Preliminary steps of a protocol to isolate transcription factors bound to a specific DNA locus in grapevine using CRISPR-dCas9 system

Considering that global warming is changing berry ripening timing and progression, uncovering the molecular mechanisms and identifying key regulators governing berry ripening could provide important tools in maintaining high quality grapes and wine. NAC (NAM/ATAF/CUC) transcription factors represent an interesting family due to their key role in the developmental processes control, such as fruit-ripening-associated genes expression, and in the regulation of multiple stress responses. Between the 74 NAC family members, we selected 12 of them as putative regulators of berry ripening: NAC01, NAC03, NAC05, NAC11, NAC13, NAC17, NAC18, NAC26, NAC33, NAC37, NAC60 and NAC61.

In the study of terroir, a separate analysis of its many component factors can be of great help in accurately identifying a vineyard’s natural elements that impact wine quality and typicity. This research used a dedicated pluri-disciplinary approach to investigate the ecological characteristics, including geology and geographical features, of 14 vineyards that produce Gewürztraminer and Sauvignon Blanc cultivars in the alpine Alto Adige DOC wine region. Both the geopedological method using Vineyards Geological Identity (VGI) and the new Solar Radiaton Identity (SRI) topoclimatic classification method were used to provide analytical measurements and qualitative/quantitative characterisations. In addition, wide-ranging targeted and untargeted oenological and chemical analyses were carried out on grapes, musts and wines to correlate the soils’ geomineral and physical conditions with the biochemical properties of their fruits and wines. The research identified strong correlations between vineyard geo-identity and wine biofingerprint, confirming a mineral traceability of strontium rubidium ratio and some minerals distinctive to the local geology, such as K, Ca, Ag, Ba and Mn.  The study also discovered that particular geomineral and physical soil conditions of the studied vineyards are related to the different amount of amino acids, primary varietal aromas and polyphenols found in grapes, musts and wines. The research confirmed that winemaking technologies support oenological quality, although in some cases, human practices can overpower certain characteristic elements in wine, erasing the typical imprint left by the vineyards’ natural terroir, which becomes less traceable. Terroir abiotic ecological factors and vineyard identity can be classified in detail using the new VGI and SRI analysis methods to discover interrelationships between geo-pedological and topoclimatic conditions that impact wine quality. These methods are also helpful in identifying which ecological elements are exclusive to a particular vineyard or wine sub-region.

Nowadays, it is well known that oxygen significantly impacts wine quality. The amount of oxygen wine consumes during the winemaking process depends on several factors, such as storage conditions, the number of rackings, the materials used for aging, and the type of closure chosen for bottling.

Long-term use of copper fungicides causes increased accumulation of total copper in the surface layer of vineyard soils. Many of authors has researched the anthropogenic influx of copper in such soils, which can result in environmental risks.

A misconception lingers in the minds of some wine consumers that Champagne wines don’t age. It’s largely a myth, certainly as far as the best cuvees are concerned. Actually, during the so-called autolysis period of time (in the closed bottle, after the “prise de mousse”), complex chemical reactions take place when the wine remains in contact with the dead yeast cells, which progressively bring complex and very much sought-after aromas to champagne. Nevertheless, despite their remarkable impermeability to liquid and air, caps or natural cork stoppers used to cork the bottles are not 100% hermetic with regard to gas transfers. Gas species therefore very slowly diffuse through the cap or cork stopper, along their respective inverse partial pressure. After the “prise de mousse”, because the partial pressure of CO2 in the bottleneck reaches up to 6 bars (at 12 °C), gaseous CO2 progressively diffuse from the bottle to the ambient air
(where the partial pressure of gaseous CO2 is only of order of 0,0004 bar).