Plant Molecular Biology

, Volume 84, Issue 4–5, pp 565–576 | Cite as

Early lignin pathway enzymes and routes to chlorogenic acid in switchgrass (Panicum virgatum L.)

  • Luis L. Escamilla-Treviño
  • Hui Shen
  • Timothy Hernandez
  • Yanbin Yin
  • Ying Xu
  • Richard A. Dixon
Article

Abstract

Studying lignin biosynthesis in Panicum virgatum (switchgrass) has provided a basis for generating plants with reduced lignin content and increased saccharification efficiency. Chlorogenic acid (CGA, caffeoyl quinate) is the major soluble phenolic compound in switchgrass, and the lignin and CGA biosynthetic pathways potentially share intermediates and enzymes. The enzyme hydroxycinnamoyl-CoA: quinate hydroxycinnamoyltransferase (HQT) is responsible for CGA biosynthesis in tobacco, tomato and globe artichoke, but there are no close orthologs of HQT in switchgrass or in other monocotyledonous plants with complete genome sequences. We examined available transcriptomic databases for genes encoding enzymes potentially involved in CGA biosynthesis in switchgrass. The protein products of two hydroxycinnamoyl-CoA shikimate/quinate hydroxycinnamoyltransferase (HCT) genes (PvHCT1a and PvHCT2a), closely related to lignin pathway HCTs from other species, were characterized biochemically and exhibited the expected HCT activity, preferring shikimic acid as acyl acceptor. We also characterized two switchgrass coumaroyl shikimate 3′-hydroxylase (C3′H) enzymes (PvC3′H1 and PvC3′H2); both of these cytochrome P450s had the capacity to hydroxylate 4-coumaroyl shikimate or 4-coumaroyl quinate to generate caffeoyl shikimate or CGA. Another switchgrass hydroxycinnamoyl transferase, PvHCT-Like1, is phylogenetically distant from HCTs or HQTs, but exhibits HQT activity, preferring quinic acid as acyl acceptor, and could therefore function in CGA biosynthesis. The biochemical features of the recombinant enzymes, the presence of the corresponding activities in plant protein extracts, and the expression patterns of the corresponding genes, suggest preferred routes to CGA in switchgrass.

Keywords

Phenylpropanoid pathway Lignin Flavonoids Chlorogenic acid 

Supplementary material

11103_2013_152_MOESM1_ESM.docx (553 kb)
Supplementary material 1 (DOCX 552 kb)

References

  1. Ahola V, Aittokallio T, Vihinen M, Uusipaikka E (2006) A statistical score for assessing the quality of multiple sequence alignments. BMC Bioinformat 7:484CrossRefGoogle Scholar
  2. Chen F, Dixon RA (2007) Lignin modification improves fermentable sugar yields for biofuel production. Nat Biotechnol 25:759–761PubMedCrossRefGoogle Scholar
  3. Chen F, Srinivasa Reddy MS, Temple S, Jackson L, Shadle G, Dixon RA (2006) Multi-site genetic modulation of monolignol biosynthesis suggests new routes for formation of syringyl lignin and wall-bound ferulic acid in alfalfa (Medicago sativa L.). Plant J 48:113–124PubMedCrossRefGoogle Scholar
  4. Comino C, Lanteri S, Portis E, Acquadro A, Romani A, Hehn A, Larbat R, Bourgaud F (2007) Isolation and functional characterization of a cDNA coding a hydroxycinnamoyltransferase involved in phenylpropanoid biosynthesis in Cynara cardunculus L. BMC Plant Biol 7:14PubMedCentralPubMedCrossRefGoogle Scholar
  5. Comino C, Hehn A, Moglia A, Menin B, Bourgaud F, Lanteri S, Portis E (2009) The isolation and mapping of a novel hydroxycinnamoyltransferase in the globe artichoke chlorogenic acid pathway. BMC Plant Biol 9:30PubMedCentralPubMedCrossRefGoogle Scholar
  6. Davison BH, Drescher SR, Tuskan GA, Davis MF, Nghiem NP (2006) Variation of S/G ratio and lignin content in a Populus family influences the release of xylose by dilute acid hydrolysis. Appl Biochem Biotechnol 129–132:427–435PubMedCrossRefGoogle Scholar
  7. Escamilla-Trevino LL, Shen H, Uppalapati SR, Ray T, Tang Y, Hernandez T, Yin Y, Xu Y, Dixon RA (2010) Switchgrass (Panicum virgatum) possesses a divergent family of cinnamoyl CoA reductases with distinct biochemical properties. New Phytol 185:143–155PubMedCrossRefGoogle Scholar
  8. Fu C, Mielenz JR, Xiao X, Ge Y, Hamilton CY, Rodriguez M Jr, Chen F, Foston M, Ragauskas A, Bouton J, Dixon RA, Wang Z-Y (2011) Genetic manipulation of lignin reduces recalcitrance and improves ethanol production from switchgrass. Proc Natl Acad Sci USA 108:3803–3808Google Scholar
  9. Gallego-Giraldo L, Escamilla-Trevino L, Jackson LA, Dixon RA (2011) Salicylic acid mediates the reduced growth of lignin down-regulated plants. Proc Natl Acad Sci USA 108:20814–20819PubMedCrossRefGoogle Scholar
  10. Grabber JH (2005) How do lignin composition, structure, and cross-linking affect degradability? A review of cell wall model studies. Crop Sci 45:820–831CrossRefGoogle Scholar
  11. Hoffmann L, Maury S, Martz F, Geoffroy P, Legrand M (2003) Purification, cloning, and properties of an acyltransferase controlling shikimate and quinate ester intermediates in phenylpropanoid metabolism. J Biol Chem 278:95–103PubMedCrossRefGoogle Scholar
  12. Hoffmann L, Besseau S, Geoffroy P, Ritzenthaler C, Meyer D, Lapierre C, Pollet B, Legrand M (2004) Silencing of hydroxycinnamoyl-coenzyme A shikimate/quinate hydroxycinnamoyltransferase affects phenylpropanoid biosynthesis. Plant Cell 16:1446–1465PubMedCentralPubMedCrossRefGoogle Scholar
  13. Jackson LA, Shadle GL, Zhou R, Nakashima J, Chen F, Dixon RA (2008) Improving saccharification efficiency of alfalfa stems through modification of the terminal stages of monolignol biosynthesis. Bioenergy Res 1:180–192CrossRefGoogle Scholar
  14. Katoh K, Kuma K, Toh H, Miyata T (2005) MAFFT version 5: improvement in accuracy of multiple sequence alignment. Nucleic Acids Res 33:511–518PubMedCentralPubMedCrossRefGoogle Scholar
  15. Lepelley M, Cheminade G, Tremillon N, Simkin A, Caillet V, McCarthy J (2007) Chlorogenic acid synthesis in coffee: an analysis of CGA content and real-time RT-PCR expression of HCT, HQT, C3H1, and CCoAOMT1 genes during grain development in C. canephora. Plant Sci 172:978–996CrossRefGoogle Scholar
  16. Liu CJ, Huhman D, Sumner LW, Dixon RA (2003) Regiospecific hydroxylation of isoflavones by cytochrome P450 81E enzymes from Medicago truncatula. Plant J 36:471–484PubMedCrossRefGoogle Scholar
  17. Mahesh V, Million-Rousseau R, Ullmann P, Chabrillange N, Bustamante J, Mondolot L, Morant M, Noirot M, Hamon S, de Kochko A (2007) Functional characterization of two p-coumaroyl ester 3′-hydroxylase genes from coffee tree: evidence of a candidate for chlorogenic acid biosynthesis. Plant Mol Biol 64:145–159PubMedCrossRefGoogle Scholar
  18. McLaughlin SB, Adams Kszos L (2005) Development of switchgrass (Panicum virgatum) as a bioenergy feedstock in the United States. Biomass Bioenergy 28:515–535CrossRefGoogle Scholar
  19. Moglia A, Comino C, Portis E, Acquadro A, De Vos RCH, Beekwilder J, Lanteri S (2009) Isolation and mapping of a C3′H gene (CYP98A49) from globe artichoke, and its expression upon UV–C stress. Plant Cell Rep 28:963–974PubMedCrossRefGoogle Scholar
  20. Moore KJ, Moser LE, Vogel KP, Waller SS, Johnson BE, Pedersen JF (1991) Describing and quantifying growth stages of perennial forage grasses. Agron J 83:1073–1077CrossRefGoogle Scholar
  21. Morant M, Schoch GA, Ullmann P, Ertunҫ T, Little D, Olsen CE, Petersen M, Negrel J, Werck-Reichhart D (2007) Catalytic activity, duplication and evolution of the CYP98 cytochrome P450 family in wheat. Plant Mol Biol 63:1–19PubMedCrossRefGoogle Scholar
  22. Niggeweg R, Michael AJ, Martin C (2004) Engineering plants with increased levels of the antioxidant chlorogenic acid. Nat Biotechnol 22:746–754PubMedCrossRefGoogle Scholar
  23. Nuin PA, Wang Z, Tillier ER (2006) The accuracy of several multiple sequence alignment programs for proteins. BMC Bioinformat 7:471CrossRefGoogle Scholar
  24. Pompon D, Louerat B, Bronine A, Urban P (1996) Yeast expression of animal and plant P450 s in optimized redox environments. Meth Enzymol 272:51–64PubMedCrossRefGoogle Scholar
  25. Price MN, Dehal PS, Arkin AP (2010) FastTree 2-approximately maximum-likelihood trees for large alignments. PLoS ONE 5:e9490PubMedCentralPubMedCrossRefGoogle Scholar
  26. Ralph J, Akiyama T, Kim H, Lu F, Schatz PF, Marita JM, Ralph SA, Reddy MS, Chen F, Dixon RA (2006) Effects of coumarate 3′-hydroxylase down-regulation on lignin structure. J Biol Chem 281:8843–8853PubMedCrossRefGoogle Scholar
  27. Reddy MS, Chen F, Shadle G, Jackson L, Aljoe H, Dixon RA (2005) Targeted down-regulation of cytochrome P450 enzymes for forage quality improvement in alfalfa (Medicago sativa L.). Proc Natl Acad Sci USA 102:16573–16578PubMedCrossRefGoogle Scholar
  28. Schmer MR, Vogel KP, Mitchell RB, Perrin RK (2008) Net energy of cellulosic ethanol from switchgrass. Proc Natl Acad Sci USA 105:464–469PubMedCrossRefGoogle Scholar
  29. Schoch G, Goepfert S, Morant M, Hehn A, Meyer D, Ullmann P, Werck-Reichhart D (2001) CYP98A3 from Arabidopsis thaliana is a 3′-hydroxylase of phenolic esters, a missing link in the phenylpropanoid pathway. J Biol Chem 276:36566–36574PubMedCrossRefGoogle Scholar
  30. Shen H, Fu CX, Xiao XR, Ray T, Tang YH, Wang ZY, Chen F (2009) Developmental control of lignification in stems of lowland switchgrass variety Alamo and the effects on saccharification efficiency. Bioenergy Res 2:233–245CrossRefGoogle Scholar
  31. Shen H, He X, Poovaiah CR, Wuddineh WA, Ma J, Mann DGJ, Wang H, Jackson L, Tang Y, Neal Stewart C Jr (2011) Functional characterization of the switchgrass (Panicum virgatum) R2R3-MYB transcription factor PvMYB4 for improvement of lignocellulosic feedstocks. New Phytol 193:121–136PubMedCrossRefGoogle Scholar
  32. Shen H, Mazarei M, Rudis MR, Tang YH, Hisano H, Jackson L, Li G, Hernandez T, Chen F, C. Stewart Jr N, Wang Z, Dixon RA (2013). A genomic approach to deciphering pathways for lignin biosynthesis in switchgrass (Panicum virgatum L.). Plant Cell, in revisionGoogle Scholar
  33. Sonnante G, D’Amore R, Blanco E, Pierri CL, De Palma M, Luo J, Tucci M, Martin C (2010) Novel hydroxycinnamoyl-coenzyme A quinate transferase genes from artichoke are involved in the synthesis of chlorogenic acid. Plant Physiol 153:1224–1238PubMedCentralPubMedCrossRefGoogle Scholar
  34. Stockigt J, Zenk MH (1975) Chemical syntheses and properties of hydroxycinnamoyl-Coenzyme A derivatives. Z Naturforsch [C] 30:352–358Google Scholar
  35. Talukder K (2006) Low-lignin wood–a case study. Nat Biotechnol 24:395–396PubMedCrossRefGoogle Scholar
  36. van der Rest B, Danoun S, Boudet AM, Rochange SF (2006) Down-regulation of cinnamoyl-CoA reductase in tomato (Solanum lycopersicum L.) induces dramatic changes in soluble phenolic pools. J Exp Bot 57:1399–1411PubMedCrossRefGoogle Scholar
  37. Vanholme R, Cesarino I, Rataj K, Xiao Y, Sundin L, Goeminne G, Kim H, Cross J, Morreel K, Araujo P, Welsh L, Haustraete J, McClellan C, Vanholme B, Ralph J, Simpson GG, Halpin C, Boerjan W (2013) Caffeoyl shikimate esterase (CSE) is an enzyme in the lignin biosynthetic pathway. Science 341:1103–1106PubMedCrossRefGoogle Scholar
  38. Villegas RJ, Kojima M (1986) Purification and characterization of hydroxycinnamoyl d-glucose: quinate hydroxycinnamoyl transferase in the root of sweet potato, Ipomoea batatas Lam. J Biol Chem 261:8729–8733PubMedGoogle Scholar
  39. Wullschleger SD, Davis EB, Borsuk ME, Gunderson CA, Lynd LR (2010) Biomass production in switchgrass across the United States: database description and determinants of yield. Agron J 102:1158–1168CrossRefGoogle Scholar
  40. Xu B, Escamilla-Trevino LL, Noppadon S, Shen Z, Shen H, Percival Zhang YH, Dixon RA, Zhao B (2011) Silencing of 4-coumarate: coenzyme A ligase in switchgrass leads to reduced lignin content and improved fermentable sugar yields for biofuel production. New Phytol 92:611–625CrossRefGoogle Scholar
  41. Yuan JS, Tiller KH, Al-Ahmad H, Stewart NR, Stewart CN (2008) Plants to power: bioenergy to fuel the future. Trends Plant Sci 13:421–429PubMedCrossRefGoogle Scholar
  42. Zhang J-Y, Lee Y-C, Torres-Jerez I, Wang M, Yin Y, Chou W-C, He J, Shen H, Srivastava AC, Pennacchio C, Lindquist E, Grimwood J, Schmutz J, Xu Y, Sharma M, Sharma R, Bartley LE, Ronald PC, Saha MC, Dixon RA, Tang Y, Udvardi MK (2013) Development of an integrated transcript sequence database and a gene expression atlas for gene discovery and analysis in switchgrass (Panicum virgatum L.). Plant J 74:160–173PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2013

Authors and Affiliations

  • Luis L. Escamilla-Treviño
    • 1
    • 3
    • 4
  • Hui Shen
    • 1
    • 3
    • 4
  • Timothy Hernandez
    • 1
    • 3
  • Yanbin Yin
    • 2
    • 3
    • 5
  • Ying Xu
    • 2
    • 3
  • Richard A. Dixon
    • 1
    • 3
    • 4
  1. 1.Plant Biology DivisionSamuel Roberts Noble FoundationArdmoreUSA
  2. 2.Department of Biochemistry and Molecular BiologyUniversity of GeorgiaAthensUSA
  3. 3.Oak Ridge National LaboratoryBioEnergy Science Center (BESC)Oak RidgeUSA
  4. 4.Department of Biological SciencesUniversity of North TexasDentonUSA
  5. 5.Department of Biological SciencesNorthern Illinois UniversityDeKalbUSA

Personalised recommendations