Caratterizzazione delle produzioni vitivinicole dell’area del Barolo: Un’esperienza pluridisciplinare triennale (4)

New Plant Breeding Techniques (NPBTs) have the potential to revolutionize the genetic improvement of grapevine. However, the practical application of these techniques is limited by several challenges, such as the difficulty in generating embryogenic calluses, maintaining their competence during in vitro cultivation, and regenerating plants without defects. To overcome these challenges, we conducted a study to test the effect of two treatments on callus cultures derived from different grapevine varieties, with and without embryogenic competence. The tested substances were Silver Thiosulphate (STS) an ethylene inhibitor, and Salicylic Acid (SA), an elicitor with different effects depending on the concentration of use beyond the ethylene inhibitor activity.

Developments in high-throughput sequencing (HTS) technologies allow us to obtain large amounts of microbial information from wine and must samples. Thus approaches, that are aimed at characterising the microbial diversity during fermentation, can be enhanced, or possibly even replaced, with HTS-based metabarcoding. To reduce experimental biases and increase data reproducibility, we compared 3 DNA extraction methods by evaluating differences in the fungal diversity with Riesling alcoholic fermentation samples at four different vineyards. The fungal diversity profiling was done using the genetic markers ITS2 and D2 using metabarcoding. The extraction methods compared consisted of a commercial kit, a recently published protocol that includes a DNA enhancer, and a protocol based on a buffer containing common inhibitor removal reagents. All methods were able to distinguish vineyard effects on the fungal diversity, but the results differed quantitatively.

Global changes in temperature patterns necessitate the development of viticultural adaptation strategies. One promising approach involves modifying the training system and elevating trunk height. This study explored the potential of raising the vine trunk as an adaptive strategy to counteract the effects of increasing temperatures and delay ripening. Thermal conditions, radiation levels, and must composition were measured at different heights (10 and 150 cm) in a commercial vineyard of the minority variety Maturana Blanca, trained on a vertical cordon.

Grapevine grafting is a complex process that since the establishment of phylloxera has become mandatory for grapevine. Grafting success in grapevine nurseries considerably varies among years and batches with most variety/rootstock combinations reach a high success rate (between 75% and 90%), but some combinations show lower success rates of around 40-50%. The causes of this variation are unknown, although biotic stresses like those caused by some viral infections have been demonstrated to affect the process. European certification schemes for the vegetative propagation of the vine include five major viruses (Arabis mosaic virus, Grapevine Fanleaf Virus, Grapevine Fleck Virus, and Grapevine-associated Leafroll Virus 1 and 3).

Cis-acting regulatory elements are DNA sequences that can be bound by transcription factors to regulate the expression of genes in a condition-dependent and tissue-specific way. It is nowadays possible to search for DNA motives and sequences that a given transcription factor is binding or at least can, but it is still hard to have a glance at all the transcription factors that are contemporaneously located at the same locus. Inspired by an existing technique that uses the CRISPR-Cas system in mammal cells, we are trying to develop a protocol to study such regulation in Vitis vinifera. Using the highly sequence-specific binding capacity of a catalytically inactive Cas9 protein (dCas9), our idea is to set up a system to target a desired sequence and precipitate all the crosslinked proteins and distantly interacting chromatin at this locus and analyze them.