Plant Molecular Biology

, Volume 76, Issue 6, pp 475–488 | Cite as

Functional characterization of two new members of the caffeoyl CoA O-methyltransferase-like gene family from Vanilla planifolia reveals a new class of plastid-localized O-methyltransferases

  • Thomas Widiez
  • Thomas G. Hartman
  • Nativ Dudai
  • Qing Yan
  • Michael Lawton
  • Daphna Havkin-Frenkel
  • Faith C. Belanger
Article

Abstract

Caffeoyl CoA O-methyltransferases (OMTs) have been characterized from numerous plant species and have been demonstrated to be involved in lignin biosynthesis. Higher plant species are known to have additional caffeoyl CoA OMT-like genes, which have not been well characterized. Here, we identified two new caffeoyl CoA OMT-like genes by screening a cDNA library from specialized hair cells of pods of the orchid Vanilla planifolia. Characterization of the corresponding two enzymes, designated Vp-OMT4 and Vp-OMT5, revealed that in vitro both enzymes preferred as a substrate the flavone tricetin, yet their sequences and phylogenetic relationships to other enzymes are distinct from each other. Quantitative analysis of gene expression indicated a dramatic tissue-specific expression pattern for Vp-OMT4, which was highly expressed in the hair cells of the developing pod, the likely location of vanillin biosynthesis. Although Vp-OMT4 had a lower activity with the proposed vanillin precursor, 3,4-dihydroxybenzaldehyde, than with tricetin, the tissue specificity of expression suggests it may be a candidate for an enzyme involved in vanillin biosynthesis. In contrast, the Vp-OMT5 gene was mainly expressed in leaf tissue and only marginally expressed in pod hair cells. Phylogenetic analysis suggests Vp-OMT5 evolved from a cyanobacterial enzyme and it clustered within a clade in which the sequences from eukaryotic species had predicted chloroplast transit peptides. Transient expression of a GFP-fusion in tobacco demonstrated that Vp-OMT5 was localized in the plastids. This is the first flavonoid OMT demonstrated to be targeted to the plastids.

Keywords

Vanilla planifolia Flavone O-methyltransferase (OMT) Plastid localization Vanillin 

References

  1. Agati G, Tattini M (2010) Multiple functional roles of flavonoids in photoprotection. New Phytol 186:786–793PubMedCrossRefGoogle Scholar
  2. Agati G, Matteini P, Goti A, Tattini M (2007) Chloroplast-located flavonoids can scavenge singlet oxygen. New Phytol 174:77–89PubMedCrossRefGoogle Scholar
  3. Bhushan S, Kuhn C, Berglund A-K, Roth C, Glaser E (2006) The role of the N-terminal domain of chloroplast targeting peptides in organellar protein import and miss-sorting. FEBS Lett 580:3966–3972PubMedCrossRefGoogle Scholar
  4. Bouvier R, Linka N, Isner J-C, Mutterer J, Weber APM, Camara B (2006) Arabidopsis SAMT1 defines a plastid transporter regulating plastid biogenesis and plant development. Plant Cell 18:3088–3105PubMedCrossRefGoogle Scholar
  5. Dixon RA (2011) Vanillin biosynthesis–not as simple as it seems? In: Havkin-Frenkel D, Belanger FC (eds) Handbook of Vanilla science and technology. Wiley-Blackwell, Oxford, pp 292–298Google Scholar
  6. Do C-T, Pollet B, Thevenin J, Sibout R, Denoue D, Barriere Y, Lapierre C, Jouanin L (2007) Both caffeoyl Coenzyme A 3-O-methyltransferase 1 and caffeic acid O-methyltransferase 1 are involved in redundant functions for lignin, flavonoids and sinapoyl malate biosynthesis in Arabidopsis. Planta 226:1117–1129PubMedCrossRefGoogle Scholar
  7. Dunlevy JD, Soote KL, Perkins MV, Dennis EG, Keyzers RA, Kalua CM, Boss PK (2010) Two O-methyltransferases involved in the biosynthesis of methoxypyrazines: grape-derived aroma compounds important to wine flavour. Plant Mol Biol 74:77–89PubMedCrossRefGoogle Scholar
  8. Earley K, Haag JR, Pontes O, Opper K, Juehne T, Song K, Pikaard CS (2006) Gateway-compatible vectors for plant functional genomics and proteomics. Plant J 45:616–629PubMedCrossRefGoogle Scholar
  9. Emanuelsson O, Nielsen H, von Heijne G (1999) ChloroP, a neural network-based method for predicting chloroplast transit peptides and their cleavage sites. Protein Sci 8:9780984CrossRefGoogle Scholar
  10. Emanuelsson O, Brunak S, von Heijne G, Nielsen H (2007) Locating proteins in the cell using TargetP, SignalP, and related tools. Nat Protoc 2:953–971PubMedCrossRefGoogle Scholar
  11. Fellenberg C, Milkowski C, Hause B, Lange P-R, Bottcher C, Schmidt J, Vogt T (2008) Tapetum-specific location of a cation-dependent O-methyltransferase in Arabidopsis thaliana. Plant J 56:132–145PubMedCrossRefGoogle Scholar
  12. Ferrer J-L, Zubieta C, Dixon RA, Noel JP (2005) Crystal structures of alfalfa caffeoyl coenzyme A 3-O-methyltransferase. Plant Physiol 137:1009–1017PubMedCrossRefGoogle Scholar
  13. Goujon T, Sibout R, Pollet B, Maba B, Nussaume L, Bechtold N, Lu F, Ralph J, Mila I, Barriere Y, Lapierre C, Jouanin L (2003) A new Arabidopsis thaliana mutant deficient in the expression of O-methyltransferase impacts lignins and sinapoyl esters. Plant Mol Biol 51:973–989PubMedCrossRefGoogle Scholar
  14. Grienenberger E, Besseu S, Geoffroy P, Debayle D, Heintz D, Lapierre C, Pollet B, Heitz T, Legrand M (2009) A BAHD acyltransferase is expressed in the tapetum of Arabidopsis anthers and is involved in the synthesis of hydroxycinnamoyl spermidines. Plant J 58:246–259PubMedCrossRefGoogle Scholar
  15. Guo D, Chen F, Inou K, Blount JW, Dixon RA (2001) Downregulation of caffeic acid 3-O-methyltransferase and caffeoyl CoA 3-O-methyltransferase in transgenic alfalfa: impacts on lignin structure and implications for the biosynthesis of G and S lignin. Plant Cell 13:73–88PubMedCrossRefGoogle Scholar
  16. Hamberger B, Ellis M, Friedman M, de Azevedo Souz C, Barbazuk B, Douglas CJ (2007) Genome-wide analyses of phenylpropanoid-related genes in Populus trichocarpa, Arabidopsis thaliana, and Oryza sativa: the Populus lignin toolbox and conservation and diversification of angiosperm gene families. Can J Bot 85:1182–1201CrossRefGoogle Scholar
  17. Hugueney P, Provenzano S, Verries C, Ferrandino A, Meudec E, Batelli G, Merdinoglu D, Cheynier V, Schubert A, Ageorges A (2009) A novel cation-dependent O-methyltransferase involved in anthocyanin methylation in grapevine. Plant Physiol 150:2057–2070PubMedCrossRefGoogle Scholar
  18. Humphreys JM, Chapple C (2002) Rewriting the lignin roadmap. Curr Opin Plant Biol 5:224–229PubMedCrossRefGoogle Scholar
  19. Ibdah M, Zhang X-H, Schmidt J, Vogt T (2003) A novel Mg2+-dependent O-methyltransferase in the phenylpropanoid metabolism of Mesembryanthemum crystallinum. J Biol Chem 278:43961–43972PubMedCrossRefGoogle Scholar
  20. Joel DM, French JC, Graft N, Kourteva G, Dixon RA, Havkin-Frenkel D (2003) A hairy tissue produces vanillin. Israel J Plant Sci 51:157–159CrossRefGoogle Scholar
  21. Joshi CP, Chiang VL (1998) Conserved sequence motifs in plant S-adenosyl-l-methionine-dependent methyltransferases. Plant Mol Biol 37:663–674PubMedCrossRefGoogle Scholar
  22. Konieczny MPJ, Benz I, Hollinderbaumer B, Beinke C, Niederweis M, Schmidt MA (2001) Modular organization of the AIDA autotransporter translocator: the N-terminal β1-domain is surface-exposed and stabilizes the transmembrane β2-domain. Antonie van Leeuwenhoek 80:19–34PubMedCrossRefGoogle Scholar
  23. Kopycki JG, Stubbs MT, Brandt W, Hagemann M, Porzel A, Schmidt J, Schliemann W, Zenk MH, Vogt T (2008) Functional and structural characterization of a cation-dependent O-methyltransferae from the cyanobacterium Synechocystis sp. strain PCC 6803. J Biol Chem 283:20888–20896PubMedCrossRefGoogle Scholar
  24. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685PubMedCrossRefGoogle Scholar
  25. Lam KC, Ibrahim RK, Behdad B, Dayanandan S (2007) Structure, function, and evolution of plant O-methyltransferases. Genome 50:1001–1013PubMedCrossRefGoogle Scholar
  26. Lee YJ, Kim BG, Chong Y, Lim Y, Ahn J-H (2008) Cation dependent O-methyltransferases from rice. Planta 227:641–647PubMedCrossRefGoogle Scholar
  27. Li HM, Rotter D, Hartman TG, Pak FE, Havkin-Frenkel D, Belanger FC (2006) Evolution of novel O-methyltransferases from the Vanilla planifolia caffeic acid O-methyltransferase. Plant Mol Biol 61:537–552PubMedCrossRefGoogle Scholar
  28. Ligrone R, Carafa A, Duckett JG, Renzaglia KS, Ruel K (2008) Immunocytochemical detection of lignin-related epitopes in cell walls in bryophytes and the charalean alga Nitella. Plant Syst Evol 270:257–272CrossRefGoogle Scholar
  29. Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. Methods 25:402–408PubMedCrossRefGoogle Scholar
  30. Lucker J, Martens S, Lund ST (2010) Characterization of a Vitis vinifera cv. Cabernet Sauvignon 3′, 5′-O-methyltransferase showing strong preference for anthocyanins and glycosylated flavonols. Phytochemistry 71:1474–1484PubMedCrossRefGoogle Scholar
  31. Lunkenbein S, Salentijn EMJ, Coiner HA, Boone MJ, Krens FA, Schwab W (2006) Up-and down-regulation of Fragaria x ananassa O-methyltransferase: impacts on furanone and phenylpropanoid metabolism. J Exp Bot 57:2445–2453PubMedCrossRefGoogle Scholar
  32. Pak FE, Gropper S, Dai WD, Havkin-Frenkel D, Belanger FC (2004) Characterization of a multifunctional methyltransferase from the orchid Vanilla planifolia. Plant Cell Rep 22:959–966PubMedCrossRefGoogle Scholar
  33. Pakusch A-E, Matern U (1991) Kinetic characterization of caffeoyl-coenzyme A-specific 3-O-methyltransferase from elicited parsley cell suspensions. Plant Physiol 96:327–330PubMedCrossRefGoogle Scholar
  34. Paterson AH, Bowers JE, Bruggmann R, Dubchak I et al (2009) The Sorghum bicolor genome and the diversification of grasses. Nature 457:551–556PubMedCrossRefGoogle Scholar
  35. Raes J, Rohde A, Christensen JH, Van de Peer Y, Boerjan W (2003) Genome-wide characterization of the lignification toolbox in Arabidopsis. Plant Physiol 133:1051–1071PubMedCrossRefGoogle Scholar
  36. Rausher MD (2006) The evolution of flavonoids and their genes. In: Groteworld E (ed) The science of flavonoids. Springer, New York, pp 175–211CrossRefGoogle Scholar
  37. Rensing SA, Lang D, Zimmer AD, Terry A et al (2008) The Physcomitrella genome reveals evolutionary insights into the conquest of land by plants. Science 319:64–69PubMedCrossRefGoogle Scholar
  38. Rozema J, Bjorn LO, Bornman JF, Gaberscik A, Hader D-P, Trost T, Germ M, Klisch M, Groniger A, Sinha RP, Lebert M, He Y-Y, Buffoni-Hall R, de Bakker NVJ, van de Staaij J, Meijkamp BB (2002) The role of UV-B radiation in aquatic and terrestrial ecosystems: an experimental and functional analysis of the evolution of UV-absorbing compounds. J Photochem Photobiol B Biol 66:2–12CrossRefGoogle Scholar
  39. Swofford L (2002) PAUP*: phylogenetic analysis using parsimony (*and other methods). Version 4. Sinauer Associates, SunderlandGoogle Scholar
  40. The French-Italian Public Consortium for Grapevine Genome Characterization (2007) The grapevine genome sequence suggests ancestral hexaploidization in major angiosperm phyla. Nature 449:463–468CrossRefGoogle Scholar
  41. Thompson JD, Gibson TJ, Plewniak F, Jeanmougin F, Higgins DG (1997) The CLUSTAL-X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Res 25:4876–4882PubMedCrossRefGoogle Scholar
  42. Tsai C-J, Harding SA, Tschaplinski TJ, Lindroth RL, Yuan Y (2006) Genome-wide analysis of the structural genes regulating defense phenylpropanoid metabolism in Populus. New Phytol 172:47–62PubMedCrossRefGoogle Scholar
  43. 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
  44. Wein M, Lavid N, Lunkenbein S, Lewinsohn E, Schwab W, Kaldenhoff R (2002) 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. Plant J 31:755–765PubMedCrossRefGoogle Scholar
  45. Wollenweber E, Dorr M (2008) Occurrence and distribution of the flavone tricetin and its methyl derivatives as free aglycones. Nat Prod Commun 3:1293–1298Google Scholar
  46. Wu K, Chung L, Revill WP, Katz L, Reeves CD (2000) The FK520 gene cluster of Streptomyces hygroscopicus var. ascomyceticus (ATCC 14891) contains genes for biosynthesis of unusual polyketide extender units. Gene 251:81–90PubMedCrossRefGoogle Scholar
  47. Zhong R, Morrison WH III, Negrel J, Ye Z-H (1998) Dual methylation pathways in lignin biosynthesis. Plant Cell 10:2033–2045PubMedCrossRefGoogle Scholar
  48. Zhong R, Morrison WH III, Himmelsbach DS, Poole FL II, Ye Z-H (2000) Essential role of caffeoyl coenzyme A O-methyltransferase in lignin biosynthesis in woody poplar plants. Plant Physiol 124:563–577PubMedCrossRefGoogle Scholar
  49. Zhou J-M, Ibrahim RK (2010) Tricin–a potential multifunctional nutraceutical. Phytochem Rev 9:413–424CrossRefGoogle Scholar
  50. Zhou J-M, Gold ND, Martin VJJ, Wollenweber E, Ibrahim RK (2006) Sequential O-methylation of tricetin by a single gene product in wheat. Biochim Biophys Acta 1760:1115–1124PubMedGoogle Scholar
  51. Zimmer R, Gibbins AMV (1997) Construction and characterization of a large-fragment chicken bacterial artificial chromosome library. Genomics 42:217–226PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2011

Authors and Affiliations

  • Thomas Widiez
    • 1
  • Thomas G. Hartman
    • 2
  • Nativ Dudai
    • 3
  • Qing Yan
    • 1
  • Michael Lawton
    • 1
  • Daphna Havkin-Frenkel
    • 1
  • Faith C. Belanger
    • 1
  1. 1.Department of Plant Biology and Pathology, School of Environmental and Biological SciencesRutgers UniversityNew BrunswickUSA
  2. 2.Center for Advanced Food Technology, School of Environmental and Biological SciencesRutgers UniversityNew BrunswickUSA
  3. 3.Aromatic, Medicinal and Spice Crops Unit, Newe Ya’ar Research CenterAgricultural Research OrganizationRamat YishayIsrael

Personalised recommendations