REVINE project : regenerative agricultural approaches to improve ecosystem services in Mediterranean vineyards

Flow cytometry (FCM) is a powerful technique allowing the detection, characterization and quantification of microbial populations in different fields of application (medical environment, food industry, enology, etc.). Depending on the fluorescent markers and specific probes used, FCM provides information on the physiological state of the cell and allows the quantification of a microorganism of interest within a mixed population. For 15 years, the enological sector has shown growing interest in this technique, which is now used to determine the populations present (of interest or spoilage) and the physiological state of microorganisms at the different stages of winemaking.

Pinot Blanc, an important grape variety grown in some mountain areas of Northern Italy such as South Tyrol over the last decades, with its cultivation covering 10.3% of the total vineyards, has compatible climatic conditions (e.g. heat requirements) which are normally found in the geographical areas of the mountain viticulture [1,2,3,4]. Climatic changes are hastening the growth of this variety at higher elevations, particularly for the production of high quality wine.

Grapevine phenology has advanced across many regions, nationally and internationally, in recent decades under the influence of increasing temperatures, resulting in earlier
vintages (Jones and Davis, 2000, Petrie and Sadras, 2008, Tomasi et al., 2011, Webb et al., 2011. Earlier vintages have several ramifications for the wine industry. There are direct implications on quality, due to the fruit ripening during the hotter conditions of summer and early autumn, which then impacts grape composition and wine style (Sadras et al., 2013, Buttrose et al., 1971, Mira de Ordũna, 2010). There are also indirect implications where the fruit is perceived to ripen at a faster rate and the crop reach optimum maturity over a shorter period (Coulter et al., 2016).

Amarone is an Italian red wine produced in the Valpolicella area, in north-eastern Italy. Due to its elaboration with withered grapes, Amarone is a rather unique example of dry red wine. However, there is very limited data so far concerning the volatile composition of commercial Amarone wines, which also undergo a cask aging of 2-4 years before release. The present work aims at characterizing the aroma composition of Amarone and to elucidate the relationships between chemical composition and sensory characters.

Epidermal Patterning Factors are a class of cysteine rich peptides known to be involved in many developmental processes. The role of EPF1, EPF2 and EPFL9 in controlling leaf stomata formation has been well described in model plants and cereals, and recently also in grapevine, while little is known about their activity in other organs. The aim of our study is to investigate whether VviEPFL9-2 can have a specific biological function in grapevine roots, where it resulted to be expressed. As grapevine is cultivated in the form of a grafted plant, we focused our study on the commonly used rootstock Kober 5BB (Vitis berlandieri x Vitis riparia). VviEPFL9-2 was edited in Kober 5BB plants using Agrobacterium tumefaciens transformation of embryogenic calli and the CRISPR/Cas9 technology. The phenotypic evaluation in greenhouse indicated that, as expected, the leaves of knock-out (KO) plants have a significant lower stomatal density compared to WT, associated with a lower stomatal conductance.