Sonia Patui

Plant Flavonoids—Biosynthesis, Transport and Involvement in Stress Responses

This paper aims at analysing the synthesis of flavonoids, their import and export in plant cell compartments, as well as their involvement in the response to stress, with particular reference to grapevine (Vitis vinifera L.). A multidrug and toxic compound extrusion (MATE) as well as ABC transporters have been demonstrated in the tonoplast of grape berry, where they perform a flavonoid transport. The involvement of a glutathione S-transferase (GST) gene has also been inferred. Recently, a putative flavonoid carrier, similar to mammalian bilitranslocase (BTL), has been identified in both grape berry skin and pulp. In skin the pattern of BTL expression increases from véraison to harvest, while in the pulp its expression reaches the maximum at the early ripening stage. Moreover, the presence of BTL in vascular bundles suggests its participation in long distance transport of flavonoids. In addition, the presence of a vesicular trafficking in plants responsible for flavonoid transport is discussed. Finally, the involvement of flavonoids in the response to stress is described.

Transport and accumulation of flavonoids in grapevine (Vitis vinifera L.)

Flavonoids are a group of secondary metabolites widely distributed in plants that represent a huge portion of the soluble phenolics present in grapevine (Vitis vinifera L.). These compounds play different physiological roles and are often involved in protection against biotic and abiotic stress. Even if the flavonoid biosynthetic pathways have been largely characterized, the mechanisms of their transport and accumulation in cell wall and vacuole are still not completely understood. This review analyses the known mechanisms of flavonoid uptake and accumulation in grapevine, with reference to the transport models and membrane carrier proteins described in other plant species. The effect of different environmental factors on flavonoid biosynthesis and transporters is also discussed.

The mitochondrial permeability transition pore (PTP) — An example of multiple molecular exaptation?

The mitochondrial permeability transition (PT) is a well-recognized phenomenon that allows mitochondria to undergo a sudden increase of permeability to solutes with molecular mass ≤ 1500Da, leading to organelle swelling and structural modifications. The relevance of PT relies on its master role in the manifestation of programmed cell death (PCD). This function is performed by a mega-channel (in some cases inhibited by cyclosporin A) named permeability transition pore (PTP), whose function could derive from the assembly of different mitochondrial proteins. In this paper we examine the distribution and characteristics of PTP in mitochondria of eukaryotic organisms so far investigated in order to draw a hypothesis on the mechanism of its evolution. As a result, we suggest that PTP may have arisen as a new function linked to a multiple molecular exaptation of different mitochondrial proteins, even though they could nevertheless still play their original role. Furthermore, we suggest that the early appearance of PTP could have had a crucial role in the establishment of endosymbiosis in eukaryotic cells, by the coordinated balancing of ATP production by glycolysis (performed by the primary phagocyte) and oxidative phosphorylation (accomplished by the endosymbiont). Indeed, we argue on the possibility that this new energetic equilibrium could have opened the way to the subsequent evolution toward metazoans.

Seed storage in polyethylene bags of a recalcitrant species (Quercus ilex): analysis of some bio-energetic and oxidative parameters

Some technological and physiological aspects were analysed during seed storage of holm oak (Quercus ilex L.), a typical Mediterranean recalcitrant species. Considering the mild dormancy and desiccation sensitivity of these seeds, the influence of the storage environment on viability was examined. Acorns were maintained at low temperature and at high moisture content either inside thin polyethylene bags or mixed with peat in a bin. Storage in polyethylene bags, if compared with peat treatment, maintained optimal seed quality parameters. The effects of the two treatments on some physiological parameters were analysed during 1-year seed storage. Acorns stored in a bin with peat started to germinate early and exhibited a more rapid seed deterioration caused by metabolism-linked oxidative damage. The latter phenomenon was linked to a higher respiration and H2O2 level, induction of catalase activity, as well as lowered glutathione pool and ATP content. In addition, a more oxidized redox poise was observed. On the contrary, the plastic film of polyethylene, limiting gaseous exchanges, maintained acorn metabolic activity at low levels and allowed the accumulation of ethylene inside the storage atmosphere. These factors, inducing a dormant-like state, could have played a crucial role in prolonging seed storage.

Lipase activity and antioxidant capacity in coffee (Coffea arabica L.) seeds during germination.

In this paper, lipase activity was characterized in coffee (Coffea arabica L.) seeds to determine its involvement in lipid degradation during germination. The lipase activity, evaluated by a colorimetric method, was already present before imbibition of seeds and was further induced during the germination process. The activity showed a biphasic behaviour, which was similar in seeds either with or without endocarp (parchment), even though the phenomenon showed a delay in the former. The enzymatic activity was inhibited by tetrahydrolipstatin (THL), a selective and irreversible inhibitor of lipases, and by a polyclonal antibody raised against purified alkaline lipase from castor bean. The immunochemical analysis evidenced a protein of ca. 60 kDa, cross-reacting with an anti-lipase antibody, in coffee samples obtained from seeds of both types. Gas chromatographic analyses of free fatty acid (FFA) content confirmed the differences shown in the lipolytic activity of the samples with or without parchment, since FFA levels increased more rapidly in samples without parchment. Finally, the analyses of the antioxidant capacity showed that the presence of parchment was crucial for lowering the oxidation of the lipophylic fraction, being the seeds with parchment less prone to oxidation processes.

The Permeability Transition in Plant Mitochondria: The Missing Link

The synthesis of ATP in mitochondria is dependent on a low permeability of the inner membrane. Nevertheless, mitochondria can undergo an increased permeability to solutes, named permeability transition (PT) that is mediated by a permeability transition pore (PTP). PTP opening requires matrix Ca2+ and leads to mitochondrial swelling and release of intramembrane space proteins (e.g., cytochrome c). This feature has been initially observed in mammalian mitochondria and tentatively attributed to some components present either in the outer or inner membrane. Recent works on mammalian mitochondria point to mitochondrial ATP synthase dimers as physical basis for PT, a finding that has been substantiated in yeast and Drosophila mitochondria. In plant mitochondria, swelling and release of proteins have been linked to programmed cell death, but in isolated mitochondria PT has been observed in only a few cases and in plant cell cultures only indirect evidence is available. The possibility that mitochondrial ATP synthase dimers could function as PTP also in plants is discussed here on the basis of the current evidence. Finally, a hypothetical explanation for the origin of PTP is provided in the framework of molecular exaptation.

Immunohistochemical localisation of a putative flavonoid transporter in grape berries.

Flavonoids are a class of secondary metabolites present in large amounts in grapevine (Vitis vinifera L.), which are involved in several aspects of its physiology (e.g. protection against biotic and abiotic stress). Even if the biosynthetic pathways of flavonoid sub-classes have been largely characterised, the mechanisms of their transport and accumulation to the final target sites are still not completely understood. Unanticipated insights have been obtained by probing plant tissues with pure antibodies targeting bilitranslocase (BTL, TCDB # 2.A.65.1.1), a mammalian transporter involved in the absorption and tissue distribution of dietary flavonoids. The occurrence of a BTL homologue has also been found in grape berries, in both tegumental layers of skin and pulp vascular bundles. In the skin, the expression of this protein starts from véraison (starting of the change in colour and softening of berries) and increases up to a maximum at the harvest stage, matching the same temporal pattern of flavonoid accumulation.