Graciano and Tempranillo. As for the nitrogen treatments, phenylalanine significantly increased the stilbene concentration in Graciano, while urea treatment increased it in Tempranillo. However, the application of elicitors had a greater effect than the nitrogen compounds.. Overall, the foliar application of the elicitors could be a suitable practice for increasing the amount of stilbenes in grape and, therefore, its nutraceutical properties. © 2017 Society of Chemical Industry.

Topics: Acetates; Chromatography, High Pressure Liquid; Crop Production; Cyclopentanes; Fruit; Oxylipins; Phenylalanine; Stilbenes; Urea; Vitis; Wine

Two preharvest treatments (methyl jasmonate or cell wall yeast) of grapevines (Monastrell and Tempranillo) were applied during two vintages (2014 and 2015) to check whether these elicitors enhanced stilbene accumulation in berries at the moment of harvest and in the corresponding wines elaborated with them. The main objective was checking the effect of treatment, variety and year on stilbene composition due to the interesting health-related properties of these compounds in both grapes and wines. The results pointed to inter-varietal and inter-annual differences, and that the treatments generally enhanced the stilbene composition of grapes and, particularly, of wines. The increase was more evident in Monastrell variety than in Tempranillo variety and in their wines more than their grapes during 2014 vintage but not during 2015 vintage.

Topics: Acetates; Agriculture; Cell Wall; Cyclopentanes; Fruit; Humans; Oxylipins; Species Specificity; Stilbenes; Vitis; Wine; Yeasts

Plants employ two layers of defence that differ with respect to cell death: pathogen-associated molecular pattern (PAMP)-triggered immunity (PTI) and effector-triggered immunity (ETI). In our previous work, we have comparatively mapped the molecular events in a cell system derived from the wild American grape Vitis rupestris, where cell death-independent defence can be triggered by PAMP flg22, whereas the elicitor Harpin activates a cell death-related ETI-like response. Both defence responses overlapped with respect to early events, such as calcium influx, apoplastic alkalinisation, oxidative burst, mitogen-activated protein kinase (MAPK) signalling, activation of defence-related genes and accumulation of phytoalexins. However, timing and amplitude of early signals differed. In the current study, we address the role of jasmonates (JAs) as key signalling compounds in hypersensitive cell death. We find, in V. rupestris, that jasmonic acid and its bioactive conjugate jasmonoyl-isoleucine (JA-Ile) rapidly accumulate in response to flg22 but not in response to Harpin. However, Harpin can induce programmed cell death, whereas exogenous methyl jasmonate (MeJA) fails to do so, although both signals induce a similar response of defence genes. Also in a second cell line from V. vinifera cv. 'Pinot Noir', where Harpin cannot activate cell death and where flg22 fails to induce JA and JA-Ile, defence genes are activated in a similar manner. These findings indicate that the signal pathway culminating in cell death must act independently from the events culminating in the accumulation of toxic stilbenes.

Topics: Acetates; Bacterial Proteins; Cell Death; Cell Line; Cell Survival; Cyclopentanes; Flagellin; Gene Expression Regulation, Plant; Isoleucine; Oxylipins; Plant Proteins; Salicylic Acid; Stilbenes; Transcription, Genetic; Vitis

In this work, transgenic lines of suspension cultured cells of Vitis vinifera cv. Monastrell containing the plasmid pMOG800-sts have been obtained. The cell growth of these transgenic cell lines decreased slightly as compared to non-transgenic suspension cultured cells, while cell viability was not affected. In addition, the elicitation with cyclodextrins and methyl jasmonate enhanced the production of trans-resveratrol, observing the highest levels of this compound in sts-expressing transgenic Vitis suspension cultured cells with the sts expression cassette in the forwards orientation. Moreover, the forwards 2 (F2) transgenic cell line produced the greater levels of trans-resveratrol in comparison with the non-transgenic cell line. In fact, when suspension cultured cells were treated with both elicitors, the accumulation of trans-resveratrol outside the cells in the F2 transgenic suspension cultured cells increased twice (1458 mg.L

Topics: Acetates; Acyltransferases; Agrobacterium; Cell Line; Cell Survival; Cells, Cultured; Cyclodextrins; Cyclopentanes; Gene Expression Regulation, Plant; Oxylipins; Plant Growth Regulators; Plants, Genetically Modified; Resveratrol; Stilbenes; Transformation, Genetic; Vitis

The effect of the postharvest treatment of methyl jasmonate enantiomers in conjunction with ethanol on bioformation of resveratrol and quercetin glycosides in grapes was evaluated. The antioxidant activity of treated grape extracts as compared with untreated extracts was also assayed. Exogenous (-)-methyl jasmonate in combination with ethanol induced a significant increase in the levels of resveratrol (from 27 to 39 μg g(-1)), quercetin-3-O-glucoside (from 59 to 136 μg g(-1)), quercetin-3-O-galactoside (from 398 to 807 μg g(-1)) and quercetin-3-O-rutinoside (from 23 to 43 μg g(-1)). (+)-Methyl jasmonate with ethanol also resulted in increase of quercetin-3-O-glucoside and quercetin-3-O-rutinoside. However, no (+)-methyl jasmonate effect was observed for resveratrol and quercetin-3-O-galactoside. Both (-)- and (+)-methyl jasmonate treatments provided with extracts with higher antioxidant activity. From the results found in the present work postharvest treatment with (-)-methyl jasmonate in conjunction with ethanol is proposed as a mean to obtain polyphenol-enriched grape extracts with improved antioxidant properties. The procedure here developed is proposed as a mean to obtain functional grapes. Extracts obtained from grapes treated with (-)-methyl jasmonate with ethanol can be particularly useful for industry due to their high antioxidant capacity.

Topics: Acetates; Antioxidants; Cyclopentanes; Ethanol; Flavonoids; Food Handling; Fruit; Glucosides; Oxylipins; Polyphenols; Quercetin; Resveratrol; Rutin; Stilbenes; Vitis

Topics: Acetates; Benzofurans; beta-Cyclodextrins; Cyclopentanes; Glucosides; Oxylipins; Plant Roots; Resorcinols; Resveratrol; Stilbenes; Vitis

Blueberry (Vaccinium corymbosum) is a fruit very much appreciated by consumers for its antioxidant potential and health-promoting traits. Its beneficial potential properties are mainly due to a high content of anthocyanins and their amount can change after elicitation with methyl jasmonate. The aim of this work is to evaluate the changes in expression of several genes, accumulation of phenolic compounds and alterations in antioxidant potential in two different blueberry cultivars ('Duke' and 'Blueray') in response to methyl jasmonate (0.1 mM). Results showed that 9 h after treatment, the expression of phenylalanine ammonium lyase, chalcone synthase and anthocyanidin synthase genes was stimulated more in the 'Blueray' variety. Among the phenols measured an increase was recorded also for epicatechin and anthocyanin concentrations. 'Duke' is a richer sourche of anthocyanins compared to 'Blueray', treatment with methyl jasmonate promoted in 'Blueray' an increase in pigments as well as in the antioxidant potential, especially in fully ripe berries, but treated 'Duke' berries had greater levels, which were not induced by methyl jasmonate treatment. In conclusion, methyl jasmonate was, in some cases, an effective elicitor of phenolic metabolism and gene expression in blueberry, though with different intensity between cultivars.

Topics: Acetates; Anthocyanins; Biphenyl Compounds; Blueberry Plants; Carbohydrates; Cyclopentanes; Flavonols; Free Radical Scavengers; Gene Expression Regulation, Plant; Genes, Plant; Oxylipins; Phenols; Picrates; Plant Extracts; Propanols; Real-Time Polymerase Chain Reaction; RNA, Messenger; Stilbenes; Sucrose

This study was conducted to characterize the forms of disease resistance induced by methyl jasmonate (MeJA) in harvested grape berries and to evaluate the impact of the induced resistance on fruit quality. The results showed that MeJA treatment at concentrations from 10 to 100μmol/L could effectively induce disease resistance against Botrytis cinerea and reduce disease incidence in grape berries. The induced disease resistance was tightly associated with increased H2O2 generation, enhanced expression of the defense-related gene VvNPR1.1 and accumulation of stilbene phytoalexins such as tran-resveratrol and its oligomer (trans-)ε-viniferin. The expression of the defense-related gene and synthesis of phytoalexins in 10μmol/L MeJA-treated grape berries were only significantly enhanced upon inoculating the berries with B. cinerea, whereas the 50 or 100μmol/L of MeJA treatment directly induced these defense responses. Hence, we deduce that the low concentration of MeJA (10μmol/L) triggered a priming defense mechanism, while higher concentrations of MeJA (50 or 100μmol/L) directly activated defense responses, thus enhancing disease resistance in grape berries. Moreover, the primed grape berries maintained higher contents of soluble sugars and higher DPPH radical scavenging activity and reducing power compared with those expressing direct defense responses. These results indicate that priming of defense is a cost-effective strategy to protect harvested grape berries from B. cinerea infection in terms of minimizing quality loss.

Topics: Acetates; Benzofurans; Botrytis; Cyclopentanes; Disease Resistance; Fruit; Gene Expression Regulation; Hydrogen Peroxide; Oxylipins; Plant Growth Regulators; Resveratrol; Stilbenes; Vitis

Peanut (Arachis hypogaea) produces stilbenoids upon exposure to abiotic and biotic stresses. Among these compounds, the prenylated stilbenoids arachidin-1 and arachidin-3 have shown diverse biological activities with potential applications in human health. These compounds exhibit higher or novel biological activities in vitro when compared to their nonprenylated analogues piceatannol and resveratrol, respectively. However, assessment of these bioactivities in vivo has been challenging because of their limited availability. In this study, hairy root cultures of peanut were induced to produce stilbenoids upon treatment with elicitors. Co-treatment with 100 μM methyl jasmonate (MeJA) and 9 g/L methyl-β-cyclodextrin (CD) led to sustained high levels of resveratrol, piceatannol, arachidin-1, and arachidin-3 in the culture medium when compared to other elicitor treatments. The average yields of arachidin-1 and arachidin-3 were 56 and 148 mg/L, respectively, after co-treatment with MeJA and CD. Furthermore, MeJA and CD had a synergistic effect on resveratrol synthase gene expression, which could explain the higher yield of resveratrol when compared to treatment with either MeJA or CD alone. Peanut hairy root cultures were shown to be a controlled and sustainable axenic system for the production of the diverse types of biologically active stilbenoids.

Topics: Acetates; Arachis; beta-Cyclodextrins; Cell Culture Techniques; Culture Media; Cyclopentanes; Gene Expression Regulation, Plant; Hemiterpenes; Oxylipins; Plant Growth Regulators; Plant Proteins; Plant Roots; Resveratrol; Stilbenes

Morus alba L. has been used in Asian traditional medicine as an anti-inflammatory, anti-asthmatic, anthelmintic and as a whitening agent in cosmetic products. Mulberroside A is the major active compound from M. alba root bark. In this study, cell suspension and root cultures of M. alba were established, and the effect of the elicitors on the enhancement of mulberroside A production in M. alba was investigated. The cell suspension and root cultures of M. alba were exposed to elicitors and then mulberroside A contents were determined by an indirect competitive ELISA method. High levels of mulberroside A were obtained by addition of 100 and 200 μM salicylic acid with 24 h exposure time in cell suspension cultures (37.9 ± 1.5 and 34.0 ± 4.7 mg/g dry wt., respectively). Furthermore, addition of yeast extract at 2 mg/mL with 24 h exposure time can significantly increase mulberroside A contents from both cell suspension (3.2-fold) and root cultures (6.6-fold). Mulberroside A contents from both cell suspension and root cultures after treatment with elicitors are similar or higher than those found in the intact root and root bark of several years old M. alba. These results indicate that mulberry tissue cultures using the elicitation method are interesting alternative sources for mulberroside A production.

Topics: Acetates; Chitosan; Culture Techniques; Cyclopentanes; Disaccharides; Morus; Oxylipins; Plant Roots; Salicylic Acid; Stilbenes; Yeasts

Transcriptome profiling has been widely used to analyze transcriptomic variation in plants subjected to abiotic or biotic stresses. Although gene expression changes induced by methyl jasmonate (MeJA) have been profiled in several plant species, no information is available on the MeJA-triggered transcriptome response of Polygonum multiflorum Thunb., a species with highly valuable medicinal properties. In this study, we used transcriptome profiling to investigate transcriptome changes in roots of P. multiflorum seedlings subjected to a 0.25 mmol/L-MeJA root-irrigation treatment. A total of 18 677 differentially expressed genes (DEGs) were induced by MeJA treatment, of which 4535 were up-regulated and 14 142 were down-regulated compared with controls. These DEGs were associated with 125 metabolic pathways. In addition to various common primary and secondary metabolic pathways, several secondary metabolic pathways related to components with significant pharmacological effects were enriched by MeJA, including arachidonic acid metabolism, linoleic acid metabolism, and stilbenoid biosynthesis. The MeJA-induced transcriptome changes uncovered in this study provide a solid foundation for future study of functional genes controlling effective components in secondary metabolic pathways of P. multiflorum.

Topics: Acetates; Arachidonic Acid; Cyclopentanes; Fallopia multiflora; Gene Expression Profiling; Gene Expression Regulation, Plant; Genome, Plant; Linoleic Acid; Metabolic Networks and Pathways; Oxylipins; Plant Growth Regulators; Plant Roots; Stilbenes; Transcriptome

Stilbenoids are polyphenolic phytoalexins that exhibit potential health applications in humans. Hairy root cultures of muscadine grape (Vitis rotundifolia Michx.) were used to study the biochemical and molecular regulation of stilbenoid biosynthesis upon treatment with 100 μM methyl jasmonate (MeJA) or 10 mM hydrogen peroxide (H2O2) over a 96-h period. Resveratrol, piceid, and ε-viniferin were identified in higher concentrations in the tissue whereas resveratrol was the most abundant stilbenoid in the medium under either treatment. An earlier increase in resveratrol accumulation was observed for the MeJA-treated group showing a maximum at 12 h in the tissue and 18 h in the medium. Furthermore, the antioxidant capacity of extracts from the tissue and medium was determined by the 2,2'-azinobis[3-ethylbenzthiazoline sulfonic acid] (ABTS) and the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assays showing correlation with the stilbenoid content. Fourteen candidate reference genes for qPCR were tested under the described experimental conditions and resulted in the selection of 5 reference genes. Quantitative analyses of transcripts for phenylalanine ammonia-lyase (PAL), resveratrol synthase (RS), and two stilbene synthases (STS and STS2) showed the highest RNA level induction at 3 h for both treatments with a higher induction for the MeJA treatment. In contrast, the flavonoid-related chalcone synthase (CHS) transcripts showed induction and a decrease in expression for MeJA and H2O2 treatments, respectively. The observed responses could be related to an oxidative burst triggered by the exposure to abiotic stressor compounds with signaling function such as MeJA and H2O2 which have been previously related to the synthesis of secondary metabolites.

Topics: Acetates; Antioxidants; Benzothiazoles; Biphenyl Compounds; Culture Media; Cyclopentanes; Genes, Plant; Hydrogen Peroxide; Oxylipins; Picrates; Plant Roots; RNA, Plant; Stilbenes; Sulfonic Acids; Vitis

Stilbene-enriched wine is considered to be an interesting new food product with added value due to its potential health-promoting properties. Stilbene concentration in grape is highly variable and rather scarce. However, it can be increased by stress treatments. For this reason, numerous pre- and postharvest grape treatments, and some combinations of them, have been tested to maximize stilbene content in grapes. In the present manuscript, Syrah grapes were treated with (i) methyl jasmonate (MEJA), (ii) ultraviolet light (UVC), and (iii) methyl jasmonate and ultraviolet light (MEJA-UVC) and compared with untreated grapes. Afterward, winemaking was developed. Wine achieved by combination of both treatments (MEJA-UVC) contained significantly higher stilbene concentration (trans-resveratrol and piceatannol) than its respective control (2.5-fold). Wine quality was improved in color-related parameters (color intensity, L*, a*, b*, ΔE*, anthocyanins, and tannin). Moreover, MEJA-UVC wines obtained the highest score in sensorial analysis. To the best of our knowledge, this is the first time that pre- and postharvest treatments are combined to increase stilbenes in wine.. The effect of treatment combination (methyl jasmonate and UVC light) on grape and wine was evaluated. Our results highlight the positive effect of the treatments in stilbene content, color parameters, and sensorial analysis. Moreover, added-value by-products were achieved.

Topics: Acetates; Anthocyanins; Color; Cyclopentanes; Fruit; Humans; Odorants; Oxylipins; Plant Growth Regulators; Resveratrol; Stilbenes; Tannins; Taste; Ultraviolet Rays; Vitis; Wine

The high effectiveness of cyclic oligosaccharides like cyclodextrins in the production of trans-resveratrol in Vitis vinifera cell cultures is enhanced in the presence of methyl jasmonate. In order to dissect the basis of the interactions among the elicitation responses triggered by these two compounds, a transcriptional analysis of grapevine cell cultures treated with cyclodextrins and methyl jasmonate separately or in combination was carried out. The results showed that the activation of genes encoding enzymes from phenylpropanoid and stilbene biosynthesis induced by cyclodextrins alone was partially enhanced in the presence of methyl jasmonate, which correlated with their effects on trans-resveratrol production. In addition, protein translation and cell cycle regulation were more highly repressed in cells treated with cyclodextrins than in those treated with methyl jasmonate, and this response was enhanced in the combined treatment. Ethylene signalling was activated by all treatments, while jasmonate signalling and salicylic acid conjugation were activated only in the presence of methyl jasmonate and cyclodextrins, respectively. Moreover, the combined treatment resulted in a crosstalk between the signalling cascades activated by cyclodextrins and methyl jasmonate, which, in turn, provoked the activation of additional regulatory pathways involving the up-regulation of MYB15, NAC and WRKY transcription factors, protein kinases and calcium signal transducers. All these results suggest that both elicitors cause an activation of the secondary metabolism in detriment of basic cell processes like the primary metabolism or cell division. Crosstalk between cyclodextrins and methyl jasmonate-induced signalling provokes an intensification of these responses resulting in a greater trans-resveratrol production.

Topics: Acetates; Cyclodextrins; Cyclopentanes; Gene Expression Profiling; Gene Expression Regulation, Plant; Oxylipins; Plant Growth Regulators; Plant Proteins; Resveratrol; Stilbenes; Transcriptome; Up-Regulation; Vitis

Suspension-cultured cells of Vitis vinifera cv Monastrell were used to investigate the effects of methyljasmonate, ethylene and salicylic acid separately or in combination with cyclodextrins on both trans-resveratrol production and the induction of defense responses. The results showed that the addition of methyljasmonate or ethylene to suspension-cultured cells jointly treated with cyclodextrins and salicylic acid provoked a decrease of trans-resveratrol levels suggesting that salicylic acid has a negative and antagonistic effect with methyljasmonate or ethylene on trans-resveratrol production. Likewise, the exogenous application of these compounds induced the accumulation of pathogenesis-related proteins. Analysis of the extracellular proteome showed the presence of amino acid sequences homologous to an specific β-1,3-glucanase, class III peroxidases and a β-1,4-mannanase, which suggests that these signal molecules could play a role in mediating defense-related gene product expression in V. vinifera cv Monastrell. Apart from these inducible proteins, other proteins were found in both the control and elicited cell cultures of V. vinifera. These included class IV chitinase, polygalacturonase inhibitor protein and reticuline oxidase-like protein, suggesting that their expression is constitutive being involved in the modification of the cell wall architecture during cell culture growth and in the prevention of pathogen attack.

Topics: Acetates; Anti-Infective Agents; Cells, Cultured; Cyclodextrins; Cyclopentanes; Disease Resistance; Ethylenes; Gene Expression Regulation, Plant; Genes, Plant; Oxylipins; Plant Diseases; Plant Growth Regulators; Plant Proteins; Resveratrol; Salicylic Acid; Signal Transduction; Stilbenes; Vitis

In this work, the effect of different inducing factors on trans-resveratrol extracellular production in Monastrell grapevine suspension cultured cells is evaluated. A detailed analysis provides the optimal concentrations of cyclodextrins, methyljasmonate and UV irradiation dosage, optimal cell density, elicitation time and sucrose content in the culture media. The results indicate that trans-resveratrol production decreases as the initial cell density increases for a constant elicitor concentration in Monastrell suspension cultured cells treated with cyclodextrins individually or in combination with methyljasmonate; the decrease observed in cell cultures elicited with cyclodextrins alone is far more drastic than those observed in the combined treatment. trans-Resveratrol extracellular production observed by the joint use of cyclodextrins and methyljasmonate (1,447.8 ± 60.4 μmol trans-resveratrol g(-1) dry weight) is lower when these chemical compounds are combined with UV light short exposure (669.9 ± 45.2 μmol trans-resveratrol g(-1) dry weight). Likewise, trans-resveratrol production is dependent on levels of sucrose in the elicitation medium with the maximal levels observed with 20 g l(-1) sucrose and the joint action of cyclodextrins and 100 μM methyljasmonate. The sucrose concentration did not seem to limit the process although it affects significantly the specific productivity since the lowest sucrose concentration is 10 g l(-1), the highest productivity is reached (100.7 ± 5.8 μmol trans-resveratrol g(-1) dry weight g(-1) sucrose) using cyclodextrins and 25 μM methyljasmonate.

Topics: Acetates; Biotechnology; Cell Culture Techniques; Cells, Cultured; Culture Media; Cyclodextrins; Cyclopentanes; Dose-Response Relationship, Drug; Extracellular Space; Oxylipins; Resveratrol; Stilbenes; Sucrose; Ultraviolet Rays; Vitis

Plant secondary metabolites, such as stilbenes, have fungicidal potential and have been found in several plant species. Stilbenes in grapevine, such as resveratrol and pterostilbene, have recently attracted much attention, they are not only helping the plant to fight against pathogen attack, but they are also being widely used as ingredients of fungicide, anti-inflammatory drugs, antioxidant, and anti-infective agents. However, resveratrol O-methyltransferase gene, related with the synthesis of pterostilbene from resveratrol, has not been characterized effectively from Chinese wild Vitis pseudoreticulata. In this study, a candidate of resveratrol O-methyltransferase gene designated as VpROMT was isolated from a powdery mildew-resistant Chinese wild V. pseudoreticulata 'Baihe-35-1', and characterization studies were performed. Expression studies showed that VpROMT was predominantly expressed in developing roots yet not found in the leaves, stems, nor tendrils when the plants are not challenged. Results of qRT-PCR showed that VpROMT was rapidly induced by Erysiphe necator in V. pseudoreticulata and by methyl-jasmonate, UV-irradiation in suspension culture cells of Vitis romanetii. The expression level varies in different tissues of grapevine, which MeJA and UV-C treatment significantly upregulated the expression of VpROMT gene while UV-B treatment failed to. Co-expression of VpROMT and grapevine stilbene synthase (VpSTS) gene leads to the accumulation of pterostilbene in leaves of tobacco (Nicotiana tabacum) indicating that VpROMT was able to catalyze the biosynthesis of pterostilbene from resveratrol in over-expression transgenic tobacco plants.

Topics: Acetates; Acyltransferases; Amino Acid Sequence; Chromatography, High Pressure Liquid; Cloning, Molecular; Conserved Sequence; Cyclopentanes; Gene Expression; Gene Expression Regulation, Plant; Genes, Plant; Genetic Enhancement; Methyltransferases; Molecular Sequence Annotation; Nicotiana; Oxylipins; Phylogeny; Plant Diseases; Plant Growth Regulators; Plant Leaves; Plant Proteins; Plant Roots; Plants, Genetically Modified; Resveratrol; Saccharomycetales; Sequence Analysis, DNA; Stilbenes; Ultraviolet Rays; Vitis

Friable calli of Polygonum multiflorum Thunb have been induced in MS medium supplemented with 6-benzylaminopurine (6-BA) and kinetin (KT). Suspension cultures were initiated from friable calli by inoculating calli in liquid MS medium in shake flasks in the dark and 25 °C on an orbital shaker at 100 rpm. The maximum dry weight (DW, 7.85 g/L) and 2,3,5,4'-tetrahydroxystilbene-2-O-β-D-glycoside (THSG, 56.39 mg/L) of suspension cells was obtained in MS medium after 16 days culture. Both methyl jasmonate (MeJA) and salicylic acid (SA) could increase THSG production. The most appropriate concentration of MeJA was 100 μmol/L in MS medium, in which concentration THSG content reached the maximum value of 147.79 mg/L, which represented a 162.36% increase compared to that of the control (56.33 mg/L). The most appropriate concentration of SA was 125 μmol/L in MS medium, at which concentration THSG content reached its maximum value of 116.43 mg/L, a 106.69% increase compared to that of the control (56.33 mg/L).

Topics: Acetates; Cell Culture Techniques; Cyclopentanes; Glucosides; Oxylipins; Plant Cells; Polygonum; Salicylic Acid; Stilbenes; Suspensions

Vitis vinifera cell suspensions are a suitable system to study the metabolic regulation of a large range of polyphenols, including flavonoids and stilbenes that play important roles in plant development. Grape cv. Barbera petioles cell cultures were treated with red light and 10 μM methyl-jasmonate (MeJA), alone or in combination, to investigate their influence and/or induction effect on the production of anthocyanins, catechins and free and mono-glucosylated stilbenes. The synthesis of total anthocyanins was slightly decreased by red light alone, while MeJA and MeJA plus red light increased the levels of these metabolites. When compared to the relative controls, the red light treatment decreased the amount of catechins and increased their release in the culture medium, while MeJA alone or in combination with red light increased their production. Red light treatment generally enhanced the amount of free and mono-glucosylated stilbenes during the entire observation period, as well as the percentage of their release in the media. Treatment with MeJA strongly promoted the production of total stilbenes, which was further elicited by the MeJA plus red light treatment. During the combined treatment, the presence of the light stimulus improved the effect of MeJA by anticipating the maximum increase of stilbenes which were also largely released (up to 90%). These results demonstrate that, in grapevine, as in other plant systems, the change of conditions in which the MeJA stimulus is perceived (e.g. going from total white to red light) drastically modifies the plant response to this hormone. The present paper confirms that the jasmonate transduction pathway is integrated into an elaborate signaling network that also comprehends the red light signaling pathway.

Topics: Acetates; Anthocyanins; Catechin; Cells, Cultured; Color; Cyclopentanes; Light; Oxylipins; Plant Growth Regulators; Stilbenes; Vitis

This study examined for the first time the effect of low-energy ultrasound (US), used alone or in combination with methyl jasmonate (MeJA), on viniferin production in cell cultures of Vitis vinifera L. cv Alphonse Lavallée. Cell suspensions were exposed for 2 min to US (power 30, 60, and 90 mW cm(-3)). The highest viniferin production was obtained at 30 mW cm(-3). When sonication was performed twice, the effect on viniferin production was negligible, whereas triple sonication slightly increased production. US treatment at 30 mW cm(-3) for 5 min decreased viniferin production and induced cellular death. The combined use of MeJA and US (2 min) increased the production of δ-viniferin, the dominant stilbene, more than each elicitor used alone. These results suggest that low-energy US, alone and in combination with MeJA, can act as a physical elicitor to stimulate viniferin production in V. vinifera cell cultures.

Topics: Acetates; Benzofurans; Chromatography, High Pressure Liquid; Cyclopentanes; Mass Spectrometry; Molecular Dynamics Simulation; Oxylipins; Quantum Theory; Resorcinols; Stilbenes; Ultrasonics; Vitis

We had previously shown that Vitis vinifera cv. Gamay cell suspension accumulates extracellularly large amounts of the phytoalexin trans-resveratrol (tR) in response to elicitation with methylated cyclodextrins (MBCD), which can be triplicated when the elicitor is combined with methyl jasmonate (MeJA). In parallel, new pathogenesis-related proteins accumulated in the apoplast-like extracellular space. The aim of this study was to investigate changes in the grapevine cell proteome potentially related to tR accumulation in response to the above elicitors. The DIGE technique was used to detect statistically significant changes in the cell's proteome. A total number of 1031 unique spots were detected, 67 of which were de-regulated upon elicitation. Sixty-four spots were successfully identified by nLC-MS/MS database search analysis. The tR biosynthetic pathway enzymes were up-regulated by MBCD alone or combined with MeJA, but not by treatment with MeJA alone, in agreement with tR accumulation pattern. Seven spots contained stilbene synthase encoded by four different isogenes. Likewise, four glutathione-S-transferases, potentially involved in tR trafficking within the cell and across membranes, were up-regulated in the same fashion as stilbene synthases. The relation of other de-regulated proteins with other effects caused by elicitors on grapevine cells, namely defense response and cell growth inhibition, is discussed.

Topics: Acetates; Acyltransferases; beta-Cyclodextrins; Cells, Cultured; Cyclopentanes; Electrophoresis, Gel, Two-Dimensional; Gene Expression Profiling; Gene Expression Regulation, Plant; Oxylipins; Proteome; Resveratrol; Stilbenes; Vitis

Methyl jasmonate, jasmonic acid and chitosan were tested as elicitors on cell suspension cultures obtained from Vitis vinifera cv Italia to investigate their effect on stilbene production. Stilbene accumulation in the callus, grown under nonelicited conditions, was also investigated. Calli and cell suspensions were obtained in a B5 culture medium supplemented with 0.2 mg L(-1) NAA and 1 mg L(-1) KIN. Stilbene determination was achieved by HPLC/DAD/MS. Whereas callus biosynthesized only piceid, cell suspensions elicited with jasmonates produced several stilbenes, mainly viniferins. In suspended cells, methyl jasmonate and jasmonic acid were the most effective in stimulating stilbene biosynthesis, whereas chitosan was less effective; in fact, the amount of stilbenes obtained with this elicitor was not significantly different from that obtained for the control cells. The maximum production of total stilbenes was at day 20 of culture with 0.970 and 1.023 mg g(-1) DW for MeJA and JA, respectively.

Topics: Acetates; Benzofurans; Cells, Cultured; Chitosan; Cyclopentanes; Oxylipins; Plant Growth Regulators; Plant Stems; Resorcinols; Resveratrol; Stilbenes; Vitis

Cell cultures obtained from Vitis vinifera cvs Michele Palieri and Red Globe were cultured in order to stimulate stilbene production. In the calli, stilbene production peaked at day 22 of culture for both cultivars; the main compound was trans-piceid, followed by cis-piceid. Methyl jasmonate, which was added to cell suspensions in the first half of the exponential growth phase, enhanced stilbene accumulation, producing mainly trans-piceid and epsilon-viniferin. Other stilbenoids, though in lower quantities, were identified by liquid chromatography/positive electrospray mass spectrometry. epsilon-Viniferin and trans-resveratrol were the main compounds released into the culture medium. The total quantity of stilbenes was genotype dependent, with a better response found for the cv Red Globe.

Topics: Acetates; Chromatography, Liquid; Cyclopentanes; Mass Spectrometry; Oxylipins; Resveratrol; Stilbenes; Vitis

Perception of elicitors triggers plant defense responses via various early signal transduction pathways. Methyl jasmonate (MeJA) stimulates defense responses in grapevine (Vitis vinifera). We investigated the involvement of various partners (calcium, ROS, reversible phosphorylation) in MeJA-induced responses by using a pharmacological approach. We used specific calcium channel effectors and inhibitors of serine/threonine phosphatases, superoxide dismutase and NAD(P)H oxidase and investigated production of stilbenes (resveratrol and its glucoside, piceid, the major form), which are the grapevine phytoalexins. RNA accumulation of two genes encoding enzymes involved in stilbene synthesis (PAL and STS), three genes encoding pathogenesis-related proteins (CHIT4C, PIN and GLU) and one gene encoding an enzyme producing jasmonates (LOX) were also assessed. Calcium and its origin seemed to play a major role in MeJA-induced grapevine defense responses. Phytoalexin production was strongly affected if calcium from the influx plasma membrane was inhibited, whereas calcium from the intracellular compartments did not seem to be involved. ROS production seemed to interfere with MeJA-stimulated defense responses, and protein phosphorylation/dephosphorylation events also played a direct role.

Topics: Acetates; Calcium; Cantharidin; Cells, Cultured; Cyclopentanes; Dicarboxylic Acids; Enzyme Inhibitors; Glucosides; NADPH Oxidases; Oxylipins; Phosphoric Monoester Hydrolases; Phosphorylation; Plant Growth Regulators; Plant Proteins; Reactive Oxygen Species; Resveratrol; RNA, Plant; Signal Transduction; Stilbenes; Superoxide Dismutase; Vitis

Grapevine (Vitis vinifera L.) is subject to a number of diseases which affect yield and wine quality. After veraison, berries become strongly susceptible to pathogens due to different physiological changes including the accumulation of glucose and fructose, on the one hand, and to the decrease of anti-microbial compounds called stilbenes, on the other. To obtain berry protection, pesticides are excessively used leading to important cost to the grower and to undesirable environmental impact of the residues, especially in grape, soil and water. As a consequence, alternative strategies have to be developed. Exogenously applied biotic elicitors induce defense responses. We studied the effects of methyl jasmonate in combination with sucrose on defense-related gene expression, stilbene and anthocyanin production in grapevine cell suspensions. The methyl jasmonate/sucrose treatment was effective in stimulating phenylalanine ammonia lyase, chalcone synthase, stilbene synthase, UDP-glucose: flavonoid-O-glucosyltransferase, proteinase inhibitor and chitinase gene expression, and triggered accumulation of both piceids and anthocyanins in cells, and trans-resveratrol and piceids in the extracellular medium. Methyl jasmonate treatment might be an efficient natural strategy to protect grapevine berries in vineyard.

Topics: Acetates; Acyltransferases; Anthocyanins; Carbohydrate Metabolism; Cyclopentanes; Fructose; Fruit; Gene Expression Regulation, Enzymologic; Gene Expression Regulation, Plant; Glucose; Oxylipins; Plant Growth Regulators; Resveratrol; Stilbenes; Sucrose; Sweetening Agents; Vitis; Wine

Here the effect of jasmonic acid, methyljasmonate and Na-orthovanadate on the production of resveratrol was studied in Vitis vinifera cv. Barbera cell suspension cultures. Na-orthovanadate at 0.1 mm and 1 mm concentration was efficient in promoting the production and/or accumulation and release in the culture medium of cis-resveratrol while trans-resveratrol levels were not affected by this treatment. Methyljasmonate was highly effective in stimulating both trans- and cis-resveratrol endogenous accumulation, as well as their release into the culture medium. Cis-resveratrol was absent or detected in very low amounts in the controls. Jasmonic acid was less efficient than methyljasmonate in promoting endogenous resveratrol accumulation, but it stimulated the release in the culture medium especially of cis-resveratrol. Gel analysis was performed on control and 10 microm MeJA treated cell suspensions. Results showed an up-regulation of the stilbene synthase demonstrating that MeJA stimulated the synthesis ex-novo of this protein.

Topics: Acetates; Cells, Cultured; Cyclopentanes; Oxylipins; Plant Growth Regulators; Resveratrol; Stilbenes; Time Factors; Vanadates; Vitis