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…

Sensory analysis in oenology: the role of methodological differences in expert panel evaluations

Sensory analysis is an essential component of oenology, offering valuable insights into wine quality that influence decision-making in viticulture and winemaking.

Variabilité spatiale du gel printanier dans le vignoble champenois : application au zonage climatique

In the Champagne vineyards, spring frosts are the cause of significant variations in the volume of the harvest which are very penalizing for the trade. This variability is reflected both in time (years without frost alternating with years with severe frosts) and in space. Certain sectors of the vineyard are in fact statistically more susceptible to frost than others, but each year no municipality can consider itself immune to this climatic accident. The objective of the study is precisely to analyze the spatial distribution of frost and to determine its main mechanisms, linked to the topography of the hillsides, their orientation but also to regional meteorological variables.

Thermal conditions during the grape ripening period in viticulture geoclimate. Cool night index and thermal amplitude

Le régime thermique en période de maturation du raisin est l’une des variables déterminantes de la coloration du raisin et de la richesse en arômes, anthocyanes et polyphénols des vins.

The albarizas and the viticultural zoning of Jerez­-Xérès-Sherry and Manzanilla-Sanlúcar de Barrameda registered apellations of origin (Cadiz, Spain)

Le terme ”Albariza” (du latin “albus“, blanc) déterminait à l’origine un type particulier du terrain calcaire, mais à présent il sert aussi à définir les sols et la bibliographie géologique actuelle le cite également pour de roches sédimentaires originaires du Neogene Betic.

AN AUTOMATIC CANOPY COOLING SYSTEM TO COPE WITH THE THERMAL-RADIATIVE STRESSES IN THE PIGNOLETTO WHITE GRAPE

In recent years characterized by hot dry summers, the implementation of innovative irrigation tools in the vineyard represents a crucial challenge to ensure optimal production and to avoid excess of water consumption. It is known that the grapevine reacts to multiple stresses – i.e., high temperatures and wa- ter shortage – through adaptive mechanisms that are detrimental to the yield. Furthermore, this condi- tion is usually aggravated by high solar radiation, which could negatively affect the phenolic composi- tion of the grapes. Therefore, a cooling system has been developed aiming to reduce bunches’ sunburn damage.