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

New Plant Breeding Techniques (NPBTs) have arisen with the objective of surmounting the constraints inherent in conventional breeding methodologies, thereby enhancing plant resilience against both biotic and abiotic stresses. To date the application of genome editing in grapevine is still limited by the necessity to overcome recalcitrance to produce embryogenic calli and to regenerate plants. In our studies, we developed a smart and versatile genetic transformation system carrying all the most promising features of different genome editing approaches. In specific, we joined the GRF-GIF expression to improve regeneration, the systemic movement of the editing transcripts through tRNA-like sequences (TLS) and the cisgenic-like approach to remove transgenes.

The aim of this study was to evaluate the effects of different soil types on the chemical composition of Mediterranean red wines, through untargeted and targeted 1H-NMR metabolomics. One milliliter of raw wine was analyzed by means of a Bruker Avance II 400 spectrometer operating at 400.15 MHz. The spectra were recorded by applying the NOESYGPPS1D pulse sequency, to achieve water and ethanol signals suppression. No modification of the pH was performed to avoid any chemical alteration of the matrix. The generation of input variables for untargeted analysis was done via bucketing the spectra. The resulting dataset was preprocessed prior to perform unsupervised PCA, by means of MetaboAnalyst web-based tool suite. The identification of compounds for the targeted analysis was performed by comparison to pure compounds spectra by means of SMA plug-in of MNova 14.2.3 software. The dataset containing the concentrations (%) of identified compounds was subjected to one-way analysis of variance (ANOVA) to highlight significant differences among the wines. The untargeted analysis, carried out through the PCA, revealed a clear differentiation among the wines. The fragments of the spectra contributing mostly to the separation were attributed to flavonoids, aroma compounds and amino acids. The targeted analysis leaded to the identification of 68 compounds, whose concentrations were significant different among the wines. The results were related to soils physical-chemical analysis and showed that: 1) high concentrations of flavan-3-ols and flavonols are correlated with high clay content in soils; 2) high concentrations of anthocyanins, amino acids, and aroma compounds are correlated with neutral and moderately alkaline soil pH; 3) low concentrations of flavonoids and aroma compounds are correlated with high soil organic matter content and acidic pH. The 1H-NMR metabolomic analysis proved to be an excellent tool to discriminate between wines originating from grapes grown on different soil types and revealed that soils in the Mediterranean area exert a strong impact on the chemical composition of the wines.

The Tannat grape, native to the foothills of the Pyrenees in France, is known for producing wines with intense colour, exceptional tannic structure, and remarkable aging potential. These distinctive characteristics are attributed to its unique genome, making Tannat one of the grape varieties with the highest tannins concentration.

Grapevine is a crop of great economic importance for several countries. The intensification of grapevine production has mostly been sustained by the increasing use of water resources at the expense of the environmental water balance. Moreover, in the last decades, climate change and the consequent expansion of drought have further compromised water availability, making current agricultural systems even more fragile both ecologically and economically. Recently, many research groups have highlighted the important role of endophytes in facilitating plant growth under optimal or stressful conditions. Within the framework of the PRIMA project, we aim to investigate the possible exploitation of the natural endophyte biodiversity as a sustainable tool to make grapevine plants more resilient to water deficit environmental conditions.

Erysiphe necator Schwein is a fungus that causes grapevine powdery mildew. It is one of the most problematic pathogens attacking Vitis vinifera L. The pathogen infects all green parts of the plant and reduces grape yield and quality. The suppression on mildew-susceptible cultivars requires intensive use of fungicides against pathogen, which has negative impact on the environment and human health.