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…

Grapevine nitrogen retrieval by hyperspectral sensing at the leaf and canopy level

Grapevine nitrogen (N) monitoring is essential for efficient N management plans that optimize fruit yield and quality while reducing fertilizer costs and the risk of environmental contamination. Unlike traditional vegetative-tissue sampling methods, remote sensing technologies, including hyperspectral imaging, have the potential to allow monitoring of the N status of entire vineyards at a per-vine resolution. However, differential N partitioning, variable spectral properties, and complex canopy structures hinder the development of a robust N retrieval algorithm. The present study aimed to establish a solid understanding of vine spectroscopic response at leaf and canopy levels by evaluating the different nitrogen retrieval approaches, including the radiative transfer model.

Read More

Eugenol:  a new marker of hybrid vines? The case study of Baco Blanc in Armagnac

Nowadays, winemaking is dealing with great challenges, notably climate change, disease resistance and low pesticide inputs, desire for more sustainable agricultural productions and permanent changing of consumer preference. Trying to propose practice improvements, scientists are exploring vine hybridization a paradoxically old but still actual way to take up such challenges

Read More

THE EFFECT OF DIFFERENT TERROIRS ON AROMA COMPOUNDS OF ‘KALECIK KARASI’ WINES

Kalecik Karası is a domestic grape variety of Turkey, originating from Kalecik district, 80 km from Ankara. Although there is no definite evidence, it is known that it was used in wine production by many civilizations that lived in the Anatolian region, especially the Hittites. Compared to other black wine grapes, it stands out with its low tannin content, rich fruity aroma and complex structure. In good vintages, red fruits such as strawberries, cherries and raspberries stand out in the aroma profile. Although its structure is elegant, it has the potential to age and develop similar to the ‘Pinot Noir’ wine of the Burgundy region. This offers a complex aroma structure including red flowers, earth and ripe fruits.

Read More

Development of a GRASS-GIS application for the characterization of vineyards in the province of Trento

The physical factors that influence the grape ripening include elevation, slope, aspect, potential global radiation, sun hours and soil type of the vineyards.

Read More

The role of rootstock and its genetic background in plant mineral status

In this video recording of the IVES science meeting 2025, Marine Morel (EGFV, Univ. Bordeaux, Bordeaux Sciences Agro, INRAE, ISVV, Villenave-d’Ornon, France) speaks about the role of rootstock and its genetic background in plant mineral status. This presentation is based on an original article accessible for free on OENO One.

Read More