A Single Amino Acid Substitution in an ORANGE Protein Promotes Carotenoid Overaccumulation in Arabidopsis

• Published in: Plant physiology (Bethesda) Vol. 169; no. 1; pp. 421 - 431
• Main Authors: Yuan, Hui, Owsiany, Katherine, Sheeja, T.E., Zhou, Xiangjun, Rodriguez, Caroline, Li, Yongxi, Welsch, Ralf, Chayut, Noam, Yang, Yong, Thannhauser, Theodore W., Parthasarathy, Mandayam V., Xu, Qiang, Deng, Xiuxin, Fei, Zhangjun, Schaffer, Ari, Katzir, Nurit, Burger, Joseph, Tadmor, Yaakov, Li, Li
• Format: Journal Article
• Language: English
• Published: United States American Society of Plant Biologists 01-09-2015
• Subjects: Amino Acid Sequence; Amino Acid Substitution; Amino acids; Arabidopsis - genetics; Arabidopsis Proteins - chemistry; Arabidopsis Proteins - genetics; BIOCHEMISTRY AND METABOLISM; Biosynthesis; Biosynthetic Pathways - genetics; Callus; Carotenoids; Carotenoids - metabolism; Cauliflower; Chromoplasts; Gene Expression Regulation, Plant; Genetic mutation; HSP40 Heat-Shock Proteins - chemistry; HSP40 Heat-Shock Proteins - genetics; Molecular Sequence Data; Plants; Plants, Genetically Modified; Plastids; Plastids - metabolism; Plastids - ultrastructure; Protein Transport; Sequence Alignment; Transgenic plants
• ISSN: 0032-0889; 1532-2548
• DOI: 10.1104/pp.15.00971

Carotenoids are crucial for plant growth and human health. The finding of ORANGE (OR) protein as a pivotal regulator of carotenogenesis offers a unique opportunity to comprehensively understand the regulatory mechanisms of carotenoid accumulation and develop crops with enhanced nutritional quality. Here, we demonstrated that alteration of a single amino acid in a wild-type OR greatly enhanced its ability to promote carotenoid accumulation. Whereas overexpression ofORfrom Arabidopsis (Arabidopsis thaliana; AtOR) or from the agronomically important crop sorghum (Sorghum bicolor; SbOR) increased carotenoid levels up to 2-fold, expression ofAtORHis
(R90H) orSbORHis
(R104H) variants dramatically enhanced carotenoid accumulation by up to 7-fold in the Arabidopsis calli. Moreover, we found thatAtORAla
(R90A) functioned similarly toAtORHis
to promote carotenoid overproduction. Neither AtOR nor AtORHisgreatly affected carotenogenic gene expression. AtORHisexhibited similar interactions with phytoene synthase (PSY) as AtOR in posttranscriptionally regulating PSY protein abundance. AtORHistriggered biogenesis of membranous chromoplasts in the Arabidopsis calli, which shared structures similar to chromoplasts found in the curd of the orange cauliflower (Brassica oleracea) mutant. By contrast, AtOR did not cause plastid-type changes in comparison with the controls, but produced plastids containing larger and electron-dense plastoglobuli. The unique ability ofAtORHis
in mediating chromoplast biogenesis is responsible for its induced carotenoid overproduction. Our study demonstratesORHis/Ala
as powerful tools for carotenoid enrichment in plants, and provides insights into the mechanisms underlyingORHis
-regulated carotenoid accumulation.