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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.

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The signal localization in embryogenic and meristematic cells of the explants. a Histogram illustrating frequencies of the signal presence in specific localizations of various types of cells. b Protodermal cell of the cotyledon exhibiting features typical of embryogenic cells (EC), signal in the anticlinal and inner periclinal walls (arrowheads) and in the cytoplasm. c Protodermal cell of the cotyledon exhibiting features typical of meristematic cells (MC), scattered labelling in the cytoplasm (arrowheads). d Meristematic cells of the shoot apical meristem (M), no labelling. Upper micrographs in b, c, d show immunofluorescence images and lower micrographs show corresponding bright field and phase contrast images. CW cell wall labelling, CWC cell wall and cytoplasm labelling, C cytoplasm labelling, NL no labelling. The number of cells per group was as follows: EC 23, MC 24, M 22. Scale bars 5 μm (b–d)
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Fig4: The signal localization in embryogenic and meristematic cells of the explants. a Histogram illustrating frequencies of the signal presence in specific localizations of various types of cells. b Protodermal cell of the cotyledon exhibiting features typical of embryogenic cells (EC), signal in the anticlinal and inner periclinal walls (arrowheads) and in the cytoplasm. c Protodermal cell of the cotyledon exhibiting features typical of meristematic cells (MC), scattered labelling in the cytoplasm (arrowheads). d Meristematic cells of the shoot apical meristem (M), no labelling. Upper micrographs in b, c, d show immunofluorescence images and lower micrographs show corresponding bright field and phase contrast images. CW cell wall labelling, CWC cell wall and cytoplasm labelling, C cytoplasm labelling, NL no labelling. The number of cells per group was as follows: EC 23, MC 24, M 22. Scale bars 5 μm (b–d)

Mentions: Figure 4 shows the results of signal localization analysis in both embryogenic and meristematic cells of explants. Cells containing one large nucleolus, which is a feature typical of embryogenic cells (Verdeil et al. 2007), exhibited strong fluorescence in both cell walls and the cytoplasm (Fig. 4a, b). In the majority of meristematic cells with nuclei containing more than one nucleolus (Verdeil et al. 2007), found both in embryogenic regions of cotyledons and in the shoot apical meristem, no signal was involved (Fig. 4a, d). However, there were several cases in which the signal was observed only in cytoplasm (Fig. 4a, c). In a minority of meristematic cells, the signal occurred both in the cell wall and the cytoplasm (Fig. 4a). A Chi-square test of independence indicated a highly significant dependence (p = 0.000001) between cell type and localization of the signal within the cell.Fig. 4


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)

The signal localization in embryogenic and meristematic cells of the explants. a Histogram illustrating frequencies of the signal presence in specific localizations of various types of cells. b Protodermal cell of the cotyledon exhibiting features typical of embryogenic cells (EC), signal in the anticlinal and inner periclinal walls (arrowheads) and in the cytoplasm. c Protodermal cell of the cotyledon exhibiting features typical of meristematic cells (MC), scattered labelling in the cytoplasm (arrowheads). d Meristematic cells of the shoot apical meristem (M), no labelling. Upper micrographs in b, c, d show immunofluorescence images and lower micrographs show corresponding bright field and phase contrast images. CW cell wall labelling, CWC cell wall and cytoplasm labelling, C cytoplasm labelling, NL no labelling. The number of cells per group was as follows: EC 23, MC 24, M 22. Scale bars 5 μm (b–d)
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Fig4: The signal localization in embryogenic and meristematic cells of the explants. a Histogram illustrating frequencies of the signal presence in specific localizations of various types of cells. b Protodermal cell of the cotyledon exhibiting features typical of embryogenic cells (EC), signal in the anticlinal and inner periclinal walls (arrowheads) and in the cytoplasm. c Protodermal cell of the cotyledon exhibiting features typical of meristematic cells (MC), scattered labelling in the cytoplasm (arrowheads). d Meristematic cells of the shoot apical meristem (M), no labelling. Upper micrographs in b, c, d show immunofluorescence images and lower micrographs show corresponding bright field and phase contrast images. CW cell wall labelling, CWC cell wall and cytoplasm labelling, C cytoplasm labelling, NL no labelling. The number of cells per group was as follows: EC 23, MC 24, M 22. Scale bars 5 μm (b–d)
Mentions: Figure 4 shows the results of signal localization analysis in both embryogenic and meristematic cells of explants. Cells containing one large nucleolus, which is a feature typical of embryogenic cells (Verdeil et al. 2007), exhibited strong fluorescence in both cell walls and the cytoplasm (Fig. 4a, b). In the majority of meristematic cells with nuclei containing more than one nucleolus (Verdeil et al. 2007), found both in embryogenic regions of cotyledons and in the shoot apical meristem, no signal was involved (Fig. 4a, d). However, there were several cases in which the signal was observed only in cytoplasm (Fig. 4a, c). In a minority of meristematic cells, the signal occurred both in the cell wall and the cytoplasm (Fig. 4a). A Chi-square test of independence indicated a highly significant dependence (p = 0.000001) between cell type and localization of the signal within the cell.Fig. 4

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