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HPLC-MS/MS analyses show that the near-starchless aps1 and pgm leaves accumulate wild type levels of ADPglucose: further evidence for the occurrence of important ADPglucose biosynthetic pathway(s) alternative to the pPGI-pPGM-AGP pathway

dc.contributor.authorBahaji, Abdellatif
dc.contributor.authorBaroja Fernández, Edurne
dc.contributor.authorSánchez López, Ángela María
dc.contributor.authorMuñoz Pérez, Francisco José
dc.contributor.authorLi, Jun
dc.contributor.authorAlmagro Zabalza, Goizeder
dc.contributor.authorMontero Macarro, Manuel
dc.contributor.authorPujol, Pablo
dc.contributor.authorGalarza, Regina
dc.contributor.authorKaneko, Kentaro
dc.contributor.authorOikawa, Kazusato
dc.contributor.authorWada, Kaede
dc.contributor.authorMitsui, Toshiaki
dc.contributor.authorPozueta Romero, Javier
dc.contributor.departmentIdAB. Instituto de Agrobiotecnología / Agrobioteknologiako Institutuaes_ES
dc.contributor.funderGobierno de Navarra / Nafarroako Gobernua, IIM010491.RI1es
dc.date.accessioned2018-09-06T12:20:18Z
dc.date.available2018-09-06T12:20:18Z
dc.date.issued2014
dc.description.abstractIn leaves, it is widely assumed that starch is the end-product of a metabolic pathway exclusively taking place in the chloroplast that (a) involves plastidic phosphoglucomutase (pPGM), ADPglucose (ADPG) pyrophosphorylase (AGP) and starch synthase (SS), and (b) is linked to the Calvin-Benson cycle by means of the plastidic phosphoglucose isomerase (pPGI). This view also implies that AGP is the sole enzyme producing the starch precursor molecule, ADPG. However, mounting evidence has been compiled pointing to the occurrence of important sources, other than the pPGI-pPGM-AGP pathway, of ADPG. To further explore this possibility, in this work two independent laboratories have carried out HPLC-MS/ MS analyses of ADPG content in leaves of the near-starchless pgm and aps1 mutants impaired in pPGM and AGP, respectively, and in leaves of double aps1/pgm mutants grown under two different culture conditions. We also measured the ADPG content in wild type (WT) and aps1 leaves expressing in the plastid two different ADPG cleaving enzymes, and in aps1 leaves expressing in the plastid GlgC, a bacterial AGP. Furthermore, we measured the ADPG content in ss3/ss4/aps1 mutants impaired in starch granule initiation and chloroplastic ADPG synthesis. We found that, irrespective of their starch contents, pgm and aps1 leaves, WT and aps1 leaves expressing in the plastid ADPG cleaving enzymes, and aps1 leaves expressing in the plastid GlgC accumulate WT ADPG content. In clear contrast, ss3/ss4/aps1 leaves accumulated ca. 300 foldmore ADPG than WT leaves. The overall data showed that, in Arabidopsis leaves, (a) there are important ADPG biosynthetic pathways, other than the pPGI-pPGM-AGP pathway, (b) pPGM and AGP are not major determinants of intracellular ADPG content, and (c) the contribution of the chloroplastic ADPG pool to the total ADPG pool is low.en
dc.description.sponsorshipThis research was partially supported by the grants [BIO2010-18239] from the Comisión Interministerial de Ciencia y Tecnología and Fondo Europeo de Desarrollo Regional (Spain) and [IIM010491.RI1] from the Government of Navarra, and by Iden Biotechnology. This research was also supported by Scientific Research on Innovative Areas [22114507] and Grants-in-Aid for Scientific Research (B) [22380186] from the Ministry of Education, Culture, Sports, Science and Technology, Japan.en
dc.format.mimetypeapplication/pdfen
dc.format.mimetypeapplication/zipen
dc.identifier.doi10.1371/journal.pone.0104997
dc.identifier.issn1932-6203
dc.identifier.urihttps://academica-e.unavarra.es/handle/2454/30543
dc.language.isoengen
dc.publisherPublic Library of Scienceen
dc.relation.ispartofPlos One, 9(8): e104997en
dc.relation.publisherversionhttps://doi.org/10.1371/journal.pone.0104997
dc.rights© 2014 Bahaji et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.en
dc.rights.accessRightsinfo:eu-repo/semantics/openAccess
dc.rights.urihttps://creativecommons.org/licenses/by/4.0/
dc.subjectaps1 leavesen
dc.subjectpgm leavesen
dc.subjectADP glucoseen
dc.subjectpPGI-pPGM-AGP pathwayen
dc.titleHPLC-MS/MS analyses show that the near-starchless aps1 and pgm leaves accumulate wild type levels of ADPglucose: further evidence for the occurrence of important ADPglucose biosynthetic pathway(s) alternative to the pPGI-pPGM-AGP pathwayen
dc.typeinfo:eu-repo/semantics/article
dc.type.versioninfo:eu-repo/semantics/publishedVersionen
dc.type.versionVersión publicada / Argitaratu den bertsioaes
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