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

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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.

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Acknowledgments

We thank Drs. Jerome Verdier and Lina Gallego-Giraldo for critical reading of the manuscript. This work was supported by the BioEnergy Science Center, a US Department of Energy Bioenergy Research Center supported by the Office of Biological and Environmental Research in the DOE Office of Science.

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Author notes

  1. Luis L. Escamilla-Treviño, Hui Shen & Richard A. Dixon

    Present address: Department of Biological Sciences, University of North Texas, 1155 Union Circle #305220, Denton, TX, 6203-5017, USA

  2. Yanbin Yin

    Present address: Department of Biological Sciences, Northern Illinois University, DeKalb, IL, 60115-2857, USA

Authors and Affiliations

  1. Plant Biology Division, Samuel Roberts Noble Foundation, Ardmore, OK, 73401, USA

    Luis L. Escamilla-Treviño, Hui Shen, Timothy Hernandez & Richard A. Dixon

  2. Department of Biochemistry and Molecular Biology, University of Georgia, Athens, GA, 30602, USA

    Yanbin Yin & Ying Xu

  3. Oak Ridge National Laboratory, BioEnergy Science Center (BESC), Oak Ridge, TN, 37831, USA

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

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  1. Luis L. Escamilla-Treviño
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Correspondence to Richard A. Dixon.

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Accessions numbers

PvHCT1a: AB723827, PvHCT2a: KC696573, PvHCT-Like1: JX845714, PvC3′H1: AB723823, PvC3′H2: AB723824.

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Escamilla-Treviño, L.L., Shen, H., Hernandez, T. et al. Early lignin pathway enzymes and routes to chlorogenic acid in switchgrass (Panicum virgatum L.). Plant Mol Biol 84, 565–576 (2014). https://doi.org/10.1007/s11103-013-0152-y

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