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…

Molecular approaches for understanding and modulating wine taste

Wine consumers generally demand wines having a perception of softer tannins and less ripe, having a heaviness and richness on palate (full-body wine) with a limpid and stable color. However, polyphenol
(tannins)-rich wines have been also correlated with unpleasant taste properties such as astringency and
bitterness when perceived at high intensities. Modulating these unpleasant properties could be important for consumer’s approval of wines.

Read More

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.

Read More

Impact of soil-applied and foliar-applied nitrogen on grape and wine composition

Foliar application of urea may be an efficient way to alter grape and wine composition without increasing vine vigor. However, we know little about the impact of this practice on phenolic compounds and yeast assimilable nitrogen (YAN). Adequate YAN is required for an efficient and complete fermentation, while phenolics are particularly important for the sensory profile of red wines. The goal of this study is to test the impact of foliar urea application at veraison, compared to the traditional soil-applied nitrogen fertilization early in the season, on Syrah berry and wine composition in field conditions.

Read More

Above and below: soil moisture and soil temperature interact to alter grapevine water relations

The combined effect of soil moisture and soil temperature on grapevine physiology is gaining interest in the context of global warming.

Read More

The effect of management practices and landscape context on vineyard biodiversity

Intensification is considered one of the major drivers of biodiversity loss in farmland. The more intensive management practices that have been adopted the last decades, contributed to species declines from all taxonomic groups. Moreover, agricultural intensification has led to an important change of land use. Complex, mixed agro-ecosystems with cultivated and non-cultivated habitats have been converted to simplified, intensive and homogeneous ones with severe effects on biodiversity.

Read More