Proteomic Approach To Identify Champagne Wine Proteins as Modified by Botrytis cinerea Infection

Laboratoire d'Oenologie et Chimie Appliquée, Université de Reims, Reims Cedex, France.
Journal of Proteome Research (Impact Factor: 4.25). 04/2008; 7(3):1199-208. DOI: 10.1021/pr070419p
Source: PubMed


The presence of the fungal pathogen, Botrytis cinerea, in the vineyard causes reductions in both quality and quantity of grapes and wine. Because proteins are involved in the foam stabilization of sparkling wines, we have undertaken, for the first time, a thorough proteomic analysis of two champagne base wines prepared with either healthy or botrytized Chardonnay grapes, using two-dimensional electrophoresis (2DE) coupled with immunodetection and tandem mass spectrometry. Most of the identified proteins were from grape origin: invertase and pathogenesis-related (PR) proteins. The disappearance of numerous grape proteins was observed in the botrytized wine, suggesting that they were probably degraded or even repressed or the result of a differential expression of grape proteins upon fungal infection. On the other hand, two pectinolytic enzymes secreted by B. cinerea were found in the botrytized wine.

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Available from: Philippe Jeandet, Mar 18, 2015
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    • "However, it is worth noting that this correlation was not systematically observed [11] [12] [13]. These discrepancies could be due to environmental conditions and grape variety which can affect the protein content of grapes [14] [15]. On the other hand, the wine proteins may be modified throughout the winemaking [16] such as clarification with bentonite which has a negative influence on the foam of base wine and/or the protein content of wines [17] [18]. "
    Dataset: ACA2010 CC
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    • "Sequence coverage of amino acids and peptide fingerprinting agree with literature data for vacuolar invertase 1, GIN1 depending upon previous deglycosylation and multiple protease digestions in the case of Nglycoproteins (Cilindre et al., 2008; Jégou et al., 2009). However, no unique results about the thermo stability of vacuolar invertase 1, GIN1 from Vitis vinifera are reported in literature. "
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    ABSTRACT: Proteins and glycoproteins have shown to play a major role in heat induced haze formation in white wines despite of their low concentrations. The aim of this study was to improve thermal stability of white wine by enrichment with heat stable wine macromolecules. First, macromolecules from Sauvignon Blanc wine were separated and concentrated by sequential membrane fractionation, followed by the precipitation with acetone and then further purified by two-dimensional polyacrylamide gel electrophoresis. The main heat stable macromolecular fraction consisted of a glycosylated protein with molecular weight (MW) between 69 and 72 kDa and an isoelectric point of 3.25, which was identified by MALDI-TOF/TOF mass spectrometry as vacuolar grape invertase 1, GIN1. Using Concanavalin A affinity chromatography, high molecular weight compounds of 74 kDa and >250 kDa were retained, increasing the heat instability of wine from 48 NTU to 292 NTU. After desorption of these retained compounds followed by the enrichment of wine with glycoproteins having molecular weights of 70-100 kDa and >250 kDa, heat instability decreased by maximum 56%. However, this effect was mainly due to the desorption solution. Addition of grape invertase to Sauvignon Blanc would not improve wine stability.
    Macromolecular Chemistry: New Research, 1 edited by Valentin Gartner, 05/2013: chapter 3: pages 47-67; Nova Science Publishers, Inc.., ISBN: 978-1-62417-854-2
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    • "Chitinases (spots n° 19, 31, 44, 55, 59, 65, 121, 122, 125, 127, 128, 134, 135, 140, 141, 142, 144, 147, 148, 149, 155, 175, 176, Figure 1B) and glucanases (spots n° 22, 24, 27, 39, 40, 41, 42, 45, 46, 47, 61, 63, 64, 136, 167, 168, 169, Figure 1B) are also highly present in the AF, respectively 10.3% and 6.7% of the overall AF proteins detected on gels. They are well characterized for their role in plant defences by degrading fungal cell walls [47-49]. Identification redundancies among all these spots suggest a high rate of PTM in chitinases and glucanases, as it has been shown in wine in which several chitinases were found to be glycosylated [50]. "
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    ABSTRACT: 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.
    BMC Plant Biology 02/2013; 13(1):24. DOI:10.1186/1471-2229-13-24 · 3.81 Impact Factor
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