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

In a long-term experiment carried out in Chinon vineyard (37, France) during 23 years, the effects of several organic amendments were studied on soil, vine, grapes and wine. Four main treatments were compared on a calcareous sandy soil: control without organic amendment, dry crushed pruning wood at 2.1.t-1.ha-1.year-1 (D1), cow manure at 10 t-1. ha-1.year-1 (D1) and cow manure applied at 20 t-1.ha-1.year-1 (D2). D1 levels were calculated to fill the annual humus losses by mineralization.

Climate change is here. One of the main consequences is an increase in the frequency and severity of abiotic stresses which mostly occur in a combined manner. Grapevine, which grows in a large diversity of pedo-climatic conditions, has presumably evolved different mechanisms to allow this widespread adaptation. Harnessing the genetic diversity in these mechanisms will be central to the future of viticulture in many traditional wine growing areas. The interactions between the scion and the rootstock through grafting add an additional level of diversity and adaptive potential to explore.
At the physiological level, these mechanisms are related to processes such as root system development and functioning (water and nutrient uptake), interactions with the soil microbiome, gas exchange regulation, hydraulic properties along the soil-plant-atmosphere continuum, reserve storage, short and long distance signaling mechanisms and plasticity for some of these traits.

The use of next-generation sequencing (NGS) has helped understand microbial genetics in oenology. Current studies mainly focus on barcoded amplicon NGS but not shotgun sequencing, which is useful for functional analyses. Since the high percentage of grapevine DNA conceals the microbial DNA in must, the majority of sequencing data is wasted in bioinformatic analyses. Here we present capture depletion of grapevine whole genome DNA.

Context and purpose. In Germany, vineyards are typically intensively managed monocultural systems shaped by low structural variability.

Grapevine powdery mildew caused by Erysiphe necator and Botrytis bunch rot caused by Botrytis cinerea are two of the most important fungal diseases in California grape production.