, Volume 236, Issue 3, pp 839–849 | Cite as

Characterization of a flavonol 3-O-methyltransferase in the trichomes of the wild tomato species Solanum habrochaites

  • Adam Schmidt
  • Chao Li
  • A. Daniel Jones
  • Eran Pichersky
Original Article


The glandular trichomes of the wild tomato species Solanum habrochaites accumulate the polymethylated flavonol aglycones, 3,7,3′-O-methyl myricetin, 3,7,3′,5′-O-methyl myricetin, and 3,7,3′,4′,5′-O-methyl myricetin. Partially methylated flavonol aglycones and partially methylated flavonol glycones containing a methyl group at the 3 position have been previously reported from a variety of plants. The 3-O-methyltransferase (3-OMT) activity has been previously partially purified from plants, but a gene transcript encoding an enzyme capable of methylating flavonols at the 3 position has not yet been identified, nor have been such proteins purified and characterized. We previously identified two gene transcripts expressed in the glandular trichomes of S. habrochaites and showed that they encode enzymes capable of methylating myricetin at the 3′ and 5′ and the 7 and 4′ positions, respectively. Here we report the identification of gene transcripts expressed in S. lycopersicum (cultivated tomato) and in S. habrochaites glandular trichomes that encode enzymes capable of methylating myricetin, and its partially methylated derivatives exclusively at the 3 position. The S. habrochaites gene transcript is preferentially expressed in the glandular trichomes and it encodes a protein with high similarity to the S. habrochaites, 3′/5′ O-methyltransferase which is also present in glandular trichomes. Phylogenic analysis suggests that the 3-OMT activity has probably evolved from an ancestral 3′/5′ methyltransferase activity. The discovery and characterization of 3-OMT provides a more complete picture of the series of reactions leading to highly methylated myricetin compounds in S. habrochaites glandular trichomes.


Plant biochemistry Specialized metabolism Glandular trichomes Solanaceae Flavonoids Myricetin 

Supplementary material

425_2012_1676_MOESM1_ESM.ppt (220 kb)
Supplementary material 1 (PPT 220 kb)


  1. Bombarely A, Menda N, Tecle IY, Buels RM, Strickler S, Fischer-York T, Pujar A, Leto J, Gosselin J, Mueller LA (2011) The sol genomics network ( growing tomatoes using Perl. Nucleic Acids Res 39:D1149–D1155PubMedCrossRefGoogle Scholar
  2. Braca A, Bilia AR, Mendez J, Morelli I (2001) Myricetin glycosides from Licania densiflora. Fitoterapia 72:182–185PubMedCrossRefGoogle Scholar
  3. Buer CS, Imin N, Djordjevic MA (2010) Flavonoids: new roles for old molecules. J Integr Plant Biol 52:98–111PubMedCrossRefGoogle Scholar
  4. Castillo-Munoz N, Gomez-Alonso S, Garcia-Romero E, Hermosin-Gutierrez I (2010) Flavonol profiles of Vitis vinifera white grape cultivars. J Food Comp Anal 23:699–705CrossRefGoogle Scholar
  5. Collins FW, Deluca V, Ibrahim RK, Voirin B, Jay M (1981) Polymethylated flavonols of Chrysosplenium americanum. 1. identification and enzymatic synthesis. J Res Biosciences 36:730–736Google Scholar
  6. Cooper JE (2004) Multiple responses of rhizobia to flavonoids during legume root infection. Adv Botanical Res Incorporating Adv Plant Pathol 41(41):1–62CrossRefGoogle Scholar
  7. D’Auria JC, Chen F, Pichersky E (2002) Characterization of an acyltransferase capable of synthesizing benzylbenzoate and other volatile esters in flowers and damaged leaves of Clarkia breweri. Plant Physiol 130:466–476PubMedCrossRefGoogle Scholar
  8. De Luca V, Ibrahim RK (1985) Enzymatic synthesis of polymethylated flavonols in Chrysosplenium americanum. 2. substrate interaction and product inhibition studies of flavonol 3, 6, and 4′-O-methyltransferases. Arch Biochem Biophys 238:606–618PubMedCrossRefGoogle Scholar
  9. Dudareva N, Pichersky E (2008) Metabolic engineering of plant volatiles. Curr Opin Biotechnol 19:181–189PubMedCrossRefGoogle Scholar
  10. Felsenstein J (1985) Confidence limits on phylogenies—an approach using the bootstrap. Evolution 39:783–791CrossRefGoogle Scholar
  11. Ferrer JL, Austin MB, Stewart C Jr, Noe JP (2008) Structure and function of enzymes involved in the biosynthesis of phenylpropanoids. Plant Physiol Biochem 46:356–370PubMedCrossRefGoogle Scholar
  12. Gerats AGM, Wallroth M, Donkerkoopman W, Groot SPC, Schram AW (1983) The genetic control of the enzyme UDP-glucose 3-O-flavonoid glucosyltransferase in flowers of Petunia hybrida. Theoretical Appl Genetics 65:349–352Google Scholar
  13. Greenham J, Harbone JB, Williams CA (2003) Identification of lipophilic flavones and flavonols by comparative HPLC, TLC and UV spectral analysis. Phytochem Anal 14:100–118PubMedCrossRefGoogle Scholar
  14. Griesser M, Vitzthum F, Fink B, Bellido ML, Raasch C, Munoz-Blanco J, Schwab W (2008) Multi-substrate flavonol O-glucosyltransferases from strawberry (Fragariaxananassa) achene and receptacle. J Exp Bot 59:2611–2625PubMedCrossRefGoogle Scholar
  15. Huang TS, Anzellotti D, Dedaldechamp F, Ibrahim RK (2004) Partial purification, kinetic analysis, and amino acid sequence information of a flavonol 3-O-methyltransferase from Serratula tinctoria. Plant Physiol 134:1366–1376PubMedCrossRefGoogle Scholar
  16. Ibrahim RK (2005) A forty-year journey in plant research: original contributions to flavonoid biochemistry. Canadian J Bot 83:433–450CrossRefGoogle Scholar
  17. Ibrahim RK, Deluca V, Khouri H, Latchinian L, Brisson L, Charest PM (1987) Enzymology and compartmentation of polymethylated flavonol glucosides in Chrysosplenium americanum. Phytochem 26:1237–1245CrossRefGoogle Scholar
  18. Jez JM, Noel JP (2000) Mechanism of chalcone synthase—pK(a) of the catalytic cysteine and the role of the conserved histidine in a plant polyketide synthase. J Biol Chem 275:39640–39646PubMedCrossRefGoogle Scholar
  19. Kumari GNK, Rao LJM, Rao NSP (1984) Myricetin methyl ethers from Solanum Pubescens. Phytochem 23:2701–2702CrossRefGoogle Scholar
  20. Lee TH, Qiu F, Waller GR, Chou CH (2000) Three new flavonol galloylglycosides from leaves of Acacia confusa. J Nat Prod 63:710–712PubMedCrossRefGoogle Scholar
  21. Martens S, Preuss A, Matern U (2010) Multifunctional flavonoid dioxygenases: flavonol and anthocyanin biosynthesis in Arabidopsis thaliana L. Phytochem 71:1040–1049CrossRefGoogle Scholar
  22. McDowell RT, Kapteyn J, Schmidt A, Li C, Kang J-H, Descour A, Shi F, Larson M, Schilmiller A, An L, Howe GA, Jones AD, Pichersky E, Soderlund CA, Gang DR (2011) Comparative functional genomic analysis of Solanum glandular trichome types. Plant Physiol 155:524–539PubMedCrossRefGoogle Scholar
  23. Modolo LV, Li L, Pan H, Blount JW, Dixon RA, Wang X (2009) Crystal structures of glycosyltransferase UGT78G1 reveal the molecular basis for glycosylation and deglycosylation of (iso)flavonoids. J Mol Biol 392:1292–1302PubMedCrossRefGoogle Scholar
  24. Motta LB, Kraus JE, Salatino A, Salatino MLF (2005) Distribution of metabolites in galled and non-galled foliar tissues of Tibouchina pulchra. Biochem Syst Ecol 33:971–981CrossRefGoogle Scholar
  25. Ono E, Homma Y, Horikawa M, Kunikane-Doi S, Imai H, Takahashi S, Kawai Y, Ishiguro M, Fukui Y, Nakayama T (2010) Functional differentiation of the glycosyltransferases that contribute to the chemical diversity of bioactive flavonol glycosides in grapevines (Vitis vinifera). Plant Cell 22:2856–2871PubMedCrossRefGoogle Scholar
  26. Owens DK, McIntosh CA (2009) Identification, recombinant expression, and biochemical characterization of a flavonol 3-O-glucosyltransferase clone from Citrus paradisi. Phytochem 70:1382–1391CrossRefGoogle Scholar
  27. Pichersky E, Lewinsohn E (2011) Convergent evolution in plant specialized metabolism. Annu Rev Plant Biol 62(62):549–566PubMedCrossRefGoogle Scholar
  28. Pollastri S, Tattini M (2011) Flavonols: old compounds for old roles. Ann Bot 108:1225–1233PubMedCrossRefGoogle Scholar
  29. Pourcel L, Routaboul JM, Kerhoas L, Caboche M, Lepiniec L, Debeaujon I (2005) Transparent testa10 encodes a laccase-like enzyme involved in oxidative polymerization of flavonoids in arabidopsis seed coat. Plant Cell 17:2966–2980PubMedCrossRefGoogle Scholar
  30. Prescott AG, John P (1996) Dioxygenases: molecular structure and role in plant metabolism. Annual Rev Plant Physiol Plant Mol Biol 47:245–271CrossRefGoogle Scholar
  31. Ringli C, Bigler L, Kuhn BM, Leiber R-M, Diet A, Santelia D, Frey B, Pollmann S, Klein M (2008) The modified flavonol glycosylation profile in the Arabidopsis rol1 mutants results in alterations in plant growth and cell shape formation. Plant Cell 20:1470–1481PubMedCrossRefGoogle Scholar
  32. Roda AL, Oldham NJ, Svatos A, Baldwin IT (2003) Allometric analysis of the induced flavonols on the leaf surface of wild tobacco (Nicotiana attenuata). Phytochem 62:527–536CrossRefGoogle Scholar
  33. Saitou N, Nei M (1987) The neighbor-joining method—a new method for reconstructing phylogenetic trees. Mol Biol Evol 4:406–425PubMedGoogle Scholar
  34. Schmidt A, Li C, Shi F, Jones AD, Pichersky E (2011) Polymethylated myricetin in trichomes of the wild tomato species Solanum habrochaites and characterization of trichome-specific 3′/5′- and 7/4′-myricetin o-methyltransferases. Plant Physiol 155:1999–2009PubMedCrossRefGoogle Scholar
  35. Schmittgen TD, Livak KJ (2008) Analyzing real-time PCR data by the comparative Ct method. Nat Protoc 3:1101–1108PubMedCrossRefGoogle Scholar
  36. Stevens JF, Hart HT, Wollenweber E (1995) The systematic and evolutionary significance of exudate flavonoids in Aeonium. Phytochem 39:805–813CrossRefGoogle Scholar
  37. Stevens JF, Hart H, Elema ET, Bolck A (1996) Flavonoid variation in Eurasian Sedum and Sempervivum. Phytochem 41:503–512CrossRefGoogle Scholar
  38. Tamura K, Dudley J, Nei M, Kumar S (2007) MEGA4: molecular evolutionary genetics analysis (MEGA) software version 4.0. Mol Biol Evol 24:1596–1599PubMedCrossRefGoogle Scholar
  39. Tohge T, Ramos MS, Nunes-Nesi A, Mutwil M, Giavalisco P, Steinhauser D, Schellenberg M, Willmitzer L, Persson S, Martinoia E, Fernie AR (2011) Toward the storage metabolome: profiling the barley vacuole. Plant Physiol 157:1469–1482PubMedCrossRefGoogle Scholar
  40. Treutter D (2005) Significance of flavonoids in plant resistance and enhancement of their biosynthesis. Plant Biol 7:581–591PubMedCrossRefGoogle Scholar
  41. Vogt T, Jones P (2000) Glycosyltransferases in plant natural product synthesis: characterization of a supergene family. Trends Plant Sci 5:380–386PubMedCrossRefGoogle Scholar
  42. Wang J, Pichersky E (1999) Identification of specific residues involved in substrate discrimination in two plant O-methyltransferases. Arch Biochem Biophys 368:172–180PubMedCrossRefGoogle Scholar
  43. Winkel-Shirley B (2001) Flavonoid biosynthesis. a colorful model for genetics, biochemistry, cell biology, and biotechnology. Plant Physiol 126:485–493PubMedCrossRefGoogle Scholar
  44. Zuckerkandl E, Pauling L (1965) Evolutionary divergence and convergence in proteins. In: Bryson V, Vogel HJ (eds) Evolving genes and proteins. Academic Press, New York, pp 97–166Google Scholar

Copyright information

© Springer-Verlag 2012

Authors and Affiliations

  • Adam Schmidt
    • 1
  • Chao Li
    • 3
  • A. Daniel Jones
    • 3
    • 4
  • Eran Pichersky
    • 2
  1. 1.Institute of Biological ChemistryWashington State UniversityPullmanUSA
  2. 2.Department of Molecular, Cellular, and Developmental BiologyUniversity of MichiganAnn ArborUSA
  3. 3.Department of ChemistryMichigan State UniversityEast LansingUSA
  4. 4.Department of Biochemistry and Molecular BiologyMichigan State UniversityEast LansingUSA

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