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Cutinsomes and lipotubuloids appear to participate in cuticle formation in Ornithogalum umbellatum ovary epidermis: EM-immunogold research.

Kwiatkowska M, Wojtczak A, Popłońska K, Polit JT, Stępiński D, Domίnguez E, Heredia A - Protoplasma (2014)

Bottom Line: They were mostly found in the outer cell wall, the cuticular layer and the cuticle proper.A lower but still significant degree of labelling was also observed in lipotubuloids, cytoplasm and near plasmalemma of epidermal cells.Thus, we suggest that (1) cutinsomes could take part in the synthesis of cuticle components also in plant species other than tomato, (2) the lipotubuloids are the cytoplasmic domains connected with cuticle formation and (3) this process proceeds via cutinsomes.

View Article: PubMed Central - PubMed

Affiliation: Department of Cytophysiology, Faculty of Biology and Environmental Protection, University of Łódź, Pomorska 141/143, 90-236, Łódź, Poland, kwiat@biol.uni.lodz.pl.

ABSTRACT
The outer wall of Ornithogalum umbellatum ovary and the fruit epidermis are covered with a thick cuticle and contain lipotubuloids incorporating (3)H-palmitic acid. This was earlier evidenced by selective autoradiographic labelling of lipotubuloids. After post-incubation in a non-radioactive medium, some marked particles insoluble in organic solvents (similar to cutin matrix) moved to the cuticular layer. Hence, it was hypothesised that lipotubuloids participated in cuticle synthesis. It was previously suggested that cutinsomes, nanoparticles containing polyhydroxy fatty acids, formed the cuticle. Thus, identification of the cutinsomes in O. umbellatum ovary epidermal cells, including lipotubuloids, was undertaken in order to verify the idea of lipotubuloid participation in cuticle synthesis in this species. Electron microscopy and immunogold method with the antibodies recognizing cutinsomes were used to identify these structures. They were mostly found in the outer cell wall, the cuticular layer and the cuticle proper. A lower but still significant degree of labelling was also observed in lipotubuloids, cytoplasm and near plasmalemma of epidermal cells. It seems that cutinsomes are formed in lipotubuloids and then they leave them and move towards the cuticle in epidermal cells of O. umbellatum ovary. Thus, we suggest that (1) cutinsomes could take part in the synthesis of cuticle components also in plant species other than tomato, (2) the lipotubuloids are the cytoplasmic domains connected with cuticle formation and (3) this process proceeds via cutinsomes.

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Cross-sections of O. umbellatum ovary epidermis and parenchyma cells after immunogold reaction with anti-cutinsome antibody. a Part of an epidermal cell with lipotubuloid, the cuticular layer and lipotubuloid labelled with gold particles. Fragments of images in windows are shown to the right in higher magnification. Gold particles are highlighted with circles. b Fragment of parenchyma cells with cell wall lacking gold particles. c cytoplasm, cl cuticular layer, cp cuticle proper, lb lipid bodies, v vacuole, w cell wall. Scale bars, 0.5 μm (a), 1 μm (b)
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Fig3: Cross-sections of O. umbellatum ovary epidermis and parenchyma cells after immunogold reaction with anti-cutinsome antibody. a Part of an epidermal cell with lipotubuloid, the cuticular layer and lipotubuloid labelled with gold particles. Fragments of images in windows are shown to the right in higher magnification. Gold particles are highlighted with circles. b Fragment of parenchyma cells with cell wall lacking gold particles. c cytoplasm, cl cuticular layer, cp cuticle proper, lb lipid bodies, v vacuole, w cell wall. Scale bars, 0.5 μm (a), 1 μm (b)

Mentions: It is known that all elements involved in cuticle synthesis are localized in those epidermal cells which are covered with cuticle. We compared the numbers of gold grains which labelled cutinsomes in ovary epidermis containing lipotubuloids with those in subepidermal parenchymal cells without lipotubuloids (Fig. 3 and Table 1). Abundant labelling was observed at the territory of an epidermal cell, especially at the area of its outer wall, and of lipotubuloids (Fig. 3a), whereas all parenchyma cells were scarcely labelled (background; Fig. 3b and Table 1). Of the whole labeling, 76 % was observed in the cell wall with cuticle, 15 % in lipotubuloids and the remaining 9 % located in the other components (Table 1). Both types of cells in which the antibody to cutinsomes was omitted were not labelled (negative control, not shown).Fig. 3


Cutinsomes and lipotubuloids appear to participate in cuticle formation in Ornithogalum umbellatum ovary epidermis: EM-immunogold research.

Kwiatkowska M, Wojtczak A, Popłońska K, Polit JT, Stępiński D, Domίnguez E, Heredia A - Protoplasma (2014)

Cross-sections of O. umbellatum ovary epidermis and parenchyma cells after immunogold reaction with anti-cutinsome antibody. a Part of an epidermal cell with lipotubuloid, the cuticular layer and lipotubuloid labelled with gold particles. Fragments of images in windows are shown to the right in higher magnification. Gold particles are highlighted with circles. b Fragment of parenchyma cells with cell wall lacking gold particles. c cytoplasm, cl cuticular layer, cp cuticle proper, lb lipid bodies, v vacuole, w cell wall. Scale bars, 0.5 μm (a), 1 μm (b)
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4125816&req=5

Fig3: Cross-sections of O. umbellatum ovary epidermis and parenchyma cells after immunogold reaction with anti-cutinsome antibody. a Part of an epidermal cell with lipotubuloid, the cuticular layer and lipotubuloid labelled with gold particles. Fragments of images in windows are shown to the right in higher magnification. Gold particles are highlighted with circles. b Fragment of parenchyma cells with cell wall lacking gold particles. c cytoplasm, cl cuticular layer, cp cuticle proper, lb lipid bodies, v vacuole, w cell wall. Scale bars, 0.5 μm (a), 1 μm (b)
Mentions: It is known that all elements involved in cuticle synthesis are localized in those epidermal cells which are covered with cuticle. We compared the numbers of gold grains which labelled cutinsomes in ovary epidermis containing lipotubuloids with those in subepidermal parenchymal cells without lipotubuloids (Fig. 3 and Table 1). Abundant labelling was observed at the territory of an epidermal cell, especially at the area of its outer wall, and of lipotubuloids (Fig. 3a), whereas all parenchyma cells were scarcely labelled (background; Fig. 3b and Table 1). Of the whole labeling, 76 % was observed in the cell wall with cuticle, 15 % in lipotubuloids and the remaining 9 % located in the other components (Table 1). Both types of cells in which the antibody to cutinsomes was omitted were not labelled (negative control, not shown).Fig. 3

Bottom Line: They were mostly found in the outer cell wall, the cuticular layer and the cuticle proper.A lower but still significant degree of labelling was also observed in lipotubuloids, cytoplasm and near plasmalemma of epidermal cells.Thus, we suggest that (1) cutinsomes could take part in the synthesis of cuticle components also in plant species other than tomato, (2) the lipotubuloids are the cytoplasmic domains connected with cuticle formation and (3) this process proceeds via cutinsomes.

View Article: PubMed Central - PubMed

Affiliation: Department of Cytophysiology, Faculty of Biology and Environmental Protection, University of Łódź, Pomorska 141/143, 90-236, Łódź, Poland, kwiat@biol.uni.lodz.pl.

ABSTRACT
The outer wall of Ornithogalum umbellatum ovary and the fruit epidermis are covered with a thick cuticle and contain lipotubuloids incorporating (3)H-palmitic acid. This was earlier evidenced by selective autoradiographic labelling of lipotubuloids. After post-incubation in a non-radioactive medium, some marked particles insoluble in organic solvents (similar to cutin matrix) moved to the cuticular layer. Hence, it was hypothesised that lipotubuloids participated in cuticle synthesis. It was previously suggested that cutinsomes, nanoparticles containing polyhydroxy fatty acids, formed the cuticle. Thus, identification of the cutinsomes in O. umbellatum ovary epidermal cells, including lipotubuloids, was undertaken in order to verify the idea of lipotubuloid participation in cuticle synthesis in this species. Electron microscopy and immunogold method with the antibodies recognizing cutinsomes were used to identify these structures. They were mostly found in the outer cell wall, the cuticular layer and the cuticle proper. A lower but still significant degree of labelling was also observed in lipotubuloids, cytoplasm and near plasmalemma of epidermal cells. It seems that cutinsomes are formed in lipotubuloids and then they leave them and move towards the cuticle in epidermal cells of O. umbellatum ovary. Thus, we suggest that (1) cutinsomes could take part in the synthesis of cuticle components also in plant species other than tomato, (2) the lipotubuloids are the cytoplasmic domains connected with cuticle formation and (3) this process proceeds via cutinsomes.

Show MeSH