Metabolic engineering of aliphatic glucosinolates in Chinese cabbage plants expressing Arabidopsis MAM1, CYP79F1, and CYP83A1.

Three Arabidopsis cDNAs, MAM1, CYP79F1, and CYP83A1, required for aliphatic glucosinolate biosynthesis were introduced into Chinese cabbage by Agrobacterium tumefaciens-mediated transformation. The transgenic lines overexpressing MAM1 or CYP83A1 showed wild-type phenotypes. However, all the lines overexpressing CYP79F1 displayed phenotypes different from wild type with respect to the stem thickness as well as leaf width and shape. Glucosinolate contents of the transgenic plants were compared with those of wild type. In the MAM1 line M1-1, accumulation of aliphatic glucosinolates gluconapin and glucobrassicanapin significantly increased. In the CYP83A1 line A1-1, all the aliphatic glucosinolate levels were increased, and the levels of gluconapin and glucobrassicanapin were elevated by 4.5 and 2 fold, respectively. The three CYP79F1 transgenic lines exhibited dissimilar glucosinolate profiles. The F1-1 line accumulated higher levels of gluconapoleiferin, glucobrassicin, and 4-methoxy glucobrassicin. However, F1-2 and F1-3 lines demonstrated a decrease in the levels of gluconapin and glucobrassicanapin and an increased level of 4-hydroxy glucobrassicin.

[1]  Barbara A Halkier,et al.  Identification of a flavin-monooxygenase as the S-oxygenating enzyme in aliphatic glucosinolate biosynthesis in Arabidopsis. , 2007, The Plant journal : for cell and molecular biology.

[2]  J. Tokuhisa,et al.  A gene controlling variation in Arabidopsis glucosinolate composition is part of the methionine chain elongation pathway. , 2001, Plant physiology.

[3]  M. Reichelt,et al.  bus, a Bushy Arabidopsis CYP79F1 Knockout Mutant with Abolished Synthesis of Short-Chain Aliphatic Glucosinolates , 2001, Plant Cell.

[4]  B. Halkier,et al.  Altering glucosinolate profiles modulates disease resistance in plants. , 2006, The Plant journal : for cell and molecular biology.

[5]  L. Davin,et al.  Biochemical Genetics of Plant Secondary Metabolites in Arabidopsis thaliana: The Glucosinolates. , 1991, Plant physiology.

[6]  M. Reichelt,et al.  Gene Duplication in the Diversification of Secondary Metabolism: Tandem 2-Oxoglutarate–Dependent Dioxygenases Control Glucosinolate Biosynthesis in Arabidopsis , 2001, Plant Cell.

[7]  W. F. Thompson,et al.  Rapid isolation of high molecular weight plant DNA. , 1980, Nucleic acids research.

[8]  J. Juvik,et al.  Genetic and Environmental Variation of Glucosinolate Content in Chinese Cabbage , 2006 .

[9]  M. Mok,et al.  CYTOKININ METABOLISM AND ACTION. , 2003, Annual review of plant physiology and plant molecular biology.

[10]  D. Kim,et al.  Metabolic engineering of indole glucosinolates in Chinese cabbage plants by expression of Arabidopsis CYP79B2, CYP79B3, and CYP83B1. , 2008, Molecules and cells.

[11]  Barbara Ann Halkier,et al.  Biology and biochemistry of glucosinolates. , 2006, Annual review of plant biology.

[12]  J. Gershenzon,et al.  Genetic control of natural variation in Arabidopsis glucosinolate accumulation. , 2001, Plant physiology.

[13]  A. Osbourn.  Preformed Antimicrobial Compounds and Plant Defense against Fungal Attack. , 1996, The Plant cell.

[14]  J. Fahey,et al.  The chemical diversity and distribution of glucosinolates and isothiocyanates among plants. , 2001, Phytochemistry.

[15]  S. Bak,et al.  The involvement of two p450 enzymes, CYP83B1 and CYP83A1, in auxin homeostasis and glucosinolate biosynthesis. , 2001, Plant physiology.

[16]  J. Finley,et al.  Cruciferous Vegetables: Cancer Protective Mechanisms of Glucosinolate Hydrolysis Products and Selenium , 2004, Integrative cancer therapies.

[17]  R. Mithen,et al.  α-Keto acid elongation and glucosinolate biosynthesis in Arabidopsis thaliana , 2000, Theoretical and Applied Genetics.

[18]  S. Abel,et al.  Glucosinolate metabolism and its control. , 2006, Trends in plant science.

[19]  R. Mithen,et al.  The effect of modifying the glucosinolate content of leaves of oilseed rape (Brassica napus ssp. oleifera) on its interaction with specialist and generalist pests , 1995 .

[20]  B. Halkier,et al.  Cytochrome P450 CYP79F1 from Arabidopsis Catalyzes the Conversion of Dihomomethionine and Trihomomethionine to the Corresponding Aldoximes in the Biosynthesis of Aliphatic Glucosinolates* , 2001, The Journal of Biological Chemistry.

[21]  B. Halkier,et al.  Functional Analysis of the Tandem-Duplicated P450 Genes SPS/BUS/CYP79F1 and CYP79F2 in Glucosinolate Biosynthesis and Plant Development by Ds Transposition-Generated Double Mutants1 , 2004, Plant Physiology.

[22]  Pengfei Li,et al.  Sulforaphane, a naturally occurring isothiocyanate, induces cell cycle arrest and apoptosis in HT29 human colon cancer cells. , 2000, Cancer research.

[23]  J. Tokuhisa,et al.  MAM3 Catalyzes the Formation of All Aliphatic Glucosinolate Chain Lengths in Arabidopsis1[W][OA] , 2007, Plant Physiology.

[24]  M. R. Hemm,et al.  The Arabidopsis ref2 Mutant Is Defective in the Gene Encoding CYP83A1 and Shows Both Phenylpropanoid and Glucosinolate Phenotypes Article, publication date, and citation information can be found at www.plantcell.org/cgi/doi/10.1105/tpc.006544. , 2003, The Plant Cell Online.

[25]  M. Montagu,et al.  Transfection and transformation of Agrobacterium tumefaciens , 1978, Molecular and General Genetics MGG.

[26]  S. Vaughn,et al.  Herbicidal activity of glucosinolate-containing seedmeals , 2006, Weed Science.

[27]  H. Shibaoka.  Involvement of wall microtubules in gibberllin promotion and kinetin inhibition of stem elongation , 1974 .

[28]  R. Mithen,et al.  Glucosinolate and Amino Acid Biosynthesis in Arabidopsis1 , 2004, Plant Physiology.

[29]  T. Kensler,et al.  Sulforaphane inhibits extracellular, intracellular, and antibiotic-resistant strains of Helicobacter pylori and prevents benzo[a]pyrene-induced stomach tumors , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[30]  K. Ljung,et al.  Control of axillary bud initiation and shoot architecture in Arabidopsis through the SUPERSHOOT gene. , 2001, Genes & development.

[31]  R. Jefferson.  Assaying chimeric genes in plants: The GUS gene fusion system , 1987, Plant Molecular Biology Reporter.

[32]  P. Naur,et al.  CYP79F1 and CYP79F2 have distinct functions in the biosynthesis of aliphatic glucosinolates in Arabidopsis. , 2003, The Plant journal : for cell and molecular biology.

[33]  Joong-Hoon Ahn,et al.  Characterization of uridine-diphosphate dependent flavonoid glucosyltransferase from Oryza sativa. , 2007, Journal of biochemistry and molecular biology.