Sylvain Cordelier

Biosynthesis, metabolism, molecular engineering, and biological functions of stilbene phytoalexins in plants

Stilbenic compounds recently have become the focus of a number of studies in medicine and plant physiology as well as have emerged as promising molecules that potentially affect human health. Stilbenes are relatively simple compounds synthesized by plants and deriving from the phenyalanine/polymalonate route, the last and key enzyme of this pathway being stilbene synthase. Here, we review the biological significance of stilbenes in plants together with their biosynthesis pathway and their metabolism both by fungi and in planta. Special attention will be paid to the role of stilbenic molecules as phytoalexins.

Molecular engineering of resveratrol in plants

The grapevine phytoalexin resveratrol, the synthesis of which is achieved by stilbene synthase (STS), displays a wide range of biological effects. Most interest has centred, in recent years, on STS gene transfer experiments from grapevine to the genome of numerous plants. This work presents a comprehensive review on plant molecular engineering with the STS gene. Gene and promoter options are discussed, namely the different promoters used to drive the transgene, as well as the enhancer elements and/or heterologous promoters used to improve transcriptional activity in the transformed lines. Factors modifying transgene expression and epigenetic modifications, for instance transgene copy number, are also presented. Resveratrol synthesis in plants, together with that of its glucoside as a result of STS expression, is described, as is the incidence of these compounds on plant metabolism and development. The ectopic production of resveratrol can lead to broad-spectrum resistance against fungi in transgenic lines, and to the enhancement of the antioxidant activities of several fruits, highlighting the potential role of this compound in health promotion and plant disease control.

Modulation of phytoalexin biosynthesis in engineered plants for disease resistance.

Phytoalexins are antimicrobial substances of low molecular weight produced by plants in response to infection or stress, which form part of their active defense mechanisms. Starting in the 1950’s, research on phytoalexins has begun with biochemistry and bio-organic chemistry, resulting in the determination of their structure, their biological activity as well as mechanisms of their synthesis and their catabolism by microorganisms. Elucidation of the biosynthesis of numerous phytoalexins has permitted the use of molecular biology tools for the exploration of the genes encoding enzymes of their synthesis pathways and their regulators. Genetic manipulation of phytoalexins has been investigated to increase the disease resistance of plants. The first example of a disease resistance resulting from foreign phytoalexin expression in a novel plant has concerned a phytoalexin from grapevine which was transferred to tobacco. Transformations were then operated to investigate the potential of other phytoalexin biosynthetic genes to confer resistance to pathogens. Unexpectedly, engineering phytoalexins for disease resistance in plants seem to have been limited to exploiting only a few phytoalexin biosynthetic genes, especially those encoding stilbenes and some isoflavonoids. Research has rather focused on indirect approaches which allow modulation of the accumulation of phytoalexin employing transcriptional regulators or components of upstream regulatory pathways. Genetic approaches using gain- or less-of functions in phytoalexin engineering together with modulation of phytoalexin accumulation through molecular engineering of plant hormones and defense-related marker and elicitor genes have been reviewed.

Uncovering plant-pathogen crosstalk through apoplastic proteomic studies.

Plant pathogens have evolved by developing different strategies to infect their host, which in turn have elaborated immune responses to counter the pathogen invasion. The apoplast, including the cell wall and extracellular space outside the plasma membrane, is one of the first compartments where pathogen-host interaction occurs. The plant cell wall is composed of a complex network of polysaccharides polymers and glycoproteins and serves as a natural physical barrier against pathogen invasion. The apoplastic fluid, circulating through the cell wall and intercellular spaces, provides a means for delivering molecules and facilitating intercellular communications. Some plant-pathogen interactions lead to plant cell wall degradation allowing pathogens to penetrate into the cells. In turn, the plant immune system recognizes microbial- or damage-associated molecular patterns (MAMPs or DAMPs) and initiates a set of basal immune responses, including the strengthening of the plant cell wall. The establishment of defense requires the regulation of a wide variety of proteins that are involved at different levels, from receptor perception of the pathogen via signaling mechanisms to the strengthening of the cell wall or degradation of the pathogen itself. A fine regulation of apoplastic proteins is therefore essential for rapid and effective pathogen perception and for maintaining cell wall integrity. This review aims to provide insight into analyses using proteomic approaches of the apoplast to highlight the modulation of the apoplastic protein patterns during pathogen infection and to unravel the key players involved in plant-pathogen interaction.

Metabolic engineering of yeast and plants for the production of the biologically active hydroxystilbene, resveratrol.

Resveratrol, a stilbenic compound deriving from the phenyalanine/polymalonate route, being stilbene synthase the last and key enzyme of this pathway, recently has become the focus of a number of studies in medicine and plant physiology. Increased demand for this molecule for nutraceutical, cosmetic and possibly pharmaceutic uses, makes its production a necessity. In this context, the use of biotechnology through recombinant microorganisms and plants is particularly promising. Interesting results can indeed arise from the potential of genetically modified microorganisms as an alternative mechanism for producing resveratrol. Strategies used to tailoring yeast as they do not possess the genes that encode for the resveratrol pathway, will be described. On the other hand, most interest has centered in recent years, on STS gene transfer experiments from various origins to the genome of numerous plants. This work also presents a comprehensive review on plant molecular engineering with the STS gene, resulting in disease resistance against microorganisms and the enhancement of the antioxidant activities of several fruits in transgenic lines.

Deciphering the Role of Phytoalexins in Plant-Microorganism Interactions and Human Health

Phytoalexins are low molecular weight antimicrobial compounds that are produced by plants as a response to biotic and abiotic stresses. As such they take part in an intricate defense system which enables plants to control invading microorganisms. In this review we present the key features of this diverse group of molecules, namely their chemical structures, biosynthesis, regulatory mechanisms, biological activities, metabolism and molecular engineering.

Relationship Between Localized Acquired Resistance (LAR) and the Hypersensitive Response (HR): HR Is Necessary for LAR to Occur and Salicylic Acid Is Not Sufficient to Trigger LAR

In tobacco plants reacting hypersensitively to pathogen infection, localized acquired resistance (LAR) is induced in a sharp zone surrounding hypersensitive response (HR) lesions. Using a fungal glycoprotein inducing HR and LAR when infiltrated at 50 nM into tobacco leaves, we have shown previously that a plant signal(s) is released by HR cells and diffuses to induce LAR. Here we address two questions: does LAR occur when HR is not induced, and is salicylic acid the (or one of the) mobile LAR signal? We found that application to tobacco leaves of 0.25 nM glycoprotein triggered defense responses without HR and without an H2O2 burst. The analyzed responses include changes in expression of O-methyltransferase (OMT), 3-hydroxy-3-methylglutarylCoA reductase, pathogenesis-related (PR) proteins, and changes in levels of the signal salicylic acid. No defense responses and no increased resistance to tobacco mosaic virus infection were found beyond the elicitor-infiltrated tissue, providing strong evidence that the...

Elicitors as alternative strategy to pesticides in grapevine? Current knowledge on their mode of action from controlled conditions to vineyard

Development and optimisation of alternative strategies to reduce the use of classic chemical inputs for protection against diseases in vineyard is becoming a necessity. Among these strategies, one of the most promising consists in the stimulation and/or potentiation of the grapevine defence responses by the means of elicitors. Elicitors are highly diverse molecules both in nature and origins. This review aims at providing an overview of the current knowledge on these molecules and will highlight their potential efficacy from the laboratory in controlled conditions to vineyards. Recent findings and concepts (especially on plant innate immunity) and the new terminology (microbe-associated molecular patterns, effectors, etc.) are also discussed in this context. Other objectives of this review are to highlight the difficulty of transferring elicitors use and results from the controlled conditions to the vineyard, to determine their practical and effective use in viticulture and to propose ideas for improving their efficacy in non-controlled conditions.

Grapevine NAC1 transcription factor as a convergent node in developmental processes, abiotic stresses, and necrotrophic/biotrophic pathogen tolerance

Transcription factors of the NAC family are known to be involved in various developmental processes and in response to environmental stresses. Whereas NAC genes have been widely studied in response to abiotic stresses, little is known about their role in response to biotic stresses, especially in crops. Here, the first characterization of a Vitis vinifera L. NAC member, named VvNAC1, and involved in organ development and defence towards pathogens is reported. Expression profile analysis of VvNAC1 showed that its expression is closely associated with later stages of leaf, flower, and berry development, suggesting a role in plant senescence. Moreover, VvNAC1 expression is stimulated in Botrytis cinerea- or microbe-associated molecular pattern (MAMP)-infected berries or leaves. Furthermore, cold, wounding, and defence-related hormones such as salicylic acid, methyl jasmonate, ethylene, and abscisic acid are all able to induce VvNAC1 expression in grapevine leaves. VvNAC1-overexpressing Arabidopsis plants exhibit enhanced tolerance to osmotic, salt, and cold stresses and to B. cinerea and Hyaloperonospora arabidopsidis pathogens. These plants present a modified pattern of defence gene markers (AtPR-1, AtPDF1.2, and AtVSP1) after stress application, suggesting that VvNAC1 is an important regulatory component of the plant signalling defence cascade. Collectively, these results provide evidence that VvNAC1 could represent a node of convergence regulating grapevine development and stress responses, including defence against necrotrophic and biotrophic pathogens.

Large-scale proteomic analysis of the grapevine leaf apoplastic fluid reveals mainly stress-related proteins and cell wall modifying enzymes

BackgroundThe extracellular space or apoplast forms a path through the whole plant and acts as an interface with the environment. The apoplast is composed of plant cell wall and space within which apoplastic fluid provides a means of delivering molecules and facilitates intercellular communications. However, the apoplastic fluid extraction from in planta systems remains challenging and this is particularly true for grapevine (Vitis vinifera L.), a worldwide-cultivated fruit plant. Large-scale proteomic analysis reveals the protein content of the grapevine leaf apoplastic fluid and the free interactive proteome map considerably facilitates the study of the grapevine proteome.ResultsTo obtain a snapshot of the grapevine apoplastic fluid proteome, a vacuum-infiltration-centrifugation method was optimized to collect the apoplastic fluid from non-challenged grapevine leaves. Soluble apoplastic protein patterns were then compared to whole leaf soluble protein profiles by 2D-PAGE analyses. Subsequent MALDI-TOF/TOF mass spectrometry of tryptically digested protein spots was used to identify proteins. This large-scale proteomic analysis established a well-defined proteomic map of whole leaf and leaf apoplastic soluble proteins, with 223 and 177 analyzed spots, respectively. All data arising from proteomic, MS and MS/MS analyses were deposited in the public database world-2DPAGE. Prediction tools revealed a high proportion of (i) classical secreted proteins but also of non-classical secreted proteins namely Leaderless Secreted Proteins (LSPs) in the apoplastic protein content and (ii) proteins potentially involved in stress reactions and/or in cell wall metabolism.ConclusionsThis approach provides free online interactive reference maps annotating a large number of soluble proteins of the whole leaf and the apoplastic fluid of grapevine leaf. To our knowledge, this is the first detailed proteome study of grapevine apoplastic fluid providing a comprehensive overview of the most abundant proteins present in the apoplast of grapevine leaf that could be further characterized in order to elucidate their physiological function.