Genetic and isotope ratio mass spectrometric evidence for the occurrence of starch degradation and cycling in illuminated Arabidopsis leaves.

Although there is a great wealth of data supporting the occurrence of simultaneous synthesis and breakdown of storage carbohydrate in many organisms, previous 13CO2 pulse-chase based studies indicated that starch degradation does not operate in illuminated Arabidopsis leaves. Here we show that leave...

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Main Authors: Marouane Baslam, Edurne Baroja-Fernández, Adriana Ricarte-Bermejo, Ángela María Sánchez-López, Iker Aranjuelo, Abdellatif Bahaji, Francisco José Muñoz, Goizeder Almagro, Pablo Pujol, Regina Galarza, Pilar Teixidor, Javier Pozueta-Romero
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2017-01-01
Series:PLoS ONE
Online Access:https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0171245&type=printable
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author Marouane Baslam
Edurne Baroja-Fernández
Adriana Ricarte-Bermejo
Ángela María Sánchez-López
Iker Aranjuelo
Abdellatif Bahaji
Francisco José Muñoz
Goizeder Almagro
Pablo Pujol
Regina Galarza
Pilar Teixidor
Javier Pozueta-Romero
author_facet Marouane Baslam
Edurne Baroja-Fernández
Adriana Ricarte-Bermejo
Ángela María Sánchez-López
Iker Aranjuelo
Abdellatif Bahaji
Francisco José Muñoz
Goizeder Almagro
Pablo Pujol
Regina Galarza
Pilar Teixidor
Javier Pozueta-Romero
author_sort Marouane Baslam
collection DOAJ
description Although there is a great wealth of data supporting the occurrence of simultaneous synthesis and breakdown of storage carbohydrate in many organisms, previous 13CO2 pulse-chase based studies indicated that starch degradation does not operate in illuminated Arabidopsis leaves. Here we show that leaves of gwd, sex4, bam4, bam1/bam3 and amy3/isa3/lda starch breakdown mutants accumulate higher levels of starch than wild type (WT) leaves when cultured under continuous light (CL) conditions. We also show that leaves of CL grown dpe1 plants impaired in the plastidic disproportionating enzyme accumulate higher levels of maltotriose than WT leaves, the overall data providing evidence for the occurrence of extensive starch degradation in illuminated leaves. Moreover, we show that leaves of CL grown mex1/pglct plants impaired in the chloroplastic maltose and glucose transporters display a severe dwarf phenotype and accumulate high levels of maltose, strongly indicating that the MEX1 and pGlcT transporters are involved in the export of starch breakdown products to the cytosol to support growth during illumination. To investigate whether starch breakdown products can be recycled back to starch during illumination through a mechanism involving ADP-glucose pyrophosphorylase (AGP) we conducted kinetic analyses of the stable isotope carbon composition (δ13C) in starch of leaves of 13CO2 pulsed-chased WT and AGP lacking aps1 plants. Notably, the rate of increase of δ13C in starch of aps1 leaves during the pulse was exceedingly higher than that of WT leaves. Furthermore, δ13C decline in starch of aps1 leaves during the chase was much faster than that of WT leaves, which provides strong evidence for the occurrence of AGP-mediated cycling of starch breakdown products in illuminated Arabidopsis leaves.
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spelling doaj-art-8fa85eae5582464ba04f1de9b6c698e22025-08-20T03:04:55ZengPublic Library of Science (PLoS)PLoS ONE1932-62032017-01-01122e017124510.1371/journal.pone.0171245Genetic and isotope ratio mass spectrometric evidence for the occurrence of starch degradation and cycling in illuminated Arabidopsis leaves.Marouane BaslamEdurne Baroja-FernándezAdriana Ricarte-BermejoÁngela María Sánchez-LópezIker AranjueloAbdellatif BahajiFrancisco José MuñozGoizeder AlmagroPablo PujolRegina GalarzaPilar TeixidorJavier Pozueta-RomeroAlthough there is a great wealth of data supporting the occurrence of simultaneous synthesis and breakdown of storage carbohydrate in many organisms, previous 13CO2 pulse-chase based studies indicated that starch degradation does not operate in illuminated Arabidopsis leaves. Here we show that leaves of gwd, sex4, bam4, bam1/bam3 and amy3/isa3/lda starch breakdown mutants accumulate higher levels of starch than wild type (WT) leaves when cultured under continuous light (CL) conditions. We also show that leaves of CL grown dpe1 plants impaired in the plastidic disproportionating enzyme accumulate higher levels of maltotriose than WT leaves, the overall data providing evidence for the occurrence of extensive starch degradation in illuminated leaves. Moreover, we show that leaves of CL grown mex1/pglct plants impaired in the chloroplastic maltose and glucose transporters display a severe dwarf phenotype and accumulate high levels of maltose, strongly indicating that the MEX1 and pGlcT transporters are involved in the export of starch breakdown products to the cytosol to support growth during illumination. To investigate whether starch breakdown products can be recycled back to starch during illumination through a mechanism involving ADP-glucose pyrophosphorylase (AGP) we conducted kinetic analyses of the stable isotope carbon composition (δ13C) in starch of leaves of 13CO2 pulsed-chased WT and AGP lacking aps1 plants. Notably, the rate of increase of δ13C in starch of aps1 leaves during the pulse was exceedingly higher than that of WT leaves. Furthermore, δ13C decline in starch of aps1 leaves during the chase was much faster than that of WT leaves, which provides strong evidence for the occurrence of AGP-mediated cycling of starch breakdown products in illuminated Arabidopsis leaves.https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0171245&type=printable
spellingShingle Marouane Baslam
Edurne Baroja-Fernández
Adriana Ricarte-Bermejo
Ángela María Sánchez-López
Iker Aranjuelo
Abdellatif Bahaji
Francisco José Muñoz
Goizeder Almagro
Pablo Pujol
Regina Galarza
Pilar Teixidor
Javier Pozueta-Romero
Genetic and isotope ratio mass spectrometric evidence for the occurrence of starch degradation and cycling in illuminated Arabidopsis leaves.
PLoS ONE
title Genetic and isotope ratio mass spectrometric evidence for the occurrence of starch degradation and cycling in illuminated Arabidopsis leaves.
title_full Genetic and isotope ratio mass spectrometric evidence for the occurrence of starch degradation and cycling in illuminated Arabidopsis leaves.
title_fullStr Genetic and isotope ratio mass spectrometric evidence for the occurrence of starch degradation and cycling in illuminated Arabidopsis leaves.
title_full_unstemmed Genetic and isotope ratio mass spectrometric evidence for the occurrence of starch degradation and cycling in illuminated Arabidopsis leaves.
title_short Genetic and isotope ratio mass spectrometric evidence for the occurrence of starch degradation and cycling in illuminated Arabidopsis leaves.
title_sort genetic and isotope ratio mass spectrometric evidence for the occurrence of starch degradation and cycling in illuminated arabidopsis leaves
url https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0171245&type=printable
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