Determination of Chlorophyll Concentration in In Vitro Peanut Leaves

Autores
Faustinelli, Paola C.; Massa, Alicia N.; Soria, Néstor W.; Lopez Colomba, Eliana; Suarez, Paola Alejandra; Lamb, Marshall C.
Año de publicación
2025
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
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
Fuente
Current Protocols : 1-9 (May 2025)
Materia
Arachis hypogaea
Laboratory Experimentation
Chlorophylls
Leaves
In Vitro Experimentation
Experimentación en Laboratorio
Clorofila
Hojas
Experimentación In Vitro
Maní
Peanuts
Nivel de accesibilidad
acceso restringido
Condiciones de uso
http://creativecommons.org/licenses/by-nc-sa/4.0/
Repositorio
INTA Digital (INTA)
Institución
Instituto Nacional de Tecnología Agropecuaria
OAI Identificador
oai:localhost:20.500.12123/24824
Acceder
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network_name_str INTA Digital (INTA)
spelling Determination of Chlorophyll Concentration in In Vitro Peanut LeavesFaustinelli, Paola C.Massa, Alicia N.Soria, Néstor W.Lopez Colomba, ElianaSuarez, Paola AlejandraLamb, Marshall C.Arachis hypogaeaLaboratory ExperimentationChlorophyllsLeavesIn Vitro ExperimentationExperimentación en LaboratorioClorofilaHojasExperimentación In VitroManíPeanutsThe 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 VegetalesFil: Faustinelli, Paola C. USDA-ARS. National Peanut Research Laboratory; Estados UnidosFil: Massa, Alicia N. USDA-ARS. National Peanut Research Laboratory; Estados UnidosFil: Soria, Néstor W. Universidad Católica de Córdoba; ArgentinaFil: Lopez Colomba, Eliana. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Fisiología y Recursos Genéticos Vegetales; ArgentinaFil: Lopez Colomba, Eliana. Consejo Nacional de Investigaciones Científicas y Técnicas. Unidad de Estudios Agropecuarios (UDEA); ArgentinaFil: Suarez, Paola Alejandra. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Fisiología y Recursos Genéticos Vegetales; ArgentinaFil: Suarez, Paola Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas. Unidad de Estudios Agropecuarios (UDEA); ArgentinaFil: Lamb, Marshall C. USDA-ARS. National Peanut Research Laboratory; Estados UnidosWiley2025-12-30T15:04:35Z2025-12-30T15:04:35Z2025-05info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfhttp://hdl.handle.net/20.500.12123/24824https://currentprotocols.onlinelibrary.wiley.com/doi/10.1002/cpz1.701502691-1299https://doi.org/10.1002/cpz1.70150Current Protocols : 1-9 (May 2025)reponame:INTA Digital (INTA)instname:Instituto Nacional de Tecnología Agropecuariaenginfo:eu-repo/semantics/restrictedAccesshttp://creativecommons.org/licenses/by-nc-sa/4.0/Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0)2026-01-08T10:41:01Zoai:localhost:20.500.12123/24824instacron:INTAInstitucionalhttp://repositorio.inta.gob.ar/Organismo científico-tecnológicoNo correspondehttp://repositorio.inta.gob.ar/oai/requesttripaldi.nicolas@inta.gob.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:l2026-01-08 10:41:01.757INTA Digital (INTA) - Instituto Nacional de Tecnología Agropecuariafalse
dc.title.none.fl_str_mv Determination of Chlorophyll Concentration in In Vitro Peanut Leaves
title Determination of Chlorophyll Concentration in In Vitro Peanut Leaves
spellingShingle Determination of Chlorophyll Concentration in In Vitro Peanut Leaves
Faustinelli, Paola C.
Arachis hypogaea
Laboratory Experimentation
Chlorophylls
Leaves
In Vitro Experimentation
Experimentación en Laboratorio
Clorofila
Hojas
Experimentación In Vitro
Maní
Peanuts
title_short 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_sort Determination of Chlorophyll Concentration in In Vitro Peanut Leaves
dc.creator.none.fl_str_mv Faustinelli, Paola C.
Massa, Alicia N.
Soria, Néstor W.
Lopez Colomba, Eliana
Suarez, Paola Alejandra
Lamb, Marshall C.
author Faustinelli, Paola C.
author_facet Faustinelli, Paola C.
Massa, Alicia N.
Soria, Néstor W.
Lopez Colomba, Eliana
Suarez, Paola Alejandra
Lamb, Marshall C.
author_role author
author2 Massa, Alicia N.
Soria, Néstor W.
Lopez Colomba, Eliana
Suarez, Paola Alejandra
Lamb, Marshall C.
author2_role author
author
author
author
author
dc.subject.none.fl_str_mv Arachis hypogaea
Laboratory Experimentation
Chlorophylls
Leaves
In Vitro Experimentation
Experimentación en Laboratorio
Clorofila
Hojas
Experimentación In Vitro
Maní
Peanuts
topic Arachis hypogaea
Laboratory Experimentation
Chlorophylls
Leaves
In Vitro Experimentation
Experimentación en Laboratorio
Clorofila
Hojas
Experimentación In Vitro
Maní
Peanuts
dc.description.none.fl_txt_mv 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
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.
publishDate 2025
dc.date.none.fl_str_mv 2025-12-30T15:04:35Z
2025-12-30T15:04:35Z
2025-05
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv 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
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
identifier_str_mv 2691-1299
dc.language.none.fl_str_mv eng
language eng
dc.rights.none.fl_str_mv 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)
eu_rights_str_mv restrictedAccess
rights_invalid_str_mv http://creativecommons.org/licenses/by-nc-sa/4.0/
Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0)
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Wiley
publisher.none.fl_str_mv Wiley
dc.source.none.fl_str_mv Current Protocols : 1-9 (May 2025)
reponame:INTA Digital (INTA)
instname:Instituto Nacional de Tecnología Agropecuaria
reponame_str INTA Digital (INTA)
collection INTA Digital (INTA)
instname_str Instituto Nacional de Tecnología Agropecuaria
repository.name.fl_str_mv INTA Digital (INTA) - Instituto Nacional de Tecnología Agropecuaria
repository.mail.fl_str_mv tripaldi.nicolas@inta.gob.ar
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score 13.25844

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