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Suppression of CCR impacts metabolite profile and cell wall composition in Pinus radiata tracheary elements

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Abstract

Suppression of the lignin-related gene cinnamoyl-CoA reductase (CCR) in the Pinus radiata tracheary element (TE) system impacted both the metabolite profile and the cell wall matrix in CCR-RNAi lines. UPLC–MS/MS-based metabolite profiling identified elevated levels of p-coumaroyl hexose, caffeic acid hexoside and ferulic acid hexoside in CCR-RNAi lines, indicating a redirection of metabolite flow within phenylpropanoid metabolism. Dilignols derived from coniferyl alcohol such as G(8-5)G, G(8-O-4)G and isodihydrodehydrodiconiferyl alcohol (IDDDC) were substantially depleted, providing evidence for CCR’s involvement in coniferyl alcohol biosynthesis. Severe CCR suppression almost halved lignin content in TEs based on a depletion of both H-type and G-type lignin, providing evidence for CCR’s involvement in the biosynthesis of both lignin types. 2D-NMR studies revealed minor changes in the H:G-ratio and consequently a largely unchanged interunit linkage distribution in the lignin polymer. However, unusual cell wall components including ferulate and unsaturated fatty acids were identified in TEs by thioacidolysis, pyrolysis-GC/MS and/or 2D-NMR in CCR-RNAi lines, providing new insights into the consequences of CCR suppression in pine. Interestingly, CCR suppression substantially promoted pyrolytic breakdown of cell wall polysaccharides, a phenotype most likely caused by the incorporation of acidic compounds into the cell wall matrix in CCR-RNAi lines.

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Acknowledgments

This work was funded in part by grants C04X0207 and C04X0703 from the New Zealand Ministry of Science and Innovation. Funding for J.R. was from the DOE Great Lakes Bioenergy Research Center (DOE Office of Science BER DE-FC02-07ER64494). Y.T. was supported by a Postdoctoral Fellowship for Research Abroad provided by the Japan Society for the Promotion of Science. G.G. and W.B. thank the Research Foundation Flanders (Grant no. G.0352.05 N). The authors would like to thank Elspeth MacRae for critical reading of this manuscript.

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Authors and Affiliations

  1. Scion, Private Bag 3020, Rotorua, New Zealand

    Armin Wagner, Lorelle Phillips, Heather Flint, Diane Steward, Kirk Torr & Lloyd Donaldson

  2. Department of Biochemistry, University of Wisconsin, Madison, WI, USA

    Yuki Tobimatsu & John Ralph

  3. DOE Great Lakes Bioenergy Research Center, Wisconsin Bioenergy Initiative, Madison, WI, USA

    John Ralph

  4. Department of Plant Systems Biology, VIB, 9052, Ghent, Belgium

    Geert Goeminne & Wout Boerjan

  5. Department of Plant Biotechnology and Bioinformatics, UGhent, 9052, Ghent, Belgium

    Geert Goeminne & Wout Boerjan

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  1. Armin Wagner
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  2. Yuki Tobimatsu
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Correspondence to Armin Wagner.

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Wagner, A., Tobimatsu, Y., Goeminne, G. et al. Suppression of CCR impacts metabolite profile and cell wall composition in Pinus radiata tracheary elements. Plant Mol Biol 81, 105–117 (2013). https://doi.org/10.1007/s11103-012-9985-z

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  • DOI: https://doi.org/10.1007/s11103-012-9985-z

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