Flavan-3-ol and ascorbate accumulation and oxidation in plants, and its physiological significance

Abstract

[eng] Flavonoids have received much attention in biology and medicine because of their high in vitro antioxidant activities. However, little is known about the physiological significance of this capacities in plants under stress conditions, but for anthocyanins, which act as photoprotective screens. The final objective of the present work was to determine the role of the accumulation and oxidation of flavonoids in plants under abiotic stress conditions, and to compare it with that of a well known antioxidant: ascorbate. To accomplish this objective, a multi-disciplinary approach was applied, including plant physiology, biochemistry and molecular biology techniques. First, the total phenolic content of three Mediterranean species - Salvia officinalis (L), Melissa officinalis (L) and Cistus clusii (Dunal)- was analyzed as well as the changes they showed during a drought treatment. C. clusii plants showed the highest phenolic levels among the studied species, and these phenolics increased during the drought treatment. It is suggested that the biosynthesis of phenolics may serve as an alternative carbon, reduction equivalents and ATP sink. Second, the main antioxidant flavonoids in C. clusii leaf extreacts were identified to be (-)-epigallocatechin gallate (EGCG), (-)-epicatechin (EC) and (-)-epicatechin gallate (ECG). Levels of these flavan-3-ols increased during the drought treatment in a simmilar manner as ascorbate did, which moreover reduced the levels of its oxidized form, dehydroascorbate (DHA). It is suggested that aside ot the mentioned role as phenolics, flavan-3-ols may act as antioxidants in C. clusii plants under stress. Next, the role of these flavan-3-ols in different aged C. clusii plants in field conditions was studied. In this experiment it is shown that the accumulation of monomeric reduced flavan-3-ols is triggered by incident light (maximum radiation or photoperiod). Moreover, the accumulation of monomeric reduced, monomeric oxidized (quinones) and polymeric (proanthocyanidins) flavan-3-ols may serve also an alternative carbon, reduction equivalents and ATP sink as well as an antioxidant mechanism as plants age. Next, the oxidation of monomeric reduced flavan-3-ols to their respective quinones was studied in tea plants under severe drught stress. EC and EGCG oxidize to their respective quinones in such conditions, suggesting that EC and EGCG may act as membrane antioxidants. Finally, the role of ascorbate oxidation state in the apoplast as a cell signal was studied. Tobacco transgenic plants with shifted ascorbate oxidase activity modulated showed shifted transcript profile. Many cell preocesses were altered at gene expression levels upon shifting the ascorbate oxidation state in the apoplast, including H2O2 homeostasis, electron transport and stress responses. Moreover, Ca2+ is shown to be a key component of the ascorbate oxidation state signal transduction pathway to the nucleus. In conclusion, in this study it is demonstrated the presence of EC, EGCG y ECG in C. clusii and that the levels of these flavan-3-ols increase with drought. These flavan-3-ols may serve as an alternative C, reduction equivalents and ATP sink. Flavan-3-ols accumulate in plants as they age, especially during streass periods. Flavan-3-ols are oxidized to their respective quinones under severe drought stress, particularly in tea plants, and they may act as membrane antioxidants. The oxidation state of the ascorbate in the apoplast acts as a cell signal regulating different processes within the symplast at gene expression levels, including H2O2 homeostasis.