Functional characterization of grapevine MLO genes to define their roles in Powdery mildew susceptibility by CRISPR/Cas9 genome editing

Wine contains secondary metabolites derived from aromatic amino acids (AADC), which can determine quality, stability and bioactivity. Several yeast species, as well as some lactic acid bacteria (LAB), can contribute in the production of these aromatic compounds. Winemaking should be studied as a series of microbial interactions, that work as an interconnected network, and can determine the metabolic and analytical profiles of wine. The aim of this work was to select microorganisms (yeast and LAB) based on their potential to produce AADC compounds, such as tyrosol and hydroxytyrosol, and design a microbial consortium that could increase the production of these AADC compounds in wines.

The temperature increase related to ongoing climate changes is causing a progressive anticipation of the ripening time, negatively affecting grape quality at harvest.

AIM AND METHODS: Grape withering is one of the key steps in the production of the most renowned red wines of the Valpolicella area, namely Amarone and Recioto. This practice, which was already used since Roman times, entails important modifications in grape composition and in the chemical and sensorial characteristics of the corresponding wines, especially in terms of aromatic profile. The aim of this research is evaluating the aromatic evolution during grape withering of the three main varieties used in Valpolicella wines: Corvina, Corvinone and Rondinella.Samples of the three varieties were analyzed at harvest and at different stages of withering, namely10%, 20% and 30% of weight loss. Free and glycosidically bound compounds were extracted and analyzed using Gas Chromatography- Mass Spectrometry (GC-MS). RESULTS: For all the samples the data were normalized to eliminate the effect of concentration due to grape dehydration. Terpene content and evolution varied considerably in relationship to grape variety. Corvinone was richer in cyclic terpenes (including phellandrene, limonene, and cymene) and they decreased during withering.

Pierce’s disease (PD) is considered a potential threat to european viticulture (EPPO a2 list of pathogens since 1981). In the usa, infections caused by the vector-borne bacterium xylella fastidiosa have caused recurrent damage to vineyards in California and the southeastern states. However, vineyards in Europe have remained free of PD until recently, when it was first detected on the island of Mallorca in 2017. The reasons for the absence of PD in continental Europe have not been convincingly explained.

The advance in maturation of wine grapes is an important climate change risk related effect that could affect warm regions like Portuguese Douro Wine Region. Indeed, the climate analysis over the past years registered a decrease in the precipitation, significant higher average temperatures, and a more frequent occurrence of extreme weather events, including heat waves. In these conditions the length from anthesis until maturation is shortened and the uncoupling of technical and phenolic maturity results in berries with higher sugar concentration (and lower acidity), but lower anthocyanins, tannins, and total phenolic concentration, which produce unbalanced wines.
In this work, an innovative strategy of crop forcing, based on forcing vine regrowth after a second pruning of green shoots, was tested, aimed at delaying ripening until the temperature becomes lower and, therefore, preventing acidity loss and increasing anthocyanin-to-sugar ratio. The experiments were conducted in 2019 and 2020 in a commercial vineyard of ‘Touriga Nacional’ located in the Douro Region. Crop forcing was conducted 15 (CF1) to 30 (CF2) days after fruit set. Vines pruned with conventional methods were used as control (CF0). Results confirmed that fruit ripening was shifted from the hot season (August/September), until a cooler period (October through early-November). At harvest, grapevine berries from CF1 and CF2 presented lower pH and higher acidity, than control, with no significant differences in colour intensity and phenolic levels composition. Sugar content was lower in CF2-treated vines in both seasons. However, in CF-treated vines the number and size of clusters were significantly lower (up to 88% reduction) than in control plants. A metabolomics analysis of mature berries from CF-treated vines and control is underway. Crop forcing was indeed effective in producing a more balance berry composition but severely reduced grapevine yield,