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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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a–e Cutinsomes identified with immunogold technique in O. umbellatum ovary epidermis lipotubuloids (white arrows), sections treated with H2O2. ER rough endoplasmic reticulum, lb lipid bodies, mt microtubules. Scale bar, 100 nm
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Fig4: a–e Cutinsomes identified with immunogold technique in O. umbellatum ovary epidermis lipotubuloids (white arrows), sections treated with H2O2. ER rough endoplasmic reticulum, lb lipid bodies, mt microtubules. Scale bar, 100 nm

Mentions: Figures 3, 4, 5, 6, 7, 8 and 9 present cutinsome-like structures of epidermal cell territories, including lipotubuloids marked with colloidal gold in response to the immunogold technique. Therefore, they may be considered as cutinsomes. In Figs. 3a, 4, 6 and 9, lipid bodies of lipotubuloids are visible as white structures because, due to immunogold technique requirements, its dark osmiophilic content was bleached with hydrogen peroxide (see “Materials and methods”). Thus, the outlines of cutinsomes treated with H2O2 are less pronounced than those of the non-treated ones (Fig. 2a–c). The cutinsomes 40 nm in diameter labelled with two gold particles are present near lipid bodies entwined with microtubules (Fig. 4a). Furthermore, cutinsomes of similar sizes can be seen near a rough ER vesicle (Fig. 4d). Bigger cutinsomes located among lipid bodies are labelled with more (three to four) gold particles, which are placed on their surface (Fig. 4b, c, e). Small cutinsomes may form near the microtubule band present between lipid bodies, as is seen in Fig. 5a, b. A cutinsome labelled with five gold particles, most probably leaving the lipotubuloid and moving towards the cell wall (Fig. 6a), and cutinsomes adhering to the plasmalemma not forming vesicles on the wall side (Fig. 6b) can be observed. These cutinsomes are probably transported through the plasmalemma by means of non-vesicular lipid transport (Lev 2010; Prinz 2010; Samuels and McFarlane 2012). Cutinsomes of different sizes are present in a polysaccharide cell wall and are labelled with gold particles on their periphery (Fig. 7a–c). Gold particles connected with the cutinsomes at the border of the cuticular layer and cuticle proper as well as in the cuticle proper can be observed in Fig. 8a, b.Fig. 4


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)

a–e Cutinsomes identified with immunogold technique in O. umbellatum ovary epidermis lipotubuloids (white arrows), sections treated with H2O2. ER rough endoplasmic reticulum, lb lipid bodies, mt microtubules. Scale bar, 100 nm
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4125816&req=5

Fig4: a–e Cutinsomes identified with immunogold technique in O. umbellatum ovary epidermis lipotubuloids (white arrows), sections treated with H2O2. ER rough endoplasmic reticulum, lb lipid bodies, mt microtubules. Scale bar, 100 nm
Mentions: Figures 3, 4, 5, 6, 7, 8 and 9 present cutinsome-like structures of epidermal cell territories, including lipotubuloids marked with colloidal gold in response to the immunogold technique. Therefore, they may be considered as cutinsomes. In Figs. 3a, 4, 6 and 9, lipid bodies of lipotubuloids are visible as white structures because, due to immunogold technique requirements, its dark osmiophilic content was bleached with hydrogen peroxide (see “Materials and methods”). Thus, the outlines of cutinsomes treated with H2O2 are less pronounced than those of the non-treated ones (Fig. 2a–c). The cutinsomes 40 nm in diameter labelled with two gold particles are present near lipid bodies entwined with microtubules (Fig. 4a). Furthermore, cutinsomes of similar sizes can be seen near a rough ER vesicle (Fig. 4d). Bigger cutinsomes located among lipid bodies are labelled with more (three to four) gold particles, which are placed on their surface (Fig. 4b, c, e). Small cutinsomes may form near the microtubule band present between lipid bodies, as is seen in Fig. 5a, b. A cutinsome labelled with five gold particles, most probably leaving the lipotubuloid and moving towards the cell wall (Fig. 6a), and cutinsomes adhering to the plasmalemma not forming vesicles on the wall side (Fig. 6b) can be observed. These cutinsomes are probably transported through the plasmalemma by means of non-vesicular lipid transport (Lev 2010; Prinz 2010; Samuels and McFarlane 2012). Cutinsomes of different sizes are present in a polysaccharide cell wall and are labelled with gold particles on their periphery (Fig. 7a–c). Gold particles connected with the cutinsomes at the border of the cuticular layer and cuticle proper as well as in the cuticle proper can be observed in Fig. 8a, b.Fig. 4

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