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Metabolic engineering to simultaneously activate anthocyanin and proanthocyanidin biosynthetic pathways in Nicotiana spp

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Metabolic engineering to simultaneously activate anthocyanin and proanthocyanidin biosynthetic pathways in Nicotiana spp

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dc.contributor.author Fresquet-Corrales, Sandra es_ES
dc.contributor.author Roque Mesa, Edelin Marta es_ES
dc.contributor.author Sarrión-Perdigones , Alejandro es_ES
dc.contributor.author Rochina, Maricruz es_ES
dc.contributor.author López-Gresa, María Pilar es_ES
dc.contributor.author Díaz-Mula, Huertas M es_ES
dc.contributor.author Belles Albert, José Mª es_ES
dc.contributor.author Tomás-Barberán, Francisco es_ES
dc.contributor.author Beltran Porter, Jose Pio es_ES
dc.contributor.author Cañas Clemente, Luís Antonio es_ES
dc.date.accessioned 2019-01-25T21:04:15Z
dc.date.available 2019-01-25T21:04:15Z
dc.date.issued 2017 es_ES
dc.identifier.issn 1932-6203 es_ES
dc.identifier.uri http://hdl.handle.net/10251/116155
dc.description.abstract [EN] Proanthocyanidins (PAs), or condensed tannins, are powerful antioxidants that remove harmful free oxygen radicals from cells. To engineer the anthocyanin and proanthocyanidin biosynthetic pathways to de novo produce PAs in two Nicotiana species, we incorporated four transgenes to the plant chassis. We opted to perform a simultaneous transformation of the genes linked in a multigenic construct rather than classical breeding or retransformation approaches. We generated a GoldenBraid 2.0 multigenic construct containing two Antirrhinum majus transcription factors (AmRosea1 and AmDelila) to upregulate the anthocyanin pathway in combination with two Medicago truncatula genes (MtLAR and MtANR) to produce the enzymes that will derivate the biosynthetic pathway to PAs production. Transient and stable transformation of Nicotiana benthamiana and Nicotiana tabacum with the multigenic construct were respectively performed. Transient expression experiments in N. benthamiana showed the activation of the anthocyanin pathway producing a purple color in the agroinfiltrated leaves and also the effective production of 208.5 nmol (-) catechin/g FW and 228.5 nmol (-) epicatechin/g FW measured by the p-dimethylaminocinnamaldehyde (DMACA) method. The integration capacity of the four transgenes, their respective expression levels and their heritability in the second generation were analyzed in stably transformed N. tabacum plants. DMACA and phoroglucinolysis/HPLC-MS analyses corroborated the activation of both pathways and the effective production of PAs in T0 and T1 transgenic tobacco plants up to a maximum of 3.48 mg/g DW. The possible biotechnological applications of the GB2.0 multigenic approach in forage legumes to produce "bloatsafe" plants and to improve the efficiency of conversion of plant protein into animal protein (ruminal protein bypass) are discussed. es_ES
dc.description.sponsorship This work was supported by grants BIO2012-39849-C02-01 and BIO2016-75485-R from the Spanish Ministry of Economy and Competitiveness (MINECO) (http://www.idi.mineco.gob.es/portal/site/MICINN) to LAC and a fellowship of the JAE-CSIC program to SF. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
dc.language Inglés es_ES
dc.publisher Public Library of Science es_ES
dc.relation MINECO/BIO2012-39849-CO2-01 es_ES
dc.relation MINECO/BIO2016-75485-R es_ES
dc.relation.ispartof PLoS ONE es_ES
dc.rights Reconocimiento (by) es_ES
dc.subject.classification BIOQUIMICA Y BIOLOGIA MOLECULAR es_ES
dc.title Metabolic engineering to simultaneously activate anthocyanin and proanthocyanidin biosynthetic pathways in Nicotiana spp es_ES
dc.type Artículo es_ES
dc.identifier.doi 10.1371/journal.pone.0184839 es_ES
dc.rights.accessRights Abierto es_ES
dc.contributor.affiliation Universitat Politècnica de València. Instituto Universitario Mixto de Biología Molecular y Celular de Plantas - Institut Universitari Mixt de Biologia Molecular i Cel·lular de Plantes es_ES
dc.contributor.affiliation Universitat Politècnica de València. Departamento de Biotecnología - Departament de Biotecnologia es_ES
dc.description.bibliographicCitation Fresquet-Corrales, S.; Roque Mesa, EM.; Sarrión-Perdigones, A.; Rochina, M.; López-Gresa, MP.; Díaz-Mula, HM.; Belles Albert, JM.... (2017). Metabolic engineering to simultaneously activate anthocyanin and proanthocyanidin biosynthetic pathways in Nicotiana spp. PLoS ONE. 12(9). doi:10.1371/journal.pone.0184839 es_ES
dc.description.accrualMethod S es_ES
dc.relation.publisherversion http://doi.org/10.1371/journal.pone.0184839 es_ES
dc.description.upvformatpinicio e0184839 es_ES
dc.type.version info:eu-repo/semantics/publishedVersion es_ES
dc.description.volume 12 es_ES
dc.description.issue 9 es_ES
dc.identifier.pmid 28902886
dc.identifier.pmcid PMC5597232
dc.relation.pasarela S\351048 es_ES
dc.contributor.funder Ministerio de Economía y Competitividad es_ES


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