A synthetic phenylglycoside (beta-GlcY) that interacts specifically with arabinogalactan-proteins (AGPs), a class of plant cell surface proteoglycans, has been used to study the spatial distribution of AGPs in the xylem tissue of radiata pine. These studies demonstrated that AGPs were located in the compound middle lamella (CML) of the newly developed tracheid. Abundant, low salt extractable AGPs were purified from xylem tissue. Monosaccharide analysis showed that arabinose and galactose were the main sugars present. Linkage analysis showed that most of the arabinose was in the furanose form, at the terminal and 5-linked positions, and the majority of the galactose was in the pyranose form at the terminal 3-, 6- and 3,6-linked positions; a linkage composition typical of AGPs. The AGPs had an abundance of characteristic amino acid residues including alanine, hydroxyproline, proline, and serine. Separation of the AGPs using reversed-phase high performance liquid chromatography showed that one main fraction was eluted, which tested positive for AGPs by dot-blot analysis using anti-AGP monoclonal antibodies. Sedimentation equilibrium analysis showed that this main fraction contained a 226 kDa species. We have examined the function of AGPs in tracheid differentiation using an established radiata pine callus culture system grown on media containing beta-GlcY. The effect of beta-GlcY on the cultures was to reduce the overall tracheid differentiation rate in a concentration dependent manner, ultimately resulting in cell death. These studies provide further evidence that AGPs play an important role in tracheid differentiation, and thus may be an important biological target for improving wood quality.