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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

Localization of (1–4)-β-D-galactan (LM5 antibody in red) and homogalacturonan (CCRC-M38 antibody in green) in transverse sections of stems after 1 or 4 weeks of TW induction. Numbers in the squares in (C) and (D) indicate similar areas shown in more detail in subsequent images. (A) to (H) are images of 1 µm resin sections and (I) and (J) are images of 100 µm unembedded sections. (A) Stem after 1 week of tipping showing the distribution of (1–4)-β-D-galactan (red) in TW fibres and of homogalacturonan (green) in the more mature inner side of the stem (detail in the inset). (B) OW side of the same section showing virtually no fluorescence in xylary fibres with the LM5 and CCRC-M38 antibodies. (C) TW side of a 4-week tipped stem showing strong labelling of G-fibres with the LM5 antibody in the outer developing and maturing part of the stem. (D) OW side of the same stem in (C). Fibre cell walls are not labelled or only weakly labelled with the LM5 and CCRC-M38 antibodies. (E) Detail of area 4 in (D). There is no labelling of OW fibre cell walls, indicating the absence of (1–4)-β-D-galactan. Some labelling is seen in ray parenchyma. Homogalacturonan is detected in cell corners (arrow). (F) and (G) Detail of area 1 in (C). LM5 antibody shows differential distribution of (1–4)-β-D-galactan from area 1 in files of G-fibres. The G-layer is labelled in some fibres, whereas in others the interface between the G-layer and SCW is labelled (arrowheads). (H) Detail of area 2 in (C). LM5 is bound mainly to the SCW and CCRC-M38 labels mainly the inner lamella of the G-layer (arrowhead). (I) and (J) Similar areas to 2 and 3 in (C) of unembedded sections showing CCRC-M38 mainly bound to the inner lamella of the G-layer. GL, G-layer; v, xylem vessel; rp, ray parenchyma. Bars: A, B, C, D = 100 µm; E, F, G, H, I, J = 10 µm.
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Figure 3: Localization of (1–4)-β-D-galactan (LM5 antibody in red) and homogalacturonan (CCRC-M38 antibody in green) in transverse sections of stems after 1 or 4 weeks of TW induction. Numbers in the squares in (C) and (D) indicate similar areas shown in more detail in subsequent images. (A) to (H) are images of 1 µm resin sections and (I) and (J) are images of 100 µm unembedded sections. (A) Stem after 1 week of tipping showing the distribution of (1–4)-β-D-galactan (red) in TW fibres and of homogalacturonan (green) in the more mature inner side of the stem (detail in the inset). (B) OW side of the same section showing virtually no fluorescence in xylary fibres with the LM5 and CCRC-M38 antibodies. (C) TW side of a 4-week tipped stem showing strong labelling of G-fibres with the LM5 antibody in the outer developing and maturing part of the stem. (D) OW side of the same stem in (C). Fibre cell walls are not labelled or only weakly labelled with the LM5 and CCRC-M38 antibodies. (E) Detail of area 4 in (D). There is no labelling of OW fibre cell walls, indicating the absence of (1–4)-β-D-galactan. Some labelling is seen in ray parenchyma. Homogalacturonan is detected in cell corners (arrow). (F) and (G) Detail of area 1 in (C). LM5 antibody shows differential distribution of (1–4)-β-D-galactan from area 1 in files of G-fibres. The G-layer is labelled in some fibres, whereas in others the interface between the G-layer and SCW is labelled (arrowheads). (H) Detail of area 2 in (C). LM5 is bound mainly to the SCW and CCRC-M38 labels mainly the inner lamella of the G-layer (arrowhead). (I) and (J) Similar areas to 2 and 3 in (C) of unembedded sections showing CCRC-M38 mainly bound to the inner lamella of the G-layer. GL, G-layer; v, xylem vessel; rp, ray parenchyma. Bars: A, B, C, D = 100 µm; E, F, G, H, I, J = 10 µm.

Mentions: The overall distribution pattern of (1–4)-β-D-galactan revealed using the LM5 antibody was similar after 1 (Fig. 3A), 2, and 4 weeks of induction, but more distinct in older stems (Fig. 3C). Strongest binding of the LM5 antibody occurred in G-fibres of the developing and maturing xylem of TW in the outer half of the stem (Fig. 3C, red fluorescence). Close examination revealed labelling in the G-layers but binding was not homogeneous. Some G-fibre cell files were interspersed with rows of other fibre cells where LM5 labelling was confined to the SCW (Fig. 3F, G). There was no evidence that this was confined to any particular fibre area, such as tips. In the inner older mature G-fibres, (1–4)-β-D-galactan was detected only in the SCW and was generally absent from the G-layer (Fig. 3H). TEM observations were largely in agreement with immunofluorescence results, but more detail on the location of the β-galactan epitope was obtained. In general only one layer (S1) of SCW could be detected in G-fibres and two layers (S1 + S2) in normal fibres. In the early stages of G-layer deposition, some fibres exhibited abundant gold labelling in the G-layer itself, whereas in others the labelling was restricted to the interface between the SCW and the newly formed G-layer (Fig. 7A, B, C). Fig. 7B shows binding of the LM5 antibody in a thin layer on the inner side of the lumen side of the SCW, where initiation of G-layer deposition is possibly occurring. In more mature G-fibres, a similar pattern was noted in which the G-layers of some fibres were enriched with (1–4)-β-D-galactan (Fig. 7D, E); however, in the fully mature fibres the labelling tended to be more sparse in the G-layer or absent and generally restricted to the SCW/G-layer interface (Fig. 7F). In contrast to TW, the fibre cell walls in OW and NW in control stems were either not labelled or poorly labelled as shown by immunofluorescence (Fig. 3 B, D, E). TEM also showed almost negligible levels of labelling of fibre SCWs in NW and OW (Fig. 7G).


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)

Localization of (1–4)-β-D-galactan (LM5 antibody in red) and homogalacturonan (CCRC-M38 antibody in green) in transverse sections of stems after 1 or 4 weeks of TW induction. Numbers in the squares in (C) and (D) indicate similar areas shown in more detail in subsequent images. (A) to (H) are images of 1 µm resin sections and (I) and (J) are images of 100 µm unembedded sections. (A) Stem after 1 week of tipping showing the distribution of (1–4)-β-D-galactan (red) in TW fibres and of homogalacturonan (green) in the more mature inner side of the stem (detail in the inset). (B) OW side of the same section showing virtually no fluorescence in xylary fibres with the LM5 and CCRC-M38 antibodies. (C) TW side of a 4-week tipped stem showing strong labelling of G-fibres with the LM5 antibody in the outer developing and maturing part of the stem. (D) OW side of the same stem in (C). Fibre cell walls are not labelled or only weakly labelled with the LM5 and CCRC-M38 antibodies. (E) Detail of area 4 in (D). There is no labelling of OW fibre cell walls, indicating the absence of (1–4)-β-D-galactan. Some labelling is seen in ray parenchyma. Homogalacturonan is detected in cell corners (arrow). (F) and (G) Detail of area 1 in (C). LM5 antibody shows differential distribution of (1–4)-β-D-galactan from area 1 in files of G-fibres. The G-layer is labelled in some fibres, whereas in others the interface between the G-layer and SCW is labelled (arrowheads). (H) Detail of area 2 in (C). LM5 is bound mainly to the SCW and CCRC-M38 labels mainly the inner lamella of the G-layer (arrowhead). (I) and (J) Similar areas to 2 and 3 in (C) of unembedded sections showing CCRC-M38 mainly bound to the inner lamella of the G-layer. GL, G-layer; v, xylem vessel; rp, ray parenchyma. Bars: A, B, C, D = 100 µm; E, F, G, H, I, J = 10 µm.
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Figure 3: Localization of (1–4)-β-D-galactan (LM5 antibody in red) and homogalacturonan (CCRC-M38 antibody in green) in transverse sections of stems after 1 or 4 weeks of TW induction. Numbers in the squares in (C) and (D) indicate similar areas shown in more detail in subsequent images. (A) to (H) are images of 1 µm resin sections and (I) and (J) are images of 100 µm unembedded sections. (A) Stem after 1 week of tipping showing the distribution of (1–4)-β-D-galactan (red) in TW fibres and of homogalacturonan (green) in the more mature inner side of the stem (detail in the inset). (B) OW side of the same section showing virtually no fluorescence in xylary fibres with the LM5 and CCRC-M38 antibodies. (C) TW side of a 4-week tipped stem showing strong labelling of G-fibres with the LM5 antibody in the outer developing and maturing part of the stem. (D) OW side of the same stem in (C). Fibre cell walls are not labelled or only weakly labelled with the LM5 and CCRC-M38 antibodies. (E) Detail of area 4 in (D). There is no labelling of OW fibre cell walls, indicating the absence of (1–4)-β-D-galactan. Some labelling is seen in ray parenchyma. Homogalacturonan is detected in cell corners (arrow). (F) and (G) Detail of area 1 in (C). LM5 antibody shows differential distribution of (1–4)-β-D-galactan from area 1 in files of G-fibres. The G-layer is labelled in some fibres, whereas in others the interface between the G-layer and SCW is labelled (arrowheads). (H) Detail of area 2 in (C). LM5 is bound mainly to the SCW and CCRC-M38 labels mainly the inner lamella of the G-layer (arrowhead). (I) and (J) Similar areas to 2 and 3 in (C) of unembedded sections showing CCRC-M38 mainly bound to the inner lamella of the G-layer. GL, G-layer; v, xylem vessel; rp, ray parenchyma. Bars: A, B, C, D = 100 µm; E, F, G, H, I, J = 10 µm.
Mentions: The overall distribution pattern of (1–4)-β-D-galactan revealed using the LM5 antibody was similar after 1 (Fig. 3A), 2, and 4 weeks of induction, but more distinct in older stems (Fig. 3C). Strongest binding of the LM5 antibody occurred in G-fibres of the developing and maturing xylem of TW in the outer half of the stem (Fig. 3C, red fluorescence). Close examination revealed labelling in the G-layers but binding was not homogeneous. Some G-fibre cell files were interspersed with rows of other fibre cells where LM5 labelling was confined to the SCW (Fig. 3F, G). There was no evidence that this was confined to any particular fibre area, such as tips. In the inner older mature G-fibres, (1–4)-β-D-galactan was detected only in the SCW and was generally absent from the G-layer (Fig. 3H). TEM observations were largely in agreement with immunofluorescence results, but more detail on the location of the β-galactan epitope was obtained. In general only one layer (S1) of SCW could be detected in G-fibres and two layers (S1 + S2) in normal fibres. In the early stages of G-layer deposition, some fibres exhibited abundant gold labelling in the G-layer itself, whereas in others the labelling was restricted to the interface between the SCW and the newly formed G-layer (Fig. 7A, B, C). Fig. 7B shows binding of the LM5 antibody in a thin layer on the inner side of the lumen side of the SCW, where initiation of G-layer deposition is possibly occurring. In more mature G-fibres, a similar pattern was noted in which the G-layers of some fibres were enriched with (1–4)-β-D-galactan (Fig. 7D, E); however, in the fully mature fibres the labelling tended to be more sparse in the G-layer or absent and generally restricted to the SCW/G-layer interface (Fig. 7F). In contrast to TW, the fibre cell walls in OW and NW in control stems were either not labelled or poorly labelled as shown by immunofluorescence (Fig. 3 B, D, E). TEM also showed almost negligible levels of labelling of fibre SCWs in NW and OW (Fig. 7G).

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