title: Identification of Homogentisate Dioxygenase as a Target for Vitamin E Biofortification in Oilseeds.
abstract: Soybean (Glycine max) is a major plant source of protein and oil and produces important secondary metabolites beneficial for human health. As a tool for gene function discovery and improvement of this important crop, a mutant population was generated using fast neutron irradiation. Visual screening of mutagenized seeds identified a mutant line, designated MO12, which produced brown seeds as opposed to the yellow seeds produced by the unmodified Williams 82 parental cultivar. Using forward genetic methods combined with comparative genome hybridization analysis, we were able to establish that deletion of the GmHGO1 gene is the genetic basis of the brown seeded phenotype exhibited by the MO12 mutant line. GmHGO1 encodes a homogentisate dioxygenase (HGO), which catalyzes the committed enzymatic step in homogentisate catabolism. This report describes to our knowledge the first functional characterization of a plant HGO gene, defects of which are linked to the human genetic disease alkaptonuria. We show that reduced homogentisate catabolism in a soybean HGO mutant is an effective strategy for enhancing the production of lipid-soluble antioxidants such as vitamin E, as well as tolerance to herbicides that target pathways associated with homogentisate metabolism. Furthermore, this work demonstrates the utility of fast neutron mutagenesis in identifying novel genes that contribute to soybean agronomic traits.
authors.affiliation: Division of Plant Sciences (M.G.S., Y.C., C.T.N., K.M.C., Y.L., J.B., P.T.D., D.A.S., G.S.), Division of Biochemistry (G.S.), and DNA Core Facility (J.F.), University of Missouri, Columbia, Missouri 65211; and staceym@missouri.edu.|Department of Biochemistry (R.E.C., N.P., E.B.C.) and Department of Agronomy and Horticulture (H.T.N., S.S., T.E.C.), Center for Plant Science Innovation, University of Nebraska, Lincoln, Nebraska 68588 staceym@missouri.edu.
authors.forename: Minviluz G
authors.identifier: 0000-0002-3774-720X
authors.initials: MG
authors.lastname: Stacey
authors.affiliation: Division of Plant Sciences (M.G.S., Y.C., C.T.N., K.M.C., Y.L., J.B., P.T.D., D.A.S., G.S.), Division of Biochemistry (G.S.), and DNA Core Facility (J.F.), University of Missouri, Columbia, Missouri 65211; and.|Department of Biochemistry (R.E.C., N.P., E.B.C.) and Department of Agronomy and Horticulture (H.T.N., S.S., T.E.C.), Center for Plant Science Innovation, University of Nebraska, Lincoln, Nebraska 68588.
authors.forename: Rebecca E
authors.identifier: 0000-0003-3392-5766
authors.initials: RE
authors.lastname: Cahoon
authors.affiliation: Division of Plant Sciences (M.G.S., Y.C., C.T.N., K.M.C., Y.L., J.B., P.T.D., D.A.S., G.S.), Division of Biochemistry (G.S.), and DNA Core Facility (J.F.), University of Missouri, Columbia, Missouri 65211; and.|Department of Biochemistry (R.E.C., N.P., E.B.C.) and Department of Agronomy and Horticulture (H.T.N., S.S., T.E.C.), Center for Plant Science Innovation, University of Nebraska, Lincoln, Nebraska 68588.
authors.forename: Hanh T
authors.initials: HT
authors.lastname: Nguyen
authors.affiliation: Division of Plant Sciences (M.G.S., Y.C., C.T.N., K.M.C., Y.L., J.B., P.T.D., D.A.S., G.S.), Division of Biochemistry (G.S.), and DNA Core Facility (J.F.), University of Missouri, Columbia, Missouri 65211; and.|Department of Biochemistry (R.E.C., N.P., E.B.C.) and Department of Agronomy and Horticulture (H.T.N., S.S., T.E.C.), Center for Plant Science Innovation, University of Nebraska, Lincoln, Nebraska 68588.
authors.forename: Yaya
authors.initials: Y
authors.lastname: Cui
authors.affiliation: Division of Plant Sciences (M.G.S., Y.C., C.T.N., K.M.C., Y.L., J.B., P.T.D., D.A.S., G.S.), Division of Biochemistry (G.S.), and DNA Core Facility (J.F.), University of Missouri, Columbia, Missouri 65211; and.|Department of Biochemistry (R.E.C., N.P., E.B.C.) and Department of Agronomy and Horticulture (H.T.N., S.S., T.E.C.), Center for Plant Science Innovation, University of Nebraska, Lincoln, Nebraska 68588.
authors.forename: Shirley
authors.initials: S
authors.lastname: Sato
authors.affiliation: Division of Plant Sciences (M.G.S., Y.C., C.T.N., K.M.C., Y.L., J.B., P.T.D., D.A.S., G.S.), Division of Biochemistry (G.S.), and DNA Core Facility (J.F.), University of Missouri, Columbia, Missouri 65211; and.|Department of Biochemistry (R.E.C., N.P., E.B.C.) and Department of Agronomy and Horticulture (H.T.N., S.S., T.E.C.), Center for Plant Science Innovation, University of Nebraska, Lincoln, Nebraska 68588.
authors.forename: Cuong T
authors.identifier: 0000-0002-8700-9598
authors.initials: CT
authors.lastname: Nguyen
authors.affiliation: Division of Plant Sciences (M.G.S., Y.C., C.T.N., K.M.C., Y.L., J.B., P.T.D., D.A.S., G.S.), Division of Biochemistry (G.S.), and DNA Core Facility (J.F.), University of Missouri, Columbia, Missouri 65211; and.|Department of Biochemistry (R.E.C., N.P., E.B.C.) and Department of Agronomy and Horticulture (H.T.N., S.S., T.E.C.), Center for Plant Science Innovation, University of Nebraska, Lincoln, Nebraska 68588.
authors.forename: Nongnat
authors.identifier: 0000-0002-3579-7668
authors.initials: N
authors.lastname: Phoka
authors.affiliation: Division of Plant Sciences (M.G.S., Y.C., C.T.N., K.M.C., Y.L., J.B., P.T.D., D.A.S., G.S.), Division of Biochemistry (G.S.), and DNA Core Facility (J.F.), University of Missouri, Columbia, Missouri 65211; and.|Department of Biochemistry (R.E.C., N.P., E.B.C.) and Department of Agronomy and Horticulture (H.T.N., S.S., T.E.C.), Center for Plant Science Innovation, University of Nebraska, Lincoln, Nebraska 68588.
authors.forename: Kerry M
authors.initials: KM
authors.lastname: Clark
authors.affiliation: Division of Plant Sciences (M.G.S., Y.C., C.T.N., K.M.C., Y.L., J.B., P.T.D., D.A.S., G.S.), Division of Biochemistry (G.S.), and DNA Core Facility (J.F.), University of Missouri, Columbia, Missouri 65211; and.|Department of Biochemistry (R.E.C., N.P., E.B.C.) and Department of Agronomy and Horticulture (H.T.N., S.S., T.E.C.), Center for Plant Science Innovation, University of Nebraska, Lincoln, Nebraska 68588.
authors.forename: Yan
authors.initials: Y
authors.lastname: Liang
authors.affiliation: Division of Plant Sciences (M.G.S., Y.C., C.T.N., K.M.C., Y.L., J.B., P.T.D., D.A.S., G.S.), Division of Biochemistry (G.S.), and DNA Core Facility (J.F.), University of Missouri, Columbia, Missouri 65211; and.|Department of Biochemistry (R.E.C., N.P., E.B.C.) and Department of Agronomy and Horticulture (H.T.N., S.S., T.E.C.), Center for Plant Science Innovation, University of Nebraska, Lincoln, Nebraska 68588.
authors.forename: Joe
authors.initials: J
authors.lastname: Forrester
authors.affiliation: Division of Plant Sciences (M.G.S., Y.C., C.T.N., K.M.C., Y.L., J.B., P.T.D., D.A.S., G.S.), Division of Biochemistry (G.S.), and DNA Core Facility (J.F.), University of Missouri, Columbia, Missouri 65211; and.|Department of Biochemistry (R.E.C., N.P., E.B.C.) and Department of Agronomy and Horticulture (H.T.N., S.S., T.E.C.), Center for Plant Science Innovation, University of Nebraska, Lincoln, Nebraska 68588.
authors.forename: Josef
authors.initials: J
authors.lastname: Batek
authors.affiliation: Division of Plant Sciences (M.G.S., Y.C., C.T.N., K.M.C., Y.L., J.B., P.T.D., D.A.S., G.S.), Division of Biochemistry (G.S.), and DNA Core Facility (J.F.), University of Missouri, Columbia, Missouri 65211; and.|Department of Biochemistry (R.E.C., N.P., E.B.C.) and Department of Agronomy and Horticulture (H.T.N., S.S., T.E.C.), Center for Plant Science Innovation, University of Nebraska, Lincoln, Nebraska 68588.
authors.forename: Phat Tien
authors.initials: PT
authors.lastname: Do
authors.affiliation: Division of Plant Sciences (M.G.S., Y.C., C.T.N., K.M.C., Y.L., J.B., P.T.D., D.A.S., G.S.), Division of Biochemistry (G.S.), and DNA Core Facility (J.F.), University of Missouri, Columbia, Missouri 65211; and.|Department of Biochemistry (R.E.C., N.P., E.B.C.) and Department of Agronomy and Horticulture (H.T.N., S.S., T.E.C.), Center for Plant Science Innovation, University of Nebraska, Lincoln, Nebraska 68588.
authors.forename: David A
authors.initials: DA
authors.lastname: Sleper
authors.affiliation: Division of Plant Sciences (M.G.S., Y.C., C.T.N., K.M.C., Y.L., J.B., P.T.D., D.A.S., G.S.), Division of Biochemistry (G.S.), and DNA Core Facility (J.F.), University of Missouri, Columbia, Missouri 65211; and.|Department of Biochemistry (R.E.C., N.P., E.B.C.) and Department of Agronomy and Horticulture (H.T.N., S.S., T.E.C.), Center for Plant Science Innovation, University of Nebraska, Lincoln, Nebraska 68588.
authors.forename: Thomas E
authors.identifier: 0000-0003-3650-852X
authors.initials: TE
authors.lastname: Clemente
authors.affiliation: Division of Plant Sciences (M.G.S., Y.C., C.T.N., K.M.C., Y.L., J.B., P.T.D., D.A.S., G.S.), Division of Biochemistry (G.S.), and DNA Core Facility (J.F.), University of Missouri, Columbia, Missouri 65211; and.|Department of Biochemistry (R.E.C., N.P., E.B.C.) and Department of Agronomy and Horticulture (H.T.N., S.S., T.E.C.), Center for Plant Science Innovation, University of Nebraska, Lincoln, Nebraska 68588.
authors.forename: Edgar B
authors.identifier: 0000-0002-7277-1176
authors.initials: EB
authors.lastname: Cahoon
authors.affiliation: Division of Plant Sciences (M.G.S., Y.C., C.T.N., K.M.C., Y.L., J.B., P.T.D., D.A.S., G.S.), Division of Biochemistry (G.S.), and DNA Core Facility (J.F.), University of Missouri, Columbia, Missouri 65211; and.|Department of Biochemistry (R.E.C., N.P., E.B.C.) and Department of Agronomy and Horticulture (H.T.N., S.S., T.E.C.), Center for Plant Science Innovation, University of Nebraska, Lincoln, Nebraska 68588.
authors.forename: Gary
authors.identifier: 0000-0001-5914-2247
authors.initials: G
authors.lastname: Stacey
mesh_terms: D015111:4-Hydroxyphenylpyruvate Dioxygenase / Q000037:antagonists & inhibitors / Q000378:metabolism; D000222:Adaptation, Physiological / Q000187:drug effects; D017360:Arabidopsis / Q000235:genetics; D000072759:Biofortification; D004791:Enzyme Inhibitors / Q000633:toxicity; D017353:Gene Deletion; D018745:Genome, Plant; D006540:Herbicides / Q000633:toxicity; D050560:Homogentisate 1,2-Dioxygenase / Q000378:metabolism*; D006713:Homogentisic Acid / Q000378:metabolism; D007527:Isoenzymes / Q000378:metabolism; D053858:Metabolic Networks and Pathways / Q000187:drug effects; D009154:Mutation / Q000235:genetics; D010641:Phenotype; D059828:Plant Cells / Q000187:drug effects / Q000378:metabolism; D010938:Plant Oils / Q000378:metabolism*; D012639:Seeds / Q000201:enzymology*; D013025:Glycine max / Q000187:drug effects / Q000201:enzymology* / Q000502:physiology; D014810:Vitamin E / Q000378:metabolism*
chemical_list: D004791:Enzyme Inhibitors; D006540:Herbicides; D007527:Isoenzymes; D010938:Plant Oils; D014810:Vitamin E; D015111:4-Hydroxyphenylpyruvate Dioxygenase; D050560:Homogentisate 1,2-Dioxygenase; D006713:Homogentisic Acid