Immunoanalysis of dehydrins in Araucaria angustifolia embryos
The aim of this study was to describe the dehydrin content of mature Araucaria angustifolia embryos, a species of endangered and economically important conifers, native to southern Brazil, northeastern Argentina, and eastern Paraguay. The A. angustifolia seeds have been categorized as recalcitrant....
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2013
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| Acceso en línea: | https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_0033183X_v250_n4_p911_FariasSoares http://hdl.handle.net/20.500.12110/paper_0033183X_v250_n4_p911_FariasSoares |
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paper:paper_0033183X_v250_n4_p911_FariasSoares |
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paper:paper_0033183X_v250_n4_p911_FariasSoares2023-06-08T15:00:25Z Immunoanalysis of dehydrins in Araucaria angustifolia embryos Araucaria angustifolia seed Dehydrin in situ immunolocalization Dehydrin subcellular localization Dehydrin western blot analysis Phosphorylated dehydrins Thermosensitive dehydrins Araucaria angustifolia Coniferophyta dehydrin proteins, plant vegetable protein article chemistry conifer metabolism plant seed Western blotting Blotting, Western Coniferophyta Plant Proteins Seeds The aim of this study was to describe the dehydrin content of mature Araucaria angustifolia embryos, a species of endangered and economically important conifers, native to southern Brazil, northeastern Argentina, and eastern Paraguay. The A. angustifolia seeds have been categorized as recalcitrant. Dehydrins were studied by western blot analysis and in situ immunolocalization microscopy using antibodies raised against the K segment, a highly conserved lysine-rich 15-amino acid sequence extensively used to recognize proteins immunologically related to the dehydrin family. Western blot analysis of the heat-stable protein fraction, as estimated by 15 % SDS-PAGE, revealed three main bands of approximately 20-, 26-, and 29-kDa; when 17.5 % SDS-PAGE was used, each band resolved into two other bands. Two thermosensitive dehydrin bands of around 16 and 35 kDa were common to the axis and cotyledons, and another thermosensitive band, with molecular mass of approximately 10 kDa, was present in the cotyledons only. Following alkaline phosphatase (AP) treatment, a gel mobility shift was detected for each one of the four main bands that can be due to phosphorylation. Dehydrins were detected in all axis and cotyledon tissues using in situ immunolocalization microscopy. At the subcellular level, dehydrins were immunolocalized in the nuclei, protein bodies, and microbodies. In the nucleus, dehydrins were found to be associated with chromatin. We concluded that the gel mobility shift for the four main bands (probably due to phosphorylation), the presence of thermosensitive bands, and the specific localizations in nuclei and protein bodies provide key starting points to understand the function of dehydrins in the embryo cells of this species. © 2012 Springer-Verlag Wien. 2013 https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_0033183X_v250_n4_p911_FariasSoares http://hdl.handle.net/20.500.12110/paper_0033183X_v250_n4_p911_FariasSoares |
| institution |
Universidad de Buenos Aires |
| institution_str |
I-28 |
| repository_str |
R-134 |
| collection |
Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) |
| topic |
Araucaria angustifolia seed Dehydrin in situ immunolocalization Dehydrin subcellular localization Dehydrin western blot analysis Phosphorylated dehydrins Thermosensitive dehydrins Araucaria angustifolia Coniferophyta dehydrin proteins, plant vegetable protein article chemistry conifer metabolism plant seed Western blotting Blotting, Western Coniferophyta Plant Proteins Seeds |
| spellingShingle |
Araucaria angustifolia seed Dehydrin in situ immunolocalization Dehydrin subcellular localization Dehydrin western blot analysis Phosphorylated dehydrins Thermosensitive dehydrins Araucaria angustifolia Coniferophyta dehydrin proteins, plant vegetable protein article chemistry conifer metabolism plant seed Western blotting Blotting, Western Coniferophyta Plant Proteins Seeds Immunoanalysis of dehydrins in Araucaria angustifolia embryos |
| topic_facet |
Araucaria angustifolia seed Dehydrin in situ immunolocalization Dehydrin subcellular localization Dehydrin western blot analysis Phosphorylated dehydrins Thermosensitive dehydrins Araucaria angustifolia Coniferophyta dehydrin proteins, plant vegetable protein article chemistry conifer metabolism plant seed Western blotting Blotting, Western Coniferophyta Plant Proteins Seeds |
| description |
The aim of this study was to describe the dehydrin content of mature Araucaria angustifolia embryos, a species of endangered and economically important conifers, native to southern Brazil, northeastern Argentina, and eastern Paraguay. The A. angustifolia seeds have been categorized as recalcitrant. Dehydrins were studied by western blot analysis and in situ immunolocalization microscopy using antibodies raised against the K segment, a highly conserved lysine-rich 15-amino acid sequence extensively used to recognize proteins immunologically related to the dehydrin family. Western blot analysis of the heat-stable protein fraction, as estimated by 15 % SDS-PAGE, revealed three main bands of approximately 20-, 26-, and 29-kDa; when 17.5 % SDS-PAGE was used, each band resolved into two other bands. Two thermosensitive dehydrin bands of around 16 and 35 kDa were common to the axis and cotyledons, and another thermosensitive band, with molecular mass of approximately 10 kDa, was present in the cotyledons only. Following alkaline phosphatase (AP) treatment, a gel mobility shift was detected for each one of the four main bands that can be due to phosphorylation. Dehydrins were detected in all axis and cotyledon tissues using in situ immunolocalization microscopy. At the subcellular level, dehydrins were immunolocalized in the nuclei, protein bodies, and microbodies. In the nucleus, dehydrins were found to be associated with chromatin. We concluded that the gel mobility shift for the four main bands (probably due to phosphorylation), the presence of thermosensitive bands, and the specific localizations in nuclei and protein bodies provide key starting points to understand the function of dehydrins in the embryo cells of this species. © 2012 Springer-Verlag Wien. |
| title |
Immunoanalysis of dehydrins in Araucaria angustifolia embryos |
| title_short |
Immunoanalysis of dehydrins in Araucaria angustifolia embryos |
| title_full |
Immunoanalysis of dehydrins in Araucaria angustifolia embryos |
| title_fullStr |
Immunoanalysis of dehydrins in Araucaria angustifolia embryos |
| title_full_unstemmed |
Immunoanalysis of dehydrins in Araucaria angustifolia embryos |
| title_sort |
immunoanalysis of dehydrins in araucaria angustifolia embryos |
| publishDate |
2013 |
| url |
https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_0033183X_v250_n4_p911_FariasSoares http://hdl.handle.net/20.500.12110/paper_0033183X_v250_n4_p911_FariasSoares |
| _version_ |
1768545250691776512 |