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

Berry maturation in warm areas takes place very early, when temperatures are still high and favorable for carbohydrate synthesis and accumulation in the berries, but not as favorable for maintaining high titratable acidity or low pH, or for increasing berry polyphenol content. Different canopy management techniques have been proven to delay berry maturation at the expense of yield (severe canopy trimming, late spring pruning to induce sprouting of dormant buds, etc.). Minimal pruning delays berry ripening by highly increasing yield and by reducing the leaf area to fruit ratio.

Grapevine (Vitis vinifera L.) is a challenging plant species to transform and regenerate due to its complex genome and biological characteristics. This limits the development of cisgenic and gene-edited varieties. One hurdle is selecting the best starting tissue for the transformation process, much like isolating suitable tissue for protoplasts. One promising method involves delivering CRISPR/Cas components to protoplasts isolated from embryogenic calli, which are then induced to regenerate. However, this process is inefficient, time-consuming, and only applicable to a few genotypes. To enhance grapevine regeneration efficiency, the expression of developmental and plant growth regulators shows promise in escaping the recalcitrance encountered in traditional tissue culture methods.

Grapevine resilience to climate change has become one of the most pressing topics in the Viticulture & Enology field. Vineyard health demands understanding the mechanisms that explain the direct and indirect interactions between environmental stressors. The current climate change scenario, where drought and heat-wave are more frequent and intense, strongly demands improving our knowledge of environmental stresses. During a heatwave, the ambient temperature rises above the plant’s average tolerance threshold and, generally, above 35 oC plant’s adaptation to heat stress is activated.

The northern vineyards produce wines with a high aromatic richness. The wines of Alsace are appreciated for the diversity of their aromas, the typicality of which was for a long time judged mainly according to the grape variety of origin. Alsatian winegrowers have however widely sensed the importance of the environment of the vine on the quality of the wines. Efforts are made to try to harmonize in a reasoned way the interaction between the natural environment and the plant material with a view to developing the character of the grape variety through the fine expression of the terroir and making the quality and typicality even more inimitable. wines produced in Alsace.

In the conducted research, 30 yeast strains from red grape varieties were isolated from the Purcari wine center, and 28 yeast strains from red grape varieties were isolated from the Trifești wine center in the Republic of Moldova.