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Originally published In Press as doi:10.1074/jbc.M006915200 on August 8, 2000

J. Biol. Chem., Vol. 275, Issue 47, 36899-36909, November 24, 2000
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Modifications in Lignin and Accumulation of Phenolic Glucosides in Poplar Xylem upon Down-regulation of Caffeoyl-Coenzyme A O-Methyltransferase, an Enzyme Involved in Lignin Biosynthesis*

Hugo Meyermansa, Kris Morreelab, Catherine Lapierrec, Brigitte Polletc, André De Bruynd, Roger Bussone, Piet Herdewijne, Bart Devreesefg, Jozef Van Beeumenf, Jane M. Maritahi, John Ralphhi, Cuiying Chena, Bart Burggraevea, Marc Van Montagua, Eric Messensa, and Wout Boerjanaj

From the a Vakgroep Moleculaire Genetica & Departement Plantengenetica, Vlaams Interuniversitair Instituut voor Biotechnologie, Universiteit Gent, B-9000 Gent, Belgium, c Laboratoire de Chimie Biologique, Institut National Agronomique Paris-Grignon, F-78850 Thiverval-Grignon, France, d Vakgroep Organische Scheikunde, Universiteit Gent, B-9000 Gent, Belgium, e Laboratorium Medicinale Scheikunde, Katholieke Universiteit Leuven, B-3000 Leuven, Belgium, f Vakgroep Biochemie, Fysiologie en Microbiologie, Universiteit Gent, B-9000 Gent, Belgium, and h U.S. Dairy Forage Research Center, U.S. Department of Agriculture-Agricultural Research Service, Madison, Wisconsin 53706-1108

Caffeoyl-coenzyme A O-methyltransferase (CCoAOMT) methylates, in vitro, caffeoyl-CoA and 5-hydroxyferuloyl-CoA, two possible precursors in monolignol biosynthesis in vivo. To clarify the in vivo role of CCoAOMT in lignin biosynthesis, transgenic poplars with 10% residual CCoAOMT protein levels in the stem xylem were generated. Upon analysis of the xylem, the affected transgenic lines had a 12% reduced Klason lignin content, an 11% increased syringyl/guaiacyl ratio in the noncondensed lignin fraction, and an increase in lignin-attached p-hydroxybenzoate but otherwise a lignin composition similar to that of wild type. Stem xylem of the CCoAOMT-down-regulated lines had a pink-red coloration, which coincided with an enhanced fluorescence of mature vessel cell walls. The reduced production of CCoAOMT caused an accumulation of O3-beta -D-glucopyranosyl-caffeic acid, O4-beta -D-glucopyranosyl-vanillic acid, and O4-beta -D-glucopyranosyl-sinapic acid (GSA), as authenticated by 1H NMR. Feeding experiments showed that O3-beta -D-glucopyranosyl-caffeic acid and GSA are storage or detoxification products of caffeic and sinapic acid, respectively. The observation that down-regulation of CCoAOMT decreases lignin amount whereas GSA accumulates to 10% of soluble phenolics indicates that endogenously produced sinapic acid is not a major precursor in syringyl lignin biosynthesis. Our in vivo results support the recently obtained in vitro enzymatic data that suggest that the route from caffeic acid to sinapic acid is not used for lignin biosynthesis.

* This work was supported by Flemish government Geconcerteerde Onderzoeksacties Grant 12052293, Fund for Scientific Research Flanders Grants FWO 1.5.515.98N and G.0040.00, and European Union Grant FAIR-PL95424.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

b Recipient of a predoctoral fellowship from Vlaams Instituut voor de Bevordering van het Wetenschappelijk-Technologisch Onderzoek in de Industrie.

g Postdoctoral fellow of the Fund for Scientific Research (Flanders).

i Supported by the U.S. Department of Agriculture-Agricultural Research Service and in part through U.S. Department of Agriculture-National Resources Inventory Grant 99-2351 (Improved Utilization of Wood and Wood Fiber Section).

j To whom correspondence should be addressed: Dept. Plantengenetica, Vlaams Interuniversitair Instituut voor Biotechnologie, Universiteit Gent, K. L. Ledeganckstraat 35, B-9000 Gent, Belgium. Tel.: 32-9-2645202; Fax: 32-9-2645349; woboe@gengenp.rug.ac.be.

Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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