title: A Single Amino Acid Substitution in an ORANGE Protein Promotes Carotenoid Overaccumulation in Arabidopsis.
abstract: 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 of OR from Arabidopsis (Arabidopsis thaliana; AtOR) or from the agronomically important crop sorghum (Sorghum bicolor; SbOR) increased carotenoid levels up to 2-fold, expression of AtOR(His) (R90H) or SbOR(His) (R104H) variants dramatically enhanced carotenoid accumulation by up to 7-fold in the Arabidopsis calli. Moreover, we found that AtOR(Ala) (R90A) functioned similarly to AtOR(His) to promote carotenoid overproduction. Neither AtOR nor AtOR(His) greatly affected carotenogenic gene expression. AtOR(His) exhibited similar interactions with phytoene synthase (PSY) as AtOR in posttranscriptionally regulating PSY protein abundance. AtOR(His) triggered 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 of AtOR(His) in mediating chromoplast biogenesis is responsible for its induced carotenoid overproduction. Our study demonstrates OR(His/Ala) as powerful tools for carotenoid enrichment in plants, and provides insights into the mechanisms underlying OR(His)-regulated carotenoid accumulation.
authors.affiliation: Robert W. Holley Center for Agriculture and Health, U.S. Department of Agriculture-Agricultural Research Service (H.Y., X.Z., Y.Y., T.W.T., L.L.), Plant Breeding and Genetics Section, School of Integrative Plant Science (H.Y., K.O., T.E.S., X.Z., C.R., L.L.), Plant Biology Section, School of Integrative Plant Science (M.V.P.), and Boyce Thompson Institute for Plant Research (Z.F.), Cornell University, Ithaca, New York 14853;Key Laboratory of Horticultural Plant Biology, Ministry of Education, Huazhong Agricultural University, Wuhan 430070, People's Republic of China (Y.L., Q.X., X.D.);University of Freiburg, Faculty of Biology II, D79104 Freiburg, Germany (R.W.); andNewe Ya'ar Research Center, Agricultural Research Organization, Ramat Yishay 30095, Israel (N.C., A.S., N.K., J.B., Y.T.).
authors.forename: Hui
authors.initials: H
authors.lastname: Yuan
authors.affiliation: Robert W. Holley Center for Agriculture and Health, U.S. Department of Agriculture-Agricultural Research Service (H.Y., X.Z., Y.Y., T.W.T., L.L.), Plant Breeding and Genetics Section, School of Integrative Plant Science (H.Y., K.O., T.E.S., X.Z., C.R., L.L.), Plant Biology Section, School of Integrative Plant Science (M.V.P.), and Boyce Thompson Institute for Plant Research (Z.F.), Cornell University, Ithaca, New York 14853;Key Laboratory of Horticultural Plant Biology, Ministry of Education, Huazhong Agricultural University, Wuhan 430070, People's Republic of China (Y.L., Q.X., X.D.);University of Freiburg, Faculty of Biology II, D79104 Freiburg, Germany (R.W.); andNewe Ya'ar Research Center, Agricultural Research Organization, Ramat Yishay 30095, Israel (N.C., A.S., N.K., J.B., Y.T.).
authors.forename: Katherine
authors.initials: K
authors.lastname: Owsiany
authors.affiliation: Robert W. Holley Center for Agriculture and Health, U.S. Department of Agriculture-Agricultural Research Service (H.Y., X.Z., Y.Y., T.W.T., L.L.), Plant Breeding and Genetics Section, School of Integrative Plant Science (H.Y., K.O., T.E.S., X.Z., C.R., L.L.), Plant Biology Section, School of Integrative Plant Science (M.V.P.), and Boyce Thompson Institute for Plant Research (Z.F.), Cornell University, Ithaca, New York 14853;Key Laboratory of Horticultural Plant Biology, Ministry of Education, Huazhong Agricultural University, Wuhan 430070, People's Republic of China (Y.L., Q.X., X.D.);University of Freiburg, Faculty of Biology II, D79104 Freiburg, Germany (R.W.); andNewe Ya'ar Research Center, Agricultural Research Organization, Ramat Yishay 30095, Israel (N.C., A.S., N.K., J.B., Y.T.).
authors.forename: T E
authors.initials: TE
authors.lastname: Sheeja
authors.affiliation: Robert W. Holley Center for Agriculture and Health, U.S. Department of Agriculture-Agricultural Research Service (H.Y., X.Z., Y.Y., T.W.T., L.L.), Plant Breeding and Genetics Section, School of Integrative Plant Science (H.Y., K.O., T.E.S., X.Z., C.R., L.L.), Plant Biology Section, School of Integrative Plant Science (M.V.P.), and Boyce Thompson Institute for Plant Research (Z.F.), Cornell University, Ithaca, New York 14853;Key Laboratory of Horticultural Plant Biology, Ministry of Education, Huazhong Agricultural University, Wuhan 430070, People's Republic of China (Y.L., Q.X., X.D.);University of Freiburg, Faculty of Biology II, D79104 Freiburg, Germany (R.W.); andNewe Ya'ar Research Center, Agricultural Research Organization, Ramat Yishay 30095, Israel (N.C., A.S., N.K., J.B., Y.T.).
authors.forename: Xiangjun
authors.initials: X
authors.lastname: Zhou
authors.affiliation: Robert W. Holley Center for Agriculture and Health, U.S. Department of Agriculture-Agricultural Research Service (H.Y., X.Z., Y.Y., T.W.T., L.L.), Plant Breeding and Genetics Section, School of Integrative Plant Science (H.Y., K.O., T.E.S., X.Z., C.R., L.L.), Plant Biology Section, School of Integrative Plant Science (M.V.P.), and Boyce Thompson Institute for Plant Research (Z.F.), Cornell University, Ithaca, New York 14853;Key Laboratory of Horticultural Plant Biology, Ministry of Education, Huazhong Agricultural University, Wuhan 430070, People's Republic of China (Y.L., Q.X., X.D.);University of Freiburg, Faculty of Biology II, D79104 Freiburg, Germany (R.W.); andNewe Ya'ar Research Center, Agricultural Research Organization, Ramat Yishay 30095, Israel (N.C., A.S., N.K., J.B., Y.T.).
authors.forename: Caroline
authors.initials: C
authors.lastname: Rodriguez
authors.affiliation: Robert W. Holley Center for Agriculture and Health, U.S. Department of Agriculture-Agricultural Research Service (H.Y., X.Z., Y.Y., T.W.T., L.L.), Plant Breeding and Genetics Section, School of Integrative Plant Science (H.Y., K.O., T.E.S., X.Z., C.R., L.L.), Plant Biology Section, School of Integrative Plant Science (M.V.P.), and Boyce Thompson Institute for Plant Research (Z.F.), Cornell University, Ithaca, New York 14853;Key Laboratory of Horticultural Plant Biology, Ministry of Education, Huazhong Agricultural University, Wuhan 430070, People's Republic of China (Y.L., Q.X., X.D.);University of Freiburg, Faculty of Biology II, D79104 Freiburg, Germany (R.W.); andNewe Ya'ar Research Center, Agricultural Research Organization, Ramat Yishay 30095, Israel (N.C., A.S., N.K., J.B., Y.T.).
authors.forename: Yongxi
authors.initials: Y
authors.lastname: Li
authors.affiliation: Robert W. Holley Center for Agriculture and Health, U.S. Department of Agriculture-Agricultural Research Service (H.Y., X.Z., Y.Y., T.W.T., L.L.), Plant Breeding and Genetics Section, School of Integrative Plant Science (H.Y., K.O., T.E.S., X.Z., C.R., L.L.), Plant Biology Section, School of Integrative Plant Science (M.V.P.), and Boyce Thompson Institute for Plant Research (Z.F.), Cornell University, Ithaca, New York 14853;Key Laboratory of Horticultural Plant Biology, Ministry of Education, Huazhong Agricultural University, Wuhan 430070, People's Republic of China (Y.L., Q.X., X.D.);University of Freiburg, Faculty of Biology II, D79104 Freiburg, Germany (R.W.); andNewe Ya'ar Research Center, Agricultural Research Organization, Ramat Yishay 30095, Israel (N.C., A.S., N.K., J.B., Y.T.).
authors.forename: Ralf
authors.initials: R
authors.lastname: Welsch
authors.affiliation: Robert W. Holley Center for Agriculture and Health, U.S. Department of Agriculture-Agricultural Research Service (H.Y., X.Z., Y.Y., T.W.T., L.L.), Plant Breeding and Genetics Section, School of Integrative Plant Science (H.Y., K.O., T.E.S., X.Z., C.R., L.L.), Plant Biology Section, School of Integrative Plant Science (M.V.P.), and Boyce Thompson Institute for Plant Research (Z.F.), Cornell University, Ithaca, New York 14853;Key Laboratory of Horticultural Plant Biology, Ministry of Education, Huazhong Agricultural University, Wuhan 430070, People's Republic of China (Y.L., Q.X., X.D.);University of Freiburg, Faculty of Biology II, D79104 Freiburg, Germany (R.W.); andNewe Ya'ar Research Center, Agricultural Research Organization, Ramat Yishay 30095, Israel (N.C., A.S., N.K., J.B., Y.T.).
authors.forename: Noam
authors.initials: N
authors.lastname: Chayut
authors.affiliation: Robert W. Holley Center for Agriculture and Health, U.S. Department of Agriculture-Agricultural Research Service (H.Y., X.Z., Y.Y., T.W.T., L.L.), Plant Breeding and Genetics Section, School of Integrative Plant Science (H.Y., K.O., T.E.S., X.Z., C.R., L.L.), Plant Biology Section, School of Integrative Plant Science (M.V.P.), and Boyce Thompson Institute for Plant Research (Z.F.), Cornell University, Ithaca, New York 14853;Key Laboratory of Horticultural Plant Biology, Ministry of Education, Huazhong Agricultural University, Wuhan 430070, People's Republic of China (Y.L., Q.X., X.D.);University of Freiburg, Faculty of Biology II, D79104 Freiburg, Germany (R.W.); andNewe Ya'ar Research Center, Agricultural Research Organization, Ramat Yishay 30095, Israel (N.C., A.S., N.K., J.B., Y.T.).
authors.forename: Yong
authors.initials: Y
authors.lastname: Yang
authors.affiliation: Robert W. Holley Center for Agriculture and Health, U.S. Department of Agriculture-Agricultural Research Service (H.Y., X.Z., Y.Y., T.W.T., L.L.), Plant Breeding and Genetics Section, School of Integrative Plant Science (H.Y., K.O., T.E.S., X.Z., C.R., L.L.), Plant Biology Section, School of Integrative Plant Science (M.V.P.), and Boyce Thompson Institute for Plant Research (Z.F.), Cornell University, Ithaca, New York 14853;Key Laboratory of Horticultural Plant Biology, Ministry of Education, Huazhong Agricultural University, Wuhan 430070, People's Republic of China (Y.L., Q.X., X.D.);University of Freiburg, Faculty of Biology II, D79104 Freiburg, Germany (R.W.); andNewe Ya'ar Research Center, Agricultural Research Organization, Ramat Yishay 30095, Israel (N.C., A.S., N.K., J.B., Y.T.).
authors.forename: Theodore W
authors.initials: TW
authors.lastname: Thannhauser
authors.affiliation: Robert W. Holley Center for Agriculture and Health, U.S. Department of Agriculture-Agricultural Research Service (H.Y., X.Z., Y.Y., T.W.T., L.L.), Plant Breeding and Genetics Section, School of Integrative Plant Science (H.Y., K.O., T.E.S., X.Z., C.R., L.L.), Plant Biology Section, School of Integrative Plant Science (M.V.P.), and Boyce Thompson Institute for Plant Research (Z.F.), Cornell University, Ithaca, New York 14853;Key Laboratory of Horticultural Plant Biology, Ministry of Education, Huazhong Agricultural University, Wuhan 430070, People's Republic of China (Y.L., Q.X., X.D.);University of Freiburg, Faculty of Biology II, D79104 Freiburg, Germany (R.W.); andNewe Ya'ar Research Center, Agricultural Research Organization, Ramat Yishay 30095, Israel (N.C., A.S., N.K., J.B., Y.T.).
authors.forename: Mandayam V
authors.initials: MV
authors.lastname: Parthasarathy
authors.affiliation: Robert W. Holley Center for Agriculture and Health, U.S. Department of Agriculture-Agricultural Research Service (H.Y., X.Z., Y.Y., T.W.T., L.L.), Plant Breeding and Genetics Section, School of Integrative Plant Science (H.Y., K.O., T.E.S., X.Z., C.R., L.L.), Plant Biology Section, School of Integrative Plant Science (M.V.P.), and Boyce Thompson Institute for Plant Research (Z.F.), Cornell University, Ithaca, New York 14853;Key Laboratory of Horticultural Plant Biology, Ministry of Education, Huazhong Agricultural University, Wuhan 430070, People's Republic of China (Y.L., Q.X., X.D.);University of Freiburg, Faculty of Biology II, D79104 Freiburg, Germany (R.W.); andNewe Ya'ar Research Center, Agricultural Research Organization, Ramat Yishay 30095, Israel (N.C., A.S., N.K., J.B., Y.T.).
authors.forename: Qiang
authors.initials: Q
authors.lastname: Xu
authors.affiliation: Robert W. Holley Center for Agriculture and Health, U.S. Department of Agriculture-Agricultural Research Service (H.Y., X.Z., Y.Y., T.W.T., L.L.), Plant Breeding and Genetics Section, School of Integrative Plant Science (H.Y., K.O., T.E.S., X.Z., C.R., L.L.), Plant Biology Section, School of Integrative Plant Science (M.V.P.), and Boyce Thompson Institute for Plant Research (Z.F.), Cornell University, Ithaca, New York 14853;Key Laboratory of Horticultural Plant Biology, Ministry of Education, Huazhong Agricultural University, Wuhan 430070, People's Republic of China (Y.L., Q.X., X.D.);University of Freiburg, Faculty of Biology II, D79104 Freiburg, Germany (R.W.); andNewe Ya'ar Research Center, Agricultural Research Organization, Ramat Yishay 30095, Israel (N.C., A.S., N.K., J.B., Y.T.).
authors.forename: Xiuxin
authors.initials: X
authors.lastname: Deng
authors.affiliation: Robert W. Holley Center for Agriculture and Health, U.S. Department of Agriculture-Agricultural Research Service (H.Y., X.Z., Y.Y., T.W.T., L.L.), Plant Breeding and Genetics Section, School of Integrative Plant Science (H.Y., K.O., T.E.S., X.Z., C.R., L.L.), Plant Biology Section, School of Integrative Plant Science (M.V.P.), and Boyce Thompson Institute for Plant Research (Z.F.), Cornell University, Ithaca, New York 14853;Key Laboratory of Horticultural Plant Biology, Ministry of Education, Huazhong Agricultural University, Wuhan 430070, People's Republic of China (Y.L., Q.X., X.D.);University of Freiburg, Faculty of Biology II, D79104 Freiburg, Germany (R.W.); andNewe Ya'ar Research Center, Agricultural Research Organization, Ramat Yishay 30095, Israel (N.C., A.S., N.K., J.B., Y.T.).
authors.forename: Zhangjun
authors.initials: Z
authors.lastname: Fei
authors.affiliation: Robert W. Holley Center for Agriculture and Health, U.S. Department of Agriculture-Agricultural Research Service (H.Y., X.Z., Y.Y., T.W.T., L.L.), Plant Breeding and Genetics Section, School of Integrative Plant Science (H.Y., K.O., T.E.S., X.Z., C.R., L.L.), Plant Biology Section, School of Integrative Plant Science (M.V.P.), and Boyce Thompson Institute for Plant Research (Z.F.), Cornell University, Ithaca, New York 14853;Key Laboratory of Horticultural Plant Biology, Ministry of Education, Huazhong Agricultural University, Wuhan 430070, People's Republic of China (Y.L., Q.X., X.D.);University of Freiburg, Faculty of Biology II, D79104 Freiburg, Germany (R.W.); andNewe Ya'ar Research Center, Agricultural Research Organization, Ramat Yishay 30095, Israel (N.C., A.S., N.K., J.B., Y.T.).
authors.forename: Ari
authors.initials: A
authors.lastname: Schaffer
authors.affiliation: Robert W. Holley Center for Agriculture and Health, U.S. Department of Agriculture-Agricultural Research Service (H.Y., X.Z., Y.Y., T.W.T., L.L.), Plant Breeding and Genetics Section, School of Integrative Plant Science (H.Y., K.O., T.E.S., X.Z., C.R., L.L.), Plant Biology Section, School of Integrative Plant Science (M.V.P.), and Boyce Thompson Institute for Plant Research (Z.F.), Cornell University, Ithaca, New York 14853;Key Laboratory of Horticultural Plant Biology, Ministry of Education, Huazhong Agricultural University, Wuhan 430070, People's Republic of China (Y.L., Q.X., X.D.);University of Freiburg, Faculty of Biology II, D79104 Freiburg, Germany (R.W.); andNewe Ya'ar Research Center, Agricultural Research Organization, Ramat Yishay 30095, Israel (N.C., A.S., N.K., J.B., Y.T.).
authors.forename: Nurit
authors.initials: N
authors.lastname: Katzir
authors.affiliation: Robert W. Holley Center for Agriculture and Health, U.S. Department of Agriculture-Agricultural Research Service (H.Y., X.Z., Y.Y., T.W.T., L.L.), Plant Breeding and Genetics Section, School of Integrative Plant Science (H.Y., K.O., T.E.S., X.Z., C.R., L.L.), Plant Biology Section, School of Integrative Plant Science (M.V.P.), and Boyce Thompson Institute for Plant Research (Z.F.), Cornell University, Ithaca, New York 14853;Key Laboratory of Horticultural Plant Biology, Ministry of Education, Huazhong Agricultural University, Wuhan 430070, People's Republic of China (Y.L., Q.X., X.D.);University of Freiburg, Faculty of Biology II, D79104 Freiburg, Germany (R.W.); andNewe Ya'ar Research Center, Agricultural Research Organization, Ramat Yishay 30095, Israel (N.C., A.S., N.K., J.B., Y.T.).
authors.forename: Joseph
authors.initials: J
authors.lastname: Burger
authors.affiliation: Robert W. Holley Center for Agriculture and Health, U.S. Department of Agriculture-Agricultural Research Service (H.Y., X.Z., Y.Y., T.W.T., L.L.), Plant Breeding and Genetics Section, School of Integrative Plant Science (H.Y., K.O., T.E.S., X.Z., C.R., L.L.), Plant Biology Section, School of Integrative Plant Science (M.V.P.), and Boyce Thompson Institute for Plant Research (Z.F.), Cornell University, Ithaca, New York 14853;Key Laboratory of Horticultural Plant Biology, Ministry of Education, Huazhong Agricultural University, Wuhan 430070, People's Republic of China (Y.L., Q.X., X.D.);University of Freiburg, Faculty of Biology II, D79104 Freiburg, Germany (R.W.); andNewe Ya'ar Research Center, Agricultural Research Organization, Ramat Yishay 30095, Israel (N.C., A.S., N.K., J.B., Y.T.).
authors.forename: Yaakov
authors.initials: Y
authors.lastname: Tadmor
authors.affiliation: Robert W. Holley Center for Agriculture and Health, U.S. Department of Agriculture-Agricultural Research Service (H.Y., X.Z., Y.Y., T.W.T., L.L.), Plant Breeding and Genetics Section, School of Integrative Plant Science (H.Y., K.O., T.E.S., X.Z., C.R., L.L.), Plant Biology Section, School of Integrative Plant Science (M.V.P.), and Boyce Thompson Institute for Plant Research (Z.F.), Cornell University, Ithaca, New York 14853;Key Laboratory of Horticultural Plant Biology, Ministry of Education, Huazhong Agricultural University, Wuhan 430070, People's Republic of China (Y.L., Q.X., X.D.);University of Freiburg, Faculty of Biology II, D79104 Freiburg, Germany (R.W.); andNewe Ya'ar Research Center, Agricultural Research Organization, Ramat Yishay 30095, Israel (N.C., A.S., N.K., J.B., Y.T.) ll37@cornell.edu.
authors.forename: Li
authors.initials: L
authors.lastname: Li
mesh_terms: D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D017360:Arabidopsis / Q000235:genetics*; D029681:Arabidopsis Proteins / Q000737:chemistry / Q000235:genetics*; D053898:Biosynthetic Pathways / Q000235:genetics; D002338:Carotenoids / Q000378:metabolism*; D018506:Gene Expression Regulation, Plant; D050956:HSP40 Heat-Shock Proteins / Q000737:chemistry / Q000235:genetics*; D008969:Molecular Sequence Data; D030821:Plants, Genetically Modified; D018087:Plastids / Q000378:metabolism / Q000648:ultrastructure; D021381:Protein Transport; D016415:Sequence Alignment
chemical_list: D029681:Arabidopsis Proteins; D050956:HSP40 Heat-Shock Proteins; C000616511:ORANGE protein, Arabidopsis; D002338:Carotenoids