Plant Proteomics pp 391-405

Part of the Methods in Molecular Biology book series (MIMB, volume 1072) | Cite as

Xylem Sap Proteomics

  • Thomas Dugé de Bernonville
  • Cécile Albenne
  • Matthieu Arlat
  • Laurent Hoffmann
  • Emmanuelle Lauber
  • Elisabeth Jamet
Protocol

Abstract

Proteomic analysis of xylem sap has recently become a major field of interest to understand several biological questions related to plant development and responses to environmental clues. The xylem sap appears as a dynamic fluid undergoing changes in its proteome upon abiotic and biotic stresses. Unlike cell compartments which are amenable to purification in sufficient amount prior to proteomic analysis, the xylem sap has to be collected in particular conditions to avoid contamination by intracellular proteins and to obtain enough material. A model plant like Arabidopsis thaliana is not suitable for such an analysis because efficient harvesting of xylem sap is difficult. The analysis of the xylem sap proteome also requires specific procedures to concentrate proteins and to focus on proteins predicted to be secreted. Indeed, xylem sap proteins appear to be synthesized and secreted in the root stele or to originate from dying differentiated xylem cells. This chapter describes protocols to collect xylem sap from Brassica species and to prepare total and N-glycoprotein extracts for identification of proteins by mass spectrometry analyses and bioinformatics.

Key words

Cell wall Glycoproteomics Xylem sap 

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Copyright information

© Springer Science+Business Media, LLC 2014

Authors and Affiliations

  • Thomas Dugé de Bernonville
    • 1
  • Cécile Albenne
    • 2
  • Matthieu Arlat
    • 1
  • Laurent Hoffmann
    • 2
  • Emmanuelle Lauber
    • 3
  • Elisabeth Jamet
    • 2
  1. 1.LIPMUMR CNRS-INRACastanet-TolosanFrance
  2. 2.Pôle de Biotechnologie Végétale, LRSV, UMR 5546Université Paul SabatierToulouse 3/CNRS, AuzevilleCastanet-TolosanFrance
  3. 3.LRSV, UMR 5546 UPS/CNRSCastanet-TolosanFrance

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