A novel approach for the identification of new biomarkers of wine consumption in human urine using untargeted metabolomics

Bobal is the second most widely grown Spanish red grape variety (54,165 has), mainly cultivated in the Valencian Community and especially, in Utiel-Requena region (about 67% of 34,000 has). In this study, agronomic and enological parameters were determined in 98 biotypes selected during 2018 and 2019 in more than 50 vineyards over 50 years-old in the Utiel-Requena region. Moreover, a multi-criteria approach considering temperature and rainfall (Fig. 1A), among other parameters, was made to establish three different zones within the region (Fig. 1B), where in the future the selected biotypes will evaluated. In fact, in 2020, 4 replicates and 12 vines per biotype were planted in an experimental vineyard to preserve this important intra-cultivar diversity.

In the O Ribeiro Denomination of Origin, there is a winemaking tradition of growing vines under a high-density plantation framework (8,920 vines/ha) and maintaining its vegetative cycle under rainfed conditions.
Currently, viticulture is advancing to plantation frames in which the density is considered medium (5,555 vines/ha), thus allowing mechanized work to be carried out for vineyard management operations. Although, the application of irrigation applied proportionally to the needs of the vegetative cycle of the vine, is a factor that increasingly helps a good development of the vine compared to the summer period, with increasingly uncertain weather forecasts.

“Sulphites” and mainly sulphur dioxide (SO2) is by far the most widely used additive (E-220/INS 220) in winemaking and likely the most difficult to replace. The well-known antioxidant, antioxidasic and antimicrobial properties of SO2 make this molecule a practically essential tool, not only in winemaking, but also in the production of other food products. The current trend in winemaking is the reduction of this unfriendly additive due to its negative effects on health and environmental. In particular, it could cause headaches and intolerance/allergic reactions in sensitive individuals. Wine is considered one of the major contributors of exposure of SO2 in the adult population, when this beverage is included in the diet.

‘Tempranillo Tinto’ is a black-berried Iberian cultivar that originated from a hybridization between cvs. ‘Benedicto’ and ‘Albillo Mayor’ [1]. Today, it is the third most widely grown wine grape cultivar worldwide with more than 200,000 hectares of vineyards mostly distributed along the Iberian Peninsula, where it is also known as ‘Cencibel’, ‘Tinta de Toro’, ‘Tinta Roriz’, and ‘Aragonez’, among other synonyms. Here, we quantified the intra-varietal genomic diversity in this cultivar through the study of 35 clones or ancient vines from seven different Iberian wine-making regions. A comparative analysis after Illumina whole-genome sequencing revealed the presence of 1,120 clonal single nucleotide variants (SNVs).

The vineyard has experienced a general increase in yields mainly due to the elevated use of technology which caused a quality loss of grapes in more than one case. A large percentage of the Spanish vineyard is covered by a Denomination of Origin which limits the productive level of the vineyards as one of its regulations. The maximum production limit is a variable characteristic of each vineyard and is not usually regulated by agronomic criteria, and this explains the fact that each vineyard can reach high quality with a totally different yield from that set by the Denomination of Origin.