Grape accumulates numerous polyphenols with abundant health benefit and organoleptic properties that in planta act as key components of the plant defense system against diseases. Considerable advances have been made in the chemical characterization of wine metabolites particularly volatile and polyphenolic compounds. However, the metabotyping (metabolite-phenotype characterization) of grape varieties, from polyphenolic-rich vineyard by-product is unprecedented. As this composition might result from the complex interaction between genotype, environment and viticultural practices, a field experiment was setting up with uniform pedo-climatic factors and viticultural practices of growing vines to favor the genetic determinism of polyphenol expression. As a result, UPLC-MS-based targeted metabolomic analyses of grape stems from 8 Vitis vinifera L. cultivars allowed the determination of 42 polyphenols related to phenolic acids, flavonoids, procyanidins, and stilbenoids as resveratrol oligomers (degree of oligomerization 1–4). Using a partial least-square discriminant analysis approach, grape stem chemical profiles were discriminated according to their genotypic origin showing that polyphenol profile express a varietal signature. Furthermore, hierarchical clustering highlights various degree of polyphenol similarity between grape varieties that were in agreement with the genetic distance using clustering analyses of 22 microsatellite DNA markers. Metabolite correlation network suggested that several polyphenol subclasses were differently controlled. The present polyphenol metabotyping approach coupled to multivariate statistical analyses might assist grape selection programs to improve metabolites with both health-benefit potential and plant defense traits.

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Lochnericine is a major monoterpene indole alkaloid (MIA) in the roots of Madagascar periwinkle (). Lochnericine is derived from the stereoselective C6,C7-epoxidation of tabersonine and can be metabolized further to generate other complex MIAs. While the enzymes responsible for its downstream modifications have been characterized, those involved in lochnericine biosynthesis remain unknown. By combining gene correlation studies, functional assays, and transient gene inactivation, we identified two highly conserved P450s that efficiently catalyze the epoxidation of tabersonine: tabersonine 6,7-epoxidase isoforms 1 and 2 (TEX1 and TEX2). Both proteins are quite divergent from the previously characterized tabersonine 2,3-epoxidase and are more closely related to tabersonine 16-hydroxylase, involved in vindoline biosynthesis in leaves. Biochemical characterization of TEX1/2 revealed their strict substrate specificity for tabersonine and their inability to epoxidize 19-hydroxytabersonine, indicating that they catalyze the first step in the pathway leading to hörhammericine production. and displayed complementary expression profiles, with expressed mainly in roots and in aerial organs. Our results suggest that and originated from a gene duplication event and later acquired divergent, organ-specific regulatory elements for lochnericine biosynthesis throughout the plant, as supported by the presence of lochnericine in flowers. Finally, through the sequential expression of and up to four other MIA biosynthetic genes in yeast, we reconstituted the 19-acetylhörhammericine biosynthetic pathway and produced tailor-made MIAs by mixing enzymatic modules that are naturally spatially separated in the plant. These results lay the groundwork for the metabolic engineering of tabersonine/lochnericine derivatives of pharmaceutical interest.

While the characterization of the biosynthetic pathway of monoterpene indole alkaloids (MIAs) in leaves of Catharanthus roseus is now reaching completion, only two enzymes from the root counterpart dedicated to tabersonine metabolism have been identified to date, namely tabersonine 19-hydroxylase (T19H) and minovincine 19-O-acetyltransferase (MAT). Albeit the recombinant MAT catalyzes MIA acetylation at low efficiency in vitro, we demonstrated that MAT was inactive when expressed in yeast and in planta, suggesting an alternative function for this enzyme. Therefore, through transcriptomic analysis of periwinkle adventitious roots, several other BAHD acyltransferase candidates were identified based on the correlation of their expression profile with T19H and found to localize in small genomic clusters. Only one, named tabersonine derivative 19-O-acetyltransferase (TAT) was able to acetylate the 19-hydroxytabersonine derivatives from roots, such as minovincinine and hörhammericine, following expression in yeast. Kinetic studies also showed that the recombinant TAT was specific for root MIAs and displayed an up to 200-fold higher catalytic efficiency than MAT. In addition, gene expression analysis, protein subcellular localization and heterologous expression in Nicotiana benthamiana were in agreement with the prominent role of TAT in acetylation of root-specific MIAs, thereby redefining the molecular determinants of the root MIA biosynthetic pathway. Finally, identification of TAT provided a convenient tool for metabolic engineering of MIAs in yeast enabling efficiently mixing different biosynthetic modules spatially separated in the whole plant. This combinatorial synthesis associating several enzymes from Catharanthus roseus resulted in the conversion of tabersonine in tailor-made MIAs bearing both leaf and root-type decorations.

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BACKGROUND: Plasmopara viticola control in organic viticulture requires copper-based fungicides with harmful effects on health and the environment. Plant extracts represent a biorational eco-friendly alternative to copper. The aim of this study was to evaluate the potential of stilbenoid-rich grape cane extract (GCE) against downy mildew on three cultivars over 3 years following natural downy mildew infection. RESULTS: Over all field trials, GCE treatments showed an average reduction in disease incidence of −35% and −38% on leaves and clusters, respectively. The average reduction in disease severity was −35% and −43% on leaves and clusters, respectively. Under artificial downy mildew infection, GCE efficacy corresponded to 1 g L −1 of copper. Neither phytotoxicity nor adverse effects on auxiliary fauna were observed after treatment with GCE. CONCLUSION: Because few or no biocontrol agents are active alone against P. viticola, GCE is a promising alternative to copper-based fungicides. Grape canes, an abundant by-product of viticulture, have great potential for valorization as a biocontrol agent for sustainable viticulture.

In the context of expansion of invasive species, survival of invasive plants is conditioned by their ability to adapt. In France, the water primrose Ludwigia grandiflora, an aquatic invasive species, invades yet wet meadows, leading to a depreciation of their fodder value. Understanding its potential adaption is necessary to its management, strong differences between both morphotypes were expected. So morphological and metabolic responses to terrestrial environment were analyzed for aquatic and terrestrial morphotypes. All morphological and biomass variables were greater in the terrestrial morphotype than the aquatic morphotype, independent of conditions. In terrestrial condition, both morphotypes showed a high production of sugars in root tissues, especially in the terrestrial morphotype and both morphotypes produced a low level of amino acids in shoot tissues. All results demonstrate that the terrestrial condition seems a stressful situation for both morphotypes, which activates glycolysis and fermentation pathways to improve their survival under hypoxic stress. But, only the terrestrial morphotype has been able to adjust its metabolism and maintain efficient growth. In the future, a differential transcriptomic analysis will be carried out to confirm this result.

Grape canes are viticulture byproducts representing a sustainable source of valuable bioactive polyphenols. However, varietal origin and postharvest treatment greatly influence the effective concentration of biomolecules, thus limiting industrial development. With the aim to develop grape cane extracts with high polyphenol contents, the selection of a specific grape variety combined with optimized postharvest treatment is the major determinant. A previously described postharvest treatment comprising cutting grapevine stalks of 0.5 cm length and storage at 15–20 °C over 2 weeks was applied on a selection of 44 grape varieties representative of the genetic diversity of the whole European collection and performing the screening of polyphenol contents. Varietal rankings according to major polyphenols (catechin, epicatechin, E-resveratrol, E-piceatannol, E-ϵ-viniferin, E-miyabenol C, ampelopsin A, E-vitisin B, hopeaphenol, and isophopeaphenol) were performed with and without the postharvest treatment. We observed that postharvest treatment greatly influenced the total polyphenol composition but also the ranking of polyphenol-rich varieties. This polyphenol screening of grape canes from a large collection of European varieties revealed the importance of postharvest treatment together with the selection of varieties to develop natural extracts based on grapevine wood biomass enriched with molecules with health benefits.