Antioxidant systems from Pepper (Capsicum annuum L.): involvement in the response to temperature changes in ripe fruits.

Sweet pepper is susceptible to changes in the environmental conditions, especially temperatures below 15 °C. In this work, two sets of pepper fruits (Capsicum annuum L.) which underwent distinct temperature profiles in planta were investigated. Accordingly, two harvesting times corresponding to each set were established: Harvest 1, whose fruits developed and ripened at 14.9 °C as average temperature; and Harvest 2, with average temperature of 12.4 °C. The oxidative metabolism was analyzed in all fruits. Although total ascorbate content did not vary between Harvests, a shift from the reduced to the oxidized form (dehydroascorbate), accompanied by a higher ascorbate peroxidase activity, was observed in Harvest 2 with respect to Harvest 1. Moreover, a decrease of the ascorbate-generating enzymatic system, the γ-galactono-lactone dehydrogenase, was found at Harvest 2. The activity values of the NADP-dependent dehydrogenases analyzed seem to indicate that a lower NADPH synthesis may occur in fruits which underwent lower temperature conditions. In spite of the important changes observed in the oxidative metabolism in fruits subjected to lower temperature, no oxidative stress appears to occur, as indicated by the lipid peroxidation and protein oxidation profiles. Thus, the antioxidative systems of pepper fruits seem to be involved in the response against temperature changes.

Transcriptomic events involved in melon mature-fruit abscission comprise the sequential induction of cell-wall degrading genes coupled to a stimulation of endo and exocytosis.

BACKGROUND: Mature-fruit abscission (MFA) in fleshy-fruit is a genetically controlled process with mechanisms that, contrary to immature-fruit abscission, has not been fully characterized. Here, we use pyrosequencing to characterize the transcriptomes of melon abscission zone (AZ) at three stages during AZ-cell separation in order to understand MFA control at an early stage of AZ-activation. PRINCIPAL FINDINGS: The results show that by early induction of MFA, the melon AZ exhibits major gene induction, while by late induction of MFA, melon AZ shows major gene repression. Although some genes displayed similar regulation in both early and late induction of abscission, such as EXT1-EXT4, EGase1, IAA2, ERF1, AP2D15, FLC, MADS2, ERAF17, SAP5 and SCL13 genes, the majority had different expression patterns. This implies that time-specific events occur during MFA, and emphasizes the value of characterizing multiple time-specific abscission transcriptomes. Analysis of gene-expression from these AZs reveal that a sequential induction of cell-wall-degrading genes is associated with the upregulation of genes involved in endo and exocytosis, and a shift in plant-hormone metabolism and signaling genes during MFA. This is accompanied by transcriptional activity of small-GTPases and synthaxins together with tubulins, dynamins, V-type ATPases and kinesin-like proteins potentially involved in MFA signaling. Early events are potentially controlled by down-regulation of MADS-box, AP2/ERF and Aux/IAA transcription-factors, and up-regulation of homeobox, zinc finger, bZIP, and WRKY transcription-factors, while late events may be controlled by up-regulation of MYB transcription-factors. SIGNIFICANCE: Overall, the data provide a comprehensive view on MFA in fleshy-fruit, identifying candidate genes and pathways associated with early induction of MFA. Our comprehensive gene-expression profile will be very useful for elucidating gene regulatory networks of the MFA in fleshy-fruit.

Understanding the mechanisms of chilling injury in bell pepper fruits using the proteomic approach.

In order to advance in the understanding of CI in pepper fruits, the cell ultrastructure alterations induced by CI and the physiological and metabolic changes have been studied along with the proteomic study. When stored at low temperatures bell pepper (Capsicum annuum) fruits exhibited visual CI symptoms and important alterations within the cell ultrastructure, since peroxisomes and starch grains were not detected and the structure of the chloroplast was seriously damaged in chilled tissues. Physiological and metabolic disorders were also observed in chilled fruits, such as higher ethylene production, increased MDA content, changes in sugar and organic acids and enzymatic activities. The comparative proteomic analysis between control and chilled fruits reveals that the main alterations induced by CI in bell pepper fruits are linked to redox homeostasis and carbohydrate metabolism. Thus, protein abundance in the ascorbate-glutathione cycle is altered and catalase is down-regulated. Key proteins from glycolysis, Calvin cycle and Krebs cycle are also inhibited in chilled fruits. Enolase and GAPDH are revealed as proteins that may play a key role in the development of chilling injury. This study also provides the first evidence at the protein level that cytosolic MDH is involved in abiotic stress.

Influence of fruit ripening stage and harvest period on the antioxidant content of sweet pepper cultivars.

Pepper (Capsicum annuum L.) fruits are highly appreciated by producers and consumers for their economical and nutritional value. Four different cultivars of coloured peppers in immature and mature stages were harvested throughout the spring and examined for their content of phenolic compounds, ascorbic acid and total antioxidant capacity (TAA) as well as for lipid peroxidation and carbonyl proteins as index of oxidative stress. Ripening and harvest period influenced the antioxidants and the development of oxidative processes in the cultivars differently: lipid peroxidation increased in mature peppers except in one cultivar (Y1075), while no changes in protein oxidation or in TAA were produced, except in Y1075 in which both parameters increased. Each cultivar presented differences in antioxidant compounds depending on the harvest period, but we could recommend May as the optimal if all cultivars have to be harvested at the same time, when levels of ascorbate, phenols and TAA were not decreased, fresh weight and proteins were elevated, and levels of oxidation were not as high as in June (except for Y1075). A previous study of the response of each cultivar to different environmental conditions results essential to establish a good program of selection of cultivars with high quality and productivity.

1-Methylcyclopropene affects the antioxidant system of apricots (Prunus armeniaca L. cv. Búlida) during storage at low temperature.

BACKGROUND: Apricots (Prunus armeniaca cv. Búlida) were treated with 1 µL L⁻¹ [corrected] 1-methylcyclopropene (1-MCP) immediately after harvest and stored in air at 2 degrees C for 21 days. Antioxidant levels (ascorbic acid and carotenoids), enzymatic antioxidant activities (superoxide dismutase (SOD) and unspecific peroxidase (POX)) and total antioxidant capacity (trolox equivalent antioxidant capacity (TEAC)) were determined. The level of oxidative stress was also established by measuring ion leakage during storage. The changes in the antioxidant potential of apricots were related to the capacity of 1-MCP to increase their commercial life. RESULTS: 1-MCP-treated fruits exhibited higher SOD activity, whereas POX activity was significantly higher only after 21 days at 2 degrees C. Treated fruits also exhibited better retention of ascorbate and carotenoids and higher TEAC during storage. In accordance with these observations, lower ion leakage values were detected in 1-MCP-treated apricots. CONCLUSION: Taken together, these results suggest that 1-MCP conferred a greater resistance to oxidative stress. This, along with the reduction in ethylene production, could contribute to the increase in commercial life and nutritional value observed in 1-MCP-treated apricots.

Six edible wild fruits as potential antioxidant additives or nutritional supplements.

Total antioxidant activity of six non-cultivated but traditionally collected fruits from the south of Europe was assessed by measuring their ability to reduce the hydroxyl radical (OH) and hydrogen peroxide (H(2)O(2)), and their Trolox equivalent antioxidant capacity (TEAC). This antioxidant activity was compared with that shown by the synthetic antioxidants BHA (E-320), BHT (E-321) and propyl gallate (E-310). Total phenolics, ascorbic acid and the carotenoid content of the fruits were also analyzed. All fruits showed a high ability to scavenge the OH radical, ranging from 60.61% to 81.04% inhibition for Rosa canina and Crataegus monogyna, respectively. The H(2)O(2) scavenging capacity and the TEAC value varied widely, ranging between 3.63% and 87.26% inhibition of H(2)O(2) and between 0.47 and 416.64 mM trolox g(-1) FW for Sorbus domestica and Rosa canina, respectively. The antioxidant activity of fruits was higher than that of the synthetic additives analyzed, except in the TEAC assay. The phenolic and carotenoid content of R. canina was much higher than that of the other fruits analyzed and its ascorbic acid concentration was also high, reflecting its higher efficacy towards ABTS⁻ (TEAC assay) and H(2)O(2) species(.) In spite of these associations, the correlation coefficients between total antioxidant activity and the antioxidant compounds analyzed were not very significant; only phenolics and carotenoids showed a marginal correlation with the TEAC assay. The results support the possible use of R. canina as natural antioxidant to replace the synthetic additives, as well as their use in the production of functional foods with a high antioxidant activity.

Oil quality and sensory evaluation of almond (Prunus amygdalus) stored after electron beam processing.

The changes in the lipid fraction and the deterioration of its quality were studied in almonds (Prunus amygdalus) of the variety Guara after treatment with accelerated electrons at doses of 3, 7, and 10 kGy, during a storage period of 5 months. In almond oil, the most significant difference from the nutritional point of view was seen in the fatty acid linolenic (18:3), which shows at 3 kGy a maintenance of the initial content during the whole storage period, whereas, at 7 and 10 kGy, the content in 18:3 disappears from the first moment. The quality indices of the oil (K(232), K(270)) decreased at all doses and remained stable during the time of storage. The peroxide value did not show changes at the doses of 3 and 7 kGy, in non-irradiated samples, but significantly increased when the maximum dose of 10 kGy was applied. These changes were reflected in the sensory analysis, in which the tasters did not find sensory differences between the controls and those irradiated at doses of 3 or 7 kGy, whereas almonds irradiated at 10 kGy exhibited a rancid flavor and a significant decrease in general quality.

Antioxidant evaluation in dessert spices compared with common food additives. Influence of irradiation procedure.

The antioxidant properties of seven dessert spices (anise, cinnamon, ginger, licorice, mint, nutmeg, and vanilla) were compared with those of the common food antioxidants butylated hydroxyanisole (BHA) (E-320), butylated hydroxytoluene (BHT) (E-321), and propyl gallate (E-310). The influence of irradiation process on antioxidant activity was also evaluated. Mint and cinnamon exhibited a higher percentage of inhibition of oxidation than the other spices analyzed and the food antioxidants, as tested by the lipid peroxidation assay (LOO*). Nutmeg, anise, and licorice showed the strongest protection in the deoxyribose assay (OH*). Vanilla exhibited the highest antioxidant activity in the peroxidase-based assay (H2O2). Nutmeg, propyl gallate, ginger, and licorice improved the stability of oils (sunflower, corn, and olive) and fats (butter and margarine) against oxidation (110 degrees C Rancimat). Cinnamon was a better superoxide radical scavenger than the other analyzed spices and additives. When the Trolox equivalent antioxidant capacity (TEAC) assay was used to provide a ranking order of antioxidant activity, the result in decreasing order of antioxidant capacity was cinnamon approximately equal to propyl gallate > mint > anise > BHA > licorice approximately equal to vanilla > ginger > nutmeg > BHT. Irradiated samples did not show significant differences (p < 0.05) in the antioxidant activity with respect to the non-irradiated samples (1, 3, 5, and 10 kGy) in the assays used.

Antioxidant systems and their relationship with the response of pepper fruits to storage at 20 degrees C.

Fresh peppers (Capsicum annuum L., variety California) in their green and red ripe stages were stored at 20 degrees C for 7 and 19 days to determine the effects of storage on whole fruit antioxidant capacity (TAA) and ascorbate (ASC) content, as well as on some antioxidant enzyme activities, such as catalase (CAT), superoxide dismutase (SOD), and those of the ASC-glutathione cycle. At least one Mn-SOD, two Fe-SODs, and three CuZn-SODs were detected in the fruit extract after native polyacrylamide gel electrophoresis. All of the SOD isozymes and glutathione reductase had higher activity levels in the red control fruits than in the green fruits, whereas the activities of monodehydroascorbate and dehydroascorbate reductase were higher in green fruits. Ascorbate peroxidase (APX) was found to be similar in both fruits. SODs, CAT, and APX seem to be involved in pepper fruit ripening and senescence during storage at 20 degrees C, perhaps influencing the active oxygen species levels in the fruit. TAA, as well as the ASC content, was higher in red peppers than in green, and storage increased the ASC in both green and red fruits.

Role of ethylene in the biosynthetic pathway of aliphatic ester aroma volatiles in Charentais Cantaloupe melons.

Compared to other melon types, Cantaloupe Charentais melons are highly aromatic with a major contribution to the aroma being made by aliphatic and branched esters. Using a transgenic line in which the synthesis of the plant hormone ethylene has been considerably lowered by antisense ACC oxidase mRNA (AS), the aliphatic ester pathway steps at which ethylene exerts its regulatory role were found. The data show that the production of aliphatic esters such as hexyl and butyl acetate was blocked in AS fruit and could be reversed by ethylene. Using fruit discs incubated in the presence of various precursors, the steps at which ester formation was inhibited in AS fruit was shown to be the reduction of fatty acids and aldehydes, the last step of acetyl transfer to alcohols being unaffected. However, treating AS fruit with the ethylene antagonist 1-methylcyclopropene resulted in about 50% inhibition of acetyl transfer activity, indicating that this portion of activity was ethylene-dependent and this was supported by the low residual ethylene concentration of AS fruit discs (around 2 microl l(-1)). In conclusion, the reduction of fatty acids and aldehydes appears essentially to be ethylene-dependent, whilst the last step of alcohol acetylation has ethylene-dependent and ethylene-independent components, probably corresponding to differentially regulated alcohol acetyltransferases.

Behaviour of abscisic acid and polyamines in antisense ACC oxidase melon (Cucumis melo) during ripening.

The behaviour of transgenic antisense 1-aminocyclopropane-1-carboxylic acid oxidase melon (TAM) was studied during fruit ripening. Work focussed mainly on analysis of the evolution of other plant growth regulators like ABA and polyamines in this type of melon. Physical-chemical and physiological differences from untransformed melon (UTM) (Cucumis melo L. var. cantalupensis Naud. cv. Védrantais) began to appear from the time of the latter's ethylene crisis. Ethylene synthesis was inhibited by 97.7% in TAM at the time when UTM showed maximum ethylene levels. The ripeness index of TAM was lower, despite their higher soluble solids content, due to the higher titratable acidity. Unlike UTM, TAM rind maintained its colour and cell membrane integrity, although there was no difference in the pulp in this respect. Blocking ethylene biosynthesis affected the levels of ABA and the polyamine putrescine, the latter only in the rind. The possible relationship between ethylene, ABA and polyamines, the effect of blocking ethylene biosynthesis on the evolution of these regulators, and their role in fruit ripening are discussed.