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…

Towards microbiota-based disease management: analysis of grapevine microbiota in plots with contrasted levels of downy mildew infection

Vineyards harbor a myriad of microorganisms that interact with each other and with the grapevines. Some microorganisms are plant pathogens, such as the oomycete Plasmopara viticola that causes grapevine downy mildew. Others, such as plant growth promoting bacteria and disease biocontrol agents, have a positive influence on vine health. The present study aims to (1) investigate whether vine-based culture media increase the cultivability of the grapevine microbiota, in comparison to standard culture media and (2) identify and isolate bacterial taxa naturally present in grapevine leaves and significantly more abundant in plots showing low susceptibility to downy mildew.

Read More

The influence of different fertiliser applications and canopy management practices on the potassium content and pH of juice and wine of Vitis vinifera L. cvs. Cabernet-Sauvignon and Cabernet franc

In an attempt to reduce the pH of juice and wine, different fertiliser applications and canopy management practices were evaluated in South Africa in a field trial. Fertiliser treatments entailed no, CaSO4, Ca(OH)2, and MgSO4 fertilisation.

Read More

Withering of the ‘Moscato giallo’ grapes under covered space

For the purpose of producing predicate wines in northern part of Croatia, grapes are traditionally left on the vine unpicked. However, grapes on the vine are exposed to unfavorable environmental conditions that affect rapid rotting and attacked by birds. To eliminate the mentioned risks, the grapes can be picked and placed in a protected space (loft, greenhouse, etc.) suitable for drying. This study presents the results of research on withering grapes of the ‘Moscato giallo’ variety in two tretment: sun drying (under covered terrace) and drying in the shade (loft). The following quality parameters were monitored: mass of grapes, sugar concentration, content of total acids, pH, content of organic acids.

Read More

Ultrastructural and chemical analysis of berry skin from two Champagne grapes varieties and in relation to Botrytis cinerea susceptibility

Botrytis cinerea is a necrotrophic pathogen that causes one of the most serious diseases of the grapevine (Vitis vinifera), grey mold or Botrytis bunch rot. In Champagne, the Botrytis cinerea disease leads to considerable economic losses for winemakers and wines exhibit organoleptic defaults.

Read More

Asymmetrical flow field-flow fractionation with online multidetection is a viable tool to investigate colored red wine colloids

Despite its relevance for wine quality and stability, red wine colloids have not still been
sufficiently investigated, an occurrence due to the lack of suitable analytical techniques to study them as they are present in wine.

Read More