Analysis of Guard Cell Readouts Using Arabidopsis thaliana Isolated Epidermal Peels

Autores
Pantaleno, Rosario; Schiel, Paula; García-Mata, Carlos; Scuffi, Denise
Año de publicación
2024
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Stomata are pores surrounded by a pair of specialized cells, called guard cells, that play a central role in plant physiology through the regulation of gas exchange between plants and the environment. Guard cells have features like cell-autonomous responses and easily measurable readouts that have turned them into a model system to study signal transduction mechanisms in plants. Here, we provide a detailed protocol to analyze different physiological responses specifically in guard cells. We describe, in detail, the steps and conditions to isolate epidermal peels with tweezers and to analyze i) stomatal aperture in response to different stimuli, ii) cytosolic parameters such as hydrogen peroxide (H2O2), glutathione redox potential (EGSH), and MgATP-2 in vivo dynamics using fluorescent biosensors, and iii) gene expression in guard cell–enriched samples. The importance of this protocol lies in the fact that most living cells on epidermal peels are guard cells, enabling the preparation of guard cell–enriched samples.
Fil: Pantaleno, Rosario. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; Argentina
Fil: Schiel, Paula. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; Argentina
Fil: García-Mata, Carlos. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; Argentina
Fil: Scuffi, Denise. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; Argentina
Materia
GUARD CELLS
EPIDERMAL PEELS
STOMATAL APERTURE
BIOSENSORS
RNA
ARABIDOPSIS THALIANA
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/242972
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/242972
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Analysis of Guard Cell Readouts Using Arabidopsis thaliana Isolated Epidermal PeelsPantaleno, RosarioSchiel, PaulaGarcía-Mata, CarlosScuffi, DeniseGUARD CELLSEPIDERMAL PEELSSTOMATAL APERTUREBIOSENSORSRNAARABIDOPSIS THALIANAhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Stomata are pores surrounded by a pair of specialized cells, called guard cells, that play a central role in plant physiology through the regulation of gas exchange between plants and the environment. Guard cells have features like cell-autonomous responses and easily measurable readouts that have turned them into a model system to study signal transduction mechanisms in plants. Here, we provide a detailed protocol to analyze different physiological responses specifically in guard cells. We describe, in detail, the steps and conditions to isolate epidermal peels with tweezers and to analyze i) stomatal aperture in response to different stimuli, ii) cytosolic parameters such as hydrogen peroxide (H2O2), glutathione redox potential (EGSH), and MgATP-2 in vivo dynamics using fluorescent biosensors, and iii) gene expression in guard cell–enriched samples. The importance of this protocol lies in the fact that most living cells on epidermal peels are guard cells, enabling the preparation of guard cell–enriched samples.Fil: Pantaleno, Rosario. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; ArgentinaFil: Schiel, Paula. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; ArgentinaFil: García-Mata, Carlos. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; ArgentinaFil: Scuffi, Denise. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; ArgentinaBio-protocol LLC2024-07-20info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/242972Pantaleno, Rosario; Schiel, Paula; García-Mata, Carlos; Scuffi, Denise; Analysis of Guard Cell Readouts Using Arabidopsis thaliana Isolated Epidermal Peels; Bio-protocol LLC; Bio-protocol; 14; 14; 20-7-2024; 1-132331-8325CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.21769/BioProtoc.5033info:eu-repo/semantics/altIdentifier/url/https://bio-protocol.org/en/bpdetail?id=5033&type=0info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T10:45:56Zoai:ri.conicet.gov.ar:11336/242972instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-09-29 10:45:57.016CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Analysis of Guard Cell Readouts Using Arabidopsis thaliana Isolated Epidermal Peels
title Analysis of Guard Cell Readouts Using Arabidopsis thaliana Isolated Epidermal Peels
spellingShingle Analysis of Guard Cell Readouts Using Arabidopsis thaliana Isolated Epidermal Peels
Pantaleno, Rosario
GUARD CELLS
EPIDERMAL PEELS
STOMATAL APERTURE
BIOSENSORS
RNA
ARABIDOPSIS THALIANA
title_short Analysis of Guard Cell Readouts Using Arabidopsis thaliana Isolated Epidermal Peels
title_full Analysis of Guard Cell Readouts Using Arabidopsis thaliana Isolated Epidermal Peels
title_fullStr Analysis of Guard Cell Readouts Using Arabidopsis thaliana Isolated Epidermal Peels
title_full_unstemmed Analysis of Guard Cell Readouts Using Arabidopsis thaliana Isolated Epidermal Peels
title_sort Analysis of Guard Cell Readouts Using Arabidopsis thaliana Isolated Epidermal Peels
dc.creator.none.fl_str_mv Pantaleno, Rosario
Schiel, Paula
García-Mata, Carlos
Scuffi, Denise
author Pantaleno, Rosario
author_facet Pantaleno, Rosario
Schiel, Paula
García-Mata, Carlos
Scuffi, Denise
author_role author
author2 Schiel, Paula
García-Mata, Carlos
Scuffi, Denise
author2_role author
author
author
dc.subject.none.fl_str_mv GUARD CELLS
EPIDERMAL PEELS
STOMATAL APERTURE
BIOSENSORS
RNA
ARABIDOPSIS THALIANA
topic GUARD CELLS
EPIDERMAL PEELS
STOMATAL APERTURE
BIOSENSORS
RNA
ARABIDOPSIS THALIANA
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Stomata are pores surrounded by a pair of specialized cells, called guard cells, that play a central role in plant physiology through the regulation of gas exchange between plants and the environment. Guard cells have features like cell-autonomous responses and easily measurable readouts that have turned them into a model system to study signal transduction mechanisms in plants. Here, we provide a detailed protocol to analyze different physiological responses specifically in guard cells. We describe, in detail, the steps and conditions to isolate epidermal peels with tweezers and to analyze i) stomatal aperture in response to different stimuli, ii) cytosolic parameters such as hydrogen peroxide (H2O2), glutathione redox potential (EGSH), and MgATP-2 in vivo dynamics using fluorescent biosensors, and iii) gene expression in guard cell–enriched samples. The importance of this protocol lies in the fact that most living cells on epidermal peels are guard cells, enabling the preparation of guard cell–enriched samples.
Fil: Pantaleno, Rosario. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; Argentina
Fil: Schiel, Paula. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; Argentina
Fil: García-Mata, Carlos. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; Argentina
Fil: Scuffi, Denise. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; Argentina
description Stomata are pores surrounded by a pair of specialized cells, called guard cells, that play a central role in plant physiology through the regulation of gas exchange between plants and the environment. Guard cells have features like cell-autonomous responses and easily measurable readouts that have turned them into a model system to study signal transduction mechanisms in plants. Here, we provide a detailed protocol to analyze different physiological responses specifically in guard cells. We describe, in detail, the steps and conditions to isolate epidermal peels with tweezers and to analyze i) stomatal aperture in response to different stimuli, ii) cytosolic parameters such as hydrogen peroxide (H2O2), glutathione redox potential (EGSH), and MgATP-2 in vivo dynamics using fluorescent biosensors, and iii) gene expression in guard cell–enriched samples. The importance of this protocol lies in the fact that most living cells on epidermal peels are guard cells, enabling the preparation of guard cell–enriched samples.
publishDate 2024
dc.date.none.fl_str_mv 2024-07-20
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/11336/242972
Pantaleno, Rosario; Schiel, Paula; García-Mata, Carlos; Scuffi, Denise; Analysis of Guard Cell Readouts Using Arabidopsis thaliana Isolated Epidermal Peels; Bio-protocol LLC; Bio-protocol; 14; 14; 20-7-2024; 1-13
2331-8325
CONICET Digital
CONICET
url http://hdl.handle.net/11336/242972
identifier_str_mv Pantaleno, Rosario; Schiel, Paula; García-Mata, Carlos; Scuffi, Denise; Analysis of Guard Cell Readouts Using Arabidopsis thaliana Isolated Epidermal Peels; Bio-protocol LLC; Bio-protocol; 14; 14; 20-7-2024; 1-13
2331-8325
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.21769/BioProtoc.5033
info:eu-repo/semantics/altIdentifier/url/https://bio-protocol.org/en/bpdetail?id=5033&type=0
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
application/pdf
dc.publisher.none.fl_str_mv Bio-protocol LLC
publisher.none.fl_str_mv Bio-protocol LLC
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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score 13.070432

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