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…

Application to the wine sector of European Convention on the landscapes

The landscape is defined by the European convention of the landscape (Florence, October 20, 2000) like part of the territory as perceived by the populations, whose character results from the action of natural and/or human factors and their interrelationships. This convention is based on the contribution cultural, ecological, environmental, social of the landscapes and aims at a reinforcement of the tools of protection and valorization in particular in the agricultural policies, of regional planning and town planning. Moreover, it encourages a step of identification and qualification of the landscapes and underlines the need for developing the sensitizing and the training of the actors concerned.

Read More

Moscatel vine-shoot extracts as grapevine biostimulant to increase the varietal aroma of Airén wines

There is a growing interest in the exploitation of vine-shoots waste, since they are often left or burned. Sánchez-Gómez et al. [1] have shown that vines-shoots aqueous extracts have significant contents of bioactive compounds, among which several polyphenols and volatiles are highlighted. Recent studied had demonstrated that the chemical composition of vine-shoots is enhanced when vine-shoots are toasted
[2,3]. The application of vegetable products in the vineyards has led to significant changes towards a more “Sustainable Viticulture”. An innovative foliar application for Airén vine-shoot extracts have been carried out to the vineyard. It has been shown that they act as grape biostimulants, improving certain wine quality characteristics [4].

Read More

Evaluating the greenness of wine analytical chemistry: A new metric approach

Wine is a complex matrix whose composition depends on climatic, agricultural, and winemaking factors, making quality control and authenticity assessment critical in the global market.

Read More

Soil Temperature and Climate Change: Implications for Mediterranean Vineyards 

More frequent and extreme temperatures and droughts pose challenges to the wine sector in Mediterranean Europe. Soil is crucial to sustain the equilibrium of ecosystems, economic growth and people’s prosperity worldwide. In viticulture, soils are a major component of the terroir and do influence vine’s growth, yield and berry composition. Soil temperature (ST) affects soil´s physical, chemical and biological processes and also crop growth. The impact of ST becomes even stronger when dealing with row crops such as grapevine, when considering the increased exposition to radiation. However, the impact of ST on crop performance remains poorly described, especially for extreme climatic conditions.

Read More

Effect of different winemaking techniques and grape variety on chemo-sensory parameters of white wines

AIM: Study the chemical and sensory parameters of fifty commercial white wines elaborated with different techniques (fermented in oak barrel and aged on lees (FB+AL); aged on lees (AL); and without aging (WA)) and different grape varieties (Verdejo, Sauvignon blanc and Godello).

Read More