Isolation of plant nuclei suitable for flow cytometry from species with extremely mucilaginous compounds: an example in the genus Viola L. (Violaceae)
Flow cytometry analysis has been widely applied in the determination of nuclear DNA content and ploidy level in many organisms. Despite being the most appropriate method for DNA content measurement, flow cytometry also presents some limitations. A fairly common, but little-studied problem is the eff...
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| Format: | Article |
| Language: | English |
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Consejo Superior de Investigaciones Científicas
2011-12-01
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| Series: | Anales del Jardín Botánico de Madrid |
| Subjects: | |
| Online Access: | http://rjb.revistas.csic.es/index.php/rjb/article/view/362 |
| _version_ | 1819048860598665216 |
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| author | Eduardo Cires Candela Cuesta María Ángeles Fernández Casado Herminio S. Nava Víctor M. Vázquez José Antonio Fernández Prieto |
| author_facet | Eduardo Cires Candela Cuesta María Ángeles Fernández Casado Herminio S. Nava Víctor M. Vázquez José Antonio Fernández Prieto |
| author_sort | Eduardo Cires |
| collection | DOAJ |
| description | Flow cytometry analysis has been widely applied in the determination of nuclear DNA content and ploidy level in many organisms. Despite being the most appropriate method for DNA content measurement, flow cytometry also presents some limitations. A fairly common, but little-studied problem is the effect on measurements of the presence of secondary metabolites. A good example is the genus Viola, which is composed of 525-600 species distributed worldwide. These species have proved to be problematic for flow cytometric analyses due to the release of extremely mucilaginous compounds into the nuclear suspension. In this work, the genome size of 13 species of Viola using flow cytometry are presented for the first time. Despite obtaining histograms with high coefficients of variation, we here present an optimized protocol to remove cytoplasmic compounds, particularly mucilaginous ones, from plant nuclei that pave the way for its application to estimate the genome size of other species exhibiting similar problems. Statistical analyses revealed significant differences between sections Viola and Melanium, and within each section (P < 0.001). Furthermore, statistically significant differences were not detected among samples of the same species. |
| first_indexed | 2024-12-21T11:22:58Z |
| format | Article |
| id | doaj.art-ae4d6845df8f4098ab4c2de746638b7a |
| institution | Directory Open Access Journal |
| issn | 0211-1322 1988-3196 |
| language | English |
| last_indexed | 2024-12-21T11:22:58Z |
| publishDate | 2011-12-01 |
| publisher | Consejo Superior de Investigaciones Científicas |
| record_format | Article |
| series | Anales del Jardín Botánico de Madrid |
| spelling | doaj.art-ae4d6845df8f4098ab4c2de746638b7a2022-12-21T19:05:44ZengConsejo Superior de Investigaciones CientíficasAnales del Jardín Botánico de Madrid0211-13221988-31962011-12-0168213915410.3989/ajbm.2273341Isolation of plant nuclei suitable for flow cytometry from species with extremely mucilaginous compounds: an example in the genus Viola L. (Violaceae)Eduardo Cires0Candela Cuesta1María Ángeles Fernández Casado2Herminio S. Nava3Víctor M. Vázquez4José Antonio Fernández Prieto5Área de Botánica, Universidad de Oviedo, Departamento de Biología de Organismos y SistemasDepartment of Plant Systems Biology - Área de Fisiología Vegetal, Universidad de Oviedo, Departamento de Biología de Organismos y SistemasÁrea de Botánica, Universidad de Oviedo, Departamento de Biología de Organismos y SistemasÁrea de Botánica, Universidad de Oviedo, Departamento de Biología de Organismos y SistemasReal Instituto de Estudios AsturianosÁrea de Botánica, Universidad de Oviedo, Departamento de Biología de Organismos y SistemasFlow cytometry analysis has been widely applied in the determination of nuclear DNA content and ploidy level in many organisms. Despite being the most appropriate method for DNA content measurement, flow cytometry also presents some limitations. A fairly common, but little-studied problem is the effect on measurements of the presence of secondary metabolites. A good example is the genus Viola, which is composed of 525-600 species distributed worldwide. These species have proved to be problematic for flow cytometric analyses due to the release of extremely mucilaginous compounds into the nuclear suspension. In this work, the genome size of 13 species of Viola using flow cytometry are presented for the first time. Despite obtaining histograms with high coefficients of variation, we here present an optimized protocol to remove cytoplasmic compounds, particularly mucilaginous ones, from plant nuclei that pave the way for its application to estimate the genome size of other species exhibiting similar problems. Statistical analyses revealed significant differences between sections Viola and Melanium, and within each section (P < 0.001). Furthermore, statistically significant differences were not detected among samples of the same species.http://rjb.revistas.csic.es/index.php/rjb/article/view/362chromosome numberflow cytometrygenome sizemucilaginous compoundsnuclear dna contentploidy levelssection melaniumsection viola |
| spellingShingle | Eduardo Cires Candela Cuesta María Ángeles Fernández Casado Herminio S. Nava Víctor M. Vázquez José Antonio Fernández Prieto Isolation of plant nuclei suitable for flow cytometry from species with extremely mucilaginous compounds: an example in the genus Viola L. (Violaceae) Anales del Jardín Botánico de Madrid chromosome number flow cytometry genome size mucilaginous compounds nuclear dna content ploidy levels section melanium section viola |
| title | Isolation of plant nuclei suitable for flow cytometry from species with extremely mucilaginous compounds: an example in the genus Viola L. (Violaceae) |
| title_full | Isolation of plant nuclei suitable for flow cytometry from species with extremely mucilaginous compounds: an example in the genus Viola L. (Violaceae) |
| title_fullStr | Isolation of plant nuclei suitable for flow cytometry from species with extremely mucilaginous compounds: an example in the genus Viola L. (Violaceae) |
| title_full_unstemmed | Isolation of plant nuclei suitable for flow cytometry from species with extremely mucilaginous compounds: an example in the genus Viola L. (Violaceae) |
| title_short | Isolation of plant nuclei suitable for flow cytometry from species with extremely mucilaginous compounds: an example in the genus Viola L. (Violaceae) |
| title_sort | isolation of plant nuclei suitable for flow cytometry from species with extremely mucilaginous compounds an example in the genus viola l violaceae |
| topic | chromosome number flow cytometry genome size mucilaginous compounds nuclear dna content ploidy levels section melanium section viola |
| url | http://rjb.revistas.csic.es/index.php/rjb/article/view/362 |
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