Received 31 Jan. 2019 Accepted 17 Feb. 2019 On line 31 Mar. 2019 BACHIROU SEYNI BODO, JEAN-MARIE ... more Received 31 Jan. 2019 Accepted 17 Feb. 2019 On line 31 Mar. 2019 BACHIROU SEYNI BODO, JEAN-MARIE KARIMOU AMBOUTA, OUMAROU MALAM ISSA, ADAMOU DIDIER TIDJANI, XAVIER MORVAN, ALEXANDRA CONREUX ,BEATRICE MARIN, MARIE PONTHIEU, GILLES FRONTEAU 3 (1) Département Sciences du sol / Faculté d’Agronomie / Université Abdou Moumouni de Niamey Niger, BP : 10960 Niamey, Niger Email : bachiroubodo@yahoo.fr (2) Institut de Recherche pour le Développement IRD Niger, BP : 11416 Niamey, Niger Email : oumarou.malamissa@ird.fr (3) UFR des Sciences Exactes et naturelles / Université de Reims Champagne-Ardenne Esplanade Roland Garros, 51100 REIMS, France Email : xavier.morvan@univ-reims.fr
Role of stilbene phytoalexins in the interaction between phytopathogenic micro-organisms and grapevine: biosynthesis, biological activity, fungal metabolism, chemical induction and stilbene synthase gene expression in transgenic plants
The grapevine phytoalexin resveratrol, the synthesis of which is achieved by stilbene synthase (S... more 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.
In grapevine, stimulation of defence responses was evidenced in response to various types of abio... more In grapevine, stimulation of defence responses was evidenced in response to various types of abiotic stresses in both leaves and berries, as revealed by the increasing expression of genes encoding defence-related proteins or the stimulation of their corresponding activities. However, the capability of inflorescences to respond to abiotic stresses has never been investigated. Therefore, plant defence reactions in response to UV-C irradiation were followed in inflorescences and young clusters focusing on both bunchstems (peduncle and pedicels) and developing flowers/ berries from separated floral buds stage [Biologische Bundesanstalt, Bundessortenamt and CHemical industry (BBCH) stage 57] to groat-sized berries stage (BBCH 73). For this purpose, the expression of various genes coding for pathogenesis-related (PR) proteins (class I and III chitinases, Chi1b and CH3; b-1,3-glucanase, GLUC), an enzyme of the phenylpropanoid pathway (phenylalanine ammonia-lyase, PAL), and stilbene synthase (STS) was analysed in parallel with variations of chitinase activity and the accumulation of the phytoalexin resveratrol. Multiple defence responses were induced in bunchstems of both inflorescences and clusters following UV-C treatment. First, expression of genes encoding PR proteins was stimulated and chitinase activity was enhanced. Secondly, PAL and STS expression increased in association with resveratrol accumulation. Amazingly, none of the tested defence processes was induced in grapevine flowers following UV-C exposure, whatever the stage analysed. Similarly, in berries at fruit set, induction of gene expression was weak and neither an increase in chitinase activity nor resveratrol synthesis was noticed. However, in groat-sized berries, responsiveness to UV-C increased, as revealed by the induction of CH3, PAL, and STS expression, together with resveratrol accumulation. The differential responsiveness between bunchstems, flowers, and berries is discussed.
Background The extracellular space or apoplast forms a path through the whole plant and acts as a... more Background The 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. Results To 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 analys...
Smelting activities are one of the most common sources of trace elements in the environment. The ... more Smelting activities are one of the most common sources of trace elements in the environment. The aim of this study was to determine the lead distribution in upper horizons (0-5 and 5-10 cm) of acidic soils in the vicinity of a lead-acid battery recycling plant in northern France. The combination of chemical methods (sequential extractions), physical methods (Raman microspectroscopy and scanning electron microscopy with an energy dispersive spectrometer) and multi-surface complexation modelling enabled an assessment of the behaviour of Pb. Regardless of the studied soil, none of the Pb-bearing phases commonly identified in similarly polluted environments (e.g., anglesite) were observed. Lead was mainly associated with organic matter and manganese oxides. The association of Pb with these soil constituents can be interpreted as evidence of Pb redistribution in the studied soils following smelter particle deposition.
Influence of dissolved organic matter and manganese oxides on metal speciation in soil solution: A modelling approach
Environmental Pollution, 2016
Trace element (TE) speciation modelling in soil solution is controlled by the assumptions made ab... more Trace element (TE) speciation modelling in soil solution is controlled by the assumptions made about the soil solution composition. To evaluate this influence, different assumptions using Visual MINTEQ were tested and compared to measurements of free TE concentrations. The soil column Donnan membrane technique (SC-DMT) was used to estimate the free TE (Cd, Cu, Ni, Pb and Zn) concentrations in six acidic soil solutions. A batch technique using DAX-8 resin was used to fractionate the dissolved organic matter (DOM) into four fractions: humic acids (HA), fulvic acids (FA), hydrophilic acids (Hy) and hydrophobic neutral organic matter (HON). To model TE speciation, particular attention was focused on the hydrous manganese oxides (HMO) and the Hy fraction, ligands not considered in most of the TE speciation modelling studies in soil solution. In this work, the model predictions of free ion activities agree with the experimental results. The knowledge of the FA fraction seems to be very useful, especially in the case of high DOM content, for more accurately representing experimental data. Finally, the role of the manganese oxides and of the Hy fraction on TE speciation was identified and, depending on the physicochemical conditions of the soil solution, should be considered in future studies.
Background: The extracellular space or apoplast forms a path through the whole plant and acts as ... more Background: The 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.
Background: The extracellular space or apoplast forms a path through the whole plant and acts as ... more Background: The 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.
ABSTRACT Both losses of dissolved CO2, and fluxes of gaseous CO2 escaping from a Champagne wine s... more ABSTRACT Both losses of dissolved CO2, and fluxes of gaseous CO2 escaping from a Champagne wine served in laser-etched INAO glasses showing an increasing number of bubble nucleation sites, were examined, all along the first 10 minutes following pouring. Logically, the more intense the effervescence is into a glass, the more rapid its progressive loss of dissolved CO2. Moreover, during the first minutes following pouring, the higher the number of laser beam’s points of impact is in glasses, the higher the fluxes of gaseous CO2 escaping from the champagne surface. Nevertheless, and quite counter intuitively, from approximately 3 min after pouring, the highest levels of gaseous CO2 fluxes escaping champagne were not sustained from glasses with the highest number of bubble trains. This experimental observation was discussed on the basis of a theoretical bubbling model. It was finally demonstrated that the flux of gaseous CO2 desorbing from the liquid phase is the result of a balance between the intensity of effervescence, and the dissolved CO2 concentration found in champagne.
Chitinases and thaumatin-like proteins are important grape proteins as they have a great influenc... more Chitinases and thaumatin-like proteins are important grape proteins as they have a great influence on wine quality. The quantification of these proteins in grape juices and wines, along with their purification, is therefore crucial to study their intrinsic characteristics and the exact role they play in wines. The main isoforms of these two proteins from Chardonnay grape juice were thus purified by liquid chromatography. Two fast protein liquid chromatography (FLPC) steps allowed the fractionation and purification of the juice proteins, using cation exchange and hydrophobic interaction media. A further high-performance liquid chromatography (HPLC) step was used to achieve higher purity levels. Fraction assessment was achieved by mass spectrometry. Fraction purity was determined by HPLC to detect the presence of protein contaminants, and by nuclear magnetic resonance (NMR) spectroscopy to detect the presence of organic contaminants. Once pure fractions of lyophilized chitinase and thaumatin-like protein were obtained, ultra-HPLC (UHPLC) and enzyme-linked immunosorbent assay (ELISA) calibration curves were constructed. The quantification of these proteins in different grape juice and wine samples was thus achieved for the first time with both techniques through comparison with the purified protein calibration curve. UHPLC and ELISA showed very consistent results (less than 16% deviation for both proteins) and either could be considered to provide an accurate and reliable quantification of proteins in the oenology field.
Invertase is a major protein of grape juice and wine. Accordingly, in order to study the biochemi... more Invertase is a major protein of grape juice and wine. Accordingly, in order to study the biochemical and structural characteristics of this protein and for a better understanding of its physico-chemical properties, large amounts of the pure protein are needed. A simple method for the purification of the grape vacuolar invertase in a preparative-scale is described in this work. The grape protein was isolated and purified from must by ultrafiltration and anion exchange chromatography. The identification and purity determination of the grape invertase fraction were assessed by SDS-PAGE, and were then confirmed using nanoLC-chip-MS/MS analysis. The laboratory fractionation procedure presented in this work generated large quantities of pure grape vacuolar invertase from must.
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