title: Introduction of sense constructs of cinnamate 4-hydroxylase (CYP73A24) in transgenic tomato plants shows opposite effects on flux into stem lignin and fruit flavonoids.
abstract: Understanding regulation of phenolic metabolism underpins attempts to engineer plants for diverse properties such as increased levels of antioxidant flavonoids for dietary improvements or reduction of lignin for improvements to fibre resources for industrial use. Previous attempts to alter phenolic metabolism at the level of the second enzyme of the pathway, cinnamate 4-hydroxylase have employed antisense expression of heterologous sequences in tobacco. The present study describes the consequences of homologous sense expression of tomato CYP73A24 on the lignin content of stems and the flavonoid content of fruits. An extensive number of lines were produced and displayed four developmental variants besides a normal phenotype. These aberrant phenotypes were classified as dwarf plants, plants with distorted (curly) leaves, plants with long internodes and plants with thickened waxy leaves. Nevertheless, some of the lines showed the desired increase in the level of rutin and naringenin in fruit in a normal phenotype background. However this could not be correlated directly to increased levels of PAL and C4H expression as other lines showed less accumulation, although all lines tested showed increases in leaf chlorogenic acid which is typical of Solanaceous plants when engineered in the phenylpropanoid pathway. Almost all transgenic lines analysed showed a considerable reduction in stem lignin and in the lines that were specifically examined, this was correlated with partial sense suppression of C4H. Although not the primary purpose of the study, these reductions in lignin were amongst the greatest seen in plants modified for lignin by manipulation of structural genes. The lignin showed higher syringyl to coniferyl monomeric content contrary to that previously seen in tobacco engineered for downregulation of cinnamate 4-hydroxylase. These outcomes are consistent with placing CYP73A24 more in the lignin pathway and having a role in flux control, while more complex regulatory processes are likely to be involved in flavonoid and chlorogenic acid accumulation.
authors.affiliation: School of Biological Sciences, Royal Holloway, University of London, Egham, Surrey TW20 0EX, UK.
authors.forename: David J
authors.initials: DJ
authors.lastname: Millar
authors.forename: Marianne
authors.initials: M
authors.lastname: Long
authors.forename: Georgina
authors.initials: G
authors.lastname: Donovan
authors.forename: Paul D
authors.initials: PD
authors.lastname: Fraser
authors.forename: Alain-Michel
authors.initials: AM
authors.lastname: Boudet
authors.forename: Saida
authors.initials: S
authors.lastname: Danoun
authors.forename: Peter M
authors.initials: PM
authors.lastname: Bramley
authors.forename: G Paul
authors.initials: GP
authors.lastname: Bolwell
mesh_terms: D000595:Amino Acid Sequence; D005419:Flavonoids / Q000378:metabolism*; D005638:Fruit / Q000235:genetics / Q000378:metabolism; D020868:Gene Silencing; D008031:Lignin / Q000378:metabolism*; D018551:Solanum lycopersicum / Q000235:genetics*; D008969:Molecular Sequence Data; D010636:Phenols / Q000378:metabolism; D010641:Phenotype; D010940:Plant Proteins / Q000235:genetics* / Q000378:metabolism; D018547:Plant Stems / Q000235:genetics / Q000378:metabolism; D030821:Plants, Genetically Modified / Q000033:anatomy & histology / Q000378:metabolism*; D016415:Sequence Alignment; D050564:Trans-Cinnamate 4-Monooxygenase / Q000235:genetics* / Q000378:metabolism; D014170:Transformation, Genetic
chemical_list: D005419:Flavonoids; D010636:Phenols; D010940:Plant Proteins; D008031:Lignin; D050564:Trans-Cinnamate 4-Monooxygenase