Enhancing hydric stress tolerance by editing the VviMYB60 promoter with CRISPR/Cas9 

Water tolerance in plants is often associated with the accumulation of osmotic protectants, which are secondary metabolites that can help the plant to cope with water stress. One of the key osmotic protectants is a sugar called Raffinose, which is synthesized by a family of enzymes called Raffinose synthases. In this work, we focused on one of these enzymes, VviRAF2, which is a gene that shows different expression levels and genetic variants (SNPs) among different grapevine cultivars, ranging from tolerant to susceptible to water stress, and the transcription factors that may regulate the expression of this gene family.

Enzymatic browning (BE) of must is caused by polyphenol oxidases (PPOs), tyrosinase and laccase. Both PPOs can oxidize diphenols such as hydroxycinnamic acids (HA) to quinones, which can later polymerize to form melanins [1], which are responsible of BE in white wines and of oxidasic haze in red wines. SO₂ is the main tool used to protect must from BE thanks to its capacity to inhibit PPOs [2]. However, the current trend in winemaking is to reduce and even eliminate this unfriendly additive. Among the different possible alternatives for protecting must against BE, the inoculation with a selected Metschnikowia pulcherrima MP1 is without any doubt one of the most promising ones.

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.

The aromatic perception is one of the main factors that influence the
consumer when determining the wine’s quality and acceptance. Numerous factors (soil, climate,
winemaking style, cultivar) can influence the volatile compounds. Some of these compounds are released directly from the grape berries while others are formed during the fermentation and aging processes. However, little is known about the quality and aromatic formation of Syrah variety in the winter cycle cultivated in São Paulo.

In response to the growing demand for rapid, precise, and efficient methods of quantifying volatile compounds in alcoholic beverages, this study presents a novel approach for the determination of higher alcohols in wine, port wine, and brandy.