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…

Effects of the addition of yeast derived products during aging in chardonnay sparkling winemaking

From the beginning of the yeast autolysis process, several interesting intracellular and cell wall constituyents are released to the media providing different characteristics to the wine, being this process extensively studied in sparkling wines due to their important contribution to their properties (1-2). Yeast derived products (YDs) try to emulate the natural yeast autolysis compounds release enhancing the organoleptic characteristics of resulting wines (2-3). This study is a comprehensive evaluation of the impact of the addition of different YDs added to base wine on the chemical, physical and sensory characteristics of the resulting sparkling wines. METHODS: Chardonnay base wine was employed to carry out this study. Three experimental YDs were added at 5 and 10 g/hL to the tirage liqueur: a yeast autolysate (YA), a yeast protein extract (PE) and an inactivated dry yeast from Torulaspora delbrueckii, (TD), and two commercial specific inactivated dry yeast: OPTIMUM WHITE® (OW) and PURE-LONGEVITY®(PL). After second fermentation, measurements were carried out after 3, 6, 9 and 18 months of aging on lees. General enological parameters, proteins, polysaccharides (HPLC-DAD-RID), volatile compounds profile (GC-MS), foaming characteristics (Mosalux), and descriptive sensory analyses were carried out.

Read More

Grape composition and wine quality of Muscat Hamburg cultivar after a specific inactivated dry yeast application as adaptation strategy to climate change

In a climate change context, the management of Mediterranean vineyards should be adapted to the new environmental conditions. Predictive models underline that in the future the most of the Mediterranean vineyard regions is expected to experience further warming events producing challenges in ripening balanced fruit. It is already registered that in warm and dry summers, the ripening process is faster and the balance between phenolic and technological (sugar) maturity may not be the desirable. This study investigates the use of specific inactivated yeast derivatives sprayed on the entire canopies of field grown cv Muscat Hamburg vines.

Read More

Balearic varieties of grapevine: study of genetic variability in the response to water stress

The photosynthetic characteristics of twenty varieties of grapevine (Vitis vinifera L.) from Mallorca (Balearic Islands, Spain) and two widespread varieties

Read More

Egg allergens in wine. Validation of a new automated method for ovalbumin quantification

Ovalbumin (ova), a natural clarifying protein, is particularly suitable for clarifying red wines. It helps improve the tannic and polyphenolic stability of the wine by removing the most astringent tannins and contributing to soften and refine the structure. Ova binds to suspended particles, proteins, polysaccharides, and, to a lesser extent, tannins through electrostatic and hydrophobic interactions, forming large complexes that can be removed from the wine through fining and/or filtration before bottling.

Read More

Landscapes of Vines and Wines Patrimony – Stakes – Valorisation

The interaction between wine and landscapes is of an unsuspected richness. On the one side, the vineyards form part of the landscapes which they model. On the other side, the wines are related in their perception to the image of a region, a landscape and are at the origin of a cultural richness.

Read More