|
Plant Physiol, December 2002, Vol. 130, pp. 1754-1763
Rapid Structural Phenotyping of Plant Cell Wall Mutants by
Enzymatic Oligosaccharide Fingerprinting1
Olivier
Lerouxel,2
Tze Siang
Choo,2
Martial
Séveno,
Björn
Usadel,
Loïc
Faye,
Patrice
Lerouge, and
Markus
Pauly*
Centre National de la Recherche Scientifique Unité Mixte de
Recherche 6037, Institute Federative de Recherche Multidisciplinaire
sur les Peptides 23, University of Rouen, 76821 Mont Saint Aignan,
France (O.L., M.S., L.F., P.L.); and Max-Planck-Institut of Molecular
Plant Physiology, Am Mühlenberg 1, 14476 Golm, Potsdam, Germany
(T.S.C., B.U., M.P.)
Various biochemical, chemical, and microspectroscopic
methods have been developed throughout the years for the
screening and identification of mutants with altered cell wall
structure. However, these procedures fail to provide the insight into
structural aspects of the cell wall polymers. In this paper, we present
various methods for rapidly screening Arabidopsis cell wall mutants.
The enzymatic fingerprinting procedures using high-performance
anion-exchange-pulsed-amperometric detection liquid chromatography,
fluorophore-assisted carbohydrate electrophoresis, and matrix-assisted
laser-desorption ionization time of flight (MALDI-TOF) mass
spectrometry (MS) were exemplified by the structural analysis of the
hemicellulose xyloglucan. All three techniques are able to identify
structural alterations of wall xyloglucans in mur1,
mur2, and mur3, which in comparison with
the wild type have side chain defects in their xyloglucan structure.
The quickest analysis was provided by MALDI-TOF MS. Although MALDI-TOF
MS per se is not quantitative, it is possible to reproducibly obtain
relative abundance information of the various oligosaccharides present
in the extract. The lack of absolute quantitation by MALDI-TOF MS was
compensated for with a xyloglucan-specific endoglucanase and simple
colorimetric assay. In view of the potential for mass screening using
MALDI-TOF MS, a PERL-based program was developed to process the spectra
obtained from MALDI-TOF MS automatically. Outliers can be identified
very rapidly according to a set of defined parameters based on data
collected from the wild-type plants. The methods presented here can
easily be adopted for the analysis of other wall polysaccharides.
MALDI-TOF MS offers a powerful tool to screen and identify cell wall
mutants rapidly and efficiently and, more importantly, is able to give
initial insights into the structural composition and/or modification
that occurs in these mutants.
1
This work was supported in Germany by the
Ministry of Research and Education within the Genom Analyse im
Biologischen System Pflanze (GABI) and GABI-Genoplante initiatives and
in France by the Centre National de la Recherche Scientifique, the
University of Rouen, and Genoplante and GABI-Genoplante programs.
2
The authors contributed equally to this work.
*
Corresponding author; e-mail pauly{at}mpimp-golm.mpg.de; fax
49-331-5678250.
© 2002 American Society of Plant Biologists
This article has been cited by other articles:
|
|
|
|
 
J. Rosti, C. J. Barton, S. Albrecht, P. Dupree, M. Pauly, K. Findlay, K. Roberts, and G. J. Seifert
UDP-Glucose 4-Epimerase Isoforms UGE2 and UGE4 Cooperate in Providing UDP-Galactose for Cell Wall Biosynthesis and Growth of Arabidopsis thaliana
PLANT CELL,
May 1, 2007;
19(5):
1565 - 1579.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
V. Lionetti, A. Raiola, L. Camardella, A. Giovane, N. Obel, M. Pauly, F. Favaron, F. Cervone, and D. Bellincampi
Overexpression of Pectin Methylesterase Inhibitors in Arabidopsis Restricts Fungal Infection by Botrytis cinerea
Plant Physiology,
April 1, 2007;
143(4):
1871 - 1880.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
S. Bosca, C. J. Barton, N. G. Taylor, P. Ryden, L. Neumetzler, M. Pauly, K. Roberts, and G. J. Seifert
Interactions between MUR10/CesA7-Dependent Secondary Cellulose Biosynthesis and Primary Cell Wall Structure
Plant Physiology,
December 1, 2006;
142(4):
1353 - 1363.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
D. M. Cavalier and K. Keegstra
Two Xyloglucan Xylosyltransferases Catalyze the Addition of Multiple Xylosyl Residues to Cellohexaose
J. Biol. Chem.,
November 10, 2006;
281(45):
34197 - 34207.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
S. Bauer, P. Vasu, S. Persson, A. J. Mort, and C. R. Somerville
From the Cover: Development and application of a suite of polysaccharide-degrading enzymes for analyzing plant cell walls
PNAS,
July 25, 2006;
103(30):
11417 - 11422.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
G. Mouille, H. Witucka-Wall, M.-P. Bruyant, O. Loudet, S. Pelletier, C. Rihouey, O. Lerouxel, P. Lerouge, H. Hofte, and M. Pauly
Quantitative Trait Loci Analysis of Primary Cell Wall Composition in Arabidopsis
Plant Physiology,
July 1, 2006;
141(3):
1035 - 1044.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
J. P. Moore, E. Nguema-Ona, L. Chevalier, G. G. Lindsey, W. F. Brandt, P. Lerouge, J. M. Farrant, and A. Driouich
Response of the Leaf Cell Wall to Desiccation in the Resurrection Plant Myrothamnus flabellifolius
Plant Physiology,
June 1, 2006;
141(2):
651 - 662.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
E. Nguema-Ona, C. Andeme-Onzighi, S. Aboughe-Angone, M. Bardor, T. Ishii, P. Lerouge, and A. Driouich
The reb1-1 Mutation of Arabidopsis. Effect on the Structure and Localization of Galactose-Containing Cell Wall Polysaccharides
Plant Physiology,
April 1, 2006;
140(4):
1406 - 1417.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
K. Vissenberg, S. C. Fry, M. Pauly, H. Hofte, and J.-P. Verbelen
XTH acts at the microfibril-matrix interface during cell elongation
J. Exp. Bot.,
February 1, 2005;
56(412):
673 - 683.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
