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

The use of grapevine genetic diversity is a way to mitigate the negative impacts of climate change on viticulture systems. Leaf epidermal flavonoids (including flavonols and anthocyanins) are involved in plant defense mechanisms against environmental stresses, like high temperatures or excessive solar radiation [1,2]. Among other factors, they modulate light absorption, which reduces photoinhibition processes in photosynthetic tissues [1]. Therefore, the identification of grapevine cultivars with an increased content on leaf epidermal flavonoids arises as a potential avenue to improve grapevine tolerance to some detrimental environmental stresses.

Some wood substances such as ellagitannins (vescalagin, castalagin, grandinin, roburins (A, B, C, D, E)…) can be extracted during wine ageing in oak barrels. The level of these hydrolysable tannins in wine depends of the species and origin of oak wood as well as its treatment during barrel realization.

In contrast to fruit and grape berry ripening, the biological processes causing ripening disorders are often much less understood, although shriveling disorders of fruits are manifold and contribute to yield losses and reduced fruit quality worldwide. Shrinking berries are a common feature for all shriveling disorders in grapevine although their timing of appearance during the berry ripening process and their underlying induction processes distinct them from each other. The sugar accumulation disorder Berry Shrivel (BS) is characterized by a suppression of sugar accumulation short after veraison resulting in berries low in sugar content and anthocyanins in berry skins, while the organic acid content is similar. Recent studies analyzed the biochemical, morphological and molecular processes affected in BS berries and linked early changes to the period of ripening onset [1,2].

The objective of the work described here is the elaboration of a map of the different types of vineyard soils that to guide the famers in the choice of the most productive vine rootstocks and varieties. 90 vineyard soils profiles were analysed in the entire territory of the Origen Denominations of Valdepeñas. The sampling was carried out in 2018 (June to October) by making a sampling grid, followed by photointerpretation and control in the field. The studied soils can be grouped into 9 different soil types (according to FAO 2006 classification): Leptosols, Regosols, Fluvisols, Gleysols, Cambisols, Calcisols, Luvisols and Anthrosols. A map showing the soil distribution with different type of soils has been made with the ArcGIS program. Regarding to the choice of rootstock, Calcisoles are soils with a high active limestone content, so the rootstocks used in these soils must be resistant to this parameter; Luvisols are deep soils with high clay content, so they will support vigorous rootstocks. Because the cartographic units are composed of two or more subgroups, with are associated in variable proportions, 9 different soil associations have been established; Unit 1: Leptosols, Cambisols and Luvisols (80%, 15% and 5% respectively); Unit 2: Cambisols with Regosols and Luvisols (40%, 30% and 30% respectively); Unit 3: Cambisols and Gleysols with Regosols (40%, 40% and 20% respectively); Unit 4: Regosols with Cambisols, Leptosols and Calcisols (40%, 30%, 15% and 15% respectively); Unit 5: Cambisols, Leptosols, Calcisols and Regosols (25% each of them); Unit 6: Luvisols with Cambisol and Calcisols (80%, 10% and 10% respectively); Unit 7: Luvisols and Calcisols with Cambisols (40%, 40% and 20% respectively); Unit 8: Calcisols with, Cambisols and Luvisols (80%, 10% and 10% respectively); Unit 9: Anthrosols. These study allow to elaborate the first map of vineyard soils of this Protected Designation of Origin in Castilla-La Mancha.

In wine growing regions around the world, climate change has the potential to affect vine transpiration and overall vineyard water use due to related changes in atmospheric demand and soil water deficits. Grapevines control their transpiration in response to a changing environment by regulating conductance of water through the soil-plant-atmosphere continuum. Most vineyard water use models currently estimate vine transpiration by applying generic crop coefficients to estimates of reference evapotranspiration, but this does not account for changes in vine conductance associated with water stress, nor differences thought to exist between varieties. The response of bulk stomatal conductance to daily weather variability and seasonal drought stress was studied on Cabernet-Sauvignon, Merlot, Tempranillo, Ugni blanc, and Semillon vines in a non-irrigated vineyard in Bordeaux France. Whole vine sap flow, temperature and humidity in the vine canopy, and net radiation absorbed by the vine canopy were measured on 15-minute intervals from early July through mid-September 2020, together with periodic measurement of leaf area, canopy porosity, and predawn leaf water potential. From this data, bulk stomatal conductance was calculated on 15-minute intervals, and multiple regression analysis was performed to identify key variables and their relative effect on conductance. Attention was focused on addressing multicollinearity and time-dependency in the explanatory variables and developing regression models that were readily interpretable. Variability of vapor pressure deficit over the day, and predawn water potential over the season explained much of the variability in conductance, with relative differences in response coefficients observed across the five varieties. By characterizing this conductance response, the dynamics of vine transpiration can be better parameterized in vineyard water use modeling of current and future climate scenarios.