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.

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…

Growth in global table grape production and consumption is fueled by the introduction of new seedless varieties

Table grape consumption worldwide has experienced a remarkable growth in the first two decades of the 21st century, becoming the third most consumed fresh fruit in some countries, after bananas and apples. This increase has been attributed to several reasons, including the availability of seedless grapes, which has been a key factor in the increase in consumption.

Sensory and physicochemical impact of proanthocyanidic tannins on red wine fruity aroma

AIM: Previous research on the fruity character of red wines highlighted the role of esters [1]. Literature provides evidence that, besides these esters, other compounds that are not necessarily volatiles may have an important impact on the overall fruity aroma of wine, contributing to a masking effect [2][3]. The goal of this work was to assess the olfactory consequences of a mixture between esters and proanthocyanidic tannins, through sensory and physico-chemical approaches.

Influence of harvest time and withering length combination on reinforced Nebbiolo wines: phenolic composition, colour traits, and sensory profile

Sforzato di Valtellina DOCG is a reinforced dry red wine produced in the mountain area of Valtellina alpine valley (North Italy), using ‘Nebbiolo’ grapes that undergo a withering process. This process impacts on the grape composition due to a sugar concentration and changes in secondary metabolism influencing volatile organic compounds (VOCs) and polyphenols.

Study and valorization of vineyards “terroirs” of A.O.C. Cahors (Lot, France)

In the current context of market competition and consumption evolution, it is necessary to produce wines of a genuine typicity. The Terroir represents an unique and irreproducible inheritance that can be valorized through the origin and the sensory characteristics of the wines.

Adapting the vineyard to climate change in warm climate regions with cultural practices

Since the 1980s global regime shift, grape growers have been steadily adapting to a changing climate. These adaptations have preserved the region-climate-cultivar rapports that have established the global trade of wine with lucrative economic benefits since the middle of 17th century. The advent of using fractions of crop and actual evapotranspiration replacement in vineyards with the use of supplemental irrigation has furthered the adaptation of wine grape cultivation. The shift in trellis systems, as well as pruning methods from positioned shoot systems to sprawling canopies, as well as adapting the bearing surface from head-trained, cane-pruned to cordon-trained, spur-pruned systems have also aided in the adaptation of grapevine to warmer temperatures. In warm climates, the use of shade cloth or over-head shade films not only have aided in arresting the damage of heat waves, but also identified opportunities to reduce the evapotranspiration from vineyards, reducing environmental footprint of vineyard. Our increase in knowledge on how best to understand the response of grapevine to climate change was aided with the identification of solar radiation exposure biomarker that is now used for phenotyping cultivars in their adaptability to harsh environments. Using fruit-based metrics such as sugar-flavonoid relationships were shown to be better indicators of losses in berry integrity associated with a warming climate, rather than solely focusing on region-climate-cultivar rapports. The resilience of wine grape was further enhanced by exploitation of rootstock × scion combinations that can resist untoward droughts and warm temperatures by making more resilient grapevine combinations. Our understanding of soil-plant-atmosphere continuum in the vineyard has increased within the last 50 years in such a manner that growers are able to use no-till systems with the aid of arbuscular mycorrhiza fungi inoculation with permanent cover cropping making the vineyard more resilient to droughts and heat waves. In premium wine grape regions viticulture has successfully adapted to a rapidly changing climate thus far, but berry based metrics are raising a concern that we may be approaching a tipping point.