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G-fibre cell wall development in willow stems during tension wood induction.

Gritsch C, Wan Y, Mitchell RA, Shewry PR, Hanley SJ, Karp A - J. Exp. Bot. (2015)

Bottom Line: In addition, the expression patterns of an FLA gene (SxFLA12) and a COBRA-like gene (SxCOBL4) were compared using RNA in situ hybridization.Deposition of the non-cellulosic polysaccharides (1-4)-β-D-galactan, mannan and de-esterified homogalacturonan was found to be highly associated with TW, often with the G-layer itself.SxFLA12 and SxCOBL4 transcripts were specifically expressed in developing TW, confirming their importance.

View Article: PubMed Central - PubMed

Affiliation: Rothamsted Research, West Common, Harpenden, Hertfordshire AL5 2JQ, UK.

No MeSH data available.


Related in: MedlinePlus

TEM immunogold labelling. (A–G) Localization of (1–4)-β-D-galactan with the LM5 antibody in TW (A–F) and OW (G) after 4 weeks of inclined growth. (A) Early G-layer formation. Binding occurs in the G-layer itself or in the interface between the SCW and the G-layer (arrow). The area in the square is magnified in (C). (B) Detail of two adjacent developing G-fibres at early development. In the G-fibre on the left, gold labelling shows that early deposition of (1–4)-β-D-galactan occurs on the inner side of the SCW before the G-layer itself is formed (arrow). The G-layer is already being formed on the adjacent cell and it is labelled. In addition, electron-dense areas in the lumen are also labelled (arrowheads). (C) Detail of the two adjacent fibres in the square in (A) showing differential LM5 labelling occurring in the interface between SCW and the G-layer (arrowhead) or in the G-layer itself. (D) Maturing G-fibres showing thicker G-layers strongly labelled with LM5 antibody next to a ray parenchyma cell with unlabelled SCW. (E) Mature G-fibres showing differential labelling with LM5. Labelling in one of the fibres is mainly in the interface between the SCW and the G-layer (arrowhead). The other two fibres exhibit strong labelling of the G-layer. (F) Fully mature G-fibre. Labelling in these fibres is mainly confined to the SCW-GL interface. (G) OW mature fibres. Very sparse gold labelling is observed in the SCWs of xylary fibres. (H–K) Localization of de-esterified homogalacturonan with the CCRC-M38 antibody in TW after 4 weeks of inclined growth (H–J) and NW in a vertically grown stem (K). (H) Developing G-fibres. Homogalacturonan is localized by the CCRC-M38 antibody in the compound middle lamella (arrowhead) and in the intercellular spaces, which are filled with fibrous material. (I) Homogalacturonan labelling in the intercellular spaces between parenchyma ray cells and G-fibres. (J) Mature G-fibre from the inner part of the stem where CCRC-M38 labelling is mainly confined to the inner lamella of the G-layer (arrowhead). (K) NW fibres walls in a straight stem. CCRC-M38 antibody labelling is present only in cell corners and compound middle lamella. (L–O) Localization of xylan with the LM10 antibody in TW (L–N) after 4 weeks of inclined growth and OW (O). (L) Developing G-fibres. Xylan is restricted to SCWs, but labelling is not very strong. (M) LM10 is bound to the SCWs of ray parenchyma and mature G-fibres. No labelling is detected in the G-layer. (N) OW. Labelling is also confined to the SCWs of fibres and parenchyma cells. (O) Negative control section probed with only the secondary 10nm gold anti-rat antibody. Straight stem fibres showing no gold labelling in any of the cell walls. GF, G-fibre; GL, G-layer; sw, secondary cell wall; rp, ray parenchyma; TW, tension wood; OW, opposite wood; NW, normal wood. Bars: 0.5 µm
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Figure 7: TEM immunogold labelling. (A–G) Localization of (1–4)-β-D-galactan with the LM5 antibody in TW (A–F) and OW (G) after 4 weeks of inclined growth. (A) Early G-layer formation. Binding occurs in the G-layer itself or in the interface between the SCW and the G-layer (arrow). The area in the square is magnified in (C). (B) Detail of two adjacent developing G-fibres at early development. In the G-fibre on the left, gold labelling shows that early deposition of (1–4)-β-D-galactan occurs on the inner side of the SCW before the G-layer itself is formed (arrow). The G-layer is already being formed on the adjacent cell and it is labelled. In addition, electron-dense areas in the lumen are also labelled (arrowheads). (C) Detail of the two adjacent fibres in the square in (A) showing differential LM5 labelling occurring in the interface between SCW and the G-layer (arrowhead) or in the G-layer itself. (D) Maturing G-fibres showing thicker G-layers strongly labelled with LM5 antibody next to a ray parenchyma cell with unlabelled SCW. (E) Mature G-fibres showing differential labelling with LM5. Labelling in one of the fibres is mainly in the interface between the SCW and the G-layer (arrowhead). The other two fibres exhibit strong labelling of the G-layer. (F) Fully mature G-fibre. Labelling in these fibres is mainly confined to the SCW-GL interface. (G) OW mature fibres. Very sparse gold labelling is observed in the SCWs of xylary fibres. (H–K) Localization of de-esterified homogalacturonan with the CCRC-M38 antibody in TW after 4 weeks of inclined growth (H–J) and NW in a vertically grown stem (K). (H) Developing G-fibres. Homogalacturonan is localized by the CCRC-M38 antibody in the compound middle lamella (arrowhead) and in the intercellular spaces, which are filled with fibrous material. (I) Homogalacturonan labelling in the intercellular spaces between parenchyma ray cells and G-fibres. (J) Mature G-fibre from the inner part of the stem where CCRC-M38 labelling is mainly confined to the inner lamella of the G-layer (arrowhead). (K) NW fibres walls in a straight stem. CCRC-M38 antibody labelling is present only in cell corners and compound middle lamella. (L–O) Localization of xylan with the LM10 antibody in TW (L–N) after 4 weeks of inclined growth and OW (O). (L) Developing G-fibres. Xylan is restricted to SCWs, but labelling is not very strong. (M) LM10 is bound to the SCWs of ray parenchyma and mature G-fibres. No labelling is detected in the G-layer. (N) OW. Labelling is also confined to the SCWs of fibres and parenchyma cells. (O) Negative control section probed with only the secondary 10nm gold anti-rat antibody. Straight stem fibres showing no gold labelling in any of the cell walls. GF, G-fibre; GL, G-layer; sw, secondary cell wall; rp, ray parenchyma; TW, tension wood; OW, opposite wood; NW, normal wood. Bars: 0.5 µm

Mentions: Distinct patterns of distribution of cell wall pectic and hemicellulosic epitopes were observed during TW wood formation after 1, 2, and 4 weeks of TW induction. Patterns were similar for all stages but much more marked after 2 and 4 weeks of induction; therefore, in most cases only images of 4-week induced stems are shown. Immunolabelling results for tissues from OW and control straight stems were similar; therefore, mostly images of OW are shown. Negative control sections probed solely with secondary antibodies showed absence of labelling (Fig. 7O for TEM and Supplementary Fig. S2 for confocal immunofluorescence). Control sections without the primary antibodies showed no significant autofluoresence from lignin using the same confocal gain settings as for experimental sections (see also Supplementary Fig. S2).


G-fibre cell wall development in willow stems during tension wood induction.

Gritsch C, Wan Y, Mitchell RA, Shewry PR, Hanley SJ, Karp A - J. Exp. Bot. (2015)

TEM immunogold labelling. (A–G) Localization of (1–4)-β-D-galactan with the LM5 antibody in TW (A–F) and OW (G) after 4 weeks of inclined growth. (A) Early G-layer formation. Binding occurs in the G-layer itself or in the interface between the SCW and the G-layer (arrow). The area in the square is magnified in (C). (B) Detail of two adjacent developing G-fibres at early development. In the G-fibre on the left, gold labelling shows that early deposition of (1–4)-β-D-galactan occurs on the inner side of the SCW before the G-layer itself is formed (arrow). The G-layer is already being formed on the adjacent cell and it is labelled. In addition, electron-dense areas in the lumen are also labelled (arrowheads). (C) Detail of the two adjacent fibres in the square in (A) showing differential LM5 labelling occurring in the interface between SCW and the G-layer (arrowhead) or in the G-layer itself. (D) Maturing G-fibres showing thicker G-layers strongly labelled with LM5 antibody next to a ray parenchyma cell with unlabelled SCW. (E) Mature G-fibres showing differential labelling with LM5. Labelling in one of the fibres is mainly in the interface between the SCW and the G-layer (arrowhead). The other two fibres exhibit strong labelling of the G-layer. (F) Fully mature G-fibre. Labelling in these fibres is mainly confined to the SCW-GL interface. (G) OW mature fibres. Very sparse gold labelling is observed in the SCWs of xylary fibres. (H–K) Localization of de-esterified homogalacturonan with the CCRC-M38 antibody in TW after 4 weeks of inclined growth (H–J) and NW in a vertically grown stem (K). (H) Developing G-fibres. Homogalacturonan is localized by the CCRC-M38 antibody in the compound middle lamella (arrowhead) and in the intercellular spaces, which are filled with fibrous material. (I) Homogalacturonan labelling in the intercellular spaces between parenchyma ray cells and G-fibres. (J) Mature G-fibre from the inner part of the stem where CCRC-M38 labelling is mainly confined to the inner lamella of the G-layer (arrowhead). (K) NW fibres walls in a straight stem. CCRC-M38 antibody labelling is present only in cell corners and compound middle lamella. (L–O) Localization of xylan with the LM10 antibody in TW (L–N) after 4 weeks of inclined growth and OW (O). (L) Developing G-fibres. Xylan is restricted to SCWs, but labelling is not very strong. (M) LM10 is bound to the SCWs of ray parenchyma and mature G-fibres. No labelling is detected in the G-layer. (N) OW. Labelling is also confined to the SCWs of fibres and parenchyma cells. (O) Negative control section probed with only the secondary 10nm gold anti-rat antibody. Straight stem fibres showing no gold labelling in any of the cell walls. GF, G-fibre; GL, G-layer; sw, secondary cell wall; rp, ray parenchyma; TW, tension wood; OW, opposite wood; NW, normal wood. Bars: 0.5 µm
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Related In: Results  -  Collection

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Figure 7: TEM immunogold labelling. (A–G) Localization of (1–4)-β-D-galactan with the LM5 antibody in TW (A–F) and OW (G) after 4 weeks of inclined growth. (A) Early G-layer formation. Binding occurs in the G-layer itself or in the interface between the SCW and the G-layer (arrow). The area in the square is magnified in (C). (B) Detail of two adjacent developing G-fibres at early development. In the G-fibre on the left, gold labelling shows that early deposition of (1–4)-β-D-galactan occurs on the inner side of the SCW before the G-layer itself is formed (arrow). The G-layer is already being formed on the adjacent cell and it is labelled. In addition, electron-dense areas in the lumen are also labelled (arrowheads). (C) Detail of the two adjacent fibres in the square in (A) showing differential LM5 labelling occurring in the interface between SCW and the G-layer (arrowhead) or in the G-layer itself. (D) Maturing G-fibres showing thicker G-layers strongly labelled with LM5 antibody next to a ray parenchyma cell with unlabelled SCW. (E) Mature G-fibres showing differential labelling with LM5. Labelling in one of the fibres is mainly in the interface between the SCW and the G-layer (arrowhead). The other two fibres exhibit strong labelling of the G-layer. (F) Fully mature G-fibre. Labelling in these fibres is mainly confined to the SCW-GL interface. (G) OW mature fibres. Very sparse gold labelling is observed in the SCWs of xylary fibres. (H–K) Localization of de-esterified homogalacturonan with the CCRC-M38 antibody in TW after 4 weeks of inclined growth (H–J) and NW in a vertically grown stem (K). (H) Developing G-fibres. Homogalacturonan is localized by the CCRC-M38 antibody in the compound middle lamella (arrowhead) and in the intercellular spaces, which are filled with fibrous material. (I) Homogalacturonan labelling in the intercellular spaces between parenchyma ray cells and G-fibres. (J) Mature G-fibre from the inner part of the stem where CCRC-M38 labelling is mainly confined to the inner lamella of the G-layer (arrowhead). (K) NW fibres walls in a straight stem. CCRC-M38 antibody labelling is present only in cell corners and compound middle lamella. (L–O) Localization of xylan with the LM10 antibody in TW (L–N) after 4 weeks of inclined growth and OW (O). (L) Developing G-fibres. Xylan is restricted to SCWs, but labelling is not very strong. (M) LM10 is bound to the SCWs of ray parenchyma and mature G-fibres. No labelling is detected in the G-layer. (N) OW. Labelling is also confined to the SCWs of fibres and parenchyma cells. (O) Negative control section probed with only the secondary 10nm gold anti-rat antibody. Straight stem fibres showing no gold labelling in any of the cell walls. GF, G-fibre; GL, G-layer; sw, secondary cell wall; rp, ray parenchyma; TW, tension wood; OW, opposite wood; NW, normal wood. Bars: 0.5 µm
Mentions: Distinct patterns of distribution of cell wall pectic and hemicellulosic epitopes were observed during TW wood formation after 1, 2, and 4 weeks of TW induction. Patterns were similar for all stages but much more marked after 2 and 4 weeks of induction; therefore, in most cases only images of 4-week induced stems are shown. Immunolabelling results for tissues from OW and control straight stems were similar; therefore, mostly images of OW are shown. Negative control sections probed solely with secondary antibodies showed absence of labelling (Fig. 7O for TEM and Supplementary Fig. S2 for confocal immunofluorescence). Control sections without the primary antibodies showed no significant autofluoresence from lignin using the same confocal gain settings as for experimental sections (see also Supplementary Fig. S2).

Bottom Line: In addition, the expression patterns of an FLA gene (SxFLA12) and a COBRA-like gene (SxCOBL4) were compared using RNA in situ hybridization.Deposition of the non-cellulosic polysaccharides (1-4)-β-D-galactan, mannan and de-esterified homogalacturonan was found to be highly associated with TW, often with the G-layer itself.SxFLA12 and SxCOBL4 transcripts were specifically expressed in developing TW, confirming their importance.

View Article: PubMed Central - PubMed

Affiliation: Rothamsted Research, West Common, Harpenden, Hertfordshire AL5 2JQ, UK.

No MeSH data available.


Related in: MedlinePlus