Development and application of CRISPR/Cas in grapevine

Accentuated cut edges (ACE) is a novel grape crushing technique used sequentially after a conventional crusher to increase the extraction rate and content of polyphenolics, as shown for Pinot noir wine. This inspired us to apply the technique during Shiraz and Sauvignon blanc winemaking, primarily to assess its impact on the extraction of varietal thiol precursors in grape must/juice and formation of varietal thiols in the resultant wines

Over the last decade, the use of non-Saccharomyces yeasts in the winemaking process has been re-assessed and accepted by winemakers. These yeasts can be used to achieve specific objectives such as lowering the ethanol content, preventing wine spoilage and increasing the production of specific aroma compounds. Since these species are unable to complete alcoholic fermentation, strategies of co- and sequential inoculation of non-Saccharomyces and Saccharomyces cerevisiae have been developed. However, when mixed starter cultures are used, several parameters (e.g. strain yeast, inoculation timing and nutrient competitions) impact the growth of the individual yeasts, the fermentation kinetics and the metabolites/aroma production. In particular, competition for nitrogen compounds could have a major impact, potentially leading to sluggish fermentation when the yeast assimilable nitrogen (YAN) availability is low. Moreover, many aroma compounds produced by the yeasts are directly produced and influenced by nitrogen metabolism such as higher alcohols, acetate esters and ethyl esters which participate in the organoleptic complexity of wine.

In the context of LIFE project MIDMACC (LIFE18 CCA/ES/001099), several pilots have been installed in vineyards in mid mountain areas of Catalonia (NE Spain) to test well stablished agronomic practices to increase the adaptation of Mediterranean mid mountain to climate change. Soil water content (SWC) at three different depths (15, 30 and 45cm) was measured in continuum from August 2020. One pilot (WC) included a well-established green cover (GC), a new GC (NC) and a conventional soil management (CM, tilling+herbicides). NC presented an intermediate state between WC and CM, responding similarly to CM in autumn but quickly reaching similar SWC to WC, then following the same evolution till next spring, with CM presenting lower values along autumn and winter. Then vegetation activation decreased SWC in all plots, (much slower in CM, lacking GC). Sensibility to spring rains is again intermediate for NC, which joins SWC evolution of CM by the end of spring till next autumn. It is expected that NC will resemble WC more and more as its GC develops. In the pilot combining vine training (VSP vs Gobelet) and hillside management (slope vs terrace), no clear pattern could be related with these conditions. However, both terraces seem to be more sensitive to spring rains. A third pilot included new vineyards (7 and 1 year old). In the new vineyard (N), higher canopy development, a spontaneous green cover and row straw resulted in a slower SWC dynamic, not so sensitive to rains but conserving more soil water in spring and most of summer, even with presumably a higher water extraction by vines. In the newest vineyard (VN) the deepest sensor is still sensitive to rain events all over the year and SWC is always highest at this depth, revealing small water capture by vines.

Several winemaking operations, such as filtration, pumping, and racking, are known to potentially facilitate the incorporation of atmospheric O2 into the wine. Control of grape must oxidation is one key aspect in the management of white wine aroma expression, color stability and shelf-life extension. On the one hand, controlled must oxidation may help to remove highly reactive phenolic compounds, which otherwise could contribute to premature oxidation. And on the other hand, in certain cases of extreme protection of the must from O2 (e.g. pressing under inert atmosphere), it can help to preserve varietal aromas and natural must antioxidants.

Polysaccharides and more specifically pectins, make up a significant portion of the cell wall material of the plant cells including the grapes. During the fruit ripening the associated softening is related to the breakdown of the cell wall polysaccharides. During this process, it is expected that polysaccharides that are soluble in red wine will be formed influencing its texture. Anthocyanins are responsible for the wine color and tannins for the astringency, body and bitterness of the wine. In the skins, these compounds are located in the cell vacuoles and the barrier that conditions their extractability is the skin cell wall that may determine the mechanical resistance, the texture and the ease of processing berries. The aim of this work was study the evolution of the polysaccharides and the anthocyanin and tannin extractability during the ripening period in Cabernet Sauvignon grapes, trying to correlate these variables.