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…

Oxygen consumption and CO2 production rates as selection criteria for Metschnikowia pulcherrima strains in must bioprotection

Redacted.

Read More

Variety specific thresholds for plant-based indicators of vine nitrogen status

Aim: Several plant-based indicators of vine N status are reported in the literature. Among these, yeast assimilable nitrogen in grape must (YAN) and total N concentration of petiole and leaf blades are considered to be reliable indicators and so is the chlorophyll index, measured with a device called N-tester. The N-tester index is used to measure the intensity of the green colour of the leaf blade, and therefore to estimate its chlorophyll content.

Read More

Aroma accumulation trends during berry development and selection of grape aroma candidate genes suitable for functional characterization

Grape flavour management in the vineyard requires knowledge of the derivation of individual flavour and aroma characteristics and the effects that different concentrations and interactions between these compounds have on flavour potential.

Read More

Bentonite fining in cold wines: prediction tests, reduced efficiency and possibilities to avoid additional fining treatments

Bentonite fining is widely used to prevent protein haze in white wines. Most wineries use laboratory-scale fining trials to define the appropriate amount of bentonite to be used in the cellar. Those pre-tests need to mimic as much as possible the industrial scale fining procedure to determine the exact amount of bentonite necessary for protein stability. Nevertheless it is frequent that, after fining with the recommended amount of bentonite, wines appear still unstable and need an additional fining treatment. It remains a major challenge to understand why the same wine, fined with the same dosage of the same bentonite, achieves stability in the lab, but not in the cellar.

Read More

AROMA ASSESSMENT OF COMMERCIAL SFORZATO DI VALTELLINA WINES BYINSTRUMENTAL AND SENSORY METHODOLOGIES

Sforzato di Valtellina DOCG is a special dry red wine produced from partially dehydrated Nebbiolo wine-grapes growing in the Rhaetian Alps valley of Valtellina (Lombardy, Italy). Valtellina terraced vineyards are located at an altitude of 350–800 m according to ‘heroic’ viticulture on steep slopes. The harvested grape bunches are naturally dehydrated indoors, where a slow and continuous withering occurs (about 20% w/w of weight loss), until at least 1st December when the grapes reach the desired sugar content and can be processed following a normal winemaking with maceration.

Read More