An in vitro system for the rapid functional characterization of genes involved in carotenoid biosynthesis and accumulation
MetadataShow full item record
We 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.
Is part ofThe Plant Journal, 2014, vol. 77, núm. 3, p. 464–475
European research projects
- Articles publicats (Agrotecnio Center) 
- Articles publicats (IRBLleida) 
- Articles publicats (Producció Vegetal i Ciència Forestal) 
- Articles publicats (Química) 
- Articles publicats (Ciències Mèdiques Bàsiques) 
- Publicacions de projectes de recerca del Plan Nacional 
- Publicacions de projectes finançats per la Unió Europea 
Showing items related by title, author, creator and subject.
Targeted transcriptomic and metabolic profiling reveals temporal bottlenecks in the maize carotenoid pathway that may be addressed by multigene engineering Farré Martinez, Gemma; Rivera Vélez, Sol Maiam; Alves, Rui; Vilaprinyo Terré, Ester; Sorribas Tello, Albert; Canela i Garayoa, Ramon; Naqvi, Shaista; Sandmann, Gerhard; Capell Capell, Teresa; Zhu, Changfu; Christou, Paul (Wiley, 2013)Carotenoids are a diverse group of tetraterpenoid pigments found in plants, fungi, bacteria and some animals. They play vital roles in plants and provide important health benefits to mammals, including humans. We ...
Bottlenecks in carotenoid biosynthesis and accumulation in rice endosperm are influenced by the precursor–product balance Bai, Chao; Capell Capell, Teresa; Berman Quintana, Judit; Medina Piles, Vicente; Sandmann, Gerhard; Christou, Paul; Zhu, Changfu (Wiley, 2016)The profile of secondary metabolites in plants reflects the balance of biosynthesis, degradation and storage, including the availability of precursors and products that affect the metabolic equilibrium. We investigated the ...
A novel carotenoid, 4-keto-a-carotene, as an unexpected by-product during genetic engineering of carotenogenesis in rice callus Breitenbach, Jürgen; Bai, Chao; Rivera Vélez, Sol Maiam; Canela i Garayoa, Ramon; Capell Capell, Teresa; Christou, Paul; Zhu, Changfu; Sandmann, Gerhard (Elsevier, 2014)Rice endosperm is devoid of carotenoids because the initial biosynthetic steps are absent. The early carotenogenesis reactions were constituted through co-transformation of endosperm-derived rice callus with phytoene ...