Application of high-throughput sequencing tools for characterisation of microbial communities during alcoholic fermentation

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.

Newer sequencing technologies have allowed for the addition of microbes to the story of terroir. The same environmental factors that influence the phenotypic expression of a crop also shape the composition of the microbial communities found on that crop. For fermented goods, such as wine, that microbial community ultimately influences the organoleptic properties of the final product that is delivered to customers. Recent studies have begun to study the biogeography of wine-associated microbes within different growing regions, finding that communities are distinct across landscapes. Despite this new knowledge, there are still many questions about what factors drive these differences. Our goal was to quantify differences in yeast communities due to management style between seven pairs of conventional and biodynamic vineyards (14 in total) throughout Oregon, USA. We wanted to answer the following questions: 1) are yeast communities distinct between biodynamic vineyards and conventional vineyards? 2) are these differences consistent across a large geographic region? 3) can differences in yeast communities be tied to differences in metabolite profiles of the bottled wine? To collect our data we took soil, bark, leaf, and grape samples from within each vineyard from five different vines of pinot noir. We also collected must and a 10º brix sample from each winery. Using these samples, we performed 18S amplicon sequencing to identify the yeast present. We then used metabolomics to characterize the organoleptic compounds present in the bottled wine from the blocks the year that we sampled. We are actively in the process of analysing our data from this study.

The use of rootstock of American origin has been the classic method of fighting against Phylloxera for more than 100 years. For this reason, it is interesting to establish if different rootstock modifies nutrient composition as well as trace elements content that could be important for determining the traceability of the vine products. A survey of four classic rootstocks (110-Richter, SO4, FERCAL and 1103-Paulsen) and four new ones (M1, M2, M3 and M4) provided by Agromillora Iberia. S.L.U., all of them grafted with the Tempranillo variety, has been carried out during 2019. The eight rootstocks were planted in pots of 500 cc, on three soils with very different characteristics from Castilla-La Mancha (Spain). In the month of July, the leaves were collected and dried in a forced air oven for seven days at 40ºC. Then, the samples were prepared for the analysis determination, carried out by X-Ray fluorescence spectrometry. The results obtained showed that in the case of content in mineral elements in leaf, separated by soil type, we can report the importance of few elements such as Si, Fe, Pb and, especially, Sr. The rootstock does not influence the composition of the vine leaf for the studied elements that are the most important in determining the geochemical footprint of the soil. The influence of the soil can be discriminated according to some elements such as Fe, Pb, Si and, especially, Sr.

The first demonstration of the interest of carbon isotope composition of sugars in grapevine, as an integrated indicator of vineyard water status, dates back to 2000 (Gaudillère et al., 1999; Van Leeuwen et al., 2001). Thanks to the isotopic discrimination of Carbon that takes place during plant photosynthesis, under hydric stress conditions, it is possible to accurately estimate the photosynthetic activity. Ever since, δ13C has been widely applied with success to zonation, terroir studies and vine physiology research, but is still not widely used by viticulturists. This is quite astonishing by considering the impact of global warming on viticulture and the need to improve water management, that would justify a widespread use of δ13C.
The lack of private laboratories proposing the analysis, the cost of the technology, as well as the long analytical delays, have been detrimental to its development. Some laboratories tried to overcome the analytical difficulties of isotopic analysis by using fourier transformed infrared spectroscopy, as a fast and cheap alternative to the official OIV method (IRMS). These claimed FTIR models have never been published or peer reviewed and cannot be considered robust. In this work, thanks to the recent acquisition of IRMS technology, new modern and robust applications of δ13C for viticulture are proposed. This includes the use of the analysis to make parcel separations at harvesting, the possibility to increase the precision of hydric stress cartography and the potential cost reduction when compared with Scholander pressure bomb analysis.

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.