terclim by ICS banner
IVES 9 IVES Conference Series 9 DNA-Free genome editing confers disease resistance in grapevine

DNA-Free genome editing confers disease resistance in grapevine

Abstract

Grapevine (Vitis Vinifera L.), one of the most important cultivated fruit crops, is facing significant challenges due to climate change. Specifically, increasing temperatures negatively impact the physiological traits and disrupt plant phenology. Additionally, increased virulence in pathogen attacks and pests leads to significant yield loss, requiring widespread application of plant protection products. Traditional agronomic practices offer only partial mitigation, requiring the development of precise and effective intervention strategies. The economic worth of viticulture has prompted continuous efforts in grapevine genetic improvement programs, traditionally involving conventional breeding and clonal selection that, however, are complex and time-consuming approaches. Instead, the advent of New Breeding Techniques, especially genome editing via the CRISPR/Cas9 system, presents a promising avenue for the development of tools suitable to mitigate the current viticulture challenges, including fungal diseases. We report the application of a DNA-free genome editing approach to induce targeted mutations in the VviMLO17, a gene associated with powdery mildew susceptibility in grapevine. CRISPR/Cas9 ribonucleoparticles were introduced into protoplasts isolated from embryogenic calli. Through protoplast regeneration, a homozygous edited grapevine plant mutated in the VviMLO17 gene was obtained. This mutation confers resistance to Erysiphe necator, as evidenced by phenotypic analyses that demonstrated a reduced susceptibility to pathogen attack. The success of DNA-free CRISPR/Cas9 application for the improvement of target traits establishes a foundation for promoting viticulture sustainability yet preserving the identity of the grapevine cultivars. This advancement aligns with market and legislative demands, paving the way for a resilient and environmentally conscious winegrowing system.

DOI:

Publication date: June 13, 2024

Issue: Open GPB 2024

Type: Article

Authors

Marianna Fasoli1, Edoardo Bertini2, Erica D’Incà2, Luca Cattaneo1, Stefania Zattoni1, Sara Lissandrini1, Clarissa Ciffolillo1, Annalisa Polverari1, Giovanni Battista Tornielli1,3, Sara Zenoni1*

1 Department of Biotechnology, University of Verona, Verona, Italy
2 EdiVite S.r.l., San Pietro Viminario, Padua, Italy
3 Current address: Department of Agronomy, Food, Natural Resources, Animals and the Environment., University of Padua, Padua, Italy

Contact the author*

Keywords

genome editing DNA-free, CRISPR/Cas9 system, protoplast regeneration, powdery mildew resistance, sustainable viticulture

Tags

IVES Conference Series | Open GPB | Open GPB 2024

Citation

Related articles…

ALCOHOLIC FERMENTATION DRIVES THE SELECTION OF OENOCOCCUS OENI STRAINS IN WINE

Oenococcus oeni is the predominant lactic acid bacteria species in wine and cider, where it performs the malolactic fermentation (MLF) (Lonvaud-Funel, 1999). The O. oeni strains analyzed to date form four major genetic lineages named phylogroups A, B, C and D (Lorentzen et al., 2019). Most of the strains isolated from wine, cider, or kombucha belong to phylogroups A, B+C, and D, respectively, although B and C strains were also detected in wine (Campbell-Sills et al., 2015; Coton et al., 2017; Lorentzen et al., 2019;

Climate ethnography and wine environmental futures

Globalisation and climate change have radically transformed world wine production upsetting the established order of wine ecologies. Ecological risks and the future of traditional agricultural systems are widely debated in anthropology, but very little is understood of the particular challenges posed by climate change to viticulture which is seen by many as the canary in the coalmine of global agriculture. Moreover, wine as a globalised embedded commodity provides a particularly telling example for the study of climate change having already attracted early scientific attention. Studies of climate change in viticulture have focused primarily on the production of systematic models of adaptation and vulnerability, while the human and cultural factors, which are key to adaptation and sustainable futures, are largely missing. Climate experts have been unanimous in recognising the urgent need for a better understanding of the complex dynamics that shape how climate change is experienced and responded to by human systems. Yet this call has not yet been addressed. Climate ethnography, coined by the anthropologist Susan Crate (2011), aims to bridge this growing disjuncture between climate science and everyday life through the exploration of the social meaning of climate change. It seeks to investigate the confrontation of its social salience in different locations and under different environmental guises (Goodman 2018: 340). By understanding how wine producers make sense of the world (and the environment) and act in it, it proposes to focus on the co-production of interdisciplinary knowledge by identifying and foreshadowing problems (Goodman 2018: 342; Goodman & Marshall 2018). It seeks to offer an original, transformative and contrasted perspective to climate change scenarios by investigating human agency -individual or collective- in all its social, political and cultural diversity. An anthropological approach founded on detailed ethnographies of wine production is ideally placed to address economic, social and cultural disruptions caused by the emergence of these new environmental challenges. Indeed, the community of experts in environmental change have recently called for research that will encompass the human dimension and for more broad-based, integrated through interdisciplinarity, useful knowledge (Castree & al 2014). My paper seeks to engage with climate ethnography and discuss what it brings to the study of wine environmental futures while exploring the limitations of the anthropological environmental approach.

Il ruolo dei comuni nella gestione del territorio e nella tutela dei vitigni autoctoni di qualita’

Questo simposio organizzato dall ‘Associazione nazionale Città del Vino, che mi onoro di presiedere, è per me motivo di particolare soddisfazione perché porta a compimento parte di un percorso iniziato dall’associazione da alcuni anni e che ha un obiettivo apparentemente semplice: sollecitare gli amministratori delle Città del Vino a perseguire con tenacia, tal­volta anche con la necessaria caparbietà, programmi ed interventi che abbiano al centro, sempre, la qualità della vita dei loro territori.

Influence of grapevine rootstock/scion combination on rhizosphere and root endophytic microbiomes

Soil is a reservoir of microorganisms playing important roles in biogeochemical cycles and interacting with plants whether in the rhizosphere or in the root endosphere. The composition of the microbial communities thus impacts the plant health. Rhizodeposits (such as sugar, organic and amino acids, secondary metabolites, dead root cells …) are released by the roots and influence the communities of rhizospheric microorganisms, acting as signaling compounds or carbon sources for microbes. The composition of root exudates varies depending on several factors including genotypes. As most of the cultivated grapevines worldwide are grafted plants, the aim of this study was to explore the influence of rootstock and scion genotypes on the microbial communities of the rhizosphere and the root endosphere. The work was conducted in the GreffAdapt plot (55 rootstocks x 5 scions), in which the 275 combinations have been planted into 3 blocks designed according to the soil resistivity. Samples of roots and rhizosphere of 10 scion x rootstock combinations were first collected in May among the blocks 2 and 3. The quantities of bacteria, fungi and archaea have been assessed in the rhizosphere by quantitative PCR, and by cultivable methods for bacteria and fungi. The communities of bacteria, fungi and arbuscular mycorrhizal fungi (AMF) was analyzed by Illumina sequencing of 16S rRNA gene, ITS and 28S rRNA gene, respectively. The level of mycorrhization was also evaluated using black ink coloration of newly formed roots harvested in October. The level of bacteria, fungi and archaea was dependent on rootstock and scion genotypes. A block effect was observed, suggesting that the soil characteristics strongly influenced the microorganisms from the rhizosphere and root endosphere. High-throughput sequencing of the different target genes showed different communities of bacteria, fungi and AMF associated with the scion x rootstock combinations. Finally, all the combinations were naturally mycorrhized. The root mycorrhization intensity was influenced by the rootstock genotype, but not by the scion one. Altogether, these results suggest that both rootstock and scion genotypes influence the rhizosphere and root endophytic microbiomes. It would be interesting to analyze the biochemical composition of the rhizodeposition of these genotypes for a better understanding of the processes involved in the modulation of these microbiomes. Moreover, crossing our data with the plant agronomic characteristics could provide insights into their roles on plant fitness.

Isolated Antarctic soil yeasts with fermentative capacity with potential use in the wine industry

The wine industry is currently on the search for new aromas and less browning in their products. In the improvement process of wine, lower fermentation temperatures have been considered, however, the yeasts in the market cannot tolerate such temperatures