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…

The wine country, between landscape and promoting tool. The example of Chinon and Saint-Nicolas-de-Bourgueil vineyards (France)

When talking about wine, terroirs are never too far. The National Institute of Apellation d’Origine (INAO) defines it as a system inside of which interact a group of human factors, an agricultural production and a physical environment.

Climat-roche-sol-fromage. Cartographie fonctionnelle du terroir. Exemple de l’A.O.C. Comté

La place prépondérante que prend le Massif Jurassien en Franche-Comté confère à la région un caractère montagneux qui a orienté l’agriculture vers l’élevage laitier. Cette vocation pastorale marquée et de rudes conditions climatiques sont à l’origine de la production, attestée depuis l’Antiquité, d’un fromage de réserve pour la longue période hivernale. Cette tradition fromagère, liée à des prairies naturelles, a perduré jusqu’à nos jours. La qualité et la spécificité du produit actuel, le fromage de Comté, ont été reconnues dès 1952 par l’attribution d’un label et dès 1958, par la reconnaissance d’un périmètre d’appellation d’origine contrôlée, l’A.O.C. Comté (fig. 1).

High density balsamic vinegar: application of stable isotope ratio analysis to determine watering down.

Aceto balsamico di Modena IGP (ABM) is an Italian worldwide appreciated PGI (Protected Geographical Indication) vinegar,  obtained from cooked and/or concentrated grape must (at least 20% of the volume), with the addition of at least 10% of wine vinegar and a maximum 2% of caramel for color stability (EU Reg. 583/ 2009).

Quantification of the production of hydrogen peroxide H2O2 during wine oxidation

Chemical studies aiming at assessing how a wine reacts towards oxidation usually focus on the characterization of wine constituents, such as polyphenols, or oxidation products. As an alternative, the key oxidation intermediate hydrogen peroxide H2O2 has never been quantified, although it plays a pivotal role in wine oxidation. H2O2 is obtained from molecular oxygen as the result of a first cascade of oxidation reactions involving metal ions and polyphenols. The produced H2O2 then reacts in a second cascade of oxidation to produce reactive hydroxyl radicals that can attack almost any chemical substrate in wine.

METABOLIC INTERACTIONS OF SACCHAROMYCES CEREVISIAE COCULTURES: A WAY TO EXTEND THE AROMA DIVERSITY OF CHARDONNAY WINE

Yeast co-inoculations in winemaking have been investigated in various applications, but most often in the context of modulating the aromatic profiles of wines. Our study aimed to characterize S. cerevisiae interactions and their impact on wine by taking an integrative approach. Three cocultures and corresponding pure cultures of S. cerevisiae were characterized according to their fermentative capacities, the chemical composition and aromatic profile of the associated Chardonnay wines. The various strains studied within the cocultures showed different behaviors regarding their development.