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Essaid Ait Barka, Siamak Kalantari, and Joseph Arul

Fresh fruit and vegetables are highly perishable because of their active metabolism during the postharvest phase. Previous studies showed that hormic dose of UV cause a delay in the senescence of tomato fruit by about 7 days. The objective of this study was to elucidate whether UV acts on the cell membrane in producing the phenomenon of delayed senescence, since it is known that UV radiation can provoke photooxidation of membrane lipids. Membrane lipid peroxidation was studied in tomato fruit (Lycopersicon esculentum Mill cv. Trust) treated by hormic UV dose, and was followed by assaying products of lipid oxidation during the storage period. We observed the production of lipofuscin-like compounds, malondialdehyde, aldehydes, pentane, ethane, and hydrogen peroxide within few days of the treatment. An increase in the efflux of electrolytes (total, potassium, and calcium) was also observed. An immediate increase in the level of these products of oxidation supports the hypothesis that UV radiation induces membrane lipid peroxidation. However, beyond 5 to 7 days after treatment, the production of oxidation products and electrolyte leakage were lower than the control fruits. Thereafter, the level of products of lipid oxidation associated with senescence was higher in control fruits than in treated ones. Results suggest that the initial oxidation stress by the exposure to UV led to biochemical reactions inducing the production of stress compounds, such as polyamines, which are non specific antioxidants. Consequently, a delay in the senescence was observed.

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M.V. Bhaskara Reddy and Joseph Arul

Fruit and vegetables are highly perishable and postharvest pathogens are one of the major causes in the early termination of their shelf life. Although synthetic fungicide are effective against postharvest pathogens, they face imminent problems. One alternate approach is the induction of host resistance against postharvest diseases. We tested the efficacy of chitosan, Milsana®, and Bion® on carrot, tomato, potato, green pepper, and strawberries against postharvest fungal and bacterial rots. Chitosan was not only antimicrobial but also induced resistance against postharvest infections of Alternaria, Botrytis, and Erwinia in tomato, green pepper, carrot, and potato. Preharvest sprays of chitosan induced resistance against postharvest infection of Botrytis cinerea in strawberries and improved the storage quality. Ultrastructure and cytochemical investigations showed that chitosan not only induced several morphological alterations in B. cinerea and Alternaria alternata inhibiting the production of pathogenic factors by the fungi, but also stimulated various host defense reactions, such as papilla formation and induction of defense enzymes, phytoalexins, and phenols. Application of Milsana®, a plant extract, and Bion®, an activator of host defenses, protected carrots from postharvest decay by B. cinerea. Milsana® was effective at 4 °C, while Bion® was effective at both 4 and 13 °C storage temperatures. Thus, there is great potential for defense inducers in postharvest disease control of fruit and vegetables.

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Ahmed El Ghaouth, Rathy Ponnampalam, and Joseph Arul

The effect of chitosan coating on the respiration rate, ethylene production and quality attributes of tomatoes stored at 20°C under high humidity-regular atmosphere was investigated. Chitosan coating reduced significantly the respiration rate and ethylene production of tomatoes, with a greater effect at higher concentration. In addition coating modified the internal microatmoaphere of fruits. Furthermore, coated fruits were firmer, higher in titratable acidity, less decayed and their change in color proceeded at a slower rate than the control.

In conclusion chitosan coating delayed senescence and prolonged storage life of tomatoes, without affecting their market quality by acting as diffusion barrier for gases.

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M.V. Bhaskara Reddy, Alain Asselin, and Joseph Arul

We have investigated the relationship between chitosan treatments and maceration of potato tissue by macerating enzymes secreted by Erwinia carotovora causal agent of soft rot of potato. Erwinia isolated from potato showing soft rot symptoms was used for inoculation. The bacteria secreted a wide spectrum of enzymes that degraded potato cell walls. Polygalacturonase (PG), pectate lyase (PL), pectinmethylesterase (PME), cellulase, xylanase, and protease showed the highest activity in potato tissue inoculated with the pathogen. Accordingly increased maceration and cell death were observed. On the other hand, in chitosan-treated tissue and challenged with the pathogen, significant decrease in enzymatic activity and tissue maceration were observed, more so with increasing chitosan concentration. This observation confirmed that chitosan interfered with multiplication and pathogenic powers of the bacteria, thereby improving cell texture and viability. Crude extracts obtained from treatments were subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) to assess pectinase activity. The electophoretic profiles showed significant lytic zone of pectin degradation in the control, which decreased with increase in chitosan concentrations. No lytic zone was observed at 8 mg·ml–1 chitosan concentration and was comparable to intact activity in untreated potato tissue. Pectic enzyme reaction products were analyzed to see the action pattern of pectinases in the crude extracts. Cellulose choromatographic profiles revealed monomers and dimers of polygalacturonic acid up to 6 mg·ml–1 chitosan concentrations. The results suggest that chitosan significantly inhibits bacterial growth and the production of macerating enzymes by the pathogen and thus chitosan can be a potential anti-bacterial agent.

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Marie-Therese Charles, Alain Goulet, Francois Castaigne, and Joseph Arul

Hormic dose of ultraviolet light (3.7 kJ•m-2) induced disease resistance in tomato fruit. The biochemical nature of induced resistance by UV light was investigated by histochemical techniques. Ultraviolet light induced plasmolysis of the epicarp and few mesocarp cell layers, and collapse of these cell layers led to the formation of cell wall stacking zone (CWSZ). The treatment also stimulated the biosynthesis of phenolic compounds (Prussian blue reaction) in the epicarp and mesocarp cells. Biochemical reinforcement of the cell wall through lignification (Maule test) and suberization (berberine fluorescence) was also induced. These responses originating from the activation of phenylpropanoid path were principally localized in the CWSZ and were induced before inoculation by B. cinerea. The intensity of these responses was significantly increased in UV-treated tissue in response to infection. These responses were also induced in the inoculated control tissue but were either less substantial (phenolics, lignification, and suberization) or delayed.

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Ahmed El Ghaouth, Joseph Arul, Rathy Ponnampalam, and Francois Castaigne

The effect of chitosan coating on green peppers and cucumbers stored at 13°C and 85% R.H. on weight loss, quality and respiration was assessed. Chitosan coating markedly reduced the weight loss of both green peppers and cucumbers, with greater effect at higher concentration. In addition, color loss, wilting, decay and respiration was significantly lower in coated fruits than in the control.

The results of this study indicate that the mechanism by which chitosan coating delay senescence in green peppers and cucumbers is more likely due to its ability to alleviate water stress than to modify the internal microatmosphere.

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Julien Mercier, Mebarek Baka, Baskhara Reddy, Ronan Corcuff, and Joseph Arul

Shortwave ultraviolet radiation (UV-C) was tested for controlling natural infections and inducing resistance to fungal decay caused by Botrytis cinerea Pers.: Fr. (gray mold rot) in bell pepper [Capsicum annuum L. var. annuum (Grossum Group)] fruit. All UV-C doses tested (0.22, 0.44, 0.88, or 2.20 kJ·m-2) caused a reduction in the number of natural infections occurring during storage at 13 °C. A UV-C dose of 0.88 kJ·m-2 controlled most effectively natural infections in peppers stored at both 13 or 20 °C. Although UV-C was found to be highly germicidal to B. cinerea conidia exposed on agar or on fruit wounds, it did not prevent infection of fruit inoculated with the pathogen 24 hours before exposure to UV-C. However, fruit which were exposed to UV-C 24 hours before inoculation with B. cinerea had a lower percentage of infections. For this reason, UV-C appears to act mainly as an inducer of disease resistance in this crop rather than a sanitizing agent. UV-C was effective in inducing resistance to B. cinerea in fruit at various stages of maturity, from green to red. Disease resistance was also induced in fruit which had been stored for 7 days before UV-C treatment. The effect of UV-C doses was found to be additive as two successive exposures at 0.44 kJ·m-2 had an equivalent effect as one exposure to the optimal dose of 0.88 kJ·m-2. However, two successive exposures to 0.88 kJ·m-2 were less effective than one exposure to this dose.

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M.V. Bhaskara Reddy, Paul Angers, Francois Castaigne, and Joseph Arul

Stem scar application of chitosan inhibited growth and production of pathogenic factors by blackmold rot [Alternaria alternata (Fr.:Fr.) Keissl.] in challenged tomato (Lycopersicon esculentum Mill.) fruit stored at 20 °C for 28 days. Blackmold lesions were visible within 4 days of inoculation in control fruit, compared with >7 days in chitosantreated fruit. Macerating enzyme activity (polygalacturonase, pectate lyase, and cellulase) in the tissue in the vicinity of the lesions was <50% in chitosan-treated fruit compared with control fruit. Chitosan also inhibited production of oxalic and fumaric acids (chelating agents) and host-specific toxins such as alternariol and alternariol monomethylether by the fungus. The pH of the infected tissue decreased from 4.7 to 4.0 in the control fruit, the optimum for polygalacturonase activity, while the pH of chitosan-treated fruit remained at 4.6. In addition, chitosan also induced production of rishitin (a phytoalexin) in tomato tissue. Such chitosan-pathogen-host interactions may be exploited in the control of postharvest pathogens of fresh fruit and vegetables.

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Ahmed El Ghaouth, Rathy Ponnampalam, François Castaigne, and Joseph Arul

The effect of chitosan on respiration, ethylene production, and quality attributes of tomato (Lycopersicon esculentum Mill.) fruit stored at 20C was investigated. Coating the fruit with chitosan solutions reduced the respiration rate and ethylene production, with greater effect at 2% than 1% chitosan. Coating increased the internal CO, and decreased the internal O2 levels of the tomatoes. Chitosan-coated tomatoes were firmer, higher in titratable acidity, less decayed, and exhibited less red pigmentation than the control fruit at the end of storage. Chemical name used: 2-amino-2- deoxy-p-D)-glucan (chitosan).

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M.V. Bhaskara Reddy, Essaid Ait Barka, F. Castaigne, and Joseph Arul

The antifungal activity of chitosan, a bioplymer of β-1-4 gluscosamine, against Alternaria alternata, causal agent of black mold of tomato, was investigated. Chitosan was incorporated into potato dextrose broth (PDB) at concentrations of 100, 200, 400, 800, 1600, 3200, and 6400 μg·ml–1, growth and toxin production by the fungus were assessed after a 15-day incubation period. Chitosan significantly affected both growth and toxin production at higher concentrations. However, at lower concentrations, toxin production was affected more than the growth, as evidenced by minimum inhibitory concentrations (MIC) of chitosan derived for toxin production and mycelial growth. Excess sporulation of the fungus was observed in the presence of chitosan, but the spore viability was affected. Chitosan induced aggregation of fungal cells, abnormal shape, excess branching, and hyphal contortion. It also induced leakage of proteins from the fungal cells. The virulence of the toxin in culture filtrate of the fungus from different concentrations of chitosan was assayed by administering on tomato discs. Phospholipid content, electrolyte leakage, xylanase, and pectin methylesterase activity were measured in the culture filtrate administered tomato tissue. Decreased trend in causing electrolyte leakage, phospholipid degradation, and activation of xylanase and pectin methylesterase were observed with increasing concentrations of chitosan. The results showed that chitosan inhibits fungal growth at higher concentrations than toxin production. Further toxin produced at lower concentrations of chitosan was less virulent. Thus chitosan has potential as an antifungal agent.