Response of bacterial communities to Pb smelter pollution in contrasting soils.

Anthropogenic inputs of trace elements (TE) into soils constitute a major public and environmental health problem. Bioavailability of TE is strongly related to the soil physicochemical parameters and thus to the ecosystem type. In order to test whether soil parameters influence the response of the bacterial community to TE pollution, we collected soil samples across contrasting ecosystems (hardwood, coniferous and hydromorphic soils), which have been contaminated in TE and especially lead (Pb) over several decades due to nearby industrial smelting activities. Bacterial community composition was analysed using high throughput amplicon sequencing and compared to the soil physicochemical parameters. Multivariate analyses of the pedological and biological data revealed that the bacterial community composition was affected by ecosystem type in the first place. An influence of the contamination level was also evidenced within each ecosystem. Despite the important variability in bacterial community structure, we found that specific bacterial groups such as γ-Proteobacteria, Verrucomicrobia and Chlamydiae showed a consistent response to Pb content across contrasting ecosystems. Verrucomicrobia were less abundant at high contamination level whereas Chlamydiae and γ-Proteobacteria were more abundant. We conclude that such groups and ratio's thereof can be considered as relevant bioindicators of Pb contamination.

Lead distribution in soils impacted by a secondary lead smelter: Experimental and modelling approaches.

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 5-10cm) 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.

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.

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

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 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. CONCLUSIONS: This 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.

Simultaneous monitoring of gaseous CO(2) and ethanol above champagne glasses via micro-gas chromatography (µGC).

In champagne tasting, gaseous CO(2) and volatile organic compounds progressively invade the headspace above glasses, thus progressively modifying the chemical space perceived by the consumer. In this study, a novel, rapid, and nonintrusive method aimed to simultaneously determine the content in gaseous CO(2) and ethanol above a glass poured with champagne, using a micro-gas chromatography technique coupled with a thermal conductivity detector, was presented. The simultaneous quantification of CO(2) and ethanol in the headspace of a champagne glass was monitored, in real tasting conditions, all along the first 15 min following pouring, depending on whether or not the glass shows effervescence. Both CO(2) and ethanol were found to be enhanced by the presence of ascending bubbles, thus confirming the close link between rising bubbles and the release of gaseous CO(2) and volatile organic compounds.

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.

One step purification of the grape vacuolar invertase.

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.

Low responsiveness of grapevine flowers and berries at fruit set to UV-C irradiation.

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; beta-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.

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.