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…

Where the sky is no limit — The transformation of wine marketing through text-to-video generation AI model

The introduction of ai-driven tools in digital content creation represents a significant shift in the landscape of marketing, particularly for industries reliant on rich visual storytelling such as the wine sector. The development of ai models like openai’s sora, runway’s gen-2 or google’s lumiere, which can generate realistic video content from textual descriptions, offers promising new avenues for enhancing brand narrative and consumer engagement. This research explores the potential of text-to-video (t2v) ai models to revolutionize wine marketing by creating dynamic, engaging content that captures the essence of vineyards and their products without the need for traditional video production processes.

Read More

Potential use of the yeast Starmerella bacillaris as a sustainable biocontrol agent against gray mold disease in viticulture

Pest biocontrol strategies are gaining attention as eco-friendly alternatives to the use of synthetic pesticides, including in viticulture.

Read More

Characterization of the adaptive mechanisms of grapevine rootstocks to iron deficiency induced by lime stress

Iron (Fe) deficiency is one of the important nutritional disorders for grapevine growing in alkaline and calcareous soils. Although Fe is an abundant element in soil, several factors limiting its availability, particularly the high levels of calcium carbonate or bicarbonate in soil, leading to a remarkable reduction in grapevine growth and productivity. The use of Fe chlorosis-tolerant rootstocks seems to be a cost-effective and efficient way to maintain Fe balance. Morphological and physiological changes occur in plants to cope with low Fe availability, including enhancement of ferric chelate reductase activity and altering root system by increasing lateral roots and root hairs.

Read More

Impact of climate change on the aroma of red wines: a focus on dried fruit aromas

The volatile composition of grapes (free and bound forms) contributes greatly to the varietal aroma and quality of wines. Several agronomical parameters affect grapes composition and wine quality: maturity level at harvest, water status, and the intensity of sun exposure.

Read More

Phenolic and volatile profiles of south tyrolean pinot blanc musts and young wines

AIM. Assess the impact of different vineyards and winemaking variables on the phenolic and volatile profiles of Pinot Blanc musts and young wines from South Tyrol.

Read More