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Tubulin perturbation leads to unexpected cell wall modifications and affects stomatal behaviour in Populus.

Swamy PS, Hu H, Pattathil S, Maloney VJ, Xiao H, Xue LJ, Chung JD, Johnson VE, Zhu Y, Peter GF, Hahn MG, Mansfield SD, Harding SA, Tsai CJ - J. Exp. Bot. (2015)

Bottom Line: The results suggest that pectin and xylan polysaccharides deposited early during cell wall biogenesis are more sensitive to subtle tubulin perturbation than cellulose and matrix polysaccharides deposited later.Pectins have been shown to confer cell wall flexibility critical for reversible stomatal movement, and results presented here are consistent with microtubule involvement in this process.Taken together, the data show the value of growth-compatible tubulin perturbations for discerning microtubule functions, and add to the growing body of evidence for microtubule involvement in non-cellulosic polysaccharide assembly during cell wall biogenesis.

View Article: PubMed Central - PubMed

Affiliation: School of Forestry and Natural Resources, University of Georgia, Athens, GA 30602, USA.

No MeSH data available.


Glycome profiling of xylem. Self-organizing map clustering of 127 cell wall glycan epitopes in six sequential extracts of WT and transgenic xylem samples. The number of epitopes (in parentheses) and the predominant glycan class(es) in each cluster are indicated. The average signal intensities (Supplementary Table S3) are plotted against cell wall fractions and colour-coded by plant line for visualization. I-VI, sequential cell wall extractions with ammonium oxalate (I), sodium carbonate (II), 1M KOH (III), 4M KOH (IV), sodium chloride (V) and post-chlorite 4M KOH (VI). AG, arabinogalactan; HG, homogalacturonan; RG-I, rhamnogalacturonan I; X, xylan; XG, xyloglucan.
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Figure 3: Glycome profiling of xylem. Self-organizing map clustering of 127 cell wall glycan epitopes in six sequential extracts of WT and transgenic xylem samples. The number of epitopes (in parentheses) and the predominant glycan class(es) in each cluster are indicated. The average signal intensities (Supplementary Table S3) are plotted against cell wall fractions and colour-coded by plant line for visualization. I-VI, sequential cell wall extractions with ammonium oxalate (I), sodium carbonate (II), 1M KOH (III), 4M KOH (IV), sodium chloride (V) and post-chlorite 4M KOH (VI). AG, arabinogalactan; HG, homogalacturonan; RG-I, rhamnogalacturonan I; X, xylan; XG, xyloglucan.

Mentions: To further investigate possible cell wall modifications in the transgenics, extractive-free wood meal was subjected to sequential fractionation of cell wall polysaccharides for glycome profiling (Pattathil et al., 2010; Pattathil et al., 2012). In general, mild solvent extractions by oxalate (fraction I) and carbonate (II) preferentially release arabinogalactans (AGs) and pectic components. Subsequent alkaline extractions with 1M (III) and 4M KOH (IV) remove hemicellulosic components (xylans and xyloglucans). Chlorite extraction (V) degrades lignin and associated carbohydrates, with the post-chlorite 4M KOH treatment (VI) releasing additional tightly bound polysaccharides. Total sugars of each cell wall fraction estimated by the phenol-sulfuric acid method differed little between genotypes (Supplementary Fig. S4A). Self-organizing map clustering of ELISA data from 127 cell wall glycan-directed mAbs revealed six major epitope clusters across the cell wall fractions and genotypes (Fig. 3, Supplementary Table S3). Cluster 1 consisted mainly of xyloglucan epitopes, with similar extractability between genotypes (Fig. 3). The other clusters were dominated by pectin- (clusters 2–5) and/or xylan-derived (clusters 5 and 6) epitopes, and showed a general trend of increased extractability in the transgenics (Fig. 3). Statistical analysis confirmed that the vast majority of epitopes with significant changes (P ≤0.05 and fold-change ≥1.5) in both transgenic groups (39 out of 44) were derived from pectins (Supplementary Table S3).


Tubulin perturbation leads to unexpected cell wall modifications and affects stomatal behaviour in Populus.

Swamy PS, Hu H, Pattathil S, Maloney VJ, Xiao H, Xue LJ, Chung JD, Johnson VE, Zhu Y, Peter GF, Hahn MG, Mansfield SD, Harding SA, Tsai CJ - J. Exp. Bot. (2015)

Glycome profiling of xylem. Self-organizing map clustering of 127 cell wall glycan epitopes in six sequential extracts of WT and transgenic xylem samples. The number of epitopes (in parentheses) and the predominant glycan class(es) in each cluster are indicated. The average signal intensities (Supplementary Table S3) are plotted against cell wall fractions and colour-coded by plant line for visualization. I-VI, sequential cell wall extractions with ammonium oxalate (I), sodium carbonate (II), 1M KOH (III), 4M KOH (IV), sodium chloride (V) and post-chlorite 4M KOH (VI). AG, arabinogalactan; HG, homogalacturonan; RG-I, rhamnogalacturonan I; X, xylan; XG, xyloglucan.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

License 1 - License 2
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getmorefigures.php?uid=PMC4588895&req=5

Figure 3: Glycome profiling of xylem. Self-organizing map clustering of 127 cell wall glycan epitopes in six sequential extracts of WT and transgenic xylem samples. The number of epitopes (in parentheses) and the predominant glycan class(es) in each cluster are indicated. The average signal intensities (Supplementary Table S3) are plotted against cell wall fractions and colour-coded by plant line for visualization. I-VI, sequential cell wall extractions with ammonium oxalate (I), sodium carbonate (II), 1M KOH (III), 4M KOH (IV), sodium chloride (V) and post-chlorite 4M KOH (VI). AG, arabinogalactan; HG, homogalacturonan; RG-I, rhamnogalacturonan I; X, xylan; XG, xyloglucan.
Mentions: To further investigate possible cell wall modifications in the transgenics, extractive-free wood meal was subjected to sequential fractionation of cell wall polysaccharides for glycome profiling (Pattathil et al., 2010; Pattathil et al., 2012). In general, mild solvent extractions by oxalate (fraction I) and carbonate (II) preferentially release arabinogalactans (AGs) and pectic components. Subsequent alkaline extractions with 1M (III) and 4M KOH (IV) remove hemicellulosic components (xylans and xyloglucans). Chlorite extraction (V) degrades lignin and associated carbohydrates, with the post-chlorite 4M KOH treatment (VI) releasing additional tightly bound polysaccharides. Total sugars of each cell wall fraction estimated by the phenol-sulfuric acid method differed little between genotypes (Supplementary Fig. S4A). Self-organizing map clustering of ELISA data from 127 cell wall glycan-directed mAbs revealed six major epitope clusters across the cell wall fractions and genotypes (Fig. 3, Supplementary Table S3). Cluster 1 consisted mainly of xyloglucan epitopes, with similar extractability between genotypes (Fig. 3). The other clusters were dominated by pectin- (clusters 2–5) and/or xylan-derived (clusters 5 and 6) epitopes, and showed a general trend of increased extractability in the transgenics (Fig. 3). Statistical analysis confirmed that the vast majority of epitopes with significant changes (P ≤0.05 and fold-change ≥1.5) in both transgenic groups (39 out of 44) were derived from pectins (Supplementary Table S3).

Bottom Line: The results suggest that pectin and xylan polysaccharides deposited early during cell wall biogenesis are more sensitive to subtle tubulin perturbation than cellulose and matrix polysaccharides deposited later.Pectins have been shown to confer cell wall flexibility critical for reversible stomatal movement, and results presented here are consistent with microtubule involvement in this process.Taken together, the data show the value of growth-compatible tubulin perturbations for discerning microtubule functions, and add to the growing body of evidence for microtubule involvement in non-cellulosic polysaccharide assembly during cell wall biogenesis.

View Article: PubMed Central - PubMed

Affiliation: School of Forestry and Natural Resources, University of Georgia, Athens, GA 30602, USA.

No MeSH data available.