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Multi-scale spatial heterogeneity of pectic rhamnogalacturonan I (RG-I) structural features in tobacco seed endosperm cell walls.

Lee KJ, Cornuault V, Manfield IW, Ralet MC, Knox JP - Plant J. (2013)

Bottom Line: Heterogeneous RG-I polymers are implicated in generating the mechanical properties of cell walls during cell development and plant growth, but are poorly understood in architectural, biochemical and functional terms.The analyses indicate that the features of the RG-I polymer display spatial heterogeneity at the level of the tissue and the level of single cell walls, and also heterogeneity at the biochemical level.This work has implications for understanding RG-I glycan complexity in the context of cell-wall architectures and in relation to cell-wall functions in cell and tissue development.

View Article: PubMed Central - PubMed

Affiliation: Centre for Plant Sciences, Faculty of Biological Sciences, University of Leeds, Leeds, LS2 9JT, UK.

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Indirect immunofluorescence labelling of sections through tobacco seeds with Calcofluor White, LM6 arabinan and LM21 heteromannan probes. In some cases, the equivalent sections were pre-treated with endo-β–mannanase (+EβM) to remove the abundant heteromannan. E, embryo; ME, micropylar endosperm; NME, non-micropylar endosperm. White dashed lines with an asterisk indicate that a tissue asymmetry is detected. Brown dashed lines indicate no tissue level asymmetry was detected. Scale bar = 50 μm.
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fig01: Indirect immunofluorescence labelling of sections through tobacco seeds with Calcofluor White, LM6 arabinan and LM21 heteromannan probes. In some cases, the equivalent sections were pre-treated with endo-β–mannanase (+EβM) to remove the abundant heteromannan. E, embryo; ME, micropylar endosperm; NME, non-micropylar endosperm. White dashed lines with an asterisk indicate that a tissue asymmetry is detected. Brown dashed lines indicate no tissue level asymmetry was detected. Scale bar = 50 μm.

Mentions: Calcofluor White staining of a resin-embedded medial longitudinal section through a 3 h-imbibed tobacco seed reveals strong fluorescence of all embryo cell walls and asymmetry of the fluorescence intensity of cell walls in the endosperm, with stronger fluorescence in cell walls at the ME adjacent to the embryo radicle apex, as shown in Figure 1. This asymmetry within the endosperm tissue, revealed by Calcofluor White, reflects other structural features of tobacco seed endosperm cell walls as described previously (Lee et al., 2012). Immunolabelling of an equivalent section with the LM6 monoclonal antibody, which is directed to 1,5–α–arabinan, indicates the presence of the arabinan epitope in all embryo and endosperm cell walls, with relatively greater detection of the epitope in the ME, reflecting the stronger staining by Calcofluor White (Figure 1).


Multi-scale spatial heterogeneity of pectic rhamnogalacturonan I (RG-I) structural features in tobacco seed endosperm cell walls.

Lee KJ, Cornuault V, Manfield IW, Ralet MC, Knox JP - Plant J. (2013)

Indirect immunofluorescence labelling of sections through tobacco seeds with Calcofluor White, LM6 arabinan and LM21 heteromannan probes. In some cases, the equivalent sections were pre-treated with endo-β–mannanase (+EβM) to remove the abundant heteromannan. E, embryo; ME, micropylar endosperm; NME, non-micropylar endosperm. White dashed lines with an asterisk indicate that a tissue asymmetry is detected. Brown dashed lines indicate no tissue level asymmetry was detected. Scale bar = 50 μm.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3824205&req=5

fig01: Indirect immunofluorescence labelling of sections through tobacco seeds with Calcofluor White, LM6 arabinan and LM21 heteromannan probes. In some cases, the equivalent sections were pre-treated with endo-β–mannanase (+EβM) to remove the abundant heteromannan. E, embryo; ME, micropylar endosperm; NME, non-micropylar endosperm. White dashed lines with an asterisk indicate that a tissue asymmetry is detected. Brown dashed lines indicate no tissue level asymmetry was detected. Scale bar = 50 μm.
Mentions: Calcofluor White staining of a resin-embedded medial longitudinal section through a 3 h-imbibed tobacco seed reveals strong fluorescence of all embryo cell walls and asymmetry of the fluorescence intensity of cell walls in the endosperm, with stronger fluorescence in cell walls at the ME adjacent to the embryo radicle apex, as shown in Figure 1. This asymmetry within the endosperm tissue, revealed by Calcofluor White, reflects other structural features of tobacco seed endosperm cell walls as described previously (Lee et al., 2012). Immunolabelling of an equivalent section with the LM6 monoclonal antibody, which is directed to 1,5–α–arabinan, indicates the presence of the arabinan epitope in all embryo and endosperm cell walls, with relatively greater detection of the epitope in the ME, reflecting the stronger staining by Calcofluor White (Figure 1).

Bottom Line: Heterogeneous RG-I polymers are implicated in generating the mechanical properties of cell walls during cell development and plant growth, but are poorly understood in architectural, biochemical and functional terms.The analyses indicate that the features of the RG-I polymer display spatial heterogeneity at the level of the tissue and the level of single cell walls, and also heterogeneity at the biochemical level.This work has implications for understanding RG-I glycan complexity in the context of cell-wall architectures and in relation to cell-wall functions in cell and tissue development.

View Article: PubMed Central - PubMed

Affiliation: Centre for Plant Sciences, Faculty of Biological Sciences, University of Leeds, Leeds, LS2 9JT, UK.

Show MeSH