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UDP-glucose:3-deoxyanthocyanidin 5-O-glucosyltransferase from Sinningia cardinalis

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Abstract

3-Deoxyanthocyanins are rare anthocyanin pigments produced by some mosses, ferns, and higher plants. The enzymes and genes responsible for biosynthesis of 3-deoxyanthocyanins have not been well characterized. We identified a novel gene encoding UDP-glucose:3-deoxyanthocyanidin 5-O-glucosyltransferase (dA5GT) from Sinningia cardinalis, which accumulates abundant 3-deoxyanthocyanins in its petals. Five candidate genes (ScUGT1 to ScUGT5) were isolated from an S. cardinalis flower cDNA by degenerate PCR targeted for the UGT88 clade. ScUGT1, ScUGT3, and ScUGT5 exhibited 45–47% identity with rose anthocyanidin 5,3-O-glucosyltransferase, which catalyzes glucosylation at the 5- and 3-position of 3-hydroxyanthocyanidin. Based on its temporal and spatial gene expression patterns, and enzymatic activity assays of the recombinant protein, ScUGT5 was screened as a dA5GT candidate. Recombinant ScUGT5 protein expressed in Escherichia coli was used to analyze the detailed enzymatic properties. The results demonstrated that ScUGT5 specifically transferred a glucosyl moiety to 3-deoxyanthocyanidins in the presence of UDP-glucose, but not to other flavonoid compounds, such as 3-hydroxyanthocyanidins, flavones, flavonols, or flavanones.

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Abbreviations

A5GT:

UDP-glucose:anthocyanin 5-O-glucosyltransferase

ANS:

Anthocyanidin synthase

Ap:

Apigeninidin

Ap5Glc:

Apigeninidin 5-O-glucoside

dA5GT:

UDP-glucose:3-deoxyanthocyanidin 5-O-glucosyltransferase

DFR:

Dihydroflavonol 4-reductase

FNR:

Flavanone 4-reductase

Lt:

Luteolinidin

Lt5Glc:

Luteolinidin 5-O-glucoside

PSPG:

Plant secondary product glycosyltransferase

References

  • Awika JM, Rooney LW, Waniska RD (2004) Properties of 3-deoxyanthocyanins from sorghum. J Agric Food Chem 52:4388–4394

    Article  CAS  PubMed  Google Scholar 

  • Boddu J, Svabek C, Sekhon R, Gevens A, Nicholson RL, Jones AD, Pedersen JF, L. Gustine DL, Chopra S (2004) Expression of a putative flavonoid 3′-hydroxylase in sorghum mesocotyls synthesizing 3-deoxyanthocyanidin phytoalexins. Physiol Mol Plant Pathol 65:101–113

    Article  CAS  Google Scholar 

  • Boddu J, Svabek C, Ibraheem F, Jones AD, Chopra S (2005) Characterization of a deletion allele of a sorghum Myb gene yellow seed1 showing loss of 3-deoxyflavonoids. Plant Sci 169:542–552

    Article  CAS  Google Scholar 

  • Boddu J, Jiang C, Sangar V, Olson T, Peterson T, Chopra S (2006) Comparative structural and functional characterization of sorghum and maize duplications containing orthologous myb transcription regulators of 3-deoxyflavonoid biosynthesis. Plant Mol Biol 60:185–199

    Article  CAS  PubMed  Google Scholar 

  • Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein–dye binding. Anal Biochem 72:248–254

    Article  CAS  PubMed  Google Scholar 

  • Fischer TC, Halbwirth H, Meisel B, Stich K, Forkmann G (2003) Molecular cloning, substrate specificity of the functionally expressed dihydroflavonol 4-reductases from Malus domestica and Pyrus communis cultivars and the consequences for flavonoid metabolism. Arch Biochem Biophys 412:223–230

    Article  CAS  PubMed  Google Scholar 

  • Ford CM, Boss PK, Høj PB (1998) Cloning and characterization of Vitis vinifera UDP-glucose:flavonoid 3-O-glucosyltransferase, a homologue of the enzyme encoded by the maize Bronze-1 locus that may primarily serve to glucosylate anthocyanidins in vivo. J Biol Chem 273:9224–9233

    Article  CAS  PubMed  Google Scholar 

  • Gachon CM, Langlois-Meurinne M, Saindrenan P (2005) Plant secondary metabolism glycosyltransferases: the emerging functional analysis. Trends Plant Sci 10:542–549

    Article  CAS  PubMed  Google Scholar 

  • Grotewold E, Drummond BJ, Bowen B, Peterson T (1994) The myb-homologous P gene controls phlobaphene pigmentation in maize floral organs by directly activating a flavonoid biosynthetic gene subset. Cell 76:543–553

    Article  CAS  PubMed  Google Scholar 

  • Halbwirth H, Fischer TC, Roemmelt S, Spinelli F, Schlangen K, Peterek S, Sabatini E, Messina C, Speakman JB, Andreotti C, Rademacher W, Bazzi C, Costa G, Treutter D, Forkmann G, Stich K (2003a) Induction of antimicrobial 3-deoxyflavonoids in pome fruit trees controls fire blight. Z Naturforsch 58c:765–770

    Google Scholar 

  • Halbwirth H, Martens S, Wienand U, Forkmann G, Stich K (2003b) Biochemical formation of anthocyanins in silk tissue of Zea mays. Plant Sci 164:489–495

    Article  CAS  Google Scholar 

  • Harborne JB (1966) Comparative biochemistry of flavonoids, II: 3-desoxyanthocyanins and their systematic distribution in ferns and gesnerads. Phytochemistry 5:589–600

    Article  CAS  Google Scholar 

  • Harborne JB, Grayer RJ (1988) The anthocyanins. In: Harborne JB (ed) The flavonoids. Chapman and Hall, London, pp 1–20

    Google Scholar 

  • Hirotani M, Kuroda R, Suzuki H, Yoshikawa T (2000) Cloning and expression of UDP-glucose: flavonoid 7-O-glucosyltransferase from hairy root cultures of Scutellaria baicalensis. Planta 210:1006–1013

    CAS  PubMed  Google Scholar 

  • Ibraheem F, Gaffoor I, Chopra S (2010) Flavonoid phytoalexin dependent resistance to anthracnose leaf blight requires a functional yellow seed1 in Sorghum bicolor. Genetics. doi:10.1534/genetics.109.111831

  • Li Y, Baldauf S, Lim EK, Bowles DJ (2001) Phylogenetic analysis of the UDP-glycosyltransferase multigene family of Arabidopsis thaliana. J Biol Chem 276:4338–4343

    Article  CAS  PubMed  Google Scholar 

  • Lo SC, Nicholson RL (1998) Reduction of light-induced anthocyanin accumulation in inoculated sorghum mesocotyls: implications for a compensatory role in the defense response. Plant Physiol 116:979–989

    Article  CAS  PubMed  Google Scholar 

  • Modolo LV, Blount JW, Achnine L, Naoumkina MA, Wang X, Dixon RA (2007) A functional genomics approach to (iso)flavonoid glycosylation in the model legume Medicago truncatula. Plant Mol Biol 64:499–518

    Article  CAS  PubMed  Google Scholar 

  • Nagashima S, Hirotani M, Yoshikawa T (2000) Purification and characterization of UDP-glucuronate: baicalein 7-O-glucuronosyltransferase from Scutellaria baicalensis Georgi. cell suspension cultures. Phytochemistry 53:533–538

    Article  CAS  PubMed  Google Scholar 

  • Nakatsuka T, Sato K, Takahashi H, Yamamura S, Nishihara M (2008) Cloning and characterization of the UDP-glucose:anthocyanin 5-O-glucosyltransferase gene from blue-flowered gentian. J Exp Bot 59:1241–1252

    Article  CAS  PubMed  Google Scholar 

  • Noguchi A, Saito A, Homma Y, Nakao M, Sasaki N, Nishino T, Takahashi S, Nakayama T (2007) A UDP-glucose:isoflavone 7-O-glucosyltransferase from the roots of soybean (Glycine max) seedlings. Purification, gene cloning, phylogenetics, and an implication for an alternative strategy of enzyme catalysis. J Biol Chem 282:23581–23590

    Article  CAS  PubMed  Google Scholar 

  • Noguchi A, Horikawa M, Fukui Y, Fukuchi-Mizutani M, Iuchi-Okada A, Ishiguro M, Kiso Y, Nakayama T, Ono E (2009) Local differentiation of sugar donor specificity of flavonoid glycosyltransferase in Lamiales. Plant Cell 21:1556–1572

    Article  CAS  PubMed  Google Scholar 

  • Ogata J, Kanno Y, Itoh Y, Tsugawa H, Suzuki M (2005) Plant biochemistry: anthocyanin biosynthesis in roses. Nature 435:757–758

    Article  CAS  PubMed  Google Scholar 

  • Ono E, Fukuchi-Mizutani M, Nakamura N, Fukui Y, Yonekura-Sakakibara K, Yamaguchi M, Nakayama T, Tanaka T, Kusumi T, Tanaka Y (2006) Yellow flowers generated by expression of the aurone biosynthetic pathway. Proc Natl Acad Sci USA 103:11075–11080

    Article  CAS  PubMed  Google Scholar 

  • Paquette S, Moller BL, Bak S (2003) On the origin of family 1 plant glycosyltransferases. Phytochemistry 62:399–413

    Article  CAS  PubMed  Google Scholar 

  • Richman A, Swanson A, Humphrey T, Chapman R, McGarvey B, Pocs R, Brandle J (2005) Functional genomics uncovers three glucosyltransferases involved in the synthesis of the major sweet glucosides of Stevia rebaudiana. Plant J 41:56–67

    Article  CAS  PubMed  Google Scholar 

  • Shao H, He X, Achnine L, Blount JW, Dixon RA, Wang X (2005) Crystal structures of a multifunctional triterpene/flavonoid glycosyltransferase from Medicago truncatula. Plant Cell 17:3141–3154

    Article  CAS  PubMed  Google Scholar 

  • Stich K, Forkmann G (1987) Enzymatic synthesis of 4′- and 3′, 4′-hydroxylated flavanones and flavones with flower extracts of Sinningia cardinalis. Z Naturforsch 42c:1193–1199

    Google Scholar 

  • Stich K, Forkmann G (1988) Biosynthesis of 3-deoxyanthocyanins with flower extracts from Sinningia cardinalis. Phytochemistry 27:785–789

    Article  CAS  Google Scholar 

  • Swinny EE, Bloor SJ, Wong H (2000) 1H and 13C NMR assignments for the 3-deoxyanthocyanins, luteolinidin 5-glucoside and apigeninidin 5-glucoside. Magn Reson Chem 38:1031–1033

    Article  CAS  Google Scholar 

  • Taguchi G, Yazawa T, Hayashida N, Okazaki M (2001) Molecular cloning and heterologous expression of novel glucosyltransferases from tobacco cultured cells that have broad substrate specificity and are induced by salicylic acid and auxin. Eur J Biochem 268:4086–4094

    Article  CAS  PubMed  Google Scholar 

  • Thompson JD, Higgins DG, Gibson TJ (1994) Improved sensitivity of profile searches through the use of sequence weights and gap excision. Comput Appl Biosci 10:19–29

    CAS  PubMed  Google Scholar 

  • Vogt T, Grimm R, Strack D (1999) Cloning and expression of a cDNA encoding betanidin 5-O-glucosyltransferase, a betanidin- and flavonoid-specific enzyme with high homology to inducible glucosyltransferases from the Solanaceae. Plant J 19:509–519

    Article  CAS  PubMed  Google Scholar 

  • Winefield CS, Lewis DH, Swinny EE, Zhang H, Arathoon HS, Fischer TC, Halbwirth H, Stich K, Gosch C, Forkmann G, Davies KM (2005) Investigation of the biosynthesis of 3-deoxyanthocyanins in Sinningia cardinalis. Physiol Plant 124:419–430

    Article  CAS  Google Scholar 

  • Yamazaki M, Gong Z, Fukuchi-Mizutani M, Fukui Y, Tanaka Y, Kusumi T, Saito K (1999) Molecular cloning and biochemical characterization of a novel anthocyanin 5-O-glucosyltransferase by mRNA differential display for plant forms regarding anthocyanin. J Biol Chem 274:7405–7411

    Article  CAS  PubMed  Google Scholar 

  • Yamazaki M, Yamagishi E, Gong Z, Fukuchi-Mizutani M, Fukui Y, Tanaka Y, Kusumi T, Yamaguchi M, Saito K (2002) Two flavonoid glucosyltransferases from Petunia hybrida: molecular cloning, biochemical properties and developmentally regulated expression. Plant Mol Biol 48:401–411

    Article  CAS  PubMed  Google Scholar 

  • Yonekura-Sakakibara K, Tohge T, Niida R, Saito K (2007) Identification of a flavonol 7-O-rhamnosyltransferase gene determining flavonoid pattern in Arabidopsis by transcriptome coexpression analysis and reverse genetics. J Biol Chem 282:14932–14941

    Article  CAS  PubMed  Google Scholar 

  • Yonekura-Sakakibara K, Nakayama T, Yamazaki M, Saito K (2009) Modification and stabilization of anthocyanins. In: Gould K, Davies K, Winefield C (eds) Anthocyanins, biosynthesis, functions, and applications. Springer, New York, pp 169–190

    Google Scholar 

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Acknowledgments

The authors thank Prof. Toru Nakayama, Tohoku University, Dr. Nobuhiro Sasaki, Tokyo University of Agriculture and Technology, and Dr. Norimoto Shimada, Kazusa DNA Research Institute for useful suggestions during this study. We also thank Dr. Tatsuya Sawasaki for his technical suggestion on wheat germ cell-free translation system. This research was supported by the Iwate prefecture government.

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Authors and Affiliations

  1. Iwate Biotechnology Research Center, Narita 22-174-4, Kitakami, Iwate, 024-0003, Japan

    Takashi Nakatsuka & Masahiro Nishihara

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  1. Takashi Nakatsuka
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  2. Masahiro Nishihara
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Correspondence to Masahiro Nishihara.

Additional information

The nucleotide sequences reported in this paper have been submitted to the DDBJ/GenBank/EMBL databases with accession numbers AB537178 to AB537182 (ScUGT1 to ScUGT5).

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Nakatsuka, T., Nishihara, M. UDP-glucose:3-deoxyanthocyanidin 5-O-glucosyltransferase from Sinningia cardinalis . Planta 232, 383–392 (2010). https://doi.org/10.1007/s00425-010-1175-0

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  • DOI: https://doi.org/10.1007/s00425-010-1175-0

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