Grapevines are sensitive to a wide range of fungal pathogens, including agents such as Phaeomoniella chlamydospora and Phaeoacremonium aleophilum that cause tracheomycosis. In the present study, a procedure for DNA extraction from grapevine woody tissue is first evaluated and shown to be suitable for quantitative analysis. Next, a multiplex real-time PCR method targeting the β-tubulin gene of the pathogens and the actin gene of plant material is developed and its quantitative capability is verified. This protocol was evaluated in inoculated grapevine-wood samples and in young vines from a nursery and was found to be reliable and highly specific. Results obtained from inoculated cuttings show that the fungal colonization process must be considered regardless of the wood phenotype. An analysis of samples of young vines from the nursery shows that a high rate of contamination occurs at the basis of plants and that this contamination is associated with low quantitative values. This finding provides evidence that in vine nurseries, these fungi may be efficient soil-borne pathogens.
In most vineyards worldwide, agents of grapevine trunk diseases represent a real threat for viticulture and are responsible for significant economic loss to the wine industry. The conventional microbiological isolation technique used to diagnose this disease is tedious and frequently leads to false negatives. Thus, a dire need exists for an alternative method to detect this disease. One possible way involves infrared spectroscopy, which is a rapid, nondestructive analytical tool that is commonly used for quality control of feed stuffs. In the present work, a midinfrared spectrometer was tested as a fast tool for detecting agents of grapevine trunk disease. Midinfrared spectra were collected from 70 Vitis vinifera L. cv. Cabernet-Sauvignon one year old trunk-wood samples that were infected naturally in one viticulture nursery of the south of France. The samples underwent polymerase chain reaction and morphological identification, and the results were correlated to the midinfrared spectra by using multivariate analysis to discriminate between noninfected and infected samples. Based on comparison with some control samples, the highest percentage of correct identification of fungal contamination when using the midinfrared spectroscopy method is 80%.
Purpose of the review: The involvement of ethylene in fruit ripening has been the subject of intensive molecular and biochemical studies over the last twenty years. Whereas new methods and new genes are being discovered, the differences between climacteric and nonclimacteric fruit seem to decrease. Recent studies are showing potential roles for ethylene signals in grape berry ripening.Main findings: Ethylene seems to be involved in some changes that occur during the grape berry ripening, the final step of berry development. The whole ethylene production pathway seems activated at the inception of the grape berry ripening, the veraison. Treatments with exogenous ethylene stimulate the long term expression of genes related to the anthocyanin synthesis. Ethylene signals are also involved in the regulation of vascular fluxes and acid content. The ethylene may also been involved in some steps of aroma production, modulated by alcohol dehydrogenases.Directions for future research: These findings could help to further unravel the differences between climacteric and non-climacteric fruit. In the future, whole genome sequencing and computer analyses will surely provide new tools to complete our knowledge of ethylene involvement in the ripening of these two categories of fruit. Regarding the economic importance of fruit ripening and preservation, this knowledge is crucial.
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