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Published on February 24, 2006; 10.1104/pp.105.074997

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Received December 2, 2005
Returned for revision January 14, 2006
Accepted February 14, 2006

The reb1-1 mutation of Arabidopsis: Effect on the structure and localization of galactose-containing cell wall polysaccharides

Eric Nguema-Ona , Christine Andème-Onzighi , Sophie Aboughe-Angone , Muriel Bardor , Tadashi Ishii , Patrice Lerouge , and Azeddine Driouich *

UMR CNRS 6037, IFRMP 23, Université de Rouen, 76 821 Mont Saint Aignan, Cedex, France
Forestry and Forest Products Research Institute, P. O. Box 16, Tsukuba Norin Kenkyu Danchi-nai, Ibaraki 305-8687, Japan

* Corresponding author; email: azeddine.driouich{at}univ-rouen.fr.

The Arabidopsis thaliana root epidermal bulger1-1 (reb1-1) mutant (allelic to root hair defective, rhd1) is characterized by a reduced root elongation rate and by bulging of trichoblast cells. REB1/RHD1 gene belongs to a family of UDP-D-glucose 4-epimerases involved in the synthesis of D-galactose. Our previous study showed that certain arabinogalactan-protein epitopes were not expressed in bulging trichoblasts of the mutant (Andème-Onzighi et al., 2002, Planta 215: 949-958). In this study, using a combination of microscopical and biochemical methods, we have investigated the occurrence and the structure of three major galactose-containing polysaccharides, namely xyloglucan, rhamnogalacturonan I (RG-I) and rhamnogalacturonan II (RG-II) in the mutant root cell walls. Our immunocytochemical data show that swollen trichoblasts were not stained with the mAb antibody CCRC-M1 specific for {alpha}-L-Fucp-(1?2)-{beta}-D-Galp side chains of xyloglucan, while they were stained with anti-XyG antibodies specific for xyloglucan backbone. In addition, analysis of a hemicellulosic fraction from roots demonstrates the presence of two structurally different xyloglucans in reb1-1. One is structurally similar to wild-type xyloglucan and the other is devoid of fuco-galactosylated side-chains and has the characteristic of being insoluble. Similar to anti-XyG antibodies, anti-bupleuran 2IIC, a polyclonal antibody specific for galactosyl epitopes associated with pectins, stained all root epidermal cells of both wild type and reb1-1. Similarly, anti-RG-II antibodies also stained swollen trichoblasts in the mutant. In addition, structural analysis of pectic polymers revealed no change in the galactosylation of RG-I and RG-II isolated from reb1-1 root cells. These findings demonstrate that the reb1-1 mutation affects xyloglucan structure, but not that of pectic polysaccharides, thus lending support to the hypothesis that biosynthesis of galactose as well as galactosylation of complex polysaccharides is regulated at the polymer level.




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