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dc.contributor.authorBai, Chao
dc.contributor.authorRivera Vélez, Sol Maiam
dc.contributor.authorMedina Piles, Vicente
dc.contributor.authorAlves, Rui
dc.contributor.authorVilaprinyo Terré, Ester
dc.contributor.authorSorribas Tello, Albert
dc.contributor.authorCanela i Garayoa, Ramon
dc.contributor.authorCapell Capell, Teresa
dc.contributor.authorSandmann, Gerhard
dc.contributor.authorChristou, Paul
dc.contributor.authorZhu, Changfu
dc.date.accessioned2016-10-24T12:05:25Z
dc.date.available2016-10-24T12:05:25Z
dc.date.issued2014
dc.identifier.issn0960-7412
dc.identifier.urihttp://hdl.handle.net/10459.1/58026
dc.description.abstractWe have developed an assay based on rice embryogenic callus for rapid functional characterization of metabolic genes. We validated the assay using a selection of well-characterized genes with known functions in the carotenoid biosynthesis pathway, allowing rapid visual screening of callus phenotypes based on tissue color. We then used the system to identify the functions of two uncharacterized genes: a chemically synthesized β–carotene ketolase gene optimized for maize codon usage, and a wild-type Arabidopsis thaliana ortholog of the cauliflower Orange gene. In contrast to previous reports (Lopez, A.B., Van Eck, J., Conlin, B.J., Paolillo, D.J., O'Neill, J. and Li, L. (2008) J. Exp. Bot. 59, 213–223; Lu, S., Van Eck, J., Zhou, X., Lopez, A.B., O'Halloran, D.M., Cosman, K.M., Conlin, B.J., Paolillo, D.J., Garvin, D.F., Vrebalov, J., Kochian, L.V., Küpper, H., Earle, E.D., Cao, J. and Li, L. (2006) Plant Cell 18, 3594–3605), we found that the wild-type Orange allele was sufficient to induce chromoplast differentiation. We also found that chromoplast differentiation was induced by increasing the availability of precursors and thus driving flux through the pathway, even in the absence of Orange. Remarkably, we found that diverse endosperm-specific promoters were highly active in rice callus despite their restricted activity in mature plants. Our callus system provides a unique opportunity to predict the effect of metabolic engineering in complex pathways, and provides a starting point for quantitative modeling and the rational design of engineering strategies using synthetic biology. We discuss the impact of our data on analysis and engineering of the carotenoid biosynthesis pathway.ca_ES
dc.description.sponsorshipThis study was supported by the Ministerio de Ciencia e Innovaci _on, Spain (AGL2010-15691, BFU2010-17704 and BIO2011-22525), the Generalitat de Catalunya (2009SGR809), and a European Research Council Advanced Grant (BIOFORCE). We thank Veronica Teixido and Jorge Comas (Department Cie` ncies Me` diques Ba` siques, Universitat de Lleida & Institut de Recerca Biomèdica de Lleida Edifici Recerca Biomèdica I, Avenida Alcalde Rovira Roure 80, 25198 Lleida, Spain) for assistance with the ImageJ analysis.ca_ES
dc.language.isoengca_ES
dc.publisherWileyca_ES
dc.relationMICINN/PN2008-2011/AGL2010-15691ca_ES
dc.relationMICINN/PN2008-2011/BFU2010-17704ca_ES
dc.relationMICINN/PN2008-2011/BIO2011-22525ca_ES
dc.relation.isformatofReproducció del document publicat a https://doi.org/10.1111/tpj.12384ca_ES
dc.relation.ispartofThe Plant Journal, 2014, vol. 77, núm. 3, p. 464–475ca_ES
dc.rights(c) Chao Bai et al., 2013ca_ES
dc.rights(c) Wiley. The Plant Journal, 2013ca_ES
dc.subjectCarotenoidsca_ES
dc.subjectGene functionca_ES
dc.subjectRice (Oryza sativa)ca_ES
dc.subject1–deoxy-D–xylulose 5–phosphate synthaseca_ES
dc.subjectb-carotene ketolaseca_ES
dc.subjectTechnical advanceca_ES
dc.titleAn in vitro system for the rapid functional characterization of genes involved in carotenoid biosynthesis and accumulationca_ES
dc.typearticleca_ES
dc.identifier.idgrec020816
dc.type.versionpublishedVersionca_ES
dc.rights.accessRightsinfo:eu-repo/semantics/restrictedAccessca_ES
dc.identifier.doihttps://doi.org/10.1111/tpj.12384
dc.relation.projectIDinfo:eu-repo/grantAgreement/EC/FP7/232933ca_ES
dc.date.embargoEndDate10000-01-01


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