Planta

, Volume 230, Issue 5, pp 947–957 | Cite as

Structural characterisation of the pectic polysaccharide rhamnogalacturonan II using an acidic fingerprinting methodology

  • Martial Séveno
  • Aline Voxeur
  • Christophe Rihouey
  • Ai-Min Wu
  • Tadashi Ishii
  • Christian Chevalier
  • Marie Christine Ralet
  • Azeddine Driouich
  • Alan Marchant
  • Patrice Lerouge
Original Article

Abstract

Rhamnogalacturonan II (RG-II) is a structurally complex cell wall pectic polysaccharide. Despite its complexity, both the structure of RG-II and its ability to dimerise via a borate diester are conserved in vascular plants suggesting that RG-II has a fundamental role in primary cell wall organisation and function. The selection and analysis of new mutants affected in RG-II formation represents a promising strategy to unravel these functions and to identify genes encoding enzymes involved in RG-II biosynthesis. In this paper, a novel fingerprinting strategy is described for the screening of RG-II mutants based on the mild acid hydrolysis of RG-II coupled to the analysis of the resulting fragments by mass spectrometry. This methodology was developed using RG-II fractions isolated from citrus pectins and then validated for RG-II isolated from the Arabidopsismur1 mutant and irx10 irx10-like double mutant.

Keywords

Cell wall mutants Fingerprinting Mass spectrometry Rhamnogalacturonan II 

Abbreviations

Api

d-Apiose

AceA

l-Aceric acid

Dha

2-Keto-3-deoxy-d-lyxo-heptulosaric acid

DMB

1,2 Diamino-4,5-methylene dioxybenzene

Kdo

2-Keto-3-deoxy-d-manno-octulosonic acid

ESI-MS

Electrospray ionisation mass spectrometry

EPG

Endo-α(1,4)-polygalacturonase

MALDI-TOF

Matrix-assisted laser desorption ionisation-time of flight

OLIMP

Oligosaccharide mass profiling method

2-OMeFuc

2-O-Methyl l-fucose

2-OMeXyl

2-O-Methyl d-xylose

RG-II

Rhamnogalacturonan II

TFA

Trifluoroacetic acid

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Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  • Martial Séveno
    • 1
    • 6
  • Aline Voxeur
    • 1
  • Christophe Rihouey
    • 1
  • Ai-Min Wu
    • 2
    • 7
  • Tadashi Ishii
    • 3
  • Christian Chevalier
    • 4
  • Marie Christine Ralet
    • 5
  • Azeddine Driouich
    • 1
  • Alan Marchant
    • 2
    • 7
  • Patrice Lerouge
    • 1
  1. 1.EA 4358, IFRMP 23, University of RouenMont Saint AignanFrance
  2. 2.Department of Forest Genetics and Plant PhysiologyUmeåSweden
  3. 3.Forestry and Forest Products Research InstituteIbarakiJapan
  4. 4.INRA, Unité Mixte de Recherche 619 Biologie du FruitVillenave d’OrnonFrance
  5. 5.UR1268 Biopolymères Interactions AssemblagesNantesFrance
  6. 6.CNRS UMR 5203, Plate-forme de Protéomique Fonctionnelle, Institut de Génomique Fonctionnelle, IFR3Montpellier Cedex 05France
  7. 7.School of Biological SciencesUniversity of SouthamptonSouthamptonUK

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