A method has been developed for nonvolatile compound analysis in fruits, namely, trans-resveratrol in grapes and vine leaves. This has been accomplished by the combination of laser desorption with resonance-enhanced multiphoton ionization coupled to time-of-flight mass spectrometry detection. After the optimization of the experimental conditions and the finding of the resonant wave-length of the substance, the full validation of the technique (i.e., linearity, repeatability, reproducibility, accuracy, detection limit, and quantification limit) was carried out with satisfactory results. Essential features of the method are as follows: (1) an enhanced desorption yield due to the mixing of the analyte with metal powder and (2) a high resolution and sensitivity and a low detection limit due to laser resonant ionization and mass spectrometric detection. Application to the analysis of trans-resveratrol in vine leaves and grape skin demonstrated the capabilities of the analytical method reaching detection limits of only few ppb.
As it is well-known, one of the main problems of modern agriculture is the postharvest fruit losses due to pathogen's attack and natural senescence during storage. Well established solutions to improve this situation, such as, for example, storage under controlled conditions and the use of synthetic pesticides, are not free of problems due to human health risks and environmental effects caused by chemical pesticides. A new strategy to solve these problems consists of developing methods to improve the natural plant resistance by using, upon their identification, the plant's own defense molecules, in other words, applying methods based on the plant's own natural processes of pest suppression to control spoilage. This requires the identification of components of the natural defense response in plants, which, in turn, demands highly sensitive, fast, and versatile analytical methods especially for trace, nonvolatile, compounds. In this work a laser-based technique has been applied for screening the postharvest elicitation of resveratrol by Botrytis cinerea in grapes. Besides antifungal character, resveratrol is known to present important antioxidant properties, which could also have positive effects on fruit conservation during storage. Consequently, several experiments were carried out in which exogenous application of resveratrol to several fruits maintained their postharvest quality. The quality of both resveratrol-treated and untreated fruits has been studied by the assessment of the biochemical composition and sensory analysis. Indeed, the present work demonstrates that the external application of resveratrol does not alter the sensorial and biochemical properties of the fruit.
Two modern laser-based techniques were synchronously applied to study the dynamics of the trans-resveratrol activity in Botrytis cinerea-infected grapes. Direct analysis of trans-resveratrol in both infected and noninfected grapes (Vitis vinifera, Aledo variety) was performed by using an analytical technique incorporating laser desorption coupled with laser resonant ionization and time-of-flight mass spectrometry. On the other hand, one of the most sensitive on-line methods for trace gas detection, laser photoacoustic spectroscopy, was used to investigate the involvement of the plant hormone ethylene (C 2 H 4 ) in the B. cinerea grapes interaction and its temporal relationship with the trans-resveratrol content upon infection. The trans-resveratrol content and the ethylene released by noninfected grapes showed an opposite behavior. In this case, a high trans-resveratrol content corresponds to a low ethylene emission. For the B. cinerea-infected grapes, ethylene emission rises up after 48 h when the analogous content of trans-resveratrol started to decrease irreversibly. Moreover, the activity of trans-resveratrol as natural pesticide has been investigated by exogenous application on grapes. A short submerge (5 s) of the grapes in 1.6 ϫ 10 Ϫ4 m solution of trans-resveratrol delays the increase of C 2 H 4 emission with about 48 h and produces a decrease of the C 2 H 4 concentration and its emission rate. The treatment has positive effects on fruit conservation during storage; it doubled the normal shelf-life of grapes at room temperature, maintaining their post-harvest quality within 10 d.The fungus Botrytis cinerea is a plant necrotrophic pathogen that colonizes senescent or dead plant tissues and causes softening in fruits. Fungal hyphae can penetrate through wounds or natural openings of the plant tissue and spread from previously colonized dead tissues into healthy tissues. B. cinerea attacks different plant tissues and has a broad host range. It is a major cause of post-harvest rot of perishable plant products, including grapes (Vitis vinifera) at harvest and in storage. Because it is also able to infect at low temperatures, it can result in important economic losses, either in pre-and post-harvest crops (Mansfield, 1980). Under a pathogen attack, plants evolve sophisticated systems of detection and response to decipher the pathogen signals and to induce appropriate defenses. These systems include specific networks that operate through the action of signaling molecules such as salicylate, jasmonate, and ethylene and generate the accumulation of pathogenicity-related proteins, phytoalexins, or other phenolic compounds (Elad, 1997; Dong, 1998; Feys and Parker, 2000).trans-Resveratrol (3,5,4Ј-trihydroxystilbene) is an antioxidant compound naturally produced in a huge number of plants, including grapevine where it is the major component of the phytoalexin response of the plant. It is accumulated in vine leaves and grape skin in response to various fungal infections, UV radiation, or chemicals (Langcake, 1981; Jea...
Resveratrol is known as a grapevine secondary metabolite with fungicide activity. Its exogenous application on harvested grapes resulted in the reduction of microbial flora growth, and consequently, prolonged shelf life, without affecting the nutritional quality of the fruit. Resveratrol treatment also resulted in being effective on fruit that normally does not accumulate such metabolites as, for example, tomatoes, apples, avocado pears, and peppers. As a result, all treated fruits maintained their post-harvest quality and health longer than the untreated ones. This study demonstrates the potential use of resveratrol as a natural pesticide to reduce post-harvest fungi development on a broad spectrum of fruit types.
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