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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.


High-magnification imaging of Basic Fuchsin staining. Basic Fuchsin staining reveals subcellular maxima of fluorescence intensity in the middle lamellae and cell corners and lattice-like patterning in the walls of vessel elements of B. distachyon ecotypes Bd21-3 (A, A’, A’’), Bd1-1 (B, B’, B’’), and Adi-10 (C’, C’, C’’) sampled at 4 weeks of growth in grey-scale (A, B, C) and with a pseudo colour look-up table applied (A’, B’, C’). Insets show magnification of area outlined in red in A, B, and C, showing high-intensity staining at middle lamellae (Ai, Bi, Ci; red arrows) and cell corners (A’i, B’i, C’I; black arrows). Scale bar=20 μm.
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Figure 3: High-magnification imaging of Basic Fuchsin staining. Basic Fuchsin staining reveals subcellular maxima of fluorescence intensity in the middle lamellae and cell corners and lattice-like patterning in the walls of vessel elements of B. distachyon ecotypes Bd21-3 (A, A’, A’’), Bd1-1 (B, B’, B’’), and Adi-10 (C’, C’, C’’) sampled at 4 weeks of growth in grey-scale (A, B, C) and with a pseudo colour look-up table applied (A’, B’, C’). Insets show magnification of area outlined in red in A, B, and C, showing high-intensity staining at middle lamellae (Ai, Bi, Ci; red arrows) and cell corners (A’i, B’i, C’I; black arrows). Scale bar=20 μm.

Mentions: At the subcellular level, Basic Fuchsin fluorescence was concentrated in the middle lamellae and cell corners in all three B. distachyon ecotypes (Fig. 3) where lignin polymerization is thought to initiate (Donaldson, 2001). More widespread Basic Fuchsin fluorescence was observed in Bd21-3 and Adi-10 vessel element and fibre cell walls than in Bd1-1 walls (Fig. 3). These results suggest that the differences in total Basic Fuchsin fluorescence between these ecotypes (Table 1) might arise both from differences in the number of cells within the stem that are lignified at a given developmental stage and the extent of lignin deposition within individual cell walls. Lattice-like gaps were also observed in Basic Fuchsin fluorescence adjacent to vessel elements in Bd21-3 stems that probably reflect the presence of non-lignified pits in these walls (Fig. 3).


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)

High-magnification imaging of Basic Fuchsin staining. Basic Fuchsin staining reveals subcellular maxima of fluorescence intensity in the middle lamellae and cell corners and lattice-like patterning in the walls of vessel elements of B. distachyon ecotypes Bd21-3 (A, A’, A’’), Bd1-1 (B, B’, B’’), and Adi-10 (C’, C’, C’’) sampled at 4 weeks of growth in grey-scale (A, B, C) and with a pseudo colour look-up table applied (A’, B’, C’). Insets show magnification of area outlined in red in A, B, and C, showing high-intensity staining at middle lamellae (Ai, Bi, Ci; red arrows) and cell corners (A’i, B’i, C’I; black arrows). Scale bar=20 μm.
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Related In: Results  -  Collection

License 1 - License 2
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Figure 3: High-magnification imaging of Basic Fuchsin staining. Basic Fuchsin staining reveals subcellular maxima of fluorescence intensity in the middle lamellae and cell corners and lattice-like patterning in the walls of vessel elements of B. distachyon ecotypes Bd21-3 (A, A’, A’’), Bd1-1 (B, B’, B’’), and Adi-10 (C’, C’, C’’) sampled at 4 weeks of growth in grey-scale (A, B, C) and with a pseudo colour look-up table applied (A’, B’, C’). Insets show magnification of area outlined in red in A, B, and C, showing high-intensity staining at middle lamellae (Ai, Bi, Ci; red arrows) and cell corners (A’i, B’i, C’I; black arrows). Scale bar=20 μm.
Mentions: At the subcellular level, Basic Fuchsin fluorescence was concentrated in the middle lamellae and cell corners in all three B. distachyon ecotypes (Fig. 3) where lignin polymerization is thought to initiate (Donaldson, 2001). More widespread Basic Fuchsin fluorescence was observed in Bd21-3 and Adi-10 vessel element and fibre cell walls than in Bd1-1 walls (Fig. 3). These results suggest that the differences in total Basic Fuchsin fluorescence between these ecotypes (Table 1) might arise both from differences in the number of cells within the stem that are lignified at a given developmental stage and the extent of lignin deposition within individual cell walls. Lattice-like gaps were also observed in Basic Fuchsin fluorescence adjacent to vessel elements in Bd21-3 stems that probably reflect the presence of non-lignified pits in these walls (Fig. 3).

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.