Preliminary steps of a protocol to isolate transcription factors bound to a specific DNA locus in grapevine using CRISPR-dCas9 system

In contrast to fruit and grape berry ripening, the biological processes causing ripening disorders are often much less understood, although shriveling disorders of fruits are manifold and contribute to yield losses and reduced fruit quality worldwide. Shrinking berries are a common feature for all shriveling disorders in grapevine although their timing of appearance during the berry ripening process and their underlying induction processes distinct them from each other. The sugar accumulation disorder Berry Shrivel (BS) is characterized by a suppression of sugar accumulation short after veraison resulting in berries low in sugar content and anthocyanins in berry skins, while the organic acid content is similar. Recent studies analyzed the biochemical, morphological and molecular processes affected in BS berries and linked early changes to the period of ripening onset [1,2].

Grapevine trunk diseases (GTD) occur wherever grapes are grown and are considered the main biotic factor reducing yields and shortening vineyards’ lifespan. Currently, no product is available to eradicate GTD once grapevines are infected. Therefore, prophylactic strategies based on pruning wound protection and ‘remedial surgery’, the only eradication method based on the elimination of infected wood and renewal of the vine by means of new canes or suckers, are the only effective strategies available. The Canadian grape and wine industry focusses on a sustainable production and thus, looking for alternatives to chemicals for disease management is a top priority.

Vineyard floors in warm, dry landscapes including those in South Australia, have traditionally been managed using intensive practices such as tillage and herbicides to control weeds and vegetation, thereby limiting competition with grapevines for water and nutrients in order to not compromise yields.

Following anecdotal evidence of unwanted ‘tropical’ character in red wines resulting from vineyard interventions and a subsequent yeast trial observing higher ‘red fruit’ character correlated with higher thiol concentrations, the role of polyfunctional thiols in commercial Australian red wines was investigated.
First, trials into the known tropical thiol modulation technique of foliar applications of sulfur and urea were conducted in parallel on Chardonnay and Shiraz.1 The Chardonnay wines showed expected results with elevated concentrations of 3-sulfanylhexanol (3-SH) and 3-sulfanylhexyl acetate (3-SHA), whereas the Shiraz wines lacked 3-SHA. Furthermore, the Shiraz wines were described as ‘drain’ (known as ‘reductive’ aroma character) during sensory evaluation although they did not contain thiols traditionally associated with ‘reductive’ thiols (H2S, methanethiol etc.).

Grape breeding for resistance to fungal diseases is today very dynamic throughout the world notably in France. New varieties are obtained by hybridization between susceptible varieties of the vitis vinifera species and resistant genotypes, with breeding programs generally lasting between 15 and 25 years and resulting in the registration of a few new varieties. Though these varieties can provide several benefits and can be planted by winegrowers, they are not always systematically adopted.