We cut thin cross-sections of wild type and mutant stem internodes and stained them with phloroglucinol-HCl and toluidine blue (Figure 4). Phloroglucinol-HCl stains mostly lignin while toluidine blue stains polysaccharides and lignin. Comparing, the wild type and mutant internode cross-sections, we noticed that the wild type wall is thicker than the mutant wall, shown by the presence of more staining in the wild type cross-section (Figure 4a, 4c) than in the mutant cross-section (Figure 4b, 4d) for both phloroglucinol-HCl and toluidine blue stains. This indicates that there are possibly higher levels of lignin and polysaccharide in the wild type cell wall. Since the mutant shows less staining, we can predict that our gene is involved in the assembly of cell wall components.
Figure 4. Brachypodium distachyon internal stem internode anatomy. (a.)The stem internode cross-section from a wild type plant, stained with phloroglucinol-HCl. (b.)The stem internode cross-section from a mutant plant, stained with phloroglucinol-HCl. (c.)The stem internode cross-section from a wild type plant stained with toluidine blue. (d.)The stem internode cross-section from a mutant plant stained with toluidine blue. The positions of the epidermis (Ep), cortex (Co), vascular bundles (Vb) and pith (Pi) are indicated.
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