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J. Biol. Chem., Vol. 277, Issue 21, 18272-18280, May 24, 2002
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From the Institute of Biological Chemistry, Washington State
University, Pullman, Washington 99164-6340
Transcriptional profiling of
the phenylpropanoid pathway in Pinus taeda cell suspension
cultures was carried out using quantitative real time PCR analyses of
all known genes involved in the biosynthesis of the two monolignols,
p-coumaryl and coniferyl alcohols (lignin/lignan precursors). When the cells were transferred to a medium containing 8%
sucrose and 20 mM potassium iodide, the
monolignol/phenylpropanoid pathway was induced, and transcript levels
for phenylalanine ammonia lyase, cinnamate 4-hydroxylase,
p-coumarate 3-hydroxylase, 4-coumarate:CoA ligase,
caffeoyl-CoA O-methyltransferase, cinnamoyl-CoA reductase, and cinnamyl alcohol dehydrogenase were coordinately up-regulated. Provision of increasing levels of exogenously supplied Phe to saturating levels (40 mM) to the induction medium resulted
in further up-regulation of their transcript levels in the P. taeda cell cultures; this in turn was accompanied by considerable
increases in both p-coumaryl and coniferyl alcohol
formation and excretion. By contrast, transcript levels for both
cinnamate 4-hydroxylase and p-coumarate 3-hydroxylase were
only slightly up-regulated. These data, when considered together with
metabolic profiling results and genetic manipulation of various plant
species, reveal that carbon allocation to the pathway and its
differential distribution into the two monolignols is controlled by
Phe supply and differential modulation of cinnamate 4-hydroxylase
and p-coumarate 3-hydroxylase activities, respectively. The
coordinated up-regulation of phenylalanine ammonia lyase,
4-coumarate:CoA ligase, caffeoyl-CoA
O-methyltransferase, cinnamoyl-CoA reductase and cinnamyl
alcohol dehydrogenase in the presence of increasing concentrations of
Phe also indicates that these steps are not truly rate-limiting,
because they are modulated according to metabolic demand. Finally, the
transcript profile of a putative acid/ester
O-methyltransferase, proposed as an alternative catalyst
for O-methylation leading to coniferyl alcohol, was not
up-regulated under any of the conditions employed, suggesting that it
is not, in fact, involved in monolignol biosynthesis.
The nucleotide sequence(s) reported in this paper has been submitted to the GenBankTM/EBI Data Bank with accession number(s) AF096998, AY065995, AY064170, and AY064169.
Transcriptional Control of Monolignol Biosynthesis in
Pinus taeda
FACTORS AFFECTING MONOLIGNOL RATIOS AND CARBON ALLOCATION IN
PHENYLPROPANOID METABOLISM*
*
This work was supported in part by United States Department
of Energy Grant DE-FG03-97ER20259, the National Aeronautics and Space
Administration Grant NAG21198, the Lewis B. and Dorothy Cullman and G. Thomas Hargrove Center for Land Plant Adaptation Studies, and the
United States Departments of Energy and Agriculture, National Science
Foundation Plant Biotechnology Research and Training Center.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.
To whom correspondence should be addressed. Tel.: 509-335-2682;
Fax: 509-335-8206; E-mail: lewisn@wsu.edu.
Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.
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