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…

Evaluation of interception traps for capture of Xylotrechus arvicola (Coleoptera: Cerambycidae) in vineyards varieties from Protected Denomination of Origin León

Xylotrechus arvicola (Coleoptera: Cerambycidae) is a pest in vineyards (Vitis vinifera) in the main Spain wine-producing regions with Protected Denomination of Origin (PDO). The action of the larvae, associated to the spreading of wood fungi, causes damage especially in important varieties of V. vinifera. X. arvicola females lay eggs concentrated in cracks or under the rhytidome in the wood vines, which allows the emerging larvae to get into the wood and make galleries inside the plant being then necessary to prune intensively or to pull up the bored plants (1). The objective of the study was to evaluate captures of X. arvicola insects in five varieties of V. vinifera in PDO León.

Read More

Evolution and sensory contribution of ethyl acetate in sweet wines

Ethyl acetate (EtOAc) is the main ester present in all wines, generally produced by yeasts during alcoholic fermentation and sometimes by bacteria during barrel ageing. Its odor is characterized by solvent notes, which give wines their acescent note [1].

Read More

Progetto di zonazione delle valli di Cembra e dell’Adige. Analisi del comportamento della varietà Pinot nero in ambiente subalpino

Nel 1990 la Cantina LA VIS ha intrapreso un progetto di zonazione dei terreni vitati allo scopo di acquisire le conoscenze scientifiche atte a consentire il miglioramento delle qualità dei prodotti. Tale progetto si è articolato su di una superficie di 2000 ettari ubicati lungo l’asta fluviale del fiume Adige da Trento a Salorno e del torrente Avisio da Lavis a Segonzano.

Read More

Pro-active management of grapevine trunk diseases by means of sanitation in nurseries

Several trunk diseases cause decline and premature dieback of grapevines. In vineyards, these pathogens gain entry into plants through unprotected wounds. Wounds are also frequently infected during the propagation stages. The pathogens survive in infected plants in a latent form and cause disease in older grapevines or in plants that are

Read More

PHOTOCHEMICAL DEGRADATION OF TRYPTOPHAN IN MODEL WINE: IMPACT OF HEAVY METALS AND OXYGEN ON 2-AMINOACETOPHENONE FORMATION

The wine industry worldwide faces more and more challenges due to climate change, such as increased dryness in some areas, water stress, sunburn and early harvesting during hot summer temperatures¹. One of the resulting problems for the wine quality might be a higher prevalence of the untypical aging off-flavor (ATA)². A substance, which Rapp and Versini made responsible for ATA, is the 2-aminoace-tophenone (2-AAP)³. 2-AAP in wine causes a naphthalene, wet towels, wet wool, acacia flower or just a soapy note⁴.

Read More