Limits...
Distribution of lipid transfer protein 1 (LTP1) epitopes associated with morphogenic events during somatic embryogenesis of Arabidopsis thaliana.

Potocka I, Baldwin TC, Kurczynska EU - Plant Cell Rep. (2012)

Bottom Line: The labelling was strongest in both the outer periclinal and anticlinal walls of the adaxial, protodermal cells of the proximal region of the cotyledon.The putative role(s) of lipid transfer proteins in the formation of lipid lamellae and in cell differentiation are discussed.Key message Occurrence of lipid transfer protein 1 epitopes in Arabidopsis explant cells accompanies changes in cell fate and may be correlated with the deposition of lipid substances in the cell walls.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Cell Biology, University of Silesia, Jagiellońska 28, 40-032, Katowice, Poland. izabela.potocka@us.edu.pl

ABSTRACT
Using immunocytochemical methods, at both the light and electron microscopic level, we have investigated the spatial and temporal distribution of lipid transfer protein 1 (LTP1) epitopes during the induction of somatic embryogenesis in explants of Arabidopsis thaliana. Immunofluorescence labelling demonstrated the presence of high levels of LTP1 epitopes within the proximal regions of the cotyledons (embryogenic regions) associated with particular morphogenetic events, including intense cell division activity, cotyledon swelling, cell loosening and callus formation. Precise analysis of the signal localization in protodermal and subprotodermal cells indicated that cells exhibiting features typical of embryogenic cells were strongly labelled, both in walls and the cytoplasm, while in the majority of meristematic-like cells no signal was observed. Staining with lipophilic dyes revealed a correlation between the distribution of LTP1 epitopes and lipid substances within the cell wall. Differences in label abundance and distribution between embryogenic and non-embryogenic regions of explants were studied in detail with the use of immunogold electron microscopy. The labelling was strongest in both the outer periclinal and anticlinal walls of the adaxial, protodermal cells of the proximal region of the cotyledon. The putative role(s) of lipid transfer proteins in the formation of lipid lamellae and in cell differentiation are discussed. Key message Occurrence of lipid transfer protein 1 epitopes in Arabidopsis explant cells accompanies changes in cell fate and may be correlated with the deposition of lipid substances in the cell walls.

Show MeSH

Related in: MedlinePlus

Co-localization of the lipid lamellae and the LTP1 epitopes in cells of 21-day cultured explants. Blue colouring (a, c, e) and yellow-gold fluorescence (g) of cell walls correspond to lipids stained with Sudan Black B and Nile Red, respectively, red fluorescence (b, d, f) corresponds to anti-AtLTP1 labelling. Faint green signal in g represents the autofluorescence of the tissue. a, b Detached callus cell (arrow), arrowhead points to the outer periclinal wall of the explant protoderm. c–g Surface cells of the cotyledons (arrows), note the loosened character of the tissue in c–d. h Negative control, no Cy3 fluorescence visible. Scale bars 10 μm (a–h)
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC3472069&req=5

Fig5: Co-localization of the lipid lamellae and the LTP1 epitopes in cells of 21-day cultured explants. Blue colouring (a, c, e) and yellow-gold fluorescence (g) of cell walls correspond to lipids stained with Sudan Black B and Nile Red, respectively, red fluorescence (b, d, f) corresponds to anti-AtLTP1 labelling. Faint green signal in g represents the autofluorescence of the tissue. a, b Detached callus cell (arrow), arrowhead points to the outer periclinal wall of the explant protoderm. c–g Surface cells of the cotyledons (arrows), note the loosened character of the tissue in c–d. h Negative control, no Cy3 fluorescence visible. Scale bars 10 μm (a–h)

Mentions: Figure 5 shows sections through 3-week-old explants stained with Sudan Black B (Fig. 5a, c, e) which can be directly compared to consecutive sections labelled with the anti-AtLTP1 antibody (Fig. 5b, d, f). Blue colouring demonstrating the presence of lipids was observed in callus cell walls (Fig. 5a) and in walls of single cells located at the explant periphery (Fig. 5c, e). Lipid lamellae were also present at the surface of the protodermal cells (Fig. 5a) indicating that the lipid substances detected within the explant cells were chemically similar to the cuticle. After staining with Nile Red, the Sudan-positive cells exhibited yellow-gold fluorescence in their cell walls (compare Fig. 5e with g). The lipid staining was performed in conjunction with the immunocytochemistry, which revealed the presence of LTP1 epitopes in walls of the stained cells (Fig. 5b, d, f). In both callus and surface cells, signal generated by the anti-AtLTP1 antibody was detected in an outline formed by thick continuous line (Fig. 5b) or separate dots (Fig. 5d, f). Sections with the primary antibody omitted (negative control) displayed red background fluorescence easily distinguishable from the Cy3 signal (Fig. 5h).Fig. 5


Distribution of lipid transfer protein 1 (LTP1) epitopes associated with morphogenic events during somatic embryogenesis of Arabidopsis thaliana.

Potocka I, Baldwin TC, Kurczynska EU - Plant Cell Rep. (2012)

Co-localization of the lipid lamellae and the LTP1 epitopes in cells of 21-day cultured explants. Blue colouring (a, c, e) and yellow-gold fluorescence (g) of cell walls correspond to lipids stained with Sudan Black B and Nile Red, respectively, red fluorescence (b, d, f) corresponds to anti-AtLTP1 labelling. Faint green signal in g represents the autofluorescence of the tissue. a, b Detached callus cell (arrow), arrowhead points to the outer periclinal wall of the explant protoderm. c–g Surface cells of the cotyledons (arrows), note the loosened character of the tissue in c–d. h Negative control, no Cy3 fluorescence visible. Scale bars 10 μm (a–h)
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3472069&req=5

Fig5: Co-localization of the lipid lamellae and the LTP1 epitopes in cells of 21-day cultured explants. Blue colouring (a, c, e) and yellow-gold fluorescence (g) of cell walls correspond to lipids stained with Sudan Black B and Nile Red, respectively, red fluorescence (b, d, f) corresponds to anti-AtLTP1 labelling. Faint green signal in g represents the autofluorescence of the tissue. a, b Detached callus cell (arrow), arrowhead points to the outer periclinal wall of the explant protoderm. c–g Surface cells of the cotyledons (arrows), note the loosened character of the tissue in c–d. h Negative control, no Cy3 fluorescence visible. Scale bars 10 μm (a–h)
Mentions: Figure 5 shows sections through 3-week-old explants stained with Sudan Black B (Fig. 5a, c, e) which can be directly compared to consecutive sections labelled with the anti-AtLTP1 antibody (Fig. 5b, d, f). Blue colouring demonstrating the presence of lipids was observed in callus cell walls (Fig. 5a) and in walls of single cells located at the explant periphery (Fig. 5c, e). Lipid lamellae were also present at the surface of the protodermal cells (Fig. 5a) indicating that the lipid substances detected within the explant cells were chemically similar to the cuticle. After staining with Nile Red, the Sudan-positive cells exhibited yellow-gold fluorescence in their cell walls (compare Fig. 5e with g). The lipid staining was performed in conjunction with the immunocytochemistry, which revealed the presence of LTP1 epitopes in walls of the stained cells (Fig. 5b, d, f). In both callus and surface cells, signal generated by the anti-AtLTP1 antibody was detected in an outline formed by thick continuous line (Fig. 5b) or separate dots (Fig. 5d, f). Sections with the primary antibody omitted (negative control) displayed red background fluorescence easily distinguishable from the Cy3 signal (Fig. 5h).Fig. 5

Bottom Line: The labelling was strongest in both the outer periclinal and anticlinal walls of the adaxial, protodermal cells of the proximal region of the cotyledon.The putative role(s) of lipid transfer proteins in the formation of lipid lamellae and in cell differentiation are discussed.Key message Occurrence of lipid transfer protein 1 epitopes in Arabidopsis explant cells accompanies changes in cell fate and may be correlated with the deposition of lipid substances in the cell walls.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Cell Biology, University of Silesia, Jagiellońska 28, 40-032, Katowice, Poland. izabela.potocka@us.edu.pl

ABSTRACT
Using immunocytochemical methods, at both the light and electron microscopic level, we have investigated the spatial and temporal distribution of lipid transfer protein 1 (LTP1) epitopes during the induction of somatic embryogenesis in explants of Arabidopsis thaliana. Immunofluorescence labelling demonstrated the presence of high levels of LTP1 epitopes within the proximal regions of the cotyledons (embryogenic regions) associated with particular morphogenetic events, including intense cell division activity, cotyledon swelling, cell loosening and callus formation. Precise analysis of the signal localization in protodermal and subprotodermal cells indicated that cells exhibiting features typical of embryogenic cells were strongly labelled, both in walls and the cytoplasm, while in the majority of meristematic-like cells no signal was observed. Staining with lipophilic dyes revealed a correlation between the distribution of LTP1 epitopes and lipid substances within the cell wall. Differences in label abundance and distribution between embryogenic and non-embryogenic regions of explants were studied in detail with the use of immunogold electron microscopy. The labelling was strongest in both the outer periclinal and anticlinal walls of the adaxial, protodermal cells of the proximal region of the cotyledon. The putative role(s) of lipid transfer proteins in the formation of lipid lamellae and in cell differentiation are discussed. Key message Occurrence of lipid transfer protein 1 epitopes in Arabidopsis explant cells accompanies changes in cell fate and may be correlated with the deposition of lipid substances in the cell walls.

Show MeSH
Related in: MedlinePlus