Stilbene Synthase Research Articles

The effect ofSaccharomyces cerevisiaeon antioxidant system of grape leaves infected by downy mildew

In this article, results of the comparative analysis and assessment of resistance of two grape cultivars Vostorg and Muscat blanc against downy mildew disease (Plasmopara viticola) with or without the symbioteSaccharomyces cerevisiae(vine yeast) were shown. The highly resistant cultivar Vostorg with yeast pre-treatment demonstrated a high defensive capability to the pathogen due to the fast immune response. On the first day after inoculation with downy downy mildew the rapid increase in the hydrogen peroxide, which is involved the first step of the grape’s defense system induction, was observed. At the same time, the upregulation of the relative expression of the PR2 protein (β-1,3-gluconase), a key gene involved in the plant’s resistance to pathogens. The oxidative burst was not detected in the susceptible cultivar Muscat blanc for the couple of hours after inoculation with downy mildew pathogen. The significant increase of the total phenols content and expression of stilbene synthase, which is an enzyme involved in the synthesis of phytoalexins, was observed in leaves of Muscat blanc. It was shown that pre-treatment of grape leaves with natural symbioteS. cerevisiaeenhanced the immune response of the resistant cultivar Vostorg and inducted phytoalexins synthesis in the susceptible cultivar Muscat blanc.

The influence of downy mildew tolerance of grape varieties on the biosynthesis of stilbenes in callus as potential sources of bioactive substances

Stilbenes represent a considerable practical interest in relation to their benifits to the human health. Callus culture of the grape, which is a natural producer of stilbenes, can serve as an effective source of these compounds. Grape’s resistance to biotic environmental stress conditions is associated with the synthesis and conversion of stilbenes, therefore the aim of the current study was to determine an interrelation between the tolerance of grape varieties to downy mildew and stilbene biosynthesis in the callus culture. Obtained results showed that there was no link between variety’s tolerance to downy mildew and content of stilbenes, chalcones and phenoloxydising enzymes in callus. Presence of the stress impact is a necessary condition for the initiation of the variety-specific synthesis of stilbenes. Callus of varieties, in which stilbenes content was higher, was characterized by a lowered expression of the chalcone synthase and chalcone isomerase relative to expression of genes of the stilbene synthase and phenylalanine ammonia liase, which confirms an interrelation between reduced competition for the substrate between two biosynthesis branches of chalcones and stilbenes with production of the latter.

Molecular Analysis of UV-C Induced Resveratrol Accumulation in Polygonum cuspidatum Leaves.

Resveratrol is one of the most studied plant secondary metabolites owing to its numerous health benefits. It is accumulated in some plants following biotic and abiotic stress pressures, including UV-C irradiation. Polygonum cuspidatum represents the major natural source of concentrated resveratrol but the underlying mechanisms as well as the effects of UV-C irradiation on resveratrol content have not yet been documented. Herein, we found that UV-C irradiation significantly increased by 2.6-fold and 1.6-fold the resveratrol content in irradiated leaf samples followed by a dark incubation for 6 h and 12 h, respectively, compared to the untreated samples. De novo transcriptome sequencing and assembly resulted into 165,013 unigenes with 98 unigenes mapped to the resveratrol biosynthetic pathway. Differential expression analysis showed that P. cuspidatum strongly induced the genes directly involved in the resveratrol synthesis, including phenylalanine ammonia-lyase, cinnamic acid 4-hydroxylase, 4-coumarate-CoA ligase and stilbene synthase (STS) genes, while strongly decreased the chalcone synthase (CHS) genes after exposure to UV-C. Since CHS and STS share the same substrate, P. cuspidatum tends to preferentially divert the substrate to the resveratrol synthesis pathway under UV-C treatment. We identified several members of the MYB, bHLH and ERF families as potential regulators of the resveratrol biosynthesis genes.

Distinct Metabolic Signals Underlie Clone by Environment Interplay in "Nebbiolo" Grapes Over Ripening.

Several research studies were focused to understand how grapevine cultivars respond to environment; nevertheless, the biological mechanisms tuning this phenomenon need to be further deepened. Particularly, the molecular processes underlying the interplay between clones of the same cultivar and environment were poorly investigated. To address this issue, we analyzed the transcriptome of berries from three "Nebbiolo" clones grown in different vineyards, during two ripening seasons. RNA-sequencing data were implemented with analyses of candidate genes, secondary metabolites, and agronomical parameters. This multidisciplinary approach helped to dissect the complexity of clone × environment interactions, by identifying the molecular responses controlled by genotype, vineyard, phenological phase, or a combination of these factors. Transcripts associated to sugar signalling, anthocyanin biosynthesis, and transport were differently modulated among clones, according to changes in berry agronomical features. Conversely, genes involved in defense response, such as stilbene synthase genes, were significantly affected by vineyard, consistently with stilbenoid accumulation. Thus, besides at the cultivar level, clone-specific molecular responses also contribute to shape the agronomic features of grapes in different environments. This reveals a further level of complexity in the regulation of genotype × environment interactions that has to be considered for orienting viticultural practices aimed at enhancing the quality of grape productions.

Effect of spruce PjSTS1a, PjSTS2, or PjSTS3 gene overexpression on stilbene biosynthesis in callus cultures of Vitis amurensis Rupr.

Stilbenes are natural compounds protecting plants against microbial pathogens and known to possess valuable biologically active properties. In the present study, we established transgenic grapevine callus cell cultures overexpressing three stilbene synthase (STS) genes of spruce Picea jezoensis PjSTS1a, PjSTS2, and PjSTS3. Transformation of Vitis amurensis calli with the PjSTS1a, PjSTS2, and PjSTS3 genes significantly increased total content of stilbenes in 3.6-6, 2.5-2.9, and 4.1-16.1 times, respectively, in comparison with the control calli. The most pronounced positive effect on the accumulation of stilbenes was observed for the PjSTS3-overexpressing calli where the total content of stilbenes was increased up to 3.1 mg/g DW, and the stilbene production reached 25.4 mg/L. These values were higher than those achieved for the grapevine callus cell cultures overexpressing three STS genes from V. amurensis. Thus, transformation of grapevine cell cultures with spruce STS genes with a relatively low degree of homology to the endogenous VaSTSs is a more effective strategy for induction of plant secondary metabolite biosynthesis than using the grapevine genes for the overexpression experiments.

The grapevine R2R3-type MYB transcription factor VdMYB1 positively regulates defense responses by activating the stilbene synthase gene 2 (VdSTS2)

BackgroundResveratrol is a naturally occurring plant stilbene that exhibits a wide range of valuable biological and pharmacological properties. Although the beneficial effects of trans-resveratrol to human health and plant protection against fungal pathogens and abiotic stresses are well-established, yet little is known about the molecular mechanisms regulating stilbene biosynthesis in plant defense progress.ResultsHere, we cloned and identified the Chinese wild grape (Vitis davidii) R2R3-MYB transcription factor VdMYB1, which activates defense responses against invading pathogen. VdMYB1 transcripts were significantly upregulated after inoculation with the grapevine powdery mildew fungus Erysiphe necator (Schw.) Burr. Transient expression analysis using onion epidermal cells and Arabidopsis thaliana protoplasts showed that VdMYB1 was localized in the nucleus. Yeast one-hybrid assays revealed that VdMYB1 acts as a transcriptional activator. Grapevine leaves transiently overexpressing VdMYB1 showed a lower number of fungal conidiophores compared with wild-type leaves. Overexpression of VdMYB1 in grapevine leaves did not alter the expression of genes in salicylic acid- and jasmonate-dependent pathways, but affected the expression of stilbene synthase (STS) genes, key regulators of flavonoid metabolism. Results of electrophoretic mobility shift assays and in vivo transcriptional activation assays showed that VdMYB1 binds to the MYB binding site (MYBBS) in the STS2 gene promoter, thus activating STS2 transcription. In heterologous expression assays using tobacco leaves, VdMYB1 activated STS2 gene expression and increased the accumulation of resveratrol.ConclusionsOur study showed that VdMYB1 activates STS2 gene expression to positively regulate defense responses, and increases the content of resveratrol in leaves.

Structure of the Cannabissativa olivetol-producing enzyme reveals cyclization plasticity in type III polyketide synthases.

In the native pathway to therapeutic cannabinoid biosynthesis in Cannabis sativa, the three‐step production of a key intermediate, olivetolic acid, is catalysed by the enzymes tetraketide synthase (TKS; linear tetraketide intermediate production in two stages) and olivetolic acid cyclase (OAC; final C2 → C7 aldol condensation). In the absence of OAC, a nonenzymatic C2 → C7 decarboxylative aldol condensation of the tetraketide intermediate occurs forming olivetol. TKS is a type III polyketide synthase, and the question arises why it is unable to form olivetolic acid directly, but instead forms this unwanted side product. We determined the TKS, CoA complex structure, and performed structurally guided mutagenesis studies to identify potential residues responsible for cyclization pathway discrimination in type III polyketide synthases. Prior studies suggested an ‘aldol switch’ is necessary to allow linear tetraketide intermediate release prior to cyclization, thereby enabling subsequent olivetolic acid production by OAC. However, our studies do not support the presence of a universal or predictable ‘aldol switch’ consensus sequence. Instead, we propose the mode of ordered active site water activation between type III polyketide synthases catalysing different cyclization mechanisms is subtle and homologue‐specific. Our work indicates that subtle structural variations between homologous enzymes can have a major mechanistic impact on the catalytic outcome. This highlights the importance of embedding high‐resolution structural analysis of multiple enzyme homologues with classical site‐directed mutagenesis studies when investigating highly similar enzymes with different mechanistic pathway outcomes.EnzymesTKS, http://www.chem.qmul.ac.uk/iubmb/enzyme/EC2/3/1/206.html; OAC, http://www.chem.qmul.ac.uk/iubmb/enzyme/EC4/4/1/26.html; chalcone synthase, http://www.chem.qmul.ac.uk/iubmb/enzyme/EC2/3/1/74.html; stilbene synthase, http://www.chem.qmul.ac.uk/iubmb/enzyme/EC2/3/1/95.html; 2‐PS, http://www.chem.qmul.ac.uk/iubmb/enzyme/EC2/3/1/-.html.Accession numbersThe atomic coordinates and structure factors for the crystal structure of TKS have been deposited in the Protein Data Bank with accession number http://www.rcsb.org/pdb/search/structidSearch.do?structureId=6GW3.

Assembly and Annotation of a Draft Genome of the Medicinal Plant Polygonum cuspidatum.

Polygonum cuspidatum (Japanese knotweed, also known as Huzhang in Chinese), a plant that produces bioactive components such as stilbenes and quinones, has long been recognized as important in traditional Chinese herbal medicine. To better understand the biological features of this plant and to gain genetic insight into the biosynthesis of its natural products, we assembled a draft genome of P. cuspidatum using Illumina sequencing technology. The draft genome is ca. 2.56 Gb long, with 71.54% of the genome annotated as transposable elements. Integrated gene prediction suggested that the P. cuspidatum genome encodes 55,075 functional genes, including 6,776 gene families that are conserved in the five eudicot species examined and 2,386 that are unique to P. cuspidatum. Among the functional genes identified, 4,753 are predicted to encode transcription factors. We traced the gene duplication history of P. cuspidatum and determined that it has undergone two whole-genome duplication events about 65 and 6.6 million years ago. Roots are considered the primary medicinal tissue, and transcriptome analysis identified 2,173 genes that were expressed at higher levels in roots compared to aboveground tissues. Detailed phylogenetic analysis demonstrated expansion of the gene family encoding stilbene synthase and chalcone synthase enzymes in the phenylpropanoid metabolic pathway, which is associated with the biosynthesis of resveratrol, a pharmacologically important stilbene. Analysis of the draft genome identified 7 abscisic acid and water deficit stress-induced protein-coding genes and 14 cysteine-rich transmembrane module genes predicted to be involved in stress responses. The draft de novo genome assembly produced in this study represents a valuable resource for the molecular characterization of medicinal compounds in P. cuspidatum, the improvement of this important medicinal plant, and the exploration of its abiotic stress resistance.

Expression of stilbene synthase VqSTS6 from wild Chinese Vitis quinquangularis in grapevine enhances resveratrol production and powdery mildew resistance.

In grape (Vitis), stilbene phytoalexins can either be in situ synthesized or transported to the site of response during powdery mildew infection, enhancing disease resistance. Resveratrol is a phytoprotective stilbenoid compound that is synthesized by stilbene synthase (STS) in response to biotic and abiotic stresses, and is also known to have health benefits in the human diet. We have previously shown that transgenic Vitis vinifera cv. Thompson Seedless plants overexpressing a stilbene synthase gene, VqSTS6, from wild Chinese Vitis quinquangularis had a higher stilbenoid content, leading to an enhanced resistance to powdery mildew (Uncinula necator (Schw.) Burr). However, the biosynthesis and transportation in the plant tissue under powdery mildew infection are still unclear. Here, inhibitor and micro-grafting technologies were used to study the accumulation of resveratrol following powdery mildew infection. We observed that the levels of STS expression and stilbenoids increased in response to powdery mildew infection. Powdery mildew and inhibitor treatment on detached grape branches showed that resveratrol was in situ synthesized. Experiments with grafted plantlets showed that the abundance of stilbenoid compounds increased in the shoot during VqSTS6 overexpression in the root, while VqSTS6-Flag fusion was not tranported to the scions and only expressedin thetransgenic rootstocks. Compared with wild-type Thompson Seedless plants, the non-transgenic/VqSTS6 transgenic (scion/rootstock) grafted Thompson Seedless plantlets exhibited increased resistance to powdery mildew. In addition, overexpression of VqSTS6 in roots led to increased levels of stilbenoid compounds in five other European grape varieties (V. vinifera cvs. Chardonnay, Perlette, Cabernet Sauvignon, Riesling and Muscat Hamburg). In conclusion, stilbenoid compounds can be either in situ synthesized or transported to the site of powdery mildew infection, and overexpression of VqSTS6 in the root promotes stilbenoids accumulation and disease resistance in European grapevine varieties.

Transcriptional regulation of stilbene synthases in grapevine germplasm differentially susceptible to downy mildew

BackgroundTo limit the impact of the downy mildew disease of grapevine and reduce the need to recur to chemical treatments, an effective strategy might be recovering adaptive resistance traits in both cultivated and wild V. vinifera germplasm. Considering that stilbenes represent the most important class of phytoalexins in the Vitaceae, the constitutive expression and transcriptional activation of all the functional members of the stilbene synthase gene family were analysed in a group of nine grapevine genotypes following artificial infection with the oomycete Plasmopara viticola, the causal agent of the disease. In addition, in the same genotypes we analyzed the expression of genes encoding for two transcription factors involved in the transcriptional regulation of the stilbene synthase genes, namely VvMYB14 and VvMYB15, and of genes encoding for chalcone synthases.ResultsDowny mildew incidence and severity ranged from nihil to high in the grapevine genotypes considered, being low to moderate in a subgroup of V. vinifera genotypes. The constitutive expression of the stilbene synthase genes as well as the extent of their transcriptional activation following P. viticola inoculation appeared to be inversely related to the proneness to develop disease symptoms upon infection. In a specular manner, following P. viticola inoculation all the chalcone synthase genes were up-regulated in the susceptible grapevine genotypes and down-regulated in the resistant ones. The infection brought by P. viticola appeared to elicit a co-ordinated and sequential transcriptional activation of distinct stilbene synthase genes subsets, each of which may be regulated by a distinct and specific MYB transcription factor.ConclusionsThe present results suggest that the induction of stilbene biosynthesis may contribute to the basal immunity against the downy mildew of grapevine, thus representing an adaptive resistance trait to recover, in both cultivated and wild V. vinifera germplasm. During the early stages of P. viticola infection, an antagonistic interaction between flavonol and stilbene biosynthesis might occur, whose outcome might determine the subsequent extent of disease symptoms. Further studies are needed to decipher the possible regulatory mechanisms involved in the antagonistic crosstalk between these two metabolic pathways in resistant and susceptible genotypes in response to P. viticola.

Transcriptional regulation of stilbene synthases in grapevine germplasm differentially susceptible to downy mildew

Transcriptional regulation of stilbene synthases in grapevine germplasm differentially susceptible to downy mildew

Transcription factor VqERF114 regulates stilbene synthesis in Chinese wild Vitis quinquangularis by interacting with VqMYB35.

VqERF114 regulates stilbene synthesis by interacting with VqMYB35. Resveratrol is a stilbene, an important class of secondary metabolites that accumulates in some plant species, including grapevine. In the plant, these are involved in the response to attack by plant pathogens and, as a component of the human diet, they offer a range of significant health benefits. Stilbene synthase (STS), the key enzyme responsible for resveratrol synthesis, has been characterised in a small number of plant species. However, the regulatory mechanisms for stilbene synthesis are uncertain. Here, an ERF family transcription factor from Chinese wild Vitis quinquangularis, VqERF114, was characterised as an indirect regulator of stilbene synthesis. A transient overexpression assay of VqERF114 in grapevine leaves led to increased STS expression and stilbene accumulation. However, VqERF114 did not bind to the promoters of VqSTSs but the MYB transcription factor, VqMYB35, did interact with VqERF114. This interaction was confirmed by a yeast two-hybrid assay and bimolecular fluorescence complementation. Furthermore, VqMYB35 showed activation effects on the expressions of VqSTS15, VqSTS28, VqSTS42 and VqSTS46 by binding directly to the MBS elements in their promoters. Co-overexpression of VqERF114 and VqMYB35 resulted in higher VqSTSs expression and more stilbene synthesis. These results demonstrate that VqERF114 regulates stilbene synthesis by interacting with VqMYB35.

Whole-genome sequencing and gene annotation of Georgian grape cultivars

The genomes of four Georgian grape cultivars, ‘Chkhaveri’, ‘Saperavi’, ‘Meskhetian green’, and ‘Rkatsiteli’, belonging to different haplogroups, were resequenced. Shotgun genomic libraries of grape cultivars were sequenced on an Illumina HiSeq. The sequences of all 19 chromosomes of the four Georgian grape cultivars were determined by resequencing using ‘Pinot noir’ as a reference genome. Annotation of chromosomal DNA of Georgian grape revealed 17,409 candidate genes in ‘Chkhaveri’, 17,021 in ‘Saperavi’, 18,355 in ‘Meskhetian green’ and 13,960 in ‘Rkatsiteli’. Among them, 106 predicted genes and 43 pseudogenes of terpene synthase (TPS) and 44 candidate genes of stilbene synthase (STS) were found in all studied Georgian cultivars. Several additional TPS and STS genes not present in the reference ‘Pinot noir’ genome were detected. This is the first attempt at a comparative whole-genome analysis of different haplogroups of Vitis vinifera cultivars.

VqbZIP1 isolated from Chinese wild Vitis quinquangularis is involved in the ABA signaling pathway and regulates stilbene synthesis

Resveratrol is an important phytoalexin in grapevine. Not only does it confer increased disease resistance and but as a food component it offers significant benefits in human health. Abscisic acid (ABA) is an important phytohormone involved in many biological processes in plants and can also promote the accumulation of stilbenes. Stilbene synthase (STS) is an important enzyme which catalyzes the last step of resveratrol synthesis. Our study characterizes a basic leucine zipper (bZIP) transcription factor, VqbZIP1, isolated from Chinese wild Vitis quinquangularis accession Danfeng-2. The results show that VqbZIP1 encodes 299 amino acids and belongs to the Group A subfamily of the bZIP family. VqbZIP1 showed transcriptional activation activity in yeast and is predicted to be located in the nucleus. The yeast two-hybrid assay and bimolecular fluorescence complementation (BiFC) assay together show that VqbZIP1 interacts with VqSnRK2.4 and VqSnRK2.6. VqbZIP1, the STS genes, VqSnRK2.4 and VqSnRK2.6 can all be induced by ABA treatment. A GUS activity experiment indicates VqbZIP1 can activate the GUS reporter gene driven by STS promoters. Further studies show that co-expression of VqbZIP1 with VqSnRK2.4 or VqSnRK2.6 can confer higher efficiency than expression of VqbZIP1 alone in activating the STS promoters. Overexpression of VqbZIP1 in grape leaves promoted the transcript level of the STS genes and the accumulation of stilbenes. Overexpression of VqbZIP1 in Arabidopsis thaliana can confer ABA sensitivity. In summary, our results suggest VqbZIP1 participates in the ABA signaling pathway and regulates stilbene synthesis.

Differentially expressed genes during berry ripening in de novo RNA assembly of Vitis flexuosa fruits

A comparative transcriptome analysis between unripe and ripe berries of Vitis flexuosa, a Korean native wild grape, was conducted to identify differentially expressed genes (DEGs). About 12 billion nucleotides derived from unripe and ripe berries were sequenced and 180,503 contigs were obtained from the assembly results. Expression profiles of transcriptome data revealed that 5249 DEGs showed significant differences at transcript levels between unripe and ripe berries. The top five up-regulated genes in ripe berries against unripe berries were cupin family protein, cytochrome p450 79a2, HSP20-like chaperones superfamily protein, chalcone and stilbene synthase family protein, and beta-tonoplast intrinsic protein. The five most down-regulated genes in ripe berries against unripe berries were cellulose synthase family protein, laccase 17, proline-rich protein 2, subtilase 1.3, and laccase/diphenol oxidase family protein. Among DEGs, top 10 up- and down-regulated carbohydrate, organic substance, and phenylpropanoid metabolic process-related genes were selected from transcriptome analysis and their expression profiles were validated by real-time PCR. Real-time PCR analysis revealed higher expression level of DEGs encoding xyloglucan endotransglucosylase/hydrolase 32 and pectin lyase-like superfamily protein, which are related to starch and sucrose metabolism, cruciferin 3 and AGAMOUS-like 104, which are involved in organic acid metabolism, and MYB domain protein 113 and chalcone and stilbene synthase family protein, which play a role in the phenylalanine ammonia-lyase pathway in ripe than in unripe berries. Overall, we present an overview of transcriptome changes and carbohydrate, organic substance, and phenylpropanoid metabolic process-related expression patterns in unripe and ripe berries of V. flexuosa during berry ripening.

VpSTS29/STS2 enhances fungal tolerance in grapevine through a positive feedback loop.

Accumulation of stilbene phytoalexins stimulates resistance mechanisms against the grapevine fungus Uncinula necator. However, the defensive mechanisms triggered by stilbene synthase (STS) genes, remain largely unknown. Here, we report the function and molecular mechanism of the stilbene synthase gene VpSTS29/STS2 from Vitis pseudoreticulata in the regulation of plant responses to powdery mildew. Stilbene synthesis occurred mainly in root tips and mesophyll cells of transgenic grapevines via transport through the vascular bundles. Overexpression of VpSTS29/STS2 in Vitis vinifera increased the abundance of STSs in mesophyll tissue and resulted in the accumulation of biologically active resveratrol derivatives at the invasion site. Similarly, expression of VpSTS29/STS2 in Arabidopsis increased resistance to Golovinomyces cichoracearum. The VpSTS29/STS2-expressing Arabidopsis lines showed increased piceid accumulation together with more local hypersensitive reactions, inhibition of mycelial growth, and a reduced incidence of pathogens. Transcriptome profiling analyses demonstrated that VpSTS29/STS2-induced defences led to reprograming of global gene expression and activation of salicylic acid (SA) signalling, thus increasing expression of WRKY-MYB transcription factors and other defence response genes. We propose a model for resveratrol-mediated coordination of defence responses in which SA participates in a positive feedback loop.

Evolvement of nutraceutical onion plants engineered for resveratrol biosynthetic pathway

Genetically engineered onion expressing codon-optimized VvSTS1 gene accumulated stilbenes and extended life span in yeast and can serve as potential nutraceutical. Resveratrol (RV) is a natural polyphenolic compound found in certain plant species including grapes. RV is well known for its nutraceutical properties and to assuage several disease conditions. Onion is the second most consumed vegetable worldwide and contains large quantities of precursor molecules, malonyl-CoA and para-coumaroyl-CoA that are needed for RV biosynthesis. The present study reports the development of nutraceutical onion by engineering RV biosynthetic pathway. A codon-optimized grapevine synthetic stilbene synthase gene (VvSTS1) was synthesized using native grapevine sequence. Six-week-old healthy yellowish compact nodular calli were co-cultivated with Agrobacterium tumefaciens harbouring pCAMBIA1300-hpt II-CaMV35S-VvSTS1-nos. PCR analysis revealed the presence of VvSTS1 and hpt II genes in putative transgenics. Southern blot analysis confirmed the integration of VvSTS1 gene and independent nature of transformants. LC-ESI-HRMS analysis revealed the accumulation of variable quantities of RV (24.98-50.18µg/g FW) and its glycosylated form polydatin (33.6-67.15µg/g FW) in both leaves and bulbs, respectively, indicating the successful engineering of RV biosynthetic pathway into onion. The transgenic onion bulb extracts extended the life span in haploid yeast. The transgenic onion accumulating RV and polydatin, developed for the first of its kind, may serve as a potential nutraceutical resource.

Burdock fructooligosaccharides inhibits postharvest colour development in grape skins by altering phenylpropanoid fluxes from anthocyanin to the resistance‐related phenolic substances

Background and Aims Burdock fructooligosaccharides (BFO) is an elicitor that induces biotic resistance and delays colour development in grapes postharvest. The mechanism of the delayed colour development requires further study. Methods and Results Colour development index; anthocyanin, tannin, flavonol, flavanol, trans‐resveratrol and lignin; the trans‐resveratrol, lignin and anthocyanin biosynthesis‐related transcription factors; and functional gene expression were investigated. In response to BFO induction, upregulation of the v‐myb avian myeloblastosis viral oncogene homologue 14 (MYB14)‐mediated stilbene synthase (STS) altered the phenylpropanoid fluxes from anthocyanin to trans‐resveratrol; MYBA1 and MYBA2‐mediated the downregulation of dihydroflavonol 4‐reductase (DFR), and flavonoid‐3‐O‐glycosyltransferase (UFGT) limited anthocyanin biosynthesis. Downregulation of MYB4‐mediated chalcone synthase (CHS) and upregulation of cinnamoyl CoA reductase (CCR) and cinnamyl alcohol dehydrogenase (CAD) altered the phenylpropanoid fluxes from anthocyanin to lignin. Conclusions Burdock fructooligosaccharides inhibits postharvest colour development in grapes by altering the phenylpropanoid fluxes from anthocyanin to resistance‐related phenolic substances. Significance of the Study Understanding the mechanism of the effect of BFO on grape colour development may improve our understanding of colour development in grape berries.

The effect of ultraviolet-C and precursor feeding on stilbene biosynthesis in spruce Picea jezoensis

This study examines the effects of p-coumaric (CoA) and caffeic (CaA) acids on stilbene biosynthesis in one-year-old needles of Picea jezoensis (Lindl. et Gord.) Fisch ex Carr. under control conditions and after ultraviolet (UV-C) irradiation. HPLC analysis revealed that while UV-C irradiation slightly affected the total stilbene content, CoA and CaA feeding increased the total content of stilbenes by 1.2–1.3-fold. UV-C treatment combined with CoA-feeding of the P. jezoensis cuttings exerted the most pronounced stimulatory effect on stilbene accumulation leading to the total stilbene content of 9.18 mg/g of dry weight or DW. This increase correlated with the elevated transcription of the stilbene synthase PjSTS1a and PjSTS1b genes. UV-C treatment in combination with CaA feeding of P. jezoensis did not considerably influence stilbene content. These results revealed a positive effect of UV-C radiation and phenolic precursors on the content of stilbenes in spruce needles.

Global transcriptome analysis and differentially expressed genes in grapevine after application of the yeast-derived defense inducer cerevisane.

Cerevisane, made up of cell wall derivatives from the Saccharomyces cerevisiae strain LAS117, is proposed as a resistance inducer in plants. The mode of action of cerevisane was investigated through transcriptome analysis (RNA-Seq) carried out on leaves of potted vines cv. Italia grown in the greenhouse and sprayed at 1-week intervals with cerevisane. Analyses were performed at three time points after one and three sprays as well as on vines challenged with artificial inoculation with Plasmopara viticola, Erysiphe necator and Botrytis cinerea. Cerevisane proved effective against downy mildew and caused an increase in expression levels of several genes related to defense responses to fungal pathogens and other stresses and down-regulation of genes involved in several processes related to plant growth and development. Up-regulated genes included genes encoding (i) enzymes involved in hormone metabolism (i.e. salicylic acid, jasmonate, ethylene) and related plant responses, (ii) defense compounds (i.e. pathogenesis-related proteins, phenylalanine ammonia-lyase, stilbene synthases, lipoxygenase, leucine-rich repeat receptor-like protein kinases, non-specific plant lipid transfer proteins, serine-threonine protein kinases involved in signal transduction, superoxide dismutase and glutathione S-transferase involved in response to oxidative stress), (iii) secondary metabolites (i.e. phenylpropanoids, terpenoids, lignin), and (iv) photosynthetic processes (light harvesting chlorophyll A/B-binding proteins and components of the photosystems). Cerevisane can be a useful tool in protection schedules against downy mildew on grapevine aimed at reducing the usage of synthetic fungicides and preventing fungicide resistance. The results provide the first basic knowledge on the mode of action of yeast-derived elicitors effective against P. viticola on grapevine. © 2019 Society of Chemical Industry.