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.

Download the abstract

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…

Valorization of grape marc in a biorefinery loop for producing short- and medium-chain fatty acids, hydrogen, and methane, with polyphenol recovery

Global grape production amounts to approximately 70 million tons per year, with Europe contributing 61% of the world’s wine output, primarily from Italy, France, and Spain.

Read More

Genetic and hormonal regulation of grape berry cuticle formation

The plant surface typically comprises of various epidermal cell types which synthesise and deposit a protective waxy layer known as the cuticle. The cuticle is a significant contributor to important crop traits related to drought tolerance, biotic stress, postharvest fruit quality as well as providing structural support. In this work we have investigated grape berry cuticle formation in the context of the accumulation of anti-fungal specialised metabolites and the ability of the cuticle to structurally cope with the rapid expansion of ripening berries. Metabolic QTL analysis was performed in a grapevine cross population, using chemical profiling data collected via GC-MS analysis for cuticular waxes.

Read More

Outline for the définition of “Terroirs Viticoles application to the area of El AIjarafe (Seville, Spain)

The grapes producing and wine making regions are différent in their use of agricultural, industrial or agroindustrial means. These means are quite often very original and/or specialised; and lately are also quite competitive. Such means are being defined with increased accuracy in the delimitation and definition of its characteristics (Paneque et al., 1996 a). Human action together with other Elements and Agents involved in the vine growing production (Reyner, 1989) over these means lead to agronomic systems with important characteristics. Finally, the transformation of the vine growing production, through different technologies (Fleet, 1992), results in the creation of products with a different acceptance and economical value in the market.

Read More

Chardonnay white wine bottled with different oenological tannins: effect on colour traits, volatile composition and sensory attributes during shelf-life

The use of oenological tannins during winemaking has been mostly studied for improving colour traits and stability on red wines. Their effectiveness mainly depends on the tannin composition, grape variety and winemaking approach [1].

Read More

NMR profiling of grape musts from some italian regions

With wine fraud, being a widespread problem [1], the need for more sophisticated and precise analytical methods of its detection remains ever persistent.

Read More