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…

IMPACT OF CLIMATIC CONDITIONS ON THE SEASONING QUALITY OF OAK WOOD FOR OENOLOGICAL USE (QUERCUS PETRAEA)

For coopers, seasoning and toasting are considered crucial steps in barrel making during which the oak wood develops specific organoleptic properties. Seasoning, carried out in the open air, allows reducing the moisture content of the staves to between 14 and 18% (compared to 70 to 90% after splitting) while modulating the intrinsic composition of the oak wood. Toasting consists of applying different degrees of heat to a barrel for a specific period of time. As the temperature increases, oak wood produces a wide range of chemical compounds through thermal degradation of its intrinsic composition.

Characterization of a Sémillon clonal population: exploring genetic diversity, metabolomic profiles, and phenotypic variations

Sémillon is a cultivated grape variety known for contributing to dry and sweet white wine production. However, only seven approved clones have been officially recognized in France[1]. In this study, we aimed to characterize the genetic diversity and metabolomic profiles of a Sémillon clonal population, shedding light on the potential variations within this important grape variety.

Beyond liking scores: the importance of the drinking experience to understand our consumers

The presentation will approach the understanding of wine consumers´ perception based on the experiential model suggested by Warell (2008). In this framework, wine consumption gives rise to a
variety of experiences related to the perception, understanding, and judgment of the product. These
multidimensional facets of the drinking experience can be explored by measuring affective, cognitive,
and sensory responses of consumers, which are shown to be stable regardless of the social context.

Le réseau français des partenaires de la sélection vigne : un dispositif unique au monde au service de la sauvegarde du patrimoine variétal

The French vine selection partners network is currently made up of 40 regional partners, grouped around IFV (French Institute for Vine and Wine) and INRAE (national research institute for agriculture and environment), whose missions are preservation, selection, and innovation of our varietal diversity. The originality of this device is based on a 3-level organisation: – varietal diversity preservation, with the world reference: the INRAE’s vine genetics resources centre of Vassal-Montpellier (Marseillan, France), the world’s largest ampelographic collection, which includes nearly 6 000 accessions of cultivated Vitis vinifera from 54 countries, as well as rootstocks, interspecific hybrids, wild vines (lambrusques) and wild American and Asian species.

Approche méthodologique concernant une caractérisation sensorielle de vins rouges de l’Anjou

Face à une concurrence de plus en plus rude entre pays producteurs, le vignoble de l’Anjou, déjà riche par sa diversité, souhaite renforcer sa logique de vins d’ A.O.C., notamment au travers de ses vins rouges.