Expression of heterologous xyloglucan xylosyltransferases in Arabidopsis to investigate their role in determining xyloglucan xylosylation substitution patterns.

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CONCLUSION: Putative XyG xylosyltransferases from Tropaeolum majus (nasturtium) and Solanum lycopersicum (tomato) homologous to characterized Arabidopsis genes were identified and shown to functionally complement Arabidopsis mutants lacking xyloglucan demonstrating they represent xyloglucan xylosyltransferases. Xyloglucan is a major hemicellulose in the plant cell wall and is important for the structural organization of the wall. The fine structure of xyloglucan can vary dependent on plant species and tissue type. Most vascular seed-bearing plants including Arabidopsis thaliana and nasturtium (Tropaeolum majus) have a xyloglucan structure, in which three out of four backbone glucosyl-residues are substituted with xylosyl-residues. In contrast, the xyloglucan found in plants of the Solanaceae family, which includes tomato (Solanum lycopersicum), is typically less xylosylated with only two of the four backbone glucosyl-residues substituted with xylosyl-residues. To investigate the genetics of xyloglucan xylosylation, candidate xyloglucan xylosyltransferase genes (XXTs) homologous to known A. thaliana XXTs were cloned from nasturtium and tomato. These candidate XXTs were expressed in the A. thaliana xxt1/2 double and xxt1/2/5 triple mutant, whose walls lack detectable xyloglucan. Expression of the orthologs of XXT5 resulted in no detectable xyloglucan in the transgenic A. thaliana plants, consistent with a lack of xyloglucan in the A. thaliana xxt1/2 double mutant. However, transformation of both the tomato and nasturtium orthologs of AtXXT1 and AtXXT2 resulted in the production of xyloglucan with a xylosylation pattern similar to wild type A. thaliana indicating that both SlXXT2 and TmXXT2 likely have xylosyltransferase activity. As the expression of the SlXXT2 did not result in xyloglucan with a decreased xylosylation frequency found in tomato, this gene is not responsible for the unique xylosylation pattern found in the solanaceous plants.