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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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Immunogold localization of LTP1 epitopes in the anticlinal cell walls of protoderm from embryogenic (a, b) and non-embryogenic (c, d) regions of explants cultured for 6 days. a, b The adaxial side of the proximal part of the cotyledon, showing the intense gold labelling and a gradient of labelling intensity growing towards cell surface, solid and open arrows in b indicate the apical and basal regions of the wall, respectively. c The adaxial side of the distal part of the cotyledon with few gold particles. d The abaxial side of the cotyledon, a few gold particles gathered in small clusters in the central part of the cell wall can be observed. e Negative control showing no labelling. f Quantification of immunogold label in walls of embryogenic (E) and non-embryogenic (NE) regions of the explants, data are means (±standard error and standard deviation) of ten micrographs per each region. Arrowheads (a–d) point to the cell surface. Scale bars 200 nm (a–e)
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Fig7: Immunogold localization of LTP1 epitopes in the anticlinal cell walls of protoderm from embryogenic (a, b) and non-embryogenic (c, d) regions of explants cultured for 6 days. a, b The adaxial side of the proximal part of the cotyledon, showing the intense gold labelling and a gradient of labelling intensity growing towards cell surface, solid and open arrows in b indicate the apical and basal regions of the wall, respectively. c The adaxial side of the distal part of the cotyledon with few gold particles. d The abaxial side of the cotyledon, a few gold particles gathered in small clusters in the central part of the cell wall can be observed. e Negative control showing no labelling. f Quantification of immunogold label in walls of embryogenic (E) and non-embryogenic (NE) regions of the explants, data are means (±standard error and standard deviation) of ten micrographs per each region. Arrowheads (a–d) point to the cell surface. Scale bars 200 nm (a–e)

Mentions: Localization of the LTP1 epitopes in the explant tissues was examined in more detail using immunogold labelling at the transmission electron microscope level. The TEM data demonstrated that the distribution of the LTP1 epitopes within the cell walls of embryogenic and non-embryogenic regions of the explants varied in terms of both spatial pattern and abundance. These differences were most clearly observed in both periclinal and anticlinal walls of protodermal cells (Figs. 6, 7, respectively).Fig. 6


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)

Immunogold localization of LTP1 epitopes in the anticlinal cell walls of protoderm from embryogenic (a, b) and non-embryogenic (c, d) regions of explants cultured for 6 days. a, b The adaxial side of the proximal part of the cotyledon, showing the intense gold labelling and a gradient of labelling intensity growing towards cell surface, solid and open arrows in b indicate the apical and basal regions of the wall, respectively. c The adaxial side of the distal part of the cotyledon with few gold particles. d The abaxial side of the cotyledon, a few gold particles gathered in small clusters in the central part of the cell wall can be observed. e Negative control showing no labelling. f Quantification of immunogold label in walls of embryogenic (E) and non-embryogenic (NE) regions of the explants, data are means (±standard error and standard deviation) of ten micrographs per each region. Arrowheads (a–d) point to the cell surface. Scale bars 200 nm (a–e)
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Fig7: Immunogold localization of LTP1 epitopes in the anticlinal cell walls of protoderm from embryogenic (a, b) and non-embryogenic (c, d) regions of explants cultured for 6 days. a, b The adaxial side of the proximal part of the cotyledon, showing the intense gold labelling and a gradient of labelling intensity growing towards cell surface, solid and open arrows in b indicate the apical and basal regions of the wall, respectively. c The adaxial side of the distal part of the cotyledon with few gold particles. d The abaxial side of the cotyledon, a few gold particles gathered in small clusters in the central part of the cell wall can be observed. e Negative control showing no labelling. f Quantification of immunogold label in walls of embryogenic (E) and non-embryogenic (NE) regions of the explants, data are means (±standard error and standard deviation) of ten micrographs per each region. Arrowheads (a–d) point to the cell surface. Scale bars 200 nm (a–e)
Mentions: Localization of the LTP1 epitopes in the explant tissues was examined in more detail using immunogold labelling at the transmission electron microscope level. The TEM data demonstrated that the distribution of the LTP1 epitopes within the cell walls of embryogenic and non-embryogenic regions of the explants varied in terms of both spatial pattern and abundance. These differences were most clearly observed in both periclinal and anticlinal walls of protodermal cells (Figs. 6, 7, respectively).Fig. 6

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