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…

A multidisciplinary approach to grapevine zoning G.I.S. technology based: an example of thermal data elaboration

Un grand nombre d’études ont été consacrées à l’évaluation quantitative des effets de climat sur la qualité des vignes, dans différents contextes climatiques. Généralement, la vocation viticole d’un terroire peut être étudiée par des approches mono ou multidisciplinaires.

Rootstock impact on foliar symptom expression of esca on Vitis vinifera cv. Cabernet-Sauvignon

Trunk diseases and esca in particular, represent a major threat to the sustainability of the vineyards. The percentages of unproductive vines in a plot could vary from 4% to over 20 % depending on local conditions and vintages.

Coping with heatwaves: management strategies for berry survival and vineyard resilience

Climate change is leading to an increase in average temperature and in the frequency and severity of heatwaves that is already significantly affecting grapevine phenology and berry composition (Webb et al., 2010). This is compounded by water stress, which is well known to increase the vulnerability of grapevines and berries to heatwaves. In hot climate regions like australia, grape production is only possible due to relatively secure supplies of water for irrigation. However, the upper temperature limits for berry survival of well-watered grapevines remains to be tested.

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.

Multiple description and validation of autochthone grape varieties in the Carpathian Basin

Context and Purpose of the Study. In many countries, the preservation of grape varieties with heritage value is ensured by genebanks of outstanding significance, which allow for the study of these varieties and the assessment of their future roles in response to environmental, market, and social challenges.