NTRC and thioredoxin f overexpression differentially induces starch accumulation in tobacco leaves

Thioredoxin (Trx) f and NADPH-dependent Trx reductase C (NTRC) have both been proposed as major redox regulators of starch metabolism in chloroplasts. However, little is known regarding the specific role of each protein in this complex mechanism. To shed light on this point, tobacco plants that were...

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Autores: Ancín Rípodas, María, Larraya Reta, Luis María, Fernández San Millán, Alicia, Veramendi Charola, Jon, Burch Smith, Tessa, Farrán Blanch, Inmaculada
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2019
País:España
Institución:Universidad Pública de Navarra
Repositorio:Academica-e. Repositorio Institucional de la Universidad Pública de Navarra
OAI Identifier:oai:academica-e.unavarra.es:2454/36915
Acceso en línea:https://hdl.handle.net/2454/36915
Access Level:acceso abierto
Palabra clave:Starch metabolism
Thioredoxin
NTRC
Redox regulation
Chloroplast
VIGS
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spelling NTRC and thioredoxin f overexpression differentially induces starch accumulation in tobacco leavesAncín Rípodas, MaríaLarraya Reta, Luis MaríaFernández San Millán, AliciaVeramendi Charola, JonBurch Smith, TessaFarrán Blanch, InmaculadaStarch metabolismThioredoxinNTRCRedox regulationChloroplastVIGSThioredoxin (Trx) f and NADPH-dependent Trx reductase C (NTRC) have both been proposed as major redox regulators of starch metabolism in chloroplasts. However, little is known regarding the specific role of each protein in this complex mechanism. To shed light on this point, tobacco plants that were genetically engineered to overexpress the NTRC protein from the chloroplast genome were obtained and compared to previously generated Trx f-overexpressing transplastomic plants. Likewise, we investigated the impact of NTRC and Trx f deficiency on starch metabolism by generating Nicotiana benthamiana plants that were silenced for each gene. Our results demonstrated that NTRC overexpression induced enhanced starch accumulation in tobacco leaves, as occurred with Trx f. However, only Trx f silencing leads to a significant decrease in the leaf starch content. Quantitative analysis of enzyme activities related to starch synthesis and degradation were determined in all of the genotypes. Zymographic analyses were additionally performed to compare the amylolytic enzyme profiles of both transplastomic tobacco plants. Our findings indicated that NTRC overexpression promotes the accumulation of transitory leaf starch as a consequence of a diminished starch turnover during the dark period, which seems to be related to a significant reductive activation of ADP-glucose pyrophosphorylase and/or a deactivation of a putative debranching enzyme. On the other hand, increased starch content in Trx f-overexpressing plants was connected to an increase in the capacity of soluble starch synthases during the light period. Taken together, these results suggest that NTRC and the ferredoxin/Trx system play distinct roles in starch turnover.This research was funded by the by the Spanish Ministry of Science and Innovation, grants number AGL2010-15107 and AGL2016-79868. M.A. was supported by FPU pre-doctoral fellowship from Spanish Ministry of Education, Culture and Sports. T.B.-S. was supported by NSF 1456761.MDPIInstitute for Multidisciplinary Research in Applied Biology - IMAB2019info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionapplication/pdfhttps://hdl.handle.net/2454/36915reponame:Academica-e. Repositorio Institucional de la Universidad Pública de Navarrainstname:Universidad Pública de NavarraInglésinfo:eu-repo/grantAgreement/ES/1PE/AGL2016-79868© 2019 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY)https://creativecommons.org/licenses/by/4.0/info:eu-repo/semantics/openAccessoai:academica-e.unavarra.es:2454/369152026-06-17T12:41:47Z
dc.title.none.fl_str_mv NTRC and thioredoxin f overexpression differentially induces starch accumulation in tobacco leaves
title NTRC and thioredoxin f overexpression differentially induces starch accumulation in tobacco leaves
spellingShingle NTRC and thioredoxin f overexpression differentially induces starch accumulation in tobacco leaves
Ancín Rípodas, María
Starch metabolism
Thioredoxin
NTRC
Redox regulation
Chloroplast
VIGS
title_short NTRC and thioredoxin f overexpression differentially induces starch accumulation in tobacco leaves
title_full NTRC and thioredoxin f overexpression differentially induces starch accumulation in tobacco leaves
title_fullStr NTRC and thioredoxin f overexpression differentially induces starch accumulation in tobacco leaves
title_full_unstemmed NTRC and thioredoxin f overexpression differentially induces starch accumulation in tobacco leaves
title_sort NTRC and thioredoxin f overexpression differentially induces starch accumulation in tobacco leaves
dc.creator.none.fl_str_mv Ancín Rípodas, María
Larraya Reta, Luis María
Fernández San Millán, Alicia
Veramendi Charola, Jon
Burch Smith, Tessa
Farrán Blanch, Inmaculada
author Ancín Rípodas, María
author_facet Ancín Rípodas, María
Larraya Reta, Luis María
Fernández San Millán, Alicia
Veramendi Charola, Jon
Burch Smith, Tessa
Farrán Blanch, Inmaculada
author_role author
author2 Larraya Reta, Luis María
Fernández San Millán, Alicia
Veramendi Charola, Jon
Burch Smith, Tessa
Farrán Blanch, Inmaculada
author2_role author
author
author
author
author
dc.contributor.none.fl_str_mv Institute for Multidisciplinary Research in Applied Biology - IMAB
dc.subject.none.fl_str_mv Starch metabolism
Thioredoxin
NTRC
Redox regulation
Chloroplast
VIGS
topic Starch metabolism
Thioredoxin
NTRC
Redox regulation
Chloroplast
VIGS
description Thioredoxin (Trx) f and NADPH-dependent Trx reductase C (NTRC) have both been proposed as major redox regulators of starch metabolism in chloroplasts. However, little is known regarding the specific role of each protein in this complex mechanism. To shed light on this point, tobacco plants that were genetically engineered to overexpress the NTRC protein from the chloroplast genome were obtained and compared to previously generated Trx f-overexpressing transplastomic plants. Likewise, we investigated the impact of NTRC and Trx f deficiency on starch metabolism by generating Nicotiana benthamiana plants that were silenced for each gene. Our results demonstrated that NTRC overexpression induced enhanced starch accumulation in tobacco leaves, as occurred with Trx f. However, only Trx f silencing leads to a significant decrease in the leaf starch content. Quantitative analysis of enzyme activities related to starch synthesis and degradation were determined in all of the genotypes. Zymographic analyses were additionally performed to compare the amylolytic enzyme profiles of both transplastomic tobacco plants. Our findings indicated that NTRC overexpression promotes the accumulation of transitory leaf starch as a consequence of a diminished starch turnover during the dark period, which seems to be related to a significant reductive activation of ADP-glucose pyrophosphorylase and/or a deactivation of a putative debranching enzyme. On the other hand, increased starch content in Trx f-overexpressing plants was connected to an increase in the capacity of soluble starch synthases during the light period. Taken together, these results suggest that NTRC and the ferredoxin/Trx system play distinct roles in starch turnover.
publishDate 2019
dc.date.none.fl_str_mv 2019
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv https://hdl.handle.net/2454/36915
url https://hdl.handle.net/2454/36915
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv info:eu-repo/grantAgreement/ES/1PE/AGL2016-79868
dc.rights.none.fl_str_mv https://creativecommons.org/licenses/by/4.0/
info:eu-repo/semantics/openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by/4.0/
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv MDPI
publisher.none.fl_str_mv MDPI
dc.source.none.fl_str_mv reponame:Academica-e. Repositorio Institucional de la Universidad Pública de Navarra
instname:Universidad Pública de Navarra
instname_str Universidad Pública de Navarra
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collection Academica-e. Repositorio Institucional de la Universidad Pública de Navarra
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