Glucosinolate biosynthetic genes in Brassica rapa.

Glucosinolates (GS) are a group of amino acid-derived secondary metabolites found throughout the Cruciferae family. Glucosinolates and their degradation products play important roles in pathogen and insect interactions, as well as in human health. In order to elucidate the glucosinolate biosynthetic pathway in Brassica rapa, we conducted comparative genomic analyses of Arabidopsis thaliana and B. rapa on a genome-wide level. We identified 102 putative genes in B. rapa as the orthologs of 52 GS genes in A. thaliana. All but one gene was successfully mapped on 10 chromosomes. Most GS genes exist in more than one copy in B. rapa. A high co-linearity in the glucosinolate biosynthetic pathway between A. thaliana and B. rapa was also established. The homologous GS genes in B. rapa and A. thaliana share 59-91% nucleotide sequence identity and 93% of the GS genes exhibit synteny between B. rapa and A. thaliana. Moreover, the structure and arrangement of the B. rapa GS (BrGS) genes correspond with the known evolutionary divergence of B. rapa, and may help explain the profiles and accumulation of GS in B. rapa.

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

[2]  S. Abel,et al.  Arabidopsis IQD1, a novel calmodulin-binding nuclear protein, stimulates glucosinolate accumulation and plant defense. , 2005, The Plant journal : for cell and molecular biology.

[3]  A. Ordás,et al.  Variation of glucosinolates in vegetable crops of Brassica rapa. , 2007, Phytochemistry.

[4]  S. Hecht,et al.  INHIBITION OF CARCINOGENESIS BY ISOTHIOCYANATES* , 2000, Drug metabolism reviews.

[5]  B. P. Klein,et al.  Variation of glucosinolates in vegetable crops of Brassica oleracea. , 1999, Journal of agricultural and food chemistry.

[6]  Jutta Papenbrock,et al.  The three desulfoglucosinolate sulfotransferase proteins in Arabidopsis have different substrate specificities and are differentially expressed , 2006, The FEBS journal.

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

[8]  P. Schulze-Lefert,et al.  A Glucosinolate Metabolism Pathway in Living Plant Cells Mediates Broad-Spectrum Antifungal Defense , 2009, Science.

[9]  M. Hirai,et al.  Elucidation of Gene-to-Gene and Metabolite-to-Gene Networks in Arabidopsis by Integration of Metabolomics and Transcriptomics* , 2005, Journal of Biological Chemistry.

[10]  Michael O. Kelleher,et al.  The cancer chemopreventive actions of phytochemicals derived from glucosinolates , 2008, European journal of nutrition.

[11]  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.

[12]  T. Kensler,et al.  Anticarcinogenic activities of sulforaphane and structurally related synthetic norbornyl isothiocyanates. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[13]  Hans-Peter Mock,et al.  The transcription factor HIG1/MYB51 regulates indolic glucosinolate biosynthesis in Arabidopsis thaliana. , 2007, The Plant journal : for cell and molecular biology.

[14]  P. Naur,et al.  CYP83B1 Is the Oxime-metabolizing Enzyme in the Glucosinolate Pathway in Arabidopsis * , 2001, The Journal of Biological Chemistry.

[15]  U. Flügge,et al.  HAG2/MYB76 and HAG3/MYB29 exert a specific and coordinated control on the regulation of aliphatic glucosinolate biosynthesis in Arabidopsis thaliana. , 2008, The New phytologist.

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

[17]  P. Naur,et al.  CYP83A1 and CYP83B1, Two Nonredundant Cytochrome P450 Enzymes Metabolizing Oximes in the Biosynthesis of Glucosinolates in Arabidopsis1 , 2003, Plant Physiology.

[18]  Sixue Chen,et al.  A redox-active isopropylmalate dehydrogenase functions in the biosynthesis of glucosinolates and leucine in Arabidopsis. , 2009, The Plant journal : for cell and molecular biology.

[19]  H. Yamazaki,et al.  Inhibition and inactivation of human cytochrome P450 isoforms by phenethyl isothiocyanate. , 2001, Drug metabolism and disposition: the biological fate of chemicals.

[20]  Miroslav Strnad,et al.  DOF transcription factor AtDof1.1 (OBP2) is part of a regulatory network controlling glucosinolate biosynthesis in Arabidopsis. , 2006, The Plant journal : for cell and molecular biology.

[21]  M. Koornneef,et al.  Quantitative trait loci for glucosinolate accumulation in Brassica rapa leaves. , 2008, The New phytologist.

[22]  M. Nei,et al.  MEGA4: Molecular Evolutionary Genetics Analysis (MEGA) software version 4.0. , 2007, Molecular biology and evolution.

[23]  B. Halkier,et al.  Cytochrome P450 CYP79A2 from Arabidopsis thaliana L. Catalyzes the Conversion of l-Phenylalanine to Phenylacetaldoxime in the Biosynthesis of Benzylglucosinolate* , 2000, The Journal of Biological Chemistry.

[24]  Lars Rask,et al.  Myrosinase: gene family evolution and herbivore defense in Brassicaceae , 2004, Plant Molecular Biology.

[25]  Yuji Sawada,et al.  Omics-based approaches to methionine side chain elongation in Arabidopsis: characterization of the genes encoding methylthioalkylmalate isomerase and methylthioalkylmalate dehydrogenase. , 2009, Plant & cell physiology.

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

[27]  Eran Pichersky,et al.  An Aldehyde Oxidase in Developing Seeds of Arabidopsis Converts Benzaldehyde to Benzoic Acid1[OA] , 2009, Plant Physiology.

[28]  Barbara A Halkier,et al.  Glucosinolate research in the Arabidopsis era. , 2002, Trends in plant science.

[29]  John A. Pickett,et al.  Biosynthesis of methionine-derived glucosinolates in Arabidopsis thaliana: recombinant expression and characterization of methylthioalkylmalate synthase, the condensing enzyme of the chain-elongation cycle , 2004, Planta.

[30]  B. Halkier,et al.  Glucosinolate engineering identifies a gamma-glutamyl peptidase. , 2009, Nature chemical biology.

[31]  B. Berger,et al.  The R2R3-MYB transcription factor HAG1/MYB28 is a regulator of methionine-derived glucosinolate biosynthesis in Arabidopsis thaliana. , 2007, The Plant journal : for cell and molecular biology.

[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]  D. Kliebenstein A Role for Gene Duplication and Natural Variation of Gene Expression in the Evolution of Metabolism , 2008, PloS one.

[34]  P. Reymond,et al.  The glutathione-deficient mutant pad2-1 accumulates lower amounts of glucosinolates and is more susceptible to the insect herbivore Spodoptera littoralis. , 2008, The Plant journal : for cell and molecular biology.

[35]  C. P. Hong,et al.  Physical mapping and microsynteny of Brassica rapa ssp. pekinensis genome corresponding to a 222 kbp gene-rich region of Arabidopsis chromosome 4 and partially duplicated on chromosome 5 , 2005, Molecular Genetics and Genomics.

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

[37]  I. Sønderby,et al.  Biosynthesis of glucosinolates--gene discovery and beyond. , 2010, Trends in plant science.

[38]  D. Yoon,et al.  Indole-3-carbinol induces apoptosis through p53 and activation of caspase-8 pathway in lung cancer A549 cells. , 2010, Food and chemical toxicology : an international journal published for the British Industrial Biological Research Association.

[39]  Ulf-Ingo Flügge,et al.  The Plastidic Bile Acid Transporter 5 Is Required for the Biosynthesis of Methionine-Derived Glucosinolates in Arabidopsis thaliana[W] , 2009, The Plant Cell Online.

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

[41]  H. Vogel,et al.  The Gene Controlling the Indole Glucosinolate Modifier1 Quantitative Trait Locus Alters Indole Glucosinolate Structures and Aphid Resistance in Arabidopsis[W] , 2009, The Plant Cell Online.

[42]  J. Celenza,et al.  The Arabidopsis ATR1 Myb Transcription Factor Controls Indolic Glucosinolate Homeostasis1 , 2005, Plant Physiology.

[43]  B. Halkier,et al.  Cytochrome P450 CYP79B2 from Arabidopsis Catalyzes the Conversion of Tryptophan to Indole-3-acetaldoxime, a Precursor of Indole Glucosinolates and Indole-3-acetic Acid* , 2000, The Journal of Biological Chemistry.

[44]  M. Reichelt,et al.  Arabidopsis thaliana encodes a bacterial-type heterodimeric isopropylmalate isomerase involved in both Leu biosynthesis and the Met chain elongation pathway of glucosinolate formation , 2009, Plant Molecular Biology.

[45]  Dong-Jin Lee,et al.  Variation of glucosinolates in vegetable crops of Brassica rapa L. ssp. pekinensis , 2010 .

[46]  A. Bones,et al.  The enzymic and chemically induced decomposition of glucosinolates. , 2006, Phytochemistry.

[47]  M. Hirai,et al.  Omics-based identification of Arabidopsis Myb transcription factors regulating aliphatic glucosinolate biosynthesis , 2007, Proceedings of the National Academy of Sciences.

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

[49]  Lingyun Wu,et al.  Dietary approach to attenuate oxidative stress, hypertension, and inflammation in the cardiovascular system. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[50]  E. Pichersky,et al.  Arabidopsis Chy1 null mutants are deficient in benzoic acid-containing glucosinolates in the seeds. , 2009, Plant biology.

[51]  A. Krumbein,et al.  Genotypic effects on glucosinolates and sensory properties of broccoli and cauliflower. , 2004, Die Nahrung.

[52]  P. Naur,et al.  Arabidopsis mutants in the C-S lyase of glucosinolate biosynthesis establish a critical role for indole-3-acetaldoxime in auxin homeostasis. , 2004, The Plant journal : for cell and molecular biology.

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

[54]  M. Reichelt,et al.  Disruption of Adenosine-5′-Phosphosulfate Kinase in Arabidopsis Reduces Levels of Sulfated Secondary Metabolites[W] , 2009, The Plant Cell Online.

[55]  Seung-Beom Hong,et al.  Metabolic engineering of aliphatic glucosinolates in Chinese cabbage plants expressing Arabidopsis MAM1, CYP79F1, and CYP83A1. , 2008, BMB reports.

[56]  Stefan Binder,et al.  BRANCHED-CHAIN AMINOTRANSFERASE4 Is Part of the Chain Elongation Pathway in the Biosynthesis of Methionine-Derived Glucosinolates in Arabidopsis[W] , 2006, The Plant Cell Online.

[57]  M. K. Kim,et al.  Cruciferous vegetable intake and the risk of human cancer: epidemiological evidence , 2008, Proceedings of the Nutrition Society.

[58]  K. Gunderson,et al.  Characterization of seed-specific benzoyloxyglucosinolate mutations in Arabidopsis thaliana. , 2007, The Plant journal : for cell and molecular biology.

[59]  C. Gachon,et al.  Transcriptional co-regulation of secondary metabolism enzymes in Arabidopsis: functional and evolutionary implications , 2005, Plant Molecular Biology.

[60]  T. Mitchell-Olds,et al.  The ABC's of comparative genomics in the Brassicaceae: building blocks of crucifer genomes. , 2006, Trends in plant science.

[61]  Seung-Beom Hong,et al.  Genome‐wide identification of glucosinolate synthesis genes in Brassica rapa , 2009, The FEBS journal.

[62]  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.

[63]  D. Galbraith,et al.  CYP83B1, a Cytochrome P450 at the Metabolic Branch Point in Auxin and Indole Glucosinolate Biosynthesis in Arabidopsis , 2001, Plant Cell.

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

[65]  M. Reichelt,et al.  Chapter five Glucosinolate hydrolysis and its impact on generalist and specialist insect herbivores , 2003 .

[66]  J. Celenza,et al.  Arabidopsis cytochrome P450s that catalyze the first step of tryptophan-dependent indole-3-acetic acid biosynthesis. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[67]  Bjarne Gram Hansen,et al.  Subclade of Flavin-Monooxygenases Involved in Aliphatic Glucosinolate Biosynthesis1[W] , 2008, Plant Physiology.

[68]  A. Müller,et al.  Desulfoglucosinolate Sulfotransferases from Arabidopsis thaliana Catalyze the Final Step in the Biosynthesis of the Glucosinolate Core Structure* , 2004, Journal of Biological Chemistry.

[69]  Mark G. M. Aarts,et al.  The Impact of the Absence of Aliphatic Glucosinolates on Insect Herbivory in Arabidopsis , 2008, PloS one.

[70]  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.

[71]  M. Reichelt,et al.  Arabidopsis Branched-Chain Aminotransferase 3 Functions in Both Amino Acid and Glucosinolate Biosynthesis1[W][OA] , 2007, Plant Physiology.

[72]  Frederick M. Ausubel,et al.  Glucosinolate Metabolites Required for an Arabidopsis Innate Immune Response , 2009, Science.

[73]  D. Kliebenstein,et al.  A Systems Biology Approach Identifies a R2R3 MYB Gene Subfamily with Distinct and Overlapping Functions in Regulation of Aliphatic Glucosinolates , 2007, PloS one.

[74]  Yuji Sawada,et al.  Arabidopsis bile acid:sodium symporter family protein 5 is involved in methionine-derived glucosinolate biosynthesis. , 2009, Plant & cell physiology.

[75]  T. Molinski,et al.  Arabidopsis glucosyltransferase UGT74B1 functions in glucosinolate biosynthesis and auxin homeostasis. , 2004, The Plant journal : for cell and molecular biology.

[76]  Sixue Chen,et al.  Functional specification of Arabidopsis isopropylmalate isomerases in glucosinolate and leucine biosynthesis. , 2010, Plant & cell physiology.

[77]  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.

[78]  Daniel J. Kliebenstein,et al.  Linking Metabolic QTLs with Network and cis-eQTLs Controlling Biosynthetic Pathways , 2007, PLoS genetics.

[79]  K. Guru,et al.  Consumption of Raw Cruciferous Vegetables is Inversely Associated with Bladder Cancer Risk , 2008, Cancer Epidemiology Biomarkers & Prevention.

[80]  P. Brakefield,et al.  The Male Sex Pheromone of the Butterfly Bicyclus anynana: Towards an Evolutionary Analysis , 2008, PloS one.

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

[82]  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.

[83]  Rachel E. Kerwin,et al.  A Novel 2-Oxoacid-Dependent Dioxygenase Involved in the Formation of the Goiterogenic 2-Hydroxybut-3-enyl Glucosinolate and Generalist Insect Resistance in Arabidopsis[C][W][OA] , 2008, Plant Physiology.