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…

Consistency of the hydraulic traits and stomatal responses in grapevines with contrasting hydraulic vulnerability

Different from wild species in arid and semiarid conditions, cultivated species are very sensitive to drought and, beyond some stress thresholds, food production is not possible

Read More

Effects of severe shoot trimming at different phenological stages on the composition of Merlot grapes

High concentration of sugars in grapes and alcohols in wines is one of the consequences of climate change on viticulture production in several wine regions. One of the options to alleviate this potential problem is to perform severe shoot trimming of the vines to limit the production of carbohydrates. Two different studies were performed in order to investigate the effects of severe shoot trimming on the composition of Merlot grapes; in a first study severe shoot trimming was performed at three different phenological stages (at berry set, at the beginning of veraison and at the end of veraison), while in a second study two trimming treatments (standard shoot trimming and severe shoot trimming performed at the end of veraison) were combined with two shoot densities in order to evaluate the relative impact of these treatments on Merlot grape composition.

Read More

Study of the oenological potential of varieties resistant to cryptogamic diseases and drought to anticipate varietal selection in Occitanie

In the context of climate change and the growing need to reduce the use of phytosanitary products, the exploration of disease-resistant grape varieties and/or adapted to drought conditions is becoming crucial for the wine industry in certain regions of France, such as Occitanie. Currently, exploring the oenological potential of varieties by analyzing their biochemical composition before and after winemaking comes rather late in the varietal selection process.

Read More

Possible Reduction Method Of Volatile Acid Content And Polyphenols Of Tokaj Aszú Wines With The Aid Of Citosan Bactericid Wine-Treatments

The historical Tokaj region in northeast Hungary is a UNESCO World Heritage region since 2002 owning 5.500 ha vineyards. Produced from „noble rot” grapes, Tokaji Aszú is known as one of the oldest botrytized wines all over the world. Special microclimatic conditions (due to Bodrog and Tisza rivers, Indian summer), soil circumstances (clay, loess on volcanic bedrock) and grape-varieties (Furmint, Hárslevelű) of Tokaj-region offer favourable parameters to the formation of noble rot caused by Botrytis cinerea. The special metabolic activity of Botrytis results in noble rot grapes called “aszú” berries. The grapes undergo complex chemical modifications as the joint result of the enzymatic activity of Botrytis and the physical process of concentration.

Read More

Investigating water stress-related seasonal and spatial patterns and the possible links with juice and wine compositional parameters

The mapping of spatial variability in vineyards offers the potential to implement zonal management strategies with the aim to optimize economic benefits and increase sustainability by managing natural resources, such as water used for irrigation, more optimally. This study characterized the (natural) variability in plant water status in a commercial Cabernet Sauvignon block, using remote sensing techniques, and identified the impact of this variability on the yield, and juice and wine composition. From the field data collected over two growing seasons, we demonstrated that remote sensing techniques are a practical and powerful tool for mapping spatial variability within vineyard blocks.

Read More