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See detailTuber formation and growth of Dioscorea cayenensis-D. rotundata complex. Interactions between exogenous and endogenous jasmonic acid and polyamines
Ondo Ovono, Paul ULg; Kevers, Claire ULg; Dommes, Jacques ULg

in Plant Growth Regulation (2010)

Tubers can be initiated and developed in vitro from nodal cuttings of yam (Dioscorea cayenensis - D. rotundata complex). The effect of exogenous jasmonic acid, alone or in combination with putrescine, on ... [more ▼]

Tubers can be initiated and developed in vitro from nodal cuttings of yam (Dioscorea cayenensis - D. rotundata complex). The effect of exogenous jasmonic acid, alone or in combination with putrescine, on these processes was investigated in relationship to endogenous jasmonic acid and polyamine levels. Application of exogenous jasmonic acid at various concentrations positively affected microtuber formation and growth from yam nodal cuttings. In control conditions, three weeks were needed to obtain 100% of tuberisation. Jasmonic acid at low level (0.1 µM) accelerated tuber formation (46% after one week) as did putrescine (10 µM). But endogenous levels of jasmonic acid were not significantly affected by its exogenous presence in the medium. Jasmonic acid also interacted with other growth regulators as polyamines, but the decrease in time necessary to observe tuber formation could not be correlated with endogenous modifications of PUT content. The presence of jasmonic acid (0.1 to 1 µM) as PUT (1 µM) induced also an increase of tuber length and weight. The combination of jasmonic acid (0.1 µM) and putrescine (1 µM) had no positive effect on tuber formation (precocity) but had an additive effect on further growth (length and weight). In the future, these results could help the optimising in vitro conditions for mass production of larger yam microtubers. [less ▲]

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See detailIn Vitro Preservation of Yam ( Dioscorea cayenensis-D.rotundata complex) for a Better Use of Genetic Resources
Ondo Ovono, Paul ULg; Kevers, Claire ULg; Dommes, Jacques ULg

in Notulae Botanicae Horti Agrobotanici Cluj-Napoca (2010), 38

Among the food crops, yam takes up quantitatively the first place in the gabonese diet. Unfortunately, it can stay available only 6 to 7 months in the year because of difficulties of harvest and post ... [more ▼]

Among the food crops, yam takes up quantitatively the first place in the gabonese diet. Unfortunately, it can stay available only 6 to 7 months in the year because of difficulties of harvest and post- harvest. This problem is little studied in the case of Dioscorea cayenensis-D. rotundata complex. In order to optimize the use of micro tubers for the growing in green house or field, it is important to control the duration of storage before the germination. The present study concerns microtubers obtained by in vitro culture. When microtubers were harvested (after 9 months of culture) and directly transferred on a new medium without hormone, the tubers rapidly sprouted in in vitro conditions. Harvested microtubers were also stored dry in jars in sterile conditions during 2 to 18 weeks before in vitro sprouting. In this case, microtubers stored during 18 weeks sprouted more rapidly than those stored 8 weeks. The size of the tubers used for the storage had great influence on further sprouting. The upper microtubers in 25 mm can be kept to the darkness, under 50% of relative humidity, in 25°C during at least 18 weeks. Sprouting is 100% whatever the substrate of culture. The plant tissue culture technique constitutes a serious alternative for the preservation of plant kinds and for the production of planting material. These techniques allow multiplying in a short time of thousands of copies of new varieties of newly created plants. These in vitro plants can be used on one hand, for the production planting material, and on the other hand for ex vitro storage of breeding grounds with decelerated growth, to struggle against genetic erosion. These results should allow improving in practice the multiplication of yam, while guaranteeing phytosanitary qualities. [less ▲]

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See detailEvaluation of spectrophotometric methods for antioxidant compound measurement in relation to total antioxidant capacity in beverages.
Tabart, Jessica ULg; Kevers, Claire ULg; Pincemail, Joël ULg et al

in Food Chemistry (2010), 120

The validity of different colorimetric methods used to quantify various families of antioxidant compounds was evaluated with standard compounds. The colorimetric tests for global evaluation of flavonoids ... [more ▼]

The validity of different colorimetric methods used to quantify various families of antioxidant compounds was evaluated with standard compounds. The colorimetric tests for global evaluation of flavonoids, anthocyans, and flavanols were found generally unreliable, as reactions could be different for individual compounds within a family (anthocyanins or flavonols or flavan-3-ols) and not specific to one family. In the flavonoid test, for example, flavonols reacted very well, anthocyanins did not react, and flavanons reacted only slightly. The same methods were applied also to beverages known for their antioxidant content (apple, orange, grape, and vegetable juices, ice tea, and red wine) and the data were compared with the results of HPLC analysis of specific compounds. The values obtained in a colorimetric test were generally higher than the sum of the values obtained for the corresponding individual compounds by HPLC analysis, mainly because other compounds can interfere with the colorimetric tests. For example, in wine, anthocyanin concentrations obtained by colorimetric test were 9068  1407 µ mol /100ml (mean  SEM), higher than the sum of the six main anthocyanidins detected by HPLC, only 41 µmol/100 ml. The relative antioxidant capacity values determined for beverages on the basis of colorimetric tests could exceed by far the values previously measured in radical-scavenging tests (for instance, the antioxidant capacity attributable to anthocyans in wine on the basis of the colorimetric test was 50 times higher than the total antioxidant capacity measured by the ORAC assay). In conclusion, colorimetric tests for flavonoids, anthocyans, and flavanols appeared generally unreliable for estimating their content and thus the antioxidant capacity reliable to these compounds. [less ▲]

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See detailANTIOXIDANT CAPACITY OF THREE CUBAN SPECIES OF THE GENUS Pluchea Cass. (Asteraceae)
Perera, Wilmer; Tabart, Jessica ULg; Gomez, Abel et al

in Journal of Food Biochemistry (2010), 34

Leaves of three Cuban species of the genus Pluchea: P. carolinensis, P. odorata and P. rosea were extracted with various solvents and analysed. Highest values of phenolic compounds were detected after ... [more ▼]

Leaves of three Cuban species of the genus Pluchea: P. carolinensis, P. odorata and P. rosea were extracted with various solvents and analysed. Highest values of phenolic compounds were detected after ethyl acetate (EtOAc) and n-butyl alcohol (n-BuOH) extractions. Flavonoids were detected after chloroform, EtOAc and n-BuOH extractions of the species Pluchea and three aglycone flavonol forms (quercetin, kaempferol and myricetin) were also quantified after EtOAc and n-BuOH extractions, using HPLC. The total antioxidant capacity (TAC), expressed as Trolox equivalents per gram of leaf dry weight (TE/g dw) was analyzed using 2,2-diphenyl-1-picrylhydrazyl (DPPH•), 2,2’-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS∙+), and the oxygen radical absorbance capacity (ORAC). The highest values of TAC were detected after EtOAc and n-BuOH extractions of the three species. P. carolinensis was the promising species; being the n-BuOH extraction with the most bioactive compounds : 15.3 mg TE/g dw using DPPH∙ [less ▲]

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See detailTuber formation and development of Dioscorea cayenensis-D. rotundata complex in vitro. Effect of polyamines.
Ondo Ovono, Paul ULg; Kevers, Claire ULg; Dommes, Jacques ULg

in In Vitro Cellular & Developmental Biology-Plant (2009)

Tuberisation was obtained in vitro on yam (Dioscorea cayenensis-D.rotundata complex). The effect of exogenous polyamines on tuber formation and development (length and weight of microtubers) were ... [more ▼]

Tuberisation was obtained in vitro on yam (Dioscorea cayenensis-D.rotundata complex). The effect of exogenous polyamines on tuber formation and development (length and weight of microtubers) were investigated and discussed in relation with changes in endogenous polyamines. Application of exogenous polyamines, inhibitors of their metabolism and polyamines precursors in various concentrations positively affected microtuber formation by yam nodal cuttings and their further development. In control conditions, three weeks are needed to obtain 100% of tuberisation. With low concentrations of putrescine (10 5 or 10 6 M), tuber formation occurred earlier. Polyamine endogenous level and metabolism can be significantly affected by exogenous polyamines but modifications of endogenous free polyamines could not be directly correlated to the tuber formation process. Increases in endogenous putrescine and auxins were observed in tubers showing a better development in the presence of putrescine. These results can be used for optimising in vitro conditions for mass production of larger microtubers of the Dioscorea cayenensis-D. rotundata complex. [less ▲]

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See detailEffects of reducing sugar concentration on in vitro tuber formation and sprouting in yam (Dioscorea cayenensis-D. rotundata complex).
Ondo Ovono, Paul ULg; Kevers, Claire ULg; Dommes, Jacques ULg

in Plant Cell, Tissue & Organ Culture (2009), 99(1), 55-59

The effects of reducing sucrose level on tuber formation (% of cultures with microtubers), development (length and fresh weight of microtubers) and sprouting in yam Dioscorea cayenensis-D. rotundata ... [more ▼]

The effects of reducing sucrose level on tuber formation (% of cultures with microtubers), development (length and fresh weight of microtubers) and sprouting in yam Dioscorea cayenensis-D. rotundata complex in vitro were investigated. Only 29% of the explants showed tuber formation after 3 weeks in the presence of 1% sucrose in contrast to 100% with 3%. After 120 days of culture, the length and the weight of the tubers obtained in the presence of 1% sucrose were less than with 3% sucrose. Addition of sorbitol to keep osmolarity at the same level did not restore normal rate of tuber formation. Similar results were obtained with the use of reduced fructose or glucose level. Microtuber sprouting was also affected by sucrose level incorporated into the tuberisation medium. Tubers obtained on reduced sucrose level sprouted later and the increase of osmolarity with sorbitol did not restore normal sprouting. The bigger tubers obtained on high sucrose media could contain more carbohydrate reserves that could partially explain a higher sprouting rate. These results can be used for optimising in vitro conditions for mass production of microtubers in yam and especially in Dioscorea cayenensis-D. rotundata complex, a very important species in West Africa. They specially showed the importance of tuberisation conditions on precocity of tuberisation, on tuber length and weight and on their further sprouting. [less ▲]

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See detailInsights into the Defense-Related Events Occuring in Plant Cells Following Perception of Surfactin-Type Lipopeptide from Bacillus subtilis
Jourdan, Emmanuel ULg; Henry, Guillaume; Duby, F. et al

in Molecular Plant-Microbe Interactions [=MPMI] (2009), 22(4), 456-468

Multiple strains of Bacillus subtilis were demonstrated to stimulate plant defense responses, and cyclic lipopeptides may be involved in the elicitation of this induced systemic resistance phenomenon ... [more ▼]

Multiple strains of Bacillus subtilis were demonstrated to stimulate plant defense responses, and cyclic lipopeptides may be involved in the elicitation of this induced systemic resistance phenomenon. Here, we further investigated molecular events underlying the interaction between wuch lipopeptides and plant cells. Addition of surfactin but not fengycin or iturin in the micromolar range to tobacco cell suspensions induced defense-related early events such as extracellular medium alkalinization coupled with ion fluxes and reactive oxygen species production. Surfactin also stimulated the defense enzymes phenylalanine ammonia lyase and lipoxygenase and modified the pattern of phenolics produced by the elicited cells. The occurence of these surfactin-elicited early events is closely related to Ca2+ influx and dynamic changes in protein phosphorylation but is not associed with any marked phytotoxicity or adverse effect on the integrity and growth potential of the treated tobacco cells. [less ▲]

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See detailCombining proteomics and metabolite analyses to unravel cadmium stress-response in poplar leaves
Kieffer, Pol ULg; Planchon, Sébastien; Oufir, Mouhssin et al

in Journal of Proteome Research (2009), 8

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See detailWhat we have learned about the physiology of in vitro adventitious rooting of woody plants and how it relates to improvements in the practice
Kevers, Claire ULg; Hausman, Jean-françois; Faivre-Rampant, Odile et al

in Niemi, Karoliina (Ed.) Adventitious root formation of forest trees and horticultural plants - from genes to applications (2009)

Natural auxins and synthetic analogs are the most powerful exogenous stimulators of adventitious rooting of ligneous as well as for herbaceous cuttings. For several years we have investigated the ... [more ▼]

Natural auxins and synthetic analogs are the most powerful exogenous stimulators of adventitious rooting of ligneous as well as for herbaceous cuttings. For several years we have investigated the physiology of rooting, including the metabolism of endogenous auxins and their molecular mechanisms of action. Our work has resulted in discoveries leading to practical strategies allowing faster and improved rooting of cuttings from shoots raised in vitro, and subsequently resulting in roots better adapted for acclimatization and to overcome so-called (apparent) recalcitrance-to-rooting of some species. In this chapter we describe: (i) how variation in auxin treatments during rooting can be used to regulate the endogenous levels of auxins; (ii) the importance of not maintaining auxin treatment throughout the whole rooting process, thereby the endogenous auxin level itself not always being high; (iii) the division of the organogenetic process into at least three main rooting interdependent phases, i.e. induction, initiation, and expression, with different durations depending on the species and with different requirements for each phase, and (iv) how polyamines, even if less effective from outside, and their metabolism are as important as auxins in the rooting process. We will also describe how cross-talk between hormones, manipulations with related genes and mutants, and some similarities between the induction of rooting and evocation of flowering, offer new tools for further progress in the field. [less ▲]

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See detailInvolvement of polyamines in the adventitious rooting of micropropagated shoots of the apple rootstock MM106
Naija, Sélima; Elloumi, Nadhra; Ammar, Saida et al

in In Vitro Cellular & Developmental Biology-Plant (2009), 45

Apple rootstock MM106 shoots, raised in vitro, rooted at 96.7% after culture on a medium supplemented with an auxin for 5 d in darkness followed by culture on a second medium without growth regulators for ... [more ▼]

Apple rootstock MM106 shoots, raised in vitro, rooted at 96.7% after culture on a medium supplemented with an auxin for 5 d in darkness followed by culture on a second medium without growth regulators for 25 d in light. In control conditions (in absence of auxin in the first medium), these shoots did not root. Putrescine (PUT), spermidine (SPD), cyclohexylamine (CHA), and aminoguanidine (AG) enhanced rooting when applied during the first d of culture in the absence of IBA; on the contrary, α-difluoromethylornithine (DFMO) added to the first medium with IBA inhibited rooting. The endogenous levels of indole 3-acetic acid (IAA) and indole 3-acetylaspartic acid (IAAsp) increased up to a maximum concentration at days 2 and 3, respectively, in initial rooting conditions. PUT, when added with IBA, did not affect the typical IAA and IAAsp increase; when applied alone, it provoked an increase of their levels. Similar results were recorded with CHA. SPD, AG, and DFMO did not induce an increase of IAA and IAAsp in nonrooting conditions. The levels of endogenous PUT increased to a maximum at day 2 in rooting conditions; it was slightly affected by exogenous PUT and CHA application but reduced by SPD, AG, and DFMO. In rooting conditions, if the first medium was supplemented with SPD or AG, a small increase in peroxidase activity was observed, similar to that obtained with PUT treatment. The present work indicates an involvement of polyamines in the control of rooting and an interaction with auxins during the physiological phase of rooting. The consequence of this relationship was a different rooting expression, according especially to the content of these regulators in the culture medium. [less ▲]

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See detailComparative antioxidant capacities of phenolic compounds measured by various tests
Tabart, Jessica ULg; Kevers, Claire ULg; Pincemail, Joël ULg et al

in Food Chemistry (2009), 113

The purpose of this study was to compare the antioxidant capacities of standard compounds (phenolic compounds, ascorbic acid, and glutathione) as measured by various assays. Five methods were selected so ... [more ▼]

The purpose of this study was to compare the antioxidant capacities of standard compounds (phenolic compounds, ascorbic acid, and glutathione) as measured by various assays. Five methods were selected so as to span a diversity of technical approaches: TEAC (radical 2,2′-azino-bis(3-ethylbenzothiazoline)-6 sulphonic acid), DPPH (radical 2,2-diphenyl-1-picryhydrazyl used to measure reducing capacity), ORAC (oxygen radical scavenging capacity), red blood cell haemolysis (protection of biological sample), and ESR (electron spin resonance for direct free radical evaluation). Most compounds showed significant differences in free radical scavenging activity according to the method used. Of the 25 tested compounds, only a few, such as myricetin and gallocatechin, gave comparable activities in the various tests. To standardise reporting on antioxidant capacity, it is proposed to use a weighted mean of the values obtained using the DPPH, ORAC, resistance to haemolysis, and ESR assays. This strategy was used to test the antioxidant capacity of several beverages. The highest antioxidant capacity was observed for red wine, followed by green tea, orange juice, grape juice, vegetable juice, and apple juice. [less ▲]

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See detailProteomic and enzymatic response of poplar to cadmium stress
Kieffer, Pol; Schröder, Peter; Dommes, Jacques ULg et al

in Journal of Proteomics (2009), 72

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See detailCharacterization and Cloning of Chitin Deacetylases from Rhizopus Circinans
Gauthier, Carole ULg; Clerisse, Fabienne ULg; Dommes, Jacques ULg et al

in Protein Expression & Purification (2008), 59

Chitin deacetylase catalyzes hydrolysis of the acetamido groups of N-acetylglucosamine of chitin in fungal cell walls. Here a chitin deacetylase secreted by Rhizopus circinans was purified to homogeneity ... [more ▼]

Chitin deacetylase catalyzes hydrolysis of the acetamido groups of N-acetylglucosamine of chitin in fungal cell walls. Here a chitin deacetylase secreted by Rhizopus circinans was purified to homogeneity and partially characterized. The enzyme exhibits an apparent molecular weight of approximately 75kDa. At 37 degrees C it shows optimal activity at pH 5.5-6. Its pH stability and thermal stability are good. Mn(2+) and Mg(2+) slightly enhance the activity of the enzyme and Cu(2+) strongly inhibits it. An R. circinans cDNA library was constructed and screened with a homologous probe synthesized by RT-PCR or with synthetic primers derived from the N-terminal amino-acid sequence of the native purified chitin deacetylase. Three chitin deacetylase cDNAs (RC, D2, and I3/2) were isolated from the cDNA library and sequenced. These cDNAs exhibit features characteristic of chitin deacetylase sequences: the presence of a polysaccharide deacetylase domain, a metal-binding triad, the conserved catalytic residues, and high homology with various chitin deacetylase genes. The cDNAs were cloned in a Pichia pastoris expression system and produced as polyhistidine-tagged proteins. Only one recombinant enzyme (called RC) was active under the tested conditions. It was purified to homogeneity in a single step and further characterized. The protein showed an apparent molecular mass of approximately 75kDa and, like the native enzyme, showed optimal activity at pH 5.5-6 at 37 degrees C. It was strongly inhibited by Cu(2+). The isolation of several chitin deacetylase cDNAs from the same microorganism is discussed. [less ▲]

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See detailSystemic resistance induction in tomato by Pseudomonas putida BTP1: investigation of defense pathways.
Mariutto, M.; Duby, Franceline ULg; Ongena, Marc ULg et al

in Journal of Plant Pathology [=JPP] (2008), 90

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See detailSystemic resistance and lipoxygenase-related defence response induced in tomato by Pseudomonas putida strain BTP1.
Akram, Adam; Ongena, MARC ULg; Duby, Franceline et al

in BMC Plant Biology (2008), 8

BACKGROUND: Previous studies showed the ability of Pseudomonas putida strain BTP1 to promote induced systemic resistance (ISR) in different host plants. Since ISR is long-lasting and not conducive for ... [more ▼]

BACKGROUND: Previous studies showed the ability of Pseudomonas putida strain BTP1 to promote induced systemic resistance (ISR) in different host plants. Since ISR is long-lasting and not conducive for development of resistance of the targeted pathogen, this phenomenon can take part of disease control strategies. However, in spite of the numerous examples of ISR induced by PGPR in plants, only a few biochemical studies have associated the protective effect with specific host metabolic changes. RESULTS: In this study, we showed the protective effect of this bacterium in tomato against Botrytis cinerea. Following treatment by P. putida BTP1, analyses of acid-hydrolyzed leaf extracts showed an accumulation of antifungal material after pathogen infection. The fungitoxic compounds thus mainly accumulate as conjugates from which active aglycones may be liberated through the activity of hydrolytic enzymes. These results suggest that strain BTP1 can elicit systemic phytoalexin accumulation in tomato as one defence mechanism. On another hand, we have shown that key enzymes of the lipoxygenase pathway are stimulated in plants treated with the bacteria as compared with control plants. Interestingly, this stimulation is observed only after pathogen challenge in agreement with the priming concept almost invariably associated with the ISR phenomenon. CONCLUSION: Through the demonstration of phytoalexin accumulation and LOX pathway stimulation in tomato, this work provides new insights into the diversity of defence mechanisms that are inducible by non-pathogenic bacteria in the context of ISR. [less ▲]

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See detailQuantitative changes in protein expression of cadmium-exposed poplar plants
Kieffer, Pol; Dommes, Jacques ULg; Hoffmann, Lucien et al

in Proteomics (2008), 8

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See detailAxillary proliferation and tuberisation of Dioscorea cayenensis-D-rotundata complex
Ovono, Paul O; Kevers, Claire ULg; Dommes, Jacques ULg

in Plant Cell, Tissue & Organ Culture (2007), 91(2), 107-114

Yams (Dioscorea spp) are tuber crops used as staple food in Africa because of their nutritional value. However agronomic constraints, phytosanitary problems and the lack of good healthy planting material ... [more ▼]

Yams (Dioscorea spp) are tuber crops used as staple food in Africa because of their nutritional value. However agronomic constraints, phytosanitary problems and the lack of good healthy planting material restrict their production. In contrast to the inefficiency of traditional method of planting, tissue culture techniques allow to increase the multiplication and the rapid production of pathogen- free plant material. This work was undertaken to provide farmers in African countries with healthy microplants and microtubers as seeds. In vitro nodal segments of two varieties of local yams D. cayenensis-D. rotundata complex (cv.'Singo', cv. 'Singou' and cv. 'Gnidou') were micropropagated on the modified medium of Murashige and Skoog. The morphogenesis, the growth of microplants and microtuber formation have been found to be controlled by external factors that act individually and synergistically. Addition of kinetin (2 mg l(-1)) to the culture media could reduce multiplication rate (node number) of some clones. An increase of the sucrose concentration from 3% to 5% induced no change in the multiplication and tuberisation parameters. An important reduction of the multiplication (shoot number, height and node number) and the tuberisation (tuber number and length) was observed with 8% sucrose. Multiplication (shoot and node number) was increased in the presence of jasmonic acid (10 mu M). JA also induced an increase of tuber number in the absence of Kin. Multiplication of yam by in vitro growth of nodal segments is a way for rapid clonal multiplication and could allow solving the problem of lack of seed material faced by farmers. This method could also be used for multiplication of elite cultivars, independently of the growing season. [less ▲]

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See detailEvolution of antioxidant capacity during storage of selected fruits and vegetables
Kevers, Claire ULg; Falkowski, Michael; Tabart, Jessica et al

in Journal of Agricultural and Food Chemistry (2007), 55(21), 8596-8603

Interest in the consumption of fresh fruits and vegetables is, to a large extent, due to its content of bioactive nutrients and their importance as dietary antioxidants. Among all of the selected fruits ... [more ▼]

Interest in the consumption of fresh fruits and vegetables is, to a large extent, due to its content of bioactive nutrients and their importance as dietary antioxidants. Among all of the selected fruits and vegetables, strawberries and black grapes have relatively high antioxidant capacities associated with high contents of total phenolic compounds, ascorbic acid, and flavonols. More interesting, the results of this study indicated that in most fruits and vegetables storage did not affect negatively the antioxidant capacity. Better, in some cases, an increase of the antioxidant capacity was observed in the days following their purchase, accompanied by an increase in phenolic compounds. In general, fruits and vegetables visually spoil before any significant antioxidant capacity loss occurs except in banana and broccoli. When ascorbic acid or flavonoids (aglycons of flavonols and anthocyanins) were concerned, the conclusions were similar. Their content was generally stable during storage. [less ▲]

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