ATG1 role in the immunity of tomato against Pseudomonas syringae pv. tomato

Autores
Bekier, Florencia Nicole; Rosli, Hernan Guillermo; Pombo, Marina Alejandra
Año de publicación
2019
Idioma
inglés
Tipo de recurso
documento de conferencia
Estado
versión publicada
Descripción
The bacteria Pseudomonas syringae pv. tomato DC3000 (Pst) is widely recognized as a model to study plant immunity. Plants have evolved a two-layered immunity system to defend themselves from pathogens. Pathogen- or microbe-associated molecular patterns (PAMPs or MAMPs) are detected by host´s pattern recognition receptors (PRRs) during pattern-triggered immunity (PTI) activation. This leads to changes in the intracellular calcium concentration, production of ROS, activation of MAPK cascades and transcriptional alterations. However virulent pathogens as Pst are able to bypass PTI through the delivery of effector proteins. Resistant plants recognize some of them activating effector-triggered immunity (ETI) that mainly leads to localized programmed cell death (PCD), limiting pathogen growth. Tomato Pto protein kinase interacts with the effectors AvrPto and AvrPtoB and together with Prf lead to activation of ETI response. Using previously generated RNA-seq data we identified two tomato genes (Solyc10g084930 and Solyc09g011320) whose expression is induced during ETI activation. Sequence and phylogeny analysis allowed us establishing that they encode for proteins belonging to the autophagy-related protein 1 (ATG1) group and that each of them has two orthologs in Nicotiana benthamiana, the species we use to test the role of candidate genes in immunity through virus-induced gene silencing (VIGS). Plants silenced with a construct that targets all 4 N. benthamiana orthologs (NbATG1) showed a delay in the development of PCD due to co-expression of Pto and AvrPto, as compared to non-silenced Ec1 control plants. To confirm this result, we challenged N. benthamiana 35S::pto silenced plants with Pseudomonas syringae pv. tabaci (Pstab) expressing AvrPto or an empty vector (EV). We did not observe an increase in disease symptoms with Pstab-AvrPto strain. However, we found a delay in the development of Pstab-EV disease symptoms. To test whether silencing the 4 N. benthamiana orthologs is required for the observed phenotype, we generated two constructs (6008 and 1011) to target them by pairs. We found a similar overall phenotype trend, but with a more marked delay in symptoms when silencing with 1011 and NbATG1 constructs. It is worth noticing that we have not observed growth or development abnormalities in plants silenced with any of the constructs used. We chose a well established transcriptional marker of autophagy (ATG8a) to investigate if silencing of our candidates affects this process. Using qPCR we did not observe differences in ATG8a transcript level between Ec1- and NbATG1-silenced plants, suggesting that at least in unchallenged plants, autophagy is not affected by knocking-down the genes under study. Based on our results, we believe that this group of autophagy-related proteins would be involved in two different pathways, playing a role during ETI-associated PCD and also in the development of disease symptoms.
Fil: Bekier, Florencia Nicole. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Fisiología Vegetal. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. Instituto de Fisiología Vegetal; Argentina
Fil: Rosli, Hernan Guillermo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Fisiología Vegetal. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. Instituto de Fisiología Vegetal; Argentina
Fil: Pombo, Marina Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Fisiología Vegetal. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. Instituto de Fisiología Vegetal; Argentina
LV Annual SAIB Meeting and XIV PABMB Congress
Salta
Argentina
Sociedad Argentina de Investigación Bioquímica y Biología Molecular
Pan-American Association for biochemistry and Molecular Biology
Materia
PSEUDOMONAS SYRINGAE
TOMATO
AUTOPHAGY
IMMUNITY SYSTEMS
ATG-1 PROTEIN
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:11336/271297

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repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling ATG1 role in the immunity of tomato against Pseudomonas syringae pv. tomatoBekier, Florencia NicoleRosli, Hernan GuillermoPombo, Marina AlejandraPSEUDOMONAS SYRINGAETOMATOAUTOPHAGYIMMUNITY SYSTEMSATG-1 PROTEINhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1The bacteria Pseudomonas syringae pv. tomato DC3000 (Pst) is widely recognized as a model to study plant immunity. Plants have evolved a two-layered immunity system to defend themselves from pathogens. Pathogen- or microbe-associated molecular patterns (PAMPs or MAMPs) are detected by host´s pattern recognition receptors (PRRs) during pattern-triggered immunity (PTI) activation. This leads to changes in the intracellular calcium concentration, production of ROS, activation of MAPK cascades and transcriptional alterations. However virulent pathogens as Pst are able to bypass PTI through the delivery of effector proteins. Resistant plants recognize some of them activating effector-triggered immunity (ETI) that mainly leads to localized programmed cell death (PCD), limiting pathogen growth. Tomato Pto protein kinase interacts with the effectors AvrPto and AvrPtoB and together with Prf lead to activation of ETI response. Using previously generated RNA-seq data we identified two tomato genes (Solyc10g084930 and Solyc09g011320) whose expression is induced during ETI activation. Sequence and phylogeny analysis allowed us establishing that they encode for proteins belonging to the autophagy-related protein 1 (ATG1) group and that each of them has two orthologs in Nicotiana benthamiana, the species we use to test the role of candidate genes in immunity through virus-induced gene silencing (VIGS). Plants silenced with a construct that targets all 4 N. benthamiana orthologs (NbATG1) showed a delay in the development of PCD due to co-expression of Pto and AvrPto, as compared to non-silenced Ec1 control plants. To confirm this result, we challenged N. benthamiana 35S::pto silenced plants with Pseudomonas syringae pv. tabaci (Pstab) expressing AvrPto or an empty vector (EV). We did not observe an increase in disease symptoms with Pstab-AvrPto strain. However, we found a delay in the development of Pstab-EV disease symptoms. To test whether silencing the 4 N. benthamiana orthologs is required for the observed phenotype, we generated two constructs (6008 and 1011) to target them by pairs. We found a similar overall phenotype trend, but with a more marked delay in symptoms when silencing with 1011 and NbATG1 constructs. It is worth noticing that we have not observed growth or development abnormalities in plants silenced with any of the constructs used. We chose a well established transcriptional marker of autophagy (ATG8a) to investigate if silencing of our candidates affects this process. Using qPCR we did not observe differences in ATG8a transcript level between Ec1- and NbATG1-silenced plants, suggesting that at least in unchallenged plants, autophagy is not affected by knocking-down the genes under study. Based on our results, we believe that this group of autophagy-related proteins would be involved in two different pathways, playing a role during ETI-associated PCD and also in the development of disease symptoms.Fil: Bekier, Florencia Nicole. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Fisiología Vegetal. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. Instituto de Fisiología Vegetal; ArgentinaFil: Rosli, Hernan Guillermo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Fisiología Vegetal. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. Instituto de Fisiología Vegetal; ArgentinaFil: Pombo, Marina Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Fisiología Vegetal. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. Instituto de Fisiología Vegetal; ArgentinaLV Annual SAIB Meeting and XIV PABMB CongressSaltaArgentinaSociedad Argentina de Investigación Bioquímica y Biología MolecularPan-American Association for biochemistry and Molecular BiologySociedad Argentina de Investigaciones en Bioquímica y Biología Molecular2019info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/conferenceObjectCongresoJournalhttp://purl.org/coar/resource_type/c_5794info:ar-repo/semantics/documentoDeConferenciaapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/271297ATG1 role in the immunity of tomato against Pseudomonas syringae pv. tomato; LV Annual SAIB Meeting and XIV PABMB Congress; Salta; Argentina; 2019; 124-1240327-95451667-5746CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://saib.org.ar/archivos/biocell-43.pdfInternacionalinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T09:36:24Zoai:ri.conicet.gov.ar:11336/271297instacron: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 09:36:24.524CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv ATG1 role in the immunity of tomato against Pseudomonas syringae pv. tomato
title ATG1 role in the immunity of tomato against Pseudomonas syringae pv. tomato
spellingShingle ATG1 role in the immunity of tomato against Pseudomonas syringae pv. tomato
Bekier, Florencia Nicole
PSEUDOMONAS SYRINGAE
TOMATO
AUTOPHAGY
IMMUNITY SYSTEMS
ATG-1 PROTEIN
title_short ATG1 role in the immunity of tomato against Pseudomonas syringae pv. tomato
title_full ATG1 role in the immunity of tomato against Pseudomonas syringae pv. tomato
title_fullStr ATG1 role in the immunity of tomato against Pseudomonas syringae pv. tomato
title_full_unstemmed ATG1 role in the immunity of tomato against Pseudomonas syringae pv. tomato
title_sort ATG1 role in the immunity of tomato against Pseudomonas syringae pv. tomato
dc.creator.none.fl_str_mv Bekier, Florencia Nicole
Rosli, Hernan Guillermo
Pombo, Marina Alejandra
author Bekier, Florencia Nicole
author_facet Bekier, Florencia Nicole
Rosli, Hernan Guillermo
Pombo, Marina Alejandra
author_role author
author2 Rosli, Hernan Guillermo
Pombo, Marina Alejandra
author2_role author
author
dc.subject.none.fl_str_mv PSEUDOMONAS SYRINGAE
TOMATO
AUTOPHAGY
IMMUNITY SYSTEMS
ATG-1 PROTEIN
topic PSEUDOMONAS SYRINGAE
TOMATO
AUTOPHAGY
IMMUNITY SYSTEMS
ATG-1 PROTEIN
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv The bacteria Pseudomonas syringae pv. tomato DC3000 (Pst) is widely recognized as a model to study plant immunity. Plants have evolved a two-layered immunity system to defend themselves from pathogens. Pathogen- or microbe-associated molecular patterns (PAMPs or MAMPs) are detected by host´s pattern recognition receptors (PRRs) during pattern-triggered immunity (PTI) activation. This leads to changes in the intracellular calcium concentration, production of ROS, activation of MAPK cascades and transcriptional alterations. However virulent pathogens as Pst are able to bypass PTI through the delivery of effector proteins. Resistant plants recognize some of them activating effector-triggered immunity (ETI) that mainly leads to localized programmed cell death (PCD), limiting pathogen growth. Tomato Pto protein kinase interacts with the effectors AvrPto and AvrPtoB and together with Prf lead to activation of ETI response. Using previously generated RNA-seq data we identified two tomato genes (Solyc10g084930 and Solyc09g011320) whose expression is induced during ETI activation. Sequence and phylogeny analysis allowed us establishing that they encode for proteins belonging to the autophagy-related protein 1 (ATG1) group and that each of them has two orthologs in Nicotiana benthamiana, the species we use to test the role of candidate genes in immunity through virus-induced gene silencing (VIGS). Plants silenced with a construct that targets all 4 N. benthamiana orthologs (NbATG1) showed a delay in the development of PCD due to co-expression of Pto and AvrPto, as compared to non-silenced Ec1 control plants. To confirm this result, we challenged N. benthamiana 35S::pto silenced plants with Pseudomonas syringae pv. tabaci (Pstab) expressing AvrPto or an empty vector (EV). We did not observe an increase in disease symptoms with Pstab-AvrPto strain. However, we found a delay in the development of Pstab-EV disease symptoms. To test whether silencing the 4 N. benthamiana orthologs is required for the observed phenotype, we generated two constructs (6008 and 1011) to target them by pairs. We found a similar overall phenotype trend, but with a more marked delay in symptoms when silencing with 1011 and NbATG1 constructs. It is worth noticing that we have not observed growth or development abnormalities in plants silenced with any of the constructs used. We chose a well established transcriptional marker of autophagy (ATG8a) to investigate if silencing of our candidates affects this process. Using qPCR we did not observe differences in ATG8a transcript level between Ec1- and NbATG1-silenced plants, suggesting that at least in unchallenged plants, autophagy is not affected by knocking-down the genes under study. Based on our results, we believe that this group of autophagy-related proteins would be involved in two different pathways, playing a role during ETI-associated PCD and also in the development of disease symptoms.
Fil: Bekier, Florencia Nicole. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Fisiología Vegetal. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. Instituto de Fisiología Vegetal; Argentina
Fil: Rosli, Hernan Guillermo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Fisiología Vegetal. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. Instituto de Fisiología Vegetal; Argentina
Fil: Pombo, Marina Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Fisiología Vegetal. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. Instituto de Fisiología Vegetal; Argentina
LV Annual SAIB Meeting and XIV PABMB Congress
Salta
Argentina
Sociedad Argentina de Investigación Bioquímica y Biología Molecular
Pan-American Association for biochemistry and Molecular Biology
description The bacteria Pseudomonas syringae pv. tomato DC3000 (Pst) is widely recognized as a model to study plant immunity. Plants have evolved a two-layered immunity system to defend themselves from pathogens. Pathogen- or microbe-associated molecular patterns (PAMPs or MAMPs) are detected by host´s pattern recognition receptors (PRRs) during pattern-triggered immunity (PTI) activation. This leads to changes in the intracellular calcium concentration, production of ROS, activation of MAPK cascades and transcriptional alterations. However virulent pathogens as Pst are able to bypass PTI through the delivery of effector proteins. Resistant plants recognize some of them activating effector-triggered immunity (ETI) that mainly leads to localized programmed cell death (PCD), limiting pathogen growth. Tomato Pto protein kinase interacts with the effectors AvrPto and AvrPtoB and together with Prf lead to activation of ETI response. Using previously generated RNA-seq data we identified two tomato genes (Solyc10g084930 and Solyc09g011320) whose expression is induced during ETI activation. Sequence and phylogeny analysis allowed us establishing that they encode for proteins belonging to the autophagy-related protein 1 (ATG1) group and that each of them has two orthologs in Nicotiana benthamiana, the species we use to test the role of candidate genes in immunity through virus-induced gene silencing (VIGS). Plants silenced with a construct that targets all 4 N. benthamiana orthologs (NbATG1) showed a delay in the development of PCD due to co-expression of Pto and AvrPto, as compared to non-silenced Ec1 control plants. To confirm this result, we challenged N. benthamiana 35S::pto silenced plants with Pseudomonas syringae pv. tabaci (Pstab) expressing AvrPto or an empty vector (EV). We did not observe an increase in disease symptoms with Pstab-AvrPto strain. However, we found a delay in the development of Pstab-EV disease symptoms. To test whether silencing the 4 N. benthamiana orthologs is required for the observed phenotype, we generated two constructs (6008 and 1011) to target them by pairs. We found a similar overall phenotype trend, but with a more marked delay in symptoms when silencing with 1011 and NbATG1 constructs. It is worth noticing that we have not observed growth or development abnormalities in plants silenced with any of the constructs used. We chose a well established transcriptional marker of autophagy (ATG8a) to investigate if silencing of our candidates affects this process. Using qPCR we did not observe differences in ATG8a transcript level between Ec1- and NbATG1-silenced plants, suggesting that at least in unchallenged plants, autophagy is not affected by knocking-down the genes under study. Based on our results, we believe that this group of autophagy-related proteins would be involved in two different pathways, playing a role during ETI-associated PCD and also in the development of disease symptoms.
publishDate 2019
dc.date.none.fl_str_mv 2019
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info:eu-repo/semantics/conferenceObject
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http://purl.org/coar/resource_type/c_5794
info:ar-repo/semantics/documentoDeConferencia
status_str publishedVersion
format conferenceObject
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/271297
ATG1 role in the immunity of tomato against Pseudomonas syringae pv. tomato; LV Annual SAIB Meeting and XIV PABMB Congress; Salta; Argentina; 2019; 124-124
0327-9545
1667-5746
CONICET Digital
CONICET
url http://hdl.handle.net/11336/271297
identifier_str_mv ATG1 role in the immunity of tomato against Pseudomonas syringae pv. tomato; LV Annual SAIB Meeting and XIV PABMB Congress; Salta; Argentina; 2019; 124-124
0327-9545
1667-5746
CONICET Digital
CONICET
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language eng
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dc.publisher.none.fl_str_mv Sociedad Argentina de Investigaciones en Bioquímica y Biología Molecular
publisher.none.fl_str_mv Sociedad Argentina de Investigaciones en Bioquímica y Biología Molecular
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