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Imaging with the fluorogenic dye Basic Fuchsin reveals subcellular patterning and ecotype variation of lignification in Brachypodium distachyon.

Kapp N, Barnes WJ, Richard TL, Anderson CT - J. Exp. Bot. (2015)

Bottom Line: It was found that the extent and intensity of Basic Fuchsin fluorescence increase over time in the Bd21-3 ecotype, that Basic Fuchsin staining is more widespread and intense in 4-week-old Bd21-3 and Adi-10 basal internodes than in Bd1-1 internodes, and that Basic Fuchsin staining reveals subcellular patterns of lignin in vascular and interfascicular fibre cell walls.Basic Fuchsin fluorescence did not correlate with lignin quantification by acetyl bromide analysis, indicating that whole-plant and subcellular lignin analyses provide distinct information about the extent and patterns of lignification in B. distachyon.Finally, it was found that flowering time correlated with a transient increase in total lignin, but did not correlate strongly with the patterning of stem lignification, suggesting that additional developmental pathways might regulate secondary wall formation in grasses.

View Article: PubMed Central - PubMed

Affiliation: Center for Lignocellulose Structure and Formation, The Pennsylvania State University, University Park, PA 16802, USA Department of Biology, The Pennsylvania State University, University Park, PA 16802, USA Department of Agricultural and Biological Engineering, The Pennsylvania State University, University Park, PA 16802, USA.

No MeSH data available.


Basic Fuchsin fluorescence varies across B. distachyon ecotypes. Basic Fuchsin fluorescence (A, B, C), Basic Fuchsin fluorescence with a pseudo-colour look-up table applied (A’, B’, C’), and Wiesner staining (A’’, B’’, C’’) of base internode stem sections (25 μm thickness) from B. distachyon ecotypes Bd21-3 (A, A’), Bd1-1 (B, B’), and Adi-10 (C, C’) sampled at 4 weeks of growth. See Table 1 for quantification of staining intensities for the three ecotypes. Scale bar=100 μm.
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Figure 2: Basic Fuchsin fluorescence varies across B. distachyon ecotypes. Basic Fuchsin fluorescence (A, B, C), Basic Fuchsin fluorescence with a pseudo-colour look-up table applied (A’, B’, C’), and Wiesner staining (A’’, B’’, C’’) of base internode stem sections (25 μm thickness) from B. distachyon ecotypes Bd21-3 (A, A’), Bd1-1 (B, B’), and Adi-10 (C, C’) sampled at 4 weeks of growth. See Table 1 for quantification of staining intensities for the three ecotypes. Scale bar=100 μm.

Mentions: B. distachyon ecotypes Bd1-1 and Adi-10 are both late-flowering compared with the ecotype Bd21-3, which displays synchronous growth and early flowering and is therefore often used as a control ecotype in B. distachyon studies (Vogel et al., 2009; Schwartz et al., 2010). Per cent heading was scored at 4 weeks for sets of Bd21-3, Bd1-1, and Adi-10 plants (Table 1; see Supplementary Table S1 at JXB online) to look for any correlation between flowering and lignification. In agreement with previous studies (Vogel et al., 2009; Schwartz et al., 2010), 90% of Bd21-3 plants had flowered by 4 weeks of growth, whereas 30% of Bd1-1 plants and 0% of Adi-10 plants had flowered (see Supplementary Fig. S3 at JXB online). Although Bd1-1 and Adi-10 were both late to flower, these ecotypes displayed different growth morphologies (see Supplementary Fig. S4 at JXB online) as well as different basal internode Basic Fuchsin staining intensity and patterning (Fig. 2).


Imaging with the fluorogenic dye Basic Fuchsin reveals subcellular patterning and ecotype variation of lignification in Brachypodium distachyon.

Kapp N, Barnes WJ, Richard TL, Anderson CT - J. Exp. Bot. (2015)

Basic Fuchsin fluorescence varies across B. distachyon ecotypes. Basic Fuchsin fluorescence (A, B, C), Basic Fuchsin fluorescence with a pseudo-colour look-up table applied (A’, B’, C’), and Wiesner staining (A’’, B’’, C’’) of base internode stem sections (25 μm thickness) from B. distachyon ecotypes Bd21-3 (A, A’), Bd1-1 (B, B’), and Adi-10 (C, C’) sampled at 4 weeks of growth. See Table 1 for quantification of staining intensities for the three ecotypes. Scale bar=100 μm.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

License 1 - License 2
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getmorefigures.php?uid=PMC4493785&req=5

Figure 2: Basic Fuchsin fluorescence varies across B. distachyon ecotypes. Basic Fuchsin fluorescence (A, B, C), Basic Fuchsin fluorescence with a pseudo-colour look-up table applied (A’, B’, C’), and Wiesner staining (A’’, B’’, C’’) of base internode stem sections (25 μm thickness) from B. distachyon ecotypes Bd21-3 (A, A’), Bd1-1 (B, B’), and Adi-10 (C, C’) sampled at 4 weeks of growth. See Table 1 for quantification of staining intensities for the three ecotypes. Scale bar=100 μm.
Mentions: B. distachyon ecotypes Bd1-1 and Adi-10 are both late-flowering compared with the ecotype Bd21-3, which displays synchronous growth and early flowering and is therefore often used as a control ecotype in B. distachyon studies (Vogel et al., 2009; Schwartz et al., 2010). Per cent heading was scored at 4 weeks for sets of Bd21-3, Bd1-1, and Adi-10 plants (Table 1; see Supplementary Table S1 at JXB online) to look for any correlation between flowering and lignification. In agreement with previous studies (Vogel et al., 2009; Schwartz et al., 2010), 90% of Bd21-3 plants had flowered by 4 weeks of growth, whereas 30% of Bd1-1 plants and 0% of Adi-10 plants had flowered (see Supplementary Fig. S3 at JXB online). Although Bd1-1 and Adi-10 were both late to flower, these ecotypes displayed different growth morphologies (see Supplementary Fig. S4 at JXB online) as well as different basal internode Basic Fuchsin staining intensity and patterning (Fig. 2).

Bottom Line: It was found that the extent and intensity of Basic Fuchsin fluorescence increase over time in the Bd21-3 ecotype, that Basic Fuchsin staining is more widespread and intense in 4-week-old Bd21-3 and Adi-10 basal internodes than in Bd1-1 internodes, and that Basic Fuchsin staining reveals subcellular patterns of lignin in vascular and interfascicular fibre cell walls.Basic Fuchsin fluorescence did not correlate with lignin quantification by acetyl bromide analysis, indicating that whole-plant and subcellular lignin analyses provide distinct information about the extent and patterns of lignification in B. distachyon.Finally, it was found that flowering time correlated with a transient increase in total lignin, but did not correlate strongly with the patterning of stem lignification, suggesting that additional developmental pathways might regulate secondary wall formation in grasses.

View Article: PubMed Central - PubMed

Affiliation: Center for Lignocellulose Structure and Formation, The Pennsylvania State University, University Park, PA 16802, USA Department of Biology, The Pennsylvania State University, University Park, PA 16802, USA Department of Agricultural and Biological Engineering, The Pennsylvania State University, University Park, PA 16802, USA.

No MeSH data available.