New breeding frontiers: application of the CRISPR-cas9 system in grapevine (V. vinifera L.) and improvements in plant regeneration

Trebbiano di Lugana (TdL) is a white variety of Vitis vinifera mainly cultivated in an Italian area located south near Garda lake (Verona, north of Italy). This grape cultivar, also known as “Turbiana,” is used for the production of TdL wine with recognized Protected Designation of Origin whose volatile profile was recently determined [1]. The presence of varietal thiols in TdL, namely 3-mercaptohexan-1-ol and its acetate form, conferring the tropical and citrus notes, has been documented. Winemaking strategies were also described with the purpose of protecting and maintain these desired aromas [2]. To the best of our knowledge, the varietal thiol precursors (VTPs) were not previously determined in TdL grape and must. This study aimed to quantify VTPs in both grape during the ripening and must during the pressing. Volatile C6 compounds were also measured in the must fractions.

Botrytis cinerea is a fungus that causes common infection in grapes and other fruits. In winemaking, its presence can be both considered desirable in the case of noble rot infection or undesirable when grey rot is developed. This fungus produces an extracellular enzyme known as laccase which is able to cause oxidation of phenolic compounds present in must and wine, causing most of the times a decrease in its quality and problems during the winemaking process [1]. Material and methods: Three B. cinerea strains (B0510, VA612 and RM344) were selected and grown in a liquid medium adapted from one previously described [2]. The enzyme was isolated by tangential ultrafiltration of the culture medium using a QuixStand system equipped with a 30 KDa filtration membrane.

Structure-function relationships between the polysaccharide part of S. cerevisiae Mannoprotein Pools (MPs) and their potential to interact with anthocyanins and Protein-Tannins aggregates was previously assessed [1,2].

Grapevine (Vitis vinifera L.) exhibits a high level of genetic and phenotypic diversity among the approximately 6000 cultivars recorded. This perennial crop is highly vulnerable to numerous fungal diseases, including esca, which is a complex vascular pathology that poses a significant threat to the wine sector, as there is currently no cost-efficient curative method[1]. In this context, an effective approach to mitigate the impact of such diseases is by leveraging the crop’s genetic diversity. Indeed, susceptibility to esca disease appears to vary between cultivars, under artificial or natural infection. However, the mechanisms and varietal characteristics underlying cultivar susceptibility to esca are still unknown.

Grapevine is subject to multiple stresses, either biotic or abiotic, frequently in combination. These stresses may negatively impact the health status of plants and reduce yields. For biotic stress, grapevine is affected by numerous pest and diseases such as downy and powdery mildews, grey mold, black rot, grapevine fanleaf virus and trunk diseases (namely GTDs). The interaction between grapevine and pathogens is relatively complex and linked to various pathogenicity factors including cell-wall-degrading enzymes (especially CAZymes) and phytotoxic secondary metabolites, growth regulators, effectors proteins, and fungal viruses.