Immunochemical Characterization of Antibodies to the Xyloglucan-octasaccharide and Histochemical Recognition of Xyloglucan in Plant Tissues

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An octasaccharide [XGO8, composed of n-xylose, D-galactose, and D-glucose (3: 1: 4) ] purified from the (1→4) β-D-glucanase-digest of xyloglucan of tamarind (Tamarindus indica L .) seed, was coupled with bovine serum albumin (BSA), and used as an immunogen. The polyclonal antibodies raised in rabbits interacted with XGO8-BSA conjugates and of other oligosaccharide fragments, and also with the original xyloglucan. Hapten inhibition studies using various oligosaccharides showed that the antibodies recognize β-D-galactosyl- (1→2) -α-D-xylosyl, and also α-D-xylosyl side chains attached to the β- (14) -n-glucosyl backbone chain in the xyloglucan. The antibodies were used for histological localization of the xyloglucan in plant tissues, by either the fluorescence or the peroxidase-labeled immunostaining method. By the latter method it was shown that, in the soybean cotyledon, the location of the xyloglucan is restricted to the primary cell-walls .
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