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Phenylpropanoid Biosynthesis: Thomas Vogt
Phenylpropanoid Biosynthesis: Thomas Vogt
Phenylpropanoid Biosynthesis
Thomas Vogt1
Leibniz-Institute of Plant Biochemistry, Department of Secondary Metabolism, Weinberg 3, D-06120 Halle (Saale), Germany
ABSTRACT The general phenylpropanoid metabolism generates an enormous array of secondary metabolites based on
the few intermediates of the shikimate pathway as the core unit. The resulting hydroxycinnamic acids and esters are am-
plified in several cascades by a combination of reductases, oxygenases, and transferases to result in an organ and devel-
opmentally specific pattern of metabolites, characteristic for each plant species. During the last decade, methodology
Key words: Phenylpropanoid; biosynthetic pathway; flavonoid; anthocyanin; tannin; coumarin; volatiles; lignin.
obsolute (Hartmann, 2007). The diversity and plasticity of the ª The Author 2009. Published by the Molecular Plant Shanghai Editorial
Office in association with Oxford University Press on behalf of CSPP and
resulting natural products, more specifically the phenylpropa-
IPPE, SIBS, CAS.
noid profile, are then achieved by a set of enzymes organized doi: 10.1093/mp/ssp106, Advance Access publication 24 December 2009
usually in superfamilies, like oxygenases, ligases, oxidoreduc- Received 30 September 2009; accepted 19 November 2009
Vogt d Phenylpropanoid Biosynthesis | 3
CHO
H OH COOH
H OH + H2C
CH2O P O P
Erythose-4-Phosphat PEP
COOH
CH3O
Coumarins HO OH O CH2
OH
Isochorismate
Shikimate
OH
HO O
HO O
COOH
COOH OH
OH O
OH OH O NH3+
PAL
S CoA
OH OH COOH
O O
HO
OH
HO Cinnamate Benzoic acid
p-Coumaroyl CoA
OH
Stilbenes
OH
O glucose
HO O OH CH3O
O
OH
O CH3O HO
OH HO O
CH3O
Aurones
OH HO Phenylpropanoid esters
OH
Catechin Lignans Phenylpropenes
+
Cutin Acylated Polyamines
Lignin
Suberin
Sporopollein
Proanthocyanidins
the various branches of the general phenypropanoid metab- encoding cytochrome P450s in Arabidopsis, 4-hydroxylation
olism. A crystal structure became available for a codon-opti- of trans-cinnamate to 4-coumarate is encoded by a single gene
mized, heterologously expressed enzyme from Petroselinum only, At2g30490, encoding CYP73A5. As expected, mutations
crispum (Ritter and Schulz, 2004). The structural data are con- in At2g30490 severely affect growth and development of the
sistent with the unusual deamination reaction, which is inde- plants and result in the accumulation of the quite unusual cin-
pendent of any co-factor. The required electrophilic prosthetic namoylmalate in leaves (Schilmiller et al., 2009).
group, 4-methylidene-imidazole-5-one, is auto-catalytically The subsequent step, encoded by the small gene family of
formed. These data also provide evidence for the requirements 4-CL ligases, channels the resulting aromatic CoA-esters to dif-
of antioxidative SH-protective reagent, like dithiothreitol ferent biosynthetic pathways. 4-coumaroyl CoA probably rep-
(DTT), for full enzyme activity in vitro, since an oxidized resents the most important branchpoint within the central
DTT-derivative was detected in the electron density map of phenylpropanoid biosynthesis in plants. It is either the direct
the crystal, when DTT was present into the crystallization precursor for flavonoid or H-lignin biosynthesis, or is fed into
buffer. Replacing the characteristic amino acid His89 of the ac- the production of increasingly methoxylated guaiacyl (G)- and
tive site of TAL in bacteria by phenylalanine, a characteristic syringyl (S)-monolignols. A decisive step is initiated by the en-
et al., 2004) or specifically in potato tubers by overexpressing tions of the plant. Partly due to the substrate specificity of
the StMtf1-MYB-transcription factor (Rommens et al., 2009). most 4CL-ligases earlier in the pathway, sinapoyl CoA is usually
Diversification of this small subset of phenylpropanoids like considered not to be a direct precursor for lignin monomer
coumaroyl- and caffeoyl CoA esters is achieved by an array of production—this may, however, depend on the species inves-
subsequent oxygenases, reductases, and transferases and tigated (Hamada et al., 2004). Comparison of annuals and per-
results in the observed complexity and heterogeneity of plant ennials with respect to lignin biosynthesis has revealed an
phenylpropanoid metabolite profiles. apparent functional redundancy of most participating genes
and enzymes. Although CAD is considered a key enzymatic
step in monolignol biosynthesis, a detailed analysis of 17
LIGNIN CAD encoding genes in Arabidopsis revealed that most inser-
Besides cellulose, lignin is the most prominent polymer on tion lines of individual CAD-genes only had a marginal effect
Earth. Lignin is nearly exclusively based on phenylpropanoid on the overall lignin composition (Kim et al., 2004). This sug-
units derived from the oxidative polymerization of hydroxycin- gests at least some functional redundancy among these genes
namoyl alcohol derivatives. As early as 1960, the contribution in case of Arabidopsis and single CADs are not rate-limiting.
Arabidopsis metabolome (Huang et al., 2009). Decoration of biosynthesis, where hydroxycinnamic acids are activated by
the various hydroxylated monolignol precursors in Arabidopsis CoA-esters, in sinapic ester metabolism, 1-O-acyl-glucose esters
is performed either by cation-dependent CCoAOMT1, re- serve as the energy-rich metabolites during seed maturation
quired for the 3-O-methylation of caffeoyl-CoA, or by a cation- and germination (Mock and Strack, 1992). The corresponding
independent caffeic acid OMT (COMT), which is involved in glucosyltransferases have been investigated in molecular de-
the formation of sinapoyl residues and S-lignin from the tail (Milkowski et al., 2000; Sinlapadech et al., 2007; Meissner
5-hydroxyferuloyl derivatives (Davin et al., 2008). The role of et al., 2008). Two transferases, sinapoylglucose:choline sina-
CCoAOMT1, originally considered to be only associated with poyltransferase (SCT) and sinapoylglucose:malate sinapoyl-
monolignol formation, has changed considerably in the last transferase (SMT), are involved in the accumulation of
couple of years (Do et al., 2007; Kai et al., 2008; Fellenberg sinapine in the seed or sinapoylmalate in the seedlings, respec-
et al., 2009). Its involvement in various pathways, including tively. Compared with the presumed ancestors, the serine car-
sinapoylmalate formation, scopoletin biosynthesis, and meth- boxypeptidases (SCPs), the SMT from Arabidopsis, has
ylation of acylspermidine precursors, has proven its central maintained the catalytic triad, Ser–His–Asp (Milkowski and
role in the modification of phenylpropanoids. Engineering Strack, 2004; Stehle et al., 2009). A single Asp/Glu substitution
disputed since then. The recent identification of phenylpropa- coumaric acid in lignin biosynthesis (Schoch et al., 2001).
noid polyamine conjugates in different organs of Arabidopsis The new, pathway-specific CYP98A8 and its paralog CYP98A9
wild-type and insertion lines by LC–MS analysis was accompa- belong to a monophyletic CYP98A clade and are duplications
nied by simultaneous identification of specific genes and of an ancestor of CYP98A3, which evolved rapidly via retropo-
enzymes in the acyl conjugate formation (Böttcher et al., sition and positive Darwinian selection (Matsuno et al., 2009).
2008; Matsuda et al., 2008; Luo et al., 2009). The rapid Knockout mutants of CYP98A8 display a shift in the major ac-
annotation of several genes and enzymes involved in the ylated spermidine conjugate pattern, from 5-hydroxyferuloyl
biosynthesis of these conjugates illustrates the combined to feruloyl conjugates. Likewise, AtTSM1, a cation-dependent
power of Arabidopsis knockout and knockdown insertion lines OMT with significant similarity to CCoAOMT1 in lignin biosyn-
with sensitive UPLC–MS analyses (Fellenberg et al., 2008; thesis, catalyzes the final step in acyl spermidine conjugate for-
Grienenberger et al., 2009). In the tapetum of Arabidopsis mation. The enzyme methylates a single 5-hydroxyferuloyl
flower buds, a new pathway for the biosynthesis of acylated moiety and yields N,N’-bis(5-hydroxyferuloyl)-N’’ sinapoyl sper-
spermidine conjugates was established. Hydroxycinnamic midine (Fellenberg et al., 2008). CCoAOMT1 and potentially
acids are transferred to spermidine by a BAHD-like spermidine CYP98A3 play additional roles earlier in methylation and
ANTHOCYANINS, AURONES,
(ISO)FLAVONOIDS, AND STILBENES
Due to ease of visualization and control of mutants and
genetic imbalances, anthocyanin biosynthesis was one of
the first branches of the general phenylpropanoid metabo-
lism, for which biosynthetic enzymes and corresponding tran-
scription factors were identified and has been reviewed
extensively (Allan et al., 2008). The key enzymatic step of
the whole pathway, the formation of anthocyanidins via
(2R-3R)-dihydroflavonols and oxidation of (2R, 3S, 4S)-
leucoanthocyanidins by anthocyanidin synthase (ANS) has
recently been questioned due the apparent flavonol syn-
thase activity of Arabidopsis ANS and the identification of
(+)-catechin as a substrate of Gerbera hybrida ANS (Wellmann
Figure 4. A Simplified Version of the Biosynthetic Pathway Result- et al., 2006). In a monomeric state, anthocyanins never display
ing in Acylated Spermidine Conjugates in flower buds Is Based on a blue color. Therefore, investigation of blue color formation
Recent Data by Fellenberg et al. (2008, 2009), Grienenberger et al. of anthocyanins either by pH-shift, metal complex formation,
(2009), and Matsuno et al. (2009). modification of the core unit by additional hydroxylation,
The exact sequence of the initial hydroxylation and Werner meth- methylation, or acylation and subsequent molecular stacking
ylation steps by CYP98A3 and CCoAOMT1, the subsequent hydrox-
has been a major focus of anthocyanin research in the last de-
ylation by Cyp98A9 and the transferase reaction by SHT is not
resolved yet. Sequential steps catalyzed by CYP98A9 and AtTSM1 cade (Yoshida et al., 2009). This research was partly driven by
are established. AtTSM1, localized exclusively in the tapetum, the cut-flower industry to develop blue varieties of those flow-
may serve as a tapetal marker in young flower buds of Arabidopsis. ers, which naturally do not exhibit this phenotype, like roses or
Vogt d Phenylpropanoid Biosynthesis | 9
carnations. During the process of developing a bluish tone, im- et al., 2009). UGT89C1 was linked to the late steps in flavo-
portant contributions to anthocyanin biosynthesis have been nol biosynthesis based on transcript and metabolite
made in terms of structural and regulatory genes. It was shown pattern and therefore correctly identified as Arabidopsis 7-
that roses, in contrast to most other plants, have a unique bi- O-rhamnosyltransferase (7RhaT) (Yonekura-Sakakibara et al.,
functional glucosyltransferase that catalyses the glycosylation 2007).
of both the 3- and the 5-OH group (Ogata et al., 2005). How- Several other late steps in the formation of the flavonoid
ever, they are devoid of a 5#-hydroxylase activity, which partly skeleton have only been poorly investigated. B-ring hydroxyl-
explains the lack of any natural bluish color. Expression of the ation at positions 6 and 8 of the flavonoid skeleton is limited to
Viola spp. 3#, 5#-hydroxylase and functional replacement of a few plant families only, like the Asteraceae. For Chrysanthe-
the endogenous rose dihydroflavonol reductase (DFR) for an mum segetum, Halbwirth and Stich (2006) identified a new
orthologous enzyme from Iris hollandica resulted in plants type of 8-monohydroxylase dependent on both NADPH and
that nearly exclusively accumulated delphinidin in the petals FAD, and was, therefore, clearly different from classical cyto-
and displayed a bluish, slightly purple tone (Katsumoto et al., chrome P450s. Obviously, there is a lack of enzymatic and ge-
2007). Myricetin, in addition to quercetin, was detected as netic information for this as well as other exotic plants.
transcription factors has been the topic of many reports, such the 4#- hydroxyl group of 2,7,4#-trihydroxyisoflavanone at
as Stracke et al. (2007), and several reviews (Broun, 2005; Allan an initial step of the pathway as well as towards the 3-hydroxy
et al., 2008). The complex pattern of factors regulating antho- group of the pterocarpan 6a-hydroxymaackiain resulting in
cyanin formation has recently been extended by Matsui et al. the formation of pisatin has been identified and structurally
(2008) and Zhu et al. (2009). They discovered that single repeat characterized (Liu et al., 2006). Careful analysis of both sub-
R3 Myb transcription factors may also repress anthocyanin for- strates revealed their close to identical three-dimensional
mation, probably by competing with R2R3-Mybs for the bind- structure. A single enzyme able to convert two metabolites
ing sites of bHLH proteins, thereby down-regulating structural of the same pathway suggests that substrate promiscuity does
genes and counteracting, such as the Myb-like PAP1/2 en- not only provide evolutionary advantages in response to
hancer of anthocyanin formation in Arabidopsis (Gonzales novel, exogenous phytoalexins, but may also be maintained
et al., 2008). The bHLH-domain of the maize R-gene, partici- to serve those rare cases of metabolic serendipity. This situa-
pating in anthocyanin formation, links flavonoid formation tion is somewhat different from the methylation step in
to histone modification by interacting with an EMSY-like fac- acyl-spermidine formation in Arabidopsis flowers, where not
tor, which is involved in chromatin repair (Hernandez et al., CCoAOMT1, but a similar cation-dependent OMT (AtTSM1)
transcriptional regulators R2R3 MYB-type proteins (TT2), 2009). From the sum of data, it is evident that several path-
bHLH-factors (TT8), and WD40-repeat proteins (TTG1), which ways for benzoic acid biosynthesis may co-exist in a single
act synergistically. Overexpression of those transcription fac- plant and relative fluxes from one or the other pathway
tors may create improved crop varieties with enhanced proan- may be controlled by hormones or abiotic stressors. Feeding
thocyanidin contents (Baudry et al., 2004; Mellway et al., 2009; deuterium-labeled phenylalanine showed that metabolic
Terrier et al., 2009). The reported health-promoting activities fluxes of benzaldehyde formation of the non b-oxidative
of 3-O-esterified proanthocyanidins like epigallocatechin-gal- pathway from cinnamate to benzaldehyde to benzoic acid
late (EGCG) (Rezai-Zadeh et al., 2005), a major metabolite in was about twice that of the b-oxidative pathway involving
green tea leaves, can be hampered by a low bioavailability benzoyl-CoA in P. hybrida petals (Boatright et al., 2004).
upon modification of the core structure by sulfatation, meth- The 2-hydroxyl derivative of benzoic acid, salicylic acid (SA),
ylation, and glucuronylation (Henning et al., 2008). has long been known as a key component of plant innate immu-
Similar to the proanthocyanidins (non-hydrolyzable tan- nity (Lee and Raskin, 1999; Nawrath and Metreux, 1999) and also
nins), the hydrolyzable tannins, like galloyl-glucose esters plays a central role in systemic-acquired resistance (SAR) (Vlot
and ellagitannins, characterized by oxidative coupling of et al., 2008). Biochemical evidence for a direct pathway from
O SCoA O SCoA O SCoA netic resources and metabolic diversity of plants with exotic
2-oxoglutarate
dependent phenylpropanoid-derived compounds are worth exploring
dioxygenase Rotation
Feruloyl-CoA to detect enzymes with unusual specificities for biotechnolog-
OH O
6' F6’H1 6' H ical or molecular farming approaches.
MeO MeO MeO
OH OH OH
O
MIXED POLYMERS, SUBERIN, CUTIN,
O SCoA AND SPOROPOLLENIN
O SCoA
MeO
O The contribution of the general phenylpropanoid metabolism
OH
SCoA towards the biosynthesis of mixed polymers is a challenging
O O
task for structural biochemists and physiologists. The complex-
MeO MeO ity of the pathways and robustness of cutin, suberin, or sporo-
CoAS OH OH
pollenin towards degradation requires harsh oxidative
Sporopollenin, the major part of the pollen exine, appar- and function should also consider in more detail the following
ently contains similar building blocks compared to cutin and key aspects:
suberin, namely phenylpropanoids and hydroxylated fatty
(1) A more detailed investigation of transport and shuttling of
acids. In contrast to cutin and suberin this polymer is even more
metabolites across membranes and cells.
resistant to chemical degradation, although advances in chem-
(2) Elucidation and identification of enzyme complexes and
ical analysis have enabled the complete identification and sep-
metabolon formation to understand how several pathways
aration of its constituents (Dominguez et al., 1999). Based on
may co-exist but are independently regulated.
labeling experiments, the presence of phenylpropanoid resi-
(3) A thorough understanding of the molecular mechanisms
dues is generally accepted (Schulze-Osthoff and Wiermann,
resulting in the formation of unusual phenylpropanoids in me-
1987). The mechanical strength and durability of the pollen
dicinal and exotic plants in a continuous search for new metab-
exine are probably due to a substantial amount of ether-
olites and new pathways.
linkages between fatty acid and/or phenylpropanoids. Conclu-
sive data on the assembly and the exact contribution of New analytical and biophysical methods, such as the implemen-
the phenylpropanoid conjugates are virtually non-existent. tation and application of nanotechnology to solve biological
Anterola, A.M., and Lewis, N.G. (2002). Trends in lignin modifica- Böttcher, C., vRöpenack-Lahaye, E., Schmidt, J., Schmotz, C.,
tion: a comprehensive analysis of the effect of genetic manipu- Neumann, S., Scheel, D., and Clemens, S. (2008). Metabolome
lations/mutations on lignification and vascular integrity. analysis of biosynthetic mutants reveals diversity of metabolic
Phytochemistry. 61, 221–292. changes and allows identification of a large number of new com-
Bais, H.P., Vepachedu, R., Gilroy, S., Callaway, R.M., and pounds in A. thaliana. Plant Physiol. 147, 2107–2120.
Vivanco, J.M. (2003). Allelopathy and exotic plant invasion: from Boudet, A.M. (2007). Evolution and current status of phenolic com-
molecules and genes in species interaction. Science. 301, pounds. Phytochemistry. 68, 2722–2735.
1377–1380. Boudet, A.M., Kajita, S., Grima-Pettenatti, J., and Goffner, D. (2003).
Barbehenn, R.V., Jones, C.-P., Hagerman, A.K., Karonen, M., and Lignin and lignocellulosics: a better control of synthesis for few
Salminen, J.-P. (2006). Ellagitannins have greater oxidative activ- and improved uses. Trends Pl. Sci. 8, 576–581.
ities than condensed tannins and galloyl glucose at high pH: po- Bowles, D., Lim, E.K., Poppenberger, B., and Vaijstij, F.E. (2006). Gly-
tential impact on caterpillars. J. Chem. Ecol. 32, 2253–2267. cosyltransferases of small lipophilic molecules. Ann. Rev. Plant
Barkman, T.J., Martins, T.R., Sutton, E., and Stout, J.T. (2007). Pos- Biol. 57, 567–597.
itive selection for single amino acid change promotes substrate Brand, S., Hölscher, D., Schierhorn, A., Svatos, A., Schröter, J., and
De Bolt, S., et al. (2009). Mutations in UDP-glucose sterol glucosyl- Franke, R., and Schreiber, L. (2007). Suberin: a biopolyester forming
transferase cause transparent testa phenotype and suberization apoplastic plant interfaces. Curr. Opin. Pl. Biol. 10, 252–259.
defects in seeds. Plant Physiol. 151, 78–87. o, L.F.R., Kombrink, E., and Dias, A.C.P. (2009).
Franklin, G., Conceica
Dexter, R., Qualley, A., Kish, C.M., Ma, C.J., Koeduka, T., Xanthone production in Hypericum performation cells provides
Nagegowda, D.A., Dudareva, N., Pichersky, E., and Clark, D. antioxidant and antimicrobial protection upon biotic stress. Phy-
(2007). Characterization of a Petunia acetyltransferase involved tochemistry. 70, 60–68.
in the biosynthesis of the floral volatile isoeugenol. Plant J. 49, Fraser, C.M., Thompson, M.G., Shirley, A.M., Ralph, J.,
265–275. Schönherr, J.A., Sinlapadech, T., Hall, M.C., and Chapple, C.
Dixit, R., Cyr, R., and Gilroy, S. (2006). Use of intrinsic fluorescent (2007). Related Arabidopsis serine carboxypeptidase-like sina-
proteins for plant cell imaging. Plant J. 45, 599–615. poylglucose: acyltransferases display distinct but overlapping
Dixon, R. (1999). The isoflavonoids: biochemsitry, molecular biology substrate specificities. Plant Physiol. 144, 1986–1999.
and biological functions. In Comprehensive Natural Products Gális, I., Šimek, P., Narisawa, T., Sasaki, M., Horiguchi, T., Fukuda, H.,
Chemistry, Sankawa, U., ed. (Amsterdam: Elsevier), pp. 773–823. and Matsuoka, K. (2006). A novel R2R3 MYB transcription factor
Dixon, R.A., and Reddy, M.S.S. (2003). Biosynthesis of monolignols: NtMYBJS1 is a methyl jasmonate-dependent regulator of
recruitment of an EMSY-related factor. Proc. Natl Acad. Sci. U S A. Katsumoto, Y., et al. (2007). Engineering of the rose flavonoid bio-
104, 17222–17227. synthetic pathway successfully generated blue-hued flowers ac-
Herrmann, K. (1995). The shikimate pathway as an entry to aromatic cumulating delphinidin. Plant Cell. Rep. 48, 1589–1600.
secondary metabolism. Plant Physiol. 107, 7–12. Katsuyama, Y., Kita, T., and Horinouchi, S. (2009). Identification and
Herrmann, K., and Weaver, L.M. (1999). The shikimate pathway. characterization of multiple curcumin synthases from the herb
Ann. Rev. Plant Physiol. Plant Mol Biol. 50, 472–503. Curcuma longa. FEBS Lett. 583, 2799–2803.
Hoffmann, L., Besseau, S., Geoffrey, P., Ritzenthaler, C., Meyer, D., Katsuyama, Y., Matsuzawa, M., Funa, N., and Horinuchi, S. (2007).
Lapierre, C., Pollet, B., and Legrand, M. (2004). Silencing of In vitro synthesis of curcuminoids by type III polyketide synthase
hydroxycinnamoyl-coenzyme A shikikmate/quinate hydroxycin- from rice. J. Biol Chem. 282, 37702–37709.
namoyltransferase affects phenylpropanoid biosynthesis. Plant Kim, S.J., Kim, M.R., Bedgar, D.L., Moinuddin, S.G.A., Cardenas, C.L.,
Cell. 16, 1446–1465. Davin, L.B., and Lewis, N. (2004). Functional reclassification of
Horiuchi, J., Badri, D.V., Kimball, B.A., Negre, F., Dudareva, N., the putative cinnamyl alcohol dehydrogenase multigene family
Paschke, M.W., and Vivanca, J.M. (2007). The floral volatile in Arabidopsis. Proc. Natl Acad. Sci. U S A. 101, 1455–1460.
methyl benzoate from snapdragon (Anthirrhinum majus) Kita, T., Imai, S., Sawasa, H., Kumagai, H., and Seto, H. (2008). The
the evolution of plant defence response. Plant Cell. 18, direction of the phenylpropanoid pathway to feruloylmalate in
3656–3669. Arabidopsis mutants defect in cinnamoyl-CoA reductase I.
Li-Weber, M. (2009). New therapeutic aspects of flavones: the an- Planta. 227, 943–956.
ticancer properties of Scutellaria and its main active constituents Mock, H.P., and Strack, D. (1992). Energetics of uridine 5-diphopho-
Wogonin, Baicalein and Baicalin. Cancer Treat. Rev. 35, 57–68. glucose: hydroxycinnamic acid glucosyltransferase reaction. Phy-
Long, M.C., Nagegowda, D.A., Kaminaga, Y., Ho, K.K., Kish, C.M., tochemistry. 32, 575–579.
Schnepp, J., Sherman, D., Weiner, H., Rhodes, D., and Morant, M., Jørgenson, K., Schaller, H., Pinot, F., Møller, B.L.,
Dudareva, N. (2009). Involvement of snapdragon benzaldehyde Werck-Reichardt, D., and Bak, S. (2007). CYP703 is an an-
dehydrogenase in benzoic acid biosynthesis. Plant J. 59, 256–265. cient cytochrome P450 in land plants catalyzing in-chain hy-
Louie, G.V., Baiga, T.J., Bowman, M.E., Koeduka, T., Taylor, J.H., droxylation of lauric acid to provide building blocks for
Spassova, S.M., Pichersky, E., and Noel, J.P. (2007). Structure sporopollenin synthesis in pollen. Plant Cell. 19, 1473–1487.
and reaction mechanism of basil eugenol synthase. PLoS ONE. Nagel, J., Culley, L.K., Lu, Y., Liu, E., Matthews, P.D., Stevens, J.F.,
2 (10), e993. doi:10.1371/journal.pone.0000993. and Page, J.E. (2008). EST analysis of hop glandular trichomes
Louie, G.V., Bowman, M.E., Moffitt, M.C., Baiga, T.J., Moore, B.S., identifies an O-methyltransferase that catalyzes the biosynthesis
(2009). Evolution of rosmarinic acid biosynthesis. Phytochemis- Scalliet, G., Lionnet, C., Le Bechec, M., Dutron, L., Magnard, J.L.,
try. 70, 1663–1679. Baudino, S., Bergougnoux, V., Julien, F., Chambrier, P.,
Pichersky, E., and Gang, D. (2005). Genetics and biochemistry of sec- Vergne, P., Dumas, C., Cock, J.M., and Hugueney, P. (2006). Role
ondary metabolites in plants: an evolutionary perspective. of petal specific orcinol O-methyltransferase in the evolution of
Trends Pl. Sci. 4, 439–445. rose scent. Plant Physiol. 140, 18–29.
Pollard, M., Beisson, F., Li, Y., and Ohlrogge, J.B. (2008). Building Schijlen, E., de Vos, C.H., Jjonker, H., van den Broeck, H.,
lipid barriers: biosynthesis of cutin and suberin. Trends Pl. Sci. Molthoff, J., van Tune, A., Martens, S., and Bovy, A. (2006). Path-
13, 236–246. way engineering for healthy phytochemicalsleadingtotheproduc-
Pourcel, L., Routaboul, J.M., Kerhoas, L., Caboche, M., Lepiniec, L., tion of novel flavonoids in tomato fruit. Plant Biotech. J. 4, 433–444.
and Debeaujon, I. (2005). TRANSPARENT TESTA10 encodes a Schilmiller, A.L., Stout, J., Weng, J.K., Humphreys, J., Rueger, M.O.,
laccase-like enzyme involved in oxidative polymerisation of and Chapple, C.C.S. (2009). Mutations in the CINNAMATE
flavonoids in Arabidopsis seed coat. Plant Cell. 17, 2966–2980. 4-HYDROXYLASE gene impact metabolism, growth and
Rezai-Zadeh, K., et al. (2005). Green tea epigallocatechin-3-gallate development in Arabidopsis. Plant J. doi: 10.1111/j.1365–
(EGCG) modulates amyloid precursor protein cleavage and 313X.2009.03996.x.
Stehle, F., Brandt, W., Stubbs, M.T., Milkowski, C., and Strack, D. Gerbera hybrida catalyzes the conversion of (+)-catechin to cya-
(2009). Sinapoyltransferases in the light of molecular evolution. nidin and a novel procyanidin. FEBS Lett. 580, 1642–1648.
Phytochemistry. 70, 1652–1662. Werner, I., Bacher, A., and Eisenreich, W. (1997). Retrobiosynthetic
Stevens, J., and Page, J. (2004). Xanthohumol and related prenyl- NMR studies with 13C-labelled glucose. J. Biol. Chem. 272,
flavonoids from hops and beer: to your good health. Phytochem- 25474–25482.
istry. 65, 1317–1330. Werner, R.A., Rossmann, A., Schwarz, C., Bacher, A., Schmidt, H.L.,
Stewart, J.J., Akiyama, T., Chapple, C.C.S., Ralph, J., and and Eisenreich, W. (2004). Biosynthesis of gallic acid in Rhus
Mansfield, S.D. (2009). The effects on lignin structure on over- typhina: discrimination between alternative pathways from nat-
expression of ferulate 5-hydroxylase in hybrid poplar. Plant ural oxygen isotope labelling. Phytochemistry. 65, 2809–2813.
Physiol. 150, 621–635. Wildermuth, M.C., Dewdeny, J., Wu, G., and Ausubel, F.M. (2001).
Stracke, R., Ishihara, H., Huep, G., Barsch, A., Mehrtens, F., Isochorismate synthase is required to synthesize salicylic acid for
Niehaus, K., and Weisshaar, B. (2007). Differential regulation plant defence. Nature. 414, 562–565.
of closely related R2R3-MYP transcription factors controls flavo- Xie, Z., Kapteyn, J., and Gang, D.R. (2008). A systems biology inves-
nol accumulation in different parts of the Arabidopsis thaliana tigation of the MEP/terpenoid and shikimate/phenylpropanoid