terclim by ICS banner
IVES 9 IVES Conference Series 9 DNA-free editing to improve stress resilience of wine grape genotypes recalcitrant-to-regeneration

DNA-free editing to improve stress resilience of wine grape genotypes recalcitrant-to-regeneration

Abstract

Wine viticulture, being firmly linked to the vine-terroir relationship, has always encountered significant bottlenecks to genetic innovation. Nonetheless, the development of new breeding strategies leading to the selection of stress resilient genotypes is urgent, especially in viticulture, where it would allow reducing the use of chemical treatments adopted to control fungal diseases. Genome editing represents an extremely promising breeding technique. Unfortunately, the well-known recalcitrance of several wine grape cultivars to in vitro regeneration strongly limits the exploitation of this approach, which to our knowledge has so far been developed on table grape genotypes with high regeneration potential. By targeting the phytoene desaturase gene as visual editing proof, we developed a genome editing and regeneration protocol to produce transgene-free grapevine plants exploiting the lipofectamine–mediated delivery of CRISPR–Cas9 ribonucleoproteins into protoplasts. We regenerated edited grapevines of Vitis vinifera‘Nebbiolo’, a cultivar extremely recalcitrant to in vitro regeneration and at the basis of outstanding quality wines, such as ‘Barolo’ and ‘Barbaresco’. Successful editing was confirmed by a combination of approaches: HRM, Sanger and amplicon deep sequencing, phenotype visualization. We then exploited the method to silence two micro(mi)RNAs involved in biotic stress responses: vv-miR482, which is conserved in different species, and the grapevine-specific vv-miR3623. Since NBS-LRR disease-resistance genes are the targets of those miRNAs, the objective is to regenerate vines with a broad-spectrum level of plant tolerance/resistance to different pathogens. The developed strategy could be extended to other important wine grape varieties and recalcitrant woody species.

DOI:

Publication date: June 13, 2024

Issue: Open GPB 2024

Type: Article

Authors

Giorgio Gambino1, Floriana Nuzzo1, Amedeo Moine1, Walter Chitarra1,2, Chiara Pagliarani1, Annalisa Petrelli3, Paolo Boccacci1, Andrea Delliri1, Riccardo Velasco2, Luca Nerva1,2, Irene Perrone1*

1 Institute for Sustainable Plant Protection, National Research Council (IPSP-CNR), Strada delle Cacce 73, 10135 Torino (Italy)
2 Research Centre for Viticulture and Enology, Council for Agricultural Research and Economics (CREA-VE), Via XXVIII Aprile 26, 31015 Conegliano (Italy)
3 Open Lab – Department of Veterinary Sciences, University of Turin (DSV-UNITO), Largo Paolo Braccini 2, 10095 Grugliasco, Italy

Contact the author*

Keywords

genome editing, protoplast regeneration, lipofectamines, microRNAs, biotic stress

Tags

IVES Conference Series | Open GPB | Open GPB 2024

Citation

Related articles…

Managing Grapevine Powdery Mildew with Ultraviolet-C Light in Washington State

Germicidal ultraviolet-C (UV-C) light has shown promising results for suppression of several plant-pathogenic microorganims, including Erysiphe necator, which attacks grapevine. In Washington State the majority of winegrape production is in a semi-arid steppe environment, with historically low powdery mildew disease pressure, making it a promising area to deploy UV-C as a disease management tool. Trials focusing on UVC application timing and frequency will assist in developing regionally-appropriate application recommendations for eastern Washington State.

Organic mulches slightly influence wine phenolic composition and sensorial properties

Grapevines have traditionally been grown in semi-arid areas, but viticulture is now compromised by climate change. Therefore, it is necessary to implement environmentally friendly viticulture practices to adapt grapevines to current climatic conditions. In this context, organic mulches offer many benefits, such as reduced soil erosion and increased organic matter, soil water content and crop productivity. However, these practices must not compromise grape and wine quality. Therefore, the objective of this study was to evaluate the effect on wine physicochemical and phenolic composition and sensorial properties of different soil management practices on the vine row. Over four years, five soil treatments were examined in two different vineyards.

Inhibition of Oenococcus oeni during alcoholic fermentation by a selected Lactiplantibacillus plantarum strain

The use of selected cultures of the species Lactiplantibacillus plantarum in Oenology has grown in prominence in recent years. While initial applications of this species centred very much around malolactic fermentation (MLF), there is strong evidence to show that certain strains can be harnessed for their bio-protective effects. Unwanted spontaneous MLF during alcoholic fermentation (AF), driven by rogue Oenococcus oeni, is a winemaking deviation that is very difficult to manage when it occurs. This work set out to determine the efficacy of one particular strain of Lactiplantibacillus plantarum(Viniflora® NoVA™ Protect), against this problem in Cabernet Sauvignon must. The work was carried out at commercial scale and in a winery environment and compared the bio-protective culture with the more traditional approach of reducing must pH by the addition of tartaric acid. The combination of both was also investigated. The concentration of both Oenococcus oeni and Lactiplantibacillus plantarum was determined using qPCR. The adventitious Oenococcus oeni showed the most growth during AF in the control wine, whereas in the wines treated with Lactiplantibacillus plantarum a bacteriostatic effect against this species was observed. This effect was comparable to the wines treated with tartaric acid. This has particular commercial relevance for controlling the flora in musts with high pH, or when the addition of tartaric acid is either not permitted or is prohibitive for other reasons.

Chemical activation of ABA signaling in grapevine through ABA receptor agonists

Grapevine (Vitis vinifera) and its derived products, in terms of cultivated area and economic volume, constitute the most relevant fruit crop in the world (7.5 million cultivated hectares). In the current context of climate change, the wine sector faces unprecedented challenges to satisfy a growing demand for wines of greater quality through sustainable viticulture. Global warming threatens quality wine production in Mediterranean wine regions in particular. The increase in heatwaves and drought episodes accelerate the vine phenology and alter the ripening and composition of grapes and wine. Extreme abiotic stress episodes compromise grape production and plant survival, intensifying the pressure on the use of limited resources like water. Abscisic acid (ABA) is an important hormone in the ripening of certain fruits and in plant response to abiotic stress.

Climate change projections in serbian wine-growing regions

Changes in bioclimatic indices in wine-growing region of Serbia are analyzed under the RCP 8.5 IPCC scenario.