Adrian J. Parr

Phenols in the plant and in man. The potential for possible nutritional enhancement of the diet by modifying the phenols content or profile.

There is growing recognition that many phenolic secondary metabolites present in foodstuffs may possibly exert beneficial effects on human health. This may to some degree be mediated via antioxidant actions, but a range of more specific pharmacological effects have also been proposed. Given this background, there may be favourable consequences for the general health of Western populations as a result of optimising the phenolic content of the diet. This paper reviews what is known of the function of phenolics both in the plant and in man. It also describes current understanding of the biosynthesis of phenolics in plants, with emphasis on where potential controlling steps may exist. Finally, advances in identification and isolation of the genes coding for phenolic biosynthetic enzymes or regulatory proteins are also summarised. Taken together, this information provides a basis for attempts to modify and optimise the phenolic content of food crops, using either conventional plant breeding along with manipulation of agronomic practices, or else the more targeted approaches of modern molecular biology. © 2000 Society of Chemical Industry

Transcriptional repression by AtMYB4 controls production of UV‐protecting sunscreens in Arabidopsis

An Arabidopsis thaliana line that is mutant for the R2R3 MYB gene, AtMYB4, shows enhanced levels of sinapate esters in its leaves. The mutant line is more tolerant of UV‐B irradiation than wild type. The increase in sinapate ester accumulation in the mutant is associated with an enhanced expression of the gene encoding cinnamate 4‐hydroxylase, which appears to be the principal target of AtMYB4 and an effective rate limiting step in the synthesis of sinapate ester sunscreens. AtMYB4 expression is downregulated by exposure to UV‐B light, indicating that derepression is an important mechanism for acclimation to UV‐B in A.thaliana. The response of target genes to AtMYB4 repression is dose dependent, a feature that operates under physiological conditions to reinforce the silencing effect of AtMYB4 at high activity. AtMYB4 works as a repressor of target gene expression and includes a repression domain. It belongs to a novel group of plant R2R3 MYB proteins involved in transcriptional silencing. The balance between MYB activators and repressors on common target promoters may provide extra flexibility in transcriptional control.

The AmMYB308 and AmMYB330 Transcription Factors from Antirrhinum Regulate Phenylpropanoid and Lignin Biosynthesis in Transgenic Tobacco

MYB-related transcription factors are known to regulate different branches of flavonoid metabolism in plants and are believed to play wider roles in the regulation of phenylpropanoid metabolism in general. Here, we demonstrate that overexpression of two MYB genes from Antirrhinum represses phenolic acid metabolism and lignin biosynthesis in transgenic tobacco plants. The inhibition of this branch of phenylpropanoid metabolism appears to be specific to AmMYB308 and AmMYB330, suggesting that they recognize their normal target genes in these transgenic plants. Experiments with yeast indicate that AmMYB308 can act as a very weak transcriptional activator so that overexpression may competitively inhibit the activity of stronger activators recognizing the same target motifs. The effects of the transcription factors on inhibition of phenolic acid metabolism resulted in complex modifications of the growth and development of the transgenic plants. The inhibition of monolignol production resulted in plants with at least 17% less lignin in their vascular tissue. This reduction is of importance when designing strategies for the genetic modification of woody crops.

New approaches to understanding and controlling cell separation in relation to fruit and vegetable texture

Texture is a major quality attribute of plant-based foods. The plant cell wall is a key determinant of texture in fruit and vegetables; its properties influence the way in which plant tissues undergo mechanical deformation and failure during mastication. Processes such as cooking, and physiological events such as ripening can reduce the strength of cell adhesion in many vegetables and fruit through depolymerization of pectic polysaccharides. There is considerable interest in reducing such cell separation to prevent over-softening, loss of juiciness and development of mealiness. Recent research on Chinese water chestnut has implicated diferulic acid cell-wall crosslinks in the thermal stability of cell adhesion and, therefore, maintenance of crispness after cooking. In this review, we discuss the potential for exploiting this finding in other edible plant organs, and assess the likely effects that enhanced crosslinking might have on other quality attributes.

Cell Wall Esterified Phenolic Dimers: Identification and Quantification by Reverse Phase High Performance Liquid Chromatography and Diode Array Detection

An optimized high performance liquid chromatography (HPLC) procedure has been developed for the analysis and quantification of all of the known ferulic acid dehydrodimers, and the principle phenolic aldehydes and acids, found in the cell walls of higher plants. The HPLC system uses an ODS2 reverse phase column (5 μm particle size) eluted with a methanol, acetonitrile and water gradient with detection at 280 nm. In addition to providing baseline resolution of most components, the method employs a spectrometric detector which enables the precise identification of eluted components through the analysis of their spectral properties. Analysis of the cell wall phenolics of wheat straw stem (Triticum vulgare) was carried out using this method which is highly versatile and, for certain components, more sensitive than the current gas chromatography–mass spectrometry methodology.

Structural Analyses and Dynamics of Soluble and Cell Wall-bound Phenolics in a Broad Spectrum Resistance to the Powdery Mildew Fungus in Barley

High pressure liquid chromatography profiles of barley leaf epidermal soluble and cell wall-bound phenolics were analyzed in response to challenge with the fungal pathogenErysiphe graminis f. sp. hordei. Only one soluble phenolic was found to accumulate differentially in a broad spectrum resistance reaction controlled by mlo resistance alleles in comparison to susceptible near isogenic Mlolines. Structural analysis identified this compound as a novel phenolic conjugate, p-coumaroyl-hydroxyagmatine (p-CHA).p-CHA but not the nonhydroxylated derivativep-coumaroylagmatine exhibited antifungal activity bothin vitro and in vivo. The accumulation ofp-CHA in epidermal tissue correlated tightly with fungal penetration attempts of attacked host cells. Furthermore, upon penetration, epidermal cell wall-bound phenolics became resistant to saponification at sites of attempted fungal ingress (papilla), indicating a change in, or the addition of, different chemical bonding types. The switch in saponification sensitivity occurred at least 2 h earlier in the mlo-incompatible than in theMlo-compatible interaction. Our results suggest thatp-CHA and the speed of papillae compaction play important roles in non-race-specific powdery mildew defense.

AtMYB12 regulates caffeoyl quinic acid and flavonol synthesis in tomato: expression in fruit results in very high levels of both types of polyphenol

Plant polyphenolics exhibit a broad spectrum of health-promoting effects when consumed as part of the diet, and there is considerable interest in enhancing the levels of these bioactive molecules in plants used as foods. AtMYB12 was originally identified as a flavonol-specific transcriptional activator in Arabidopsis thaliana, and this has been confirmed by ectopic expression in tobacco. AtMYB12 is able to induce the expression of additional target genes in tobacco, leading to the accumulation of very high levels of flavonols. When expressed in a tissue-specific manner in tomato, AtMYB12 activates the caffeoyl quinic acid biosynthetic pathway, in addition to the flavonol biosynthetic pathway, an activity which probably mirrors that of the orthologous MYB12-like protein in tomato. As a result of its broad specificity for transcriptional activation in tomato, AtMYB12 can be used to produce fruit with extremely high levels of multiple polyphenolic anti-oxidants. Our data indicate that transcription factors may have different specificities for target genes in different plants, which is of significance when designing strategies to improve metabolite accumulation and the anti-oxidant capacity of foods.

Secondary product formation by cultures of Beta vulgaris and Nicotiana rustica transformed with Agrobacterium rhizogenes.

Abstract‘Hairy root’ cultures of Beta vulgaris and Nicotiana rustica were established after roots were induced on plants following infection with Agrobacterium rhizogenes. The transformed cultures of B. vulgaris and N. rustica synthesised their characteristic secondary products, the betalain pigments and nicotine alkaloids respectively, at levels comparable with those of in vivo roots from the same variety. Betalains were entirely retained inside the root tissue. In contrast, a proportion of the nicotine alkaloids was secreted into the medium. The potential of this type of ‘in vitro’ plant tissue culture for the production of valuable plant secondary products is identified and confirmed.

Inhibition of Phenolic Acid Metabolism Results in Precocious Cell Death and Altered Cell Morphology in Leaves of Transgenic Tobacco Plants

Several complex phenotypic changes are induced when the transcription factor AmMYB308 is overexpressed in transgenic tobacco plants. We have previously shown that the primary effect of this transcription factor is to inhibit phenolic acid metabolism. In the plants that we produced, two morphological features were prominent: abnormal leaf palisade development and induction of premature cell death in mature leaves. Evidence from the analysis of these transgenic plants suggests that both changes resulted from the lack of phenolic intermediates. These results emphasize the importance of phenolic secondary metabolites in the normal growth and development of tobacco. We suggest that phenolic acid derivatives are important signaling molecules in the final stages of leaf palisade formation and that phenolic acid derivatives also play a prominent role in tissue senescence.

Over-expressing a yeast ornithine decarboxylase gene in transgenic roots of Nicotiana rustica can lead to enhanced nicotine accumulation.

Transformed root cultures of Nicotiana rustica have been generated in which the gene from the yeast Saccharomyces cerevisiae coding for ornithine decarboxylase has been integrated. The gene, driven by the powerful CaMV35S promoter with an upstream duplicated enhancer sequence, shows constitutive expression throughout the growth cycle of some lines, as demonstrated by the analysis of mRNA and enzyme activity. The presence of the yeast gene and enhanced ornithine decarboxylase activity is associated with an enhanced capacity of cultures to accumulate both putrescine and the putrescine-derived alkaloid, nicotine. Even, however, with the very powerful promoter used in this work the magnitude of the changes seen is typically only in the order of 2-fold, suggesting that regulatory factors exist which limit the potential increase in metabolic flux caused by these manipulations. Nevertheless, it is demonstrated that flux through a pathway to a plant secondary product can be elevated by means of genetic manipulation.