title: Functional integration of non-native carotenoids into chloroplasts by viral-derived expression of capsanthin-capsorubin synthase in Nicotiana benthamiana.
abstract: The biosynthesis of leaf carotenoids in Nicotiana benthamiana was altered by forced re-routing of the pathway to the synthesis of capsanthin, a non-native chromoplast-specific xanthophyll, using an RNA viral vector containing capsanthin-capsorubin synthase (Ccs) cDNA. The cDNA encoding Ccs was placed under the transcriptional control of a tobamovirus subgenomic promoter. Leaves from transfected plants expressing Ccs developed an orange phenotype and accumulated high levels of capsanthin (up to 36% of total carotenoids). This phenomenon was associated with thylakoid membrane distortion and reduction of grana stacking. In contrast to the situation prevailing in chromoplasts, capsanthin was not esterified and its increased level was balanced by a concomitant decrease of the major leaf xanthophylls, suggesting an autoregulatory control of chloroplast carotenoid composition. Capsanthin was exclusively recruited into the trimeric and monomeric light-harvesting complexes of photosystem II (PSII) and shown to significantly contribute to the light-harvesting capacity. On a chlorophyll basis, the concentrations of PSI and PSII reaction centres were not modified. This demonstration that higher plant antenna complexes can accommodate non-native carotenoids provides compelling evidence for functional remodelling of photosynthetic membranes toward a better photoreactivity by rational design of the incorporated carotenoid structures.
authors.affiliation: Biosource Technologies, Vacaville, CA 95688, USA.
authors.forename: M H
authors.initials: MH
authors.lastname: Kumagai
authors.forename: Y
authors.initials: Y
authors.lastname: Keller
authors.forename: F
authors.initials: F
authors.lastname: Bouvier
authors.forename: D
authors.initials: D
authors.lastname: Clary
authors.forename: B
authors.initials: B
authors.lastname: Camara
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D002338:Carotenoids / Q000031:analogs & derivatives* / Q000096:biosynthesis / Q000378:metabolism; D002736:Chloroplasts / Q000378:metabolism* / Q000648:ultrastructure; D018076:DNA, Complementary; D005822:Genetic Vectors; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D010088:Oxidoreductases / Q000235:genetics*; D010641:Phenotype; D010788:Photosynthesis; D018515:Plant Leaves / Q000201:enzymology; D010940:Plant Proteins; D010947:Plants, Toxic; D012328:RNA Viruses / Q000235:genetics*; D014026:Nicotiana / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D024341:Xanthophylls
chemical_list: D018076:DNA, Complementary; D010940:Plant Proteins; D024341:Xanthophylls; D002338:Carotenoids; C006889:capsanthin; D010088:Oxidoreductases; C086535:CCS protein, Capsicum annuum