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The effects of a reduction in water supply during fruit development and postharvest fruit ripening on the oxidative status and the antioxidant defense system were studied in the mango fruit (Mangifera indica L.) cv. Cogshall. Changes in non-enzymatic (ascorbate) and enzymatic (SOD, CAT, APX, MDHAR, DHAR and GR) antioxidants, as well as oxidative parameters (H2O2 and MDA) and major carotenoids, were measured in unripe and ripe fruits from well-irrigated and non-irrigated trees. Under non-limiting water supply conditions, ripening induced oxidation as a result of the production of ROS and decreased ascorbate content. Antioxidant enzymatic systems were activated to protect fruit tissues and to regenerate the ascorbate pool. The carotenoid pool, mainly represented by β-carotene and esterified violaxanthine isomers, accumulated naturally during mango ripening. The suppression of irrigation decreased fruit size and induced accumulation of ABA and of its storage form, ABA-GE, in fruit pulp from the earliest harvest. It also increased oxidation, which was observable by the high levels of ascorbate measured at the early stages at harvest, and by the delay in the time it took to reach the pseudo constant carotene-to-xanthophyll ratio in ripe fruits. Nevertheless, differences between the irrigation treatments on the antioxidant system in ripe fruits were not significant, mainly because of the drastic changes in this system during ripening., (Copyright © 2015 Elsevier GmbH. All rights reserved.)

Ripening of grape (Vitis vinifera L.) berry immediately precedes harvesting and the evolution of the skin tissue is important as it contains the key compounds for wine quality. Grape softening is thought to result from extensive cell wall modifications that occur during ripening. These modifications result from the activity of different cell wall-modifying enzymes. Two of the most significant pectin-degrading enzymes are pectin methylesterase (EC 3.1.1.11) and polygalacturonase (EC 3.2.1.15). In this work, the activities of both enzymes were monitored in skin tissue throughout berry development. Pectin methylesterase activity was present before the onset of veraison and increased during skin maturation. No polygalacturonase activity could be detected. The accumulation of mRNA encoding a pectin methylesterase and two polygalacturonase isoforms was examined using RT-PCR. Transcripts for pectin methylesterase were present in all stages analyzed with a maximal accumulation at the end of color change. Accumulation of VvPG1 transcript was closely correlated with berry softening, and expression of this gene was markedly increased during the color change. VvPG2 mRNA accumulation began before veraison and was low during skin ripening. A phylogenic analysis showed that this gene is classified in a different group than VvPG1. These findings suggest that both genes are associated with different mechanisms during skin development. VvPG1, in particular, is most likely to play a role in skin softening and VvPG2 in triggering the ripening process.

The characterization of proteins isolated from skin tissue is apparently an essential parameter for understanding grape ripening as this tissue contains the key compounds for wine quality. It has been particularly difficult to extract proteins from skins for analysis by two-dimensional electrophoresis gels and, therefore, a protocol for this purpose has been adapted. The focus was on the evolution of the proteome profile of grape skin during maturation. Proteome maps obtained at three stages of ripening were compared to assess the extent to which protein distribution differs in grape skin during ripening. The comparative analysis shows that numerous soluble skin proteins evolve during ripening and reveal specific distributions at different stages. Proteins involved in photosynthesis, carbohydrate metabolisms, and stress response are identified as being over-expressed at the beginning of colour-change. The end of colour-change is characterized by the over-expression of proteins involved in anthocyanin synthesis and, at harvest, the dominant proteins are involved in defence mechanisms. In particular, increases in the abundance of different chitinase and beta-1,3-glucanase isoforms were found as the berry ripens. This observation can be correlated with the increase of the activities of both of these enzymes during skin ripening. The differences observed in proteome maps clearly show that significant metabolic changes occur in grape skin during this crucial phase of ripening. This comparative analysis provides more detailed characterization of the fruit ripening process.

Yeasts can incorporate a wide variety of exogenous sterols under strict anaerobiosis. Yeasts normally require oxygen for growth when exogenous sterols are limiting, as this favours the synthesis of lipids (sterols and unsaturated fatty acids). Although much is known about the oxygen requirements of yeasts during anaerobic growth, little is known about their exact sterol requirements in such conditions. We developed a method to determine the amount of ergosterol required for the growth of several yeast strains. We found that pre-cultured yeast strains all contained similar amounts of stored sterols, but exhibited different ergosterol assimilation efficiencies in enological conditions [as measured by the ergosterol concentration required to sustain half the number of generations attributed to ergosterol assimilation (P(50))]. P(50) was correlated with the intensity of sterol synthesis. Active dry yeasts (ADYs) contained less stored sterols than their pre-cultured counterparts and displayed very different ergosterol assimilation efficiencies. We showed that five different batches of the same industrial Saccharomyces cerevisiae ADY exhibited significantly different ergosterol requirements for growth. These differences were mainly attributed to differences in initial sterol reserves. The method described here can therefore be used to quantify indirectly the sterol synthesis abilities of yeast strains and to estimate the size of sterol reserves.

During enological fermentations, superfluous oxygen consumption by yeast cells is observed. The superfluous oxygen consumed by the yeast cells is mainly related to the operation of non-respiratory oxygen consumption pathways resulting in an overall decrease in the total sterol fraction in yeast. On the other hand, yeast lees remaining at the end of alcoholic fermentations exhibit specific oxygen utilization rates ranging from 1 to 4 micromol O2 h- 10(-10) cells from the second to the thirteenth month of wine aging. This oxygen consumption capacity of yeast lees was independent of residual cell viability. In this study, we investigated the potential relationship between the oxygen added to commercial yeast strains during enological fermentation and the capacity of the corresponding yeast lees to interact with oxygen. Additions of low (7 mg l(-)) and excess (37 mg l(-1)) amounts of oxygen at the end of the cell growth phase were compared in terms of repercussions on the oxygen consumption activity of the corresponding yeast lees. As expected, the superfluous oxygen consumption by yeast cells during fermentation had a positive influence on the fermentation kinetics and increased cell biomass formation. Oxygen consumption rates and the total capacity of oxygen consumption by the corresponding yeast lees clearly decreased when oxygen was added during fermentation. This marked decrease in yeast lees reactivity towards oxygen was concomitantly related to an increase in ergosterol synthesis and to oxygen-dependent sterol degradation. Such degradation occurred when oxygen was added in excess. Therefore, oxygenation control during fermentation appears to be a potential way to optimize both the fermentation kinetics and control yeast lees reactivity towards oxygen. For practical applications, oxygenation control during alcoholic fermentation may be considered as a general tool for decreasing the highly reductive effect of yeast lees during wine aging.