Genomics-based selection and functional characterization of triterpene glycosyltransferases from the model legume Medicago truncatula.

The biosynthesis of triterpene saponins is poorly characterized in spite of the importance of these glycosylated secondary metabolites for plant defense and animal health. The model legume Medicago truncatula synthesizes more than 30 different saponins based on at least five triterpene aglycones; soyasapogenols B and E, medicagenic acid, hederagenin and bayogenin. We have employed an inducible cell culture system, DNA array-based and in silico transcript profiling, and targeted metabolite profiling, to identify triterpene glycosyltransferases (GTs) from among the more than 300 GTs expressed in M. truncatula. Two uridine diphosphate glucosyltransferases were functionally characterized; UGT73K1 with specificity for hederagenin and soyasapogenols B and E, and UGT71G1 with specificity for medicagenic acid. The latter enzyme also glycosylated certain isoflavones and the flavonol quercetin with higher efficiency than triterpenes; however, integrated transcript and metabolite profiling supported a function for UGT71G1 in terpenoid but not (iso)flavonoid biosynthesis in the elicited cell cultures.

[1]  Pedro Mendes,et al.  Methyl jasmonate and yeast elicitor induce differential transcriptional and metabolic re-programming in cell suspension cultures of the model legume Medicago truncatula , 2005, Planta.

[2]  R. Dixon,et al.  Colocalization of l-Phenylalanine Ammonia-Lyase and Cinnamate 4-Hydroxylase for Metabolic Channeling in Phenylpropanoid Biosynthesis , 2004, The Plant Cell Online.

[3]  A. Kubo,et al.  Alteration of sugar donor specificities of plant glycosyltransferases by a single point mutation. , 2004, Archives of biochemistry and biophysics.

[4]  M. Hirai,et al.  Integration of transcriptomics and metabolomics for understanding of global responses to nutritional stresses in Arabidopsis thaliana. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[5]  A. Osbourn,et al.  A gene cluster for secondary metabolism in oat: implications for the evolution of metabolic diversity in plants. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[6]  Patrik R. Jones,et al.  UGT73C6 and UGT78D1, Glycosyltransferases Involved in Flavonol Glycoside Biosynthesis in Arabidopsis thaliana* , 2003, Journal of Biological Chemistry.

[7]  M. Willits,et al.  Cloning and regiospecificity studies of two flavonoid glucosyltransferases from Allium cepa. , 2003, Phytochemistry.

[8]  Patrik R. Jones,et al.  The in vitro substrate regiospecificity of recombinant UGT85B1, the cyanohydrin glucosyltransferase from Sorghum bicolor. , 2003, Phytochemistry.

[9]  Y. Hahn,et al.  Discovery of genes for ginsenoside biosynthesis by analysis of ginseng expressed sequence tags , 2003, Plant Cell Reports.

[10]  D. Inzé,et al.  A functional genomics approach toward the understanding of secondary metabolism in plant cells , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[11]  H. Hayashi,et al.  Up-regulation of soyasaponin biosynthesis by methyl jasmonate in cultured cells of Glycyrrhiza glabra. , 2003, Plant & cell physiology.

[12]  Yi Li,et al.  Evolution of substrate recognition across a multigene family of glycosyltransferases in Arabidopsis. , 2003, Glycobiology.

[13]  Hideyuki Suzuki,et al.  A genomics approach to the early stages of triterpene saponin biosynthesis in Medicago truncatula. , 2002, The Plant journal : for cell and molecular biology.

[14]  W. Schwab,et al.  Isolation, cloning and expression of a multifunctional O-methyltransferase capable of forming 2,5-dimethyl-4-methoxy-3(2H)-furanone, one of the key aroma compounds in strawberry fruits. , 2002, The Plant journal : for cell and molecular biology.

[15]  D. Guo,et al.  Estrogen-like activity of ginsenoside Rg1 derived from Panax notoginseng. , 2002, The Journal of clinical endocrinology and metabolism.

[16]  H. Takahara,et al.  UDP-glucuronic acid:soyasapogenol glucuronosyltransferase involved in saponin biosynthesis in germinating soybean seeds , 2002, Planta.

[17]  Lloyd W Sumner,et al.  Metabolic profiling of saponins in Medicago sativa and Medicago truncatula using HPLC coupled to an electrospray ion-trap mass spectrometer. , 2002, Phytochemistry.

[18]  Yi Li,et al.  The Activity of ArabidopsisGlycosyltransferases toward Salicylic Acid, 4-Hydroxybenzoic Acid, and Other Benzoates* , 2002, Journal of Biological Chemistry.

[19]  R. Dixon,et al.  Elicitor-Induced Association of Isoflavone O-Methyltransferase with Endomembranes Prevents the Formation and 7-O-Methylation of Daidzein during Isoflavonoid Phytoalexin Biosynthesis Article, publication date, and citation information can be found at www.plantcell.org/cgi/doi/10.1105/tpc.010382. , 2001, The Plant Cell Online.

[20]  R. Dixon,et al.  The biosynthesis of monolignols: a "metabolic grid", or independent pathways to guaiacyl and syringyl units? , 2001, Phytochemistry.

[21]  R. Dixon Natural products and plant disease resistance , 2001, Nature.

[22]  Patrick Jones,et al.  Glycosyltransferases in secondary plant metabolism: tranquilizers and stimulant controllers , 2001, Planta.

[23]  J. Gutterman,et al.  Avicins: Triterpenoid saponins from Acacia victoriae (Bentham) induce apoptosis by mitochondrial perturbation , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[24]  Yue Xu Perspectives on the 21st century development of functional foods: bridging Chinese medicated diet and functional foods , 2001 .

[25]  S. Baldauf,et al.  Phylogenetic Analysis of the UDP-glycosyltransferase Multigene Family of Arabidopsis thaliana * 210 , 2001, The Journal of Biological Chemistry.

[26]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[27]  Patrik R. Jones,et al.  Glycosyltransferases in plant natural product synthesis: characterization of a supergene family. , 2000, Trends in plant science.

[28]  A. Osbourn,et al.  Compromised disease resistance in saponin-deficient plants. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[29]  Villacres‐Eriksson,et al.  Quillaja Saponin Formulations that Stimulate Proinflammatory Cytokines Elicit a Potent Acquired Cell‐Mediated Immunity , 1999, Scandinavian journal of immunology.

[30]  W. Oleszek,et al.  Determination and toxicity of saponins from Amaranthus cruentus seeds. , 1999, Journal of agricultural and food chemistry.

[31]  Y. Ebizuka,et al.  Beta-amyrin synthase--cloning of oxidosqualene cyclase that catalyzes the formation of the most popular triterpene among higher plants. , 1998, European journal of biochemistry.

[32]  E. Logemann,et al.  Extensive Reprogramming of Primary and Secondary Metabolism by Fungal Elicitor or Infection in Parsley Cells , 1998, Biological chemistry.

[33]  E. Heinz,et al.  UDP-glucose:sterol glucosyltransferase: cloning and functional expression in Escherichia coli/ , 1997, Plant Molecular Biology.

[34]  M. Friedman,et al.  Cloning and expression of solanidine UDP-glucose glucosyltransferase from potato. , 1997, The Plant journal : for cell and molecular biology.

[35]  E. Small Adaptations to herbivory in alfalfa (Medicago sativa) , 1996 .

[36]  M. Kataoka,et al.  Intracellular localization of UDPG: p-Hydroxybenzoate glucosyltransferase and its reaction product in Lithospermum cell cultures , 1995 .

[37]  R. Dixon,et al.  The isoflavonoid phytoalexin pathway: From enzymes to genes to transcription factors , 1995 .

[38]  W. Oleszek,et al.  Determination of Alfalfa (Medicago sativa) Saponins by High-Performance Liquid Chromatography , 1994 .

[39]  G. Weissenböck,et al.  Subcellular localization of luteolin glucuronides and related enzymes in rye mesophyll , 1992, Planta.

[40]  G. Church,et al.  Genomic sequencing. , 1993, Methods in molecular biology.

[41]  G. Schmid,et al.  Appearance and immunohistochemical localization of UDP-glucose: coniferyl alcohol glucosyltransferase in spruce (Picea abies (L.) Karst.) seedlings , 1982, Planta.

[42]  R. Dixon,et al.  Dose responses for Colletotrichum lindemuthianum elicitor-mediated enzyme induction in French bean cell suspension cultures , 1981, Planta.

[43]  M. M. Bradford A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. , 1976, Analytical biochemistry.

[44]  M. Jones,et al.  Two Rapid Assays for Saponin in Individual Alfalfa Plants 1 , 1969 .

[45]  D. E. Zimmer,et al.  A Bioassay for Alfalfa Saponins Using the Fungus, Trichoderma viride Pers. ex Fr.1 , 1967 .

[46]  J. W. McClure,et al.  Uridinediphosphate-glucose: Isovitexin 7-O-glucosyltransferase from barley protoplasts: Subcellular localization , 2004, Planta.

[47]  A. Osbourn Saponins in cereals. , 2003, Phytochemistry.

[48]  A. Tava,et al.  Saponins from Medicago Spp.: chemical characterization and biological activity against insects. , 1996, Advances in experimental medicine and biology.

[49]  R. Ibrahim IMMUNOLOCALIZATION OF FLAVONOID CONJUGATES AND THEIR ENZYMES , 1992 .

[50]  R. Ibrahim,et al.  Partial purification and some properties of a ring B-O-glucosyltransferase from onion bulbs , 1991 .