Determination of Chlorophyll Concentration in In Vitro Peanut Leaves
The detection of chlorophyll is a convenient method for assessing gene editing efficiency when generating mutations in the phytoene desaturase (PDS) gene, which is related to chlorophyll biosynthesis. Various instruments and protocols are available for measuring chlorophyll; however, a minimum leaf...
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| Format: | info:ar-repo/semantics/artículo |
| Language: | Inglés |
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Wiley
2025
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| Online Access: | http://hdl.handle.net/20.500.12123/24824 https://currentprotocols.onlinelibrary.wiley.com/doi/10.1002/cpz1.70150 https://doi.org/10.1002/cpz1.70150 |
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| author | Faustinelli, Paola C. Massa, Alicia N. Soria, Néstor W. Lopez Colomba, Eliana Suarez, Paola Alejandra Lamb, Marshall C. |
| author_browse | Faustinelli, Paola C. Lamb, Marshall C. Lopez Colomba, Eliana Massa, Alicia N. Soria, Néstor W. Suarez, Paola Alejandra |
| author_facet | Faustinelli, Paola C. Massa, Alicia N. Soria, Néstor W. Lopez Colomba, Eliana Suarez, Paola Alejandra Lamb, Marshall C. |
| author_sort | Faustinelli, Paola C. |
| collection | INTA Digital |
| description | The detection of chlorophyll is a convenient method for assessing gene editing efficiency when generating mutations in the phytoene desaturase (PDS) gene, which is related to chlorophyll biosynthesis. Various instruments and protocols are available for measuring chlorophyll; however, a minimum leaf area is required for precise quantifications. In vitro plant regeneration often requires several months to produce enough leaf tissue for chlorophyll content analysis. In this study, we optimized an existing chlorophyll quantification method by using a minimal amount of in vitro leaf tissue (<2.0 mg) and a microvolume of chlorophyll extraction solution (3 µl) to quantify chlorophyll a, chlorophyll b, and total chlorophyll concentration using a UV-Vis spectrophotometer. This method will serve as a basic protocol for evaluating gene editing efficiency by correlating degrees of albinism (pigment content) with changes in the PDS gene sequence. |
| format | info:ar-repo/semantics/artículo |
| id | INTA24824 |
| institution | Instituto Nacional de Tecnología Agropecuaria (INTA -Argentina) |
| language | Inglés |
| publishDate | 2025 |
| publishDateRange | 2025 |
| publishDateSort | 2025 |
| publisher | Wiley |
| publisherStr | Wiley |
| record_format | dspace |
| spelling | INTA248242025-12-30T15:07:09Z Determination of Chlorophyll Concentration in In Vitro Peanut Leaves Faustinelli, Paola C. Massa, Alicia N. Soria, Néstor W. Lopez Colomba, Eliana Suarez, Paola Alejandra Lamb, Marshall C. Arachis hypogaea Laboratory Experimentation Chlorophylls Leaves In Vitro Experimentation Experimentación en Laboratorio Clorofila Hojas Experimentación In Vitro Maní Peanuts The detection of chlorophyll is a convenient method for assessing gene editing efficiency when generating mutations in the phytoene desaturase (PDS) gene, which is related to chlorophyll biosynthesis. Various instruments and protocols are available for measuring chlorophyll; however, a minimum leaf area is required for precise quantifications. In vitro plant regeneration often requires several months to produce enough leaf tissue for chlorophyll content analysis. In this study, we optimized an existing chlorophyll quantification method by using a minimal amount of in vitro leaf tissue (<2.0 mg) and a microvolume of chlorophyll extraction solution (3 µl) to quantify chlorophyll a, chlorophyll b, and total chlorophyll concentration using a UV-Vis spectrophotometer. This method will serve as a basic protocol for evaluating gene editing efficiency by correlating degrees of albinism (pigment content) with changes in the PDS gene sequence. Instituto de Fisiología y Recursos Genéticos Vegetales Fil: Faustinelli, Paola C. USDA-ARS. National Peanut Research Laboratory; Estados Unidos Fil: Massa, Alicia N. USDA-ARS. National Peanut Research Laboratory; Estados Unidos Fil: Soria, Néstor W. Universidad Católica de Córdoba; Argentina Fil: Lopez Colomba, Eliana. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Fisiología y Recursos Genéticos Vegetales; Argentina Fil: Lopez Colomba, Eliana. Consejo Nacional de Investigaciones Científicas y Técnicas. Unidad de Estudios Agropecuarios (UDEA); Argentina Fil: Suarez, Paola Alejandra. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Fisiología y Recursos Genéticos Vegetales; Argentina Fil: Suarez, Paola Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas. Unidad de Estudios Agropecuarios (UDEA); Argentina Fil: Lamb, Marshall C. USDA-ARS. National Peanut Research Laboratory; Estados Unidos 2025-12-30T15:04:35Z 2025-12-30T15:04:35Z 2025-05 info:ar-repo/semantics/artículo info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion http://hdl.handle.net/20.500.12123/24824 https://currentprotocols.onlinelibrary.wiley.com/doi/10.1002/cpz1.70150 2691-1299 https://doi.org/10.1002/cpz1.70150 eng info:eu-repo/semantics/restrictedAccess http://creativecommons.org/licenses/by-nc-sa/4.0/ Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0) application/pdf Wiley Current Protocols : 1-9 (May 2025) |
| spellingShingle | Arachis hypogaea Laboratory Experimentation Chlorophylls Leaves In Vitro Experimentation Experimentación en Laboratorio Clorofila Hojas Experimentación In Vitro Maní Peanuts Faustinelli, Paola C. Massa, Alicia N. Soria, Néstor W. Lopez Colomba, Eliana Suarez, Paola Alejandra Lamb, Marshall C. Determination of Chlorophyll Concentration in In Vitro Peanut Leaves |
| title | Determination of Chlorophyll Concentration in In Vitro Peanut Leaves |
| title_full | Determination of Chlorophyll Concentration in In Vitro Peanut Leaves |
| title_fullStr | Determination of Chlorophyll Concentration in In Vitro Peanut Leaves |
| title_full_unstemmed | Determination of Chlorophyll Concentration in In Vitro Peanut Leaves |
| title_short | Determination of Chlorophyll Concentration in In Vitro Peanut Leaves |
| title_sort | determination of chlorophyll concentration in in vitro peanut leaves |
| topic | Arachis hypogaea Laboratory Experimentation Chlorophylls Leaves In Vitro Experimentation Experimentación en Laboratorio Clorofila Hojas Experimentación In Vitro Maní Peanuts |
| url | http://hdl.handle.net/20.500.12123/24824 https://currentprotocols.onlinelibrary.wiley.com/doi/10.1002/cpz1.70150 https://doi.org/10.1002/cpz1.70150 |
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