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

The use of selected cultures of the species Lactiplantibacillus plantarum in Oenology has grown in prominence in recent years. While initial applications of this species centred very much around malolactic fermentation (MLF), there is strong evidence to show that certain strains can be harnessed for their bio-protective effects. Unwanted spontaneous MLF during alcoholic fermentation (AF), driven by rogue Oenococcus oeni, is a winemaking deviation that is very difficult to manage when it occurs. This work set out to determine the efficacy of one particular strain of Lactiplantibacillus plantarum(Viniflora® NoVA™ Protect), against this problem in Cabernet Sauvignon must. The work was carried out at commercial scale and in a winery environment and compared the bio-protective culture with the more traditional approach of reducing must pH by the addition of tartaric acid. The combination of both was also investigated. The concentration of both Oenococcus oeni and Lactiplantibacillus plantarum was determined using qPCR. The adventitious Oenococcus oeni showed the most growth during AF in the control wine, whereas in the wines treated with Lactiplantibacillus plantarum a bacteriostatic effect against this species was observed. This effect was comparable to the wines treated with tartaric acid. This has particular commercial relevance for controlling the flora in musts with high pH, or when the addition of tartaric acid is either not permitted or is prohibitive for other reasons.

The actual climate changes, together with the strong regulation of the European Union and Spanish government, in search of sustainable viticulture, have forced the recovery of minority varieties, expanding the range of grape varieties, as well as the possible development of wines with unique profiles. In the Ribeira Sacra DO (Spain), a comparative study of the agronomic and qualitative behavior of the ‘Branco lexítimo’ variety has been carried out, compared to the majority white variety in the DO: ‘Godello’, located in the same study plot, with identic soil and climatic conditions. The study contemplated the analysis of phenology and leaf water potential, as well as the productive results and the analysis of the must quality, during four seasons: 2018 – 2021.

The present work contributes by developing a rapid sensory-directed methodology for the screening and selection of high quality wines with different sensory profiles Therefore, Verdejo and Tempranillo musts were fermented with 50 different yeasts each under controlled laboratory conditions. Resulting samples were firstly categorized according to five levels of quality by a panel of wine professionals (Sáenz-Navajas, Ballester et al. 2013). Higher quality samples were described by flash profiling by a semi-trained panel
(Valentin, Chollet et al. 2012) and most distinctive samples were screened by gas chromatography-olfactometry (GC-O) (López, Aznar et al. 2002).

While SO2 is one of the oldest and widest additive used in enology for its well-known antioxidant, anti-laccase and antimicrobial properties, it can cause health problems in some individuals.

The shelf life of wine is a major concern for the wine industry. This is particularly true for wines intended for long cellaring, which are supposed to reach their peak after an ageing period ranging from a few months to several years, or even decades. Low, controlled oxygen inputs through the closure system are generally necessary for the wine to evolve towards its optimum organoleptic characteristics. Our previous studies have already shown that the interface between the cork and the bottleneck plays a crucial role in the transfer of oxygen into the bottled wine.