Optimized protocol for high-quality RNA extraction from grape tissues using sorbitol pre-wash

Climate change is having a significant impact on the wine industry through more regular drought conditions, fires, and heat events, leading to crop loss. Furthermore, these events can reduce overall quality of the fruit, even when crop yields are not impacted. Anthocyanins are considered one of the most important classes of compounds for red wine production and are known to be sensitive to vine water status and heat events.

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.

“Sangiovese” (Vitis vinifera L. sativa cv. Sangiovese) is the main grape variety to be established in Italy, being the only country in Europe where this grape is commonly found.

Esca complex is a disease affecting grapevine trunks, characterized by the colonization of the wood by xylem-residing fungi (Phaeomoniella chlamydospora, Phaeoacremonium minimum and Fomitiporia mediterranea), and posing significant risks to vineyard longevity since no efficient treatment is available. Despite its prevalence, the mechanisms beyond symptomatic manifestations like interveinal chlorosis and leaf necrosis remain unclear. Preliminary findings indicated a more pronounced metabolic reprogramming in fruits compared to vegetative organs and a putative impact on wine quality by using fruits from symptomatic grapevines.

Increasing regulatory restrictions on herbicide use and growing interest in sustainable viticulture are driving the search for alternative weed management strategies.