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…

Characterizing the effects of nitrogen on grapevines with different scion/rootstock combinations: agronomic, metabolomic and transcriptomic approaches

Most vineyards are grafted and include a variety (Vitis vinifera) grafted over a wild Vitis rootstock (hybrids of V. berlandieri, riparia and rupestris). Grape berry quality at harvest depends on a subtle balance between acidity and the concentrations of sugars, polyphenols and precursors of aroma compounds. The mechanisms controlling the balance of sugars/acids/polyphenols are influenced by the abiotic environment, in particular nitrogen supply, and interact with the genotypes of both the scion variety and the rootstock. Previous work suggests that some of the effects of water stress are in fact linked to a nitrogen deficiency driven indirectly by the reduction of water absorption.

Les effets du terroir ou l’expression des potentiels à valoriser

Research into the effects of the Terroir is of major interest for the wine sector. The study of Terroir-Vine-Grape relations, even if it is complex, is fundamental for all viticulture: indeed, the quality of the grape must be the result of the most reasoned agro-viticultural management of the vine possible, which must first, to respect a production balance. The goal sought by the winegrower is to obtain a wine, the optimized result of the interactions Terroir-Grape variety. This link to the terroir is therefore essential to establish by taking into account on the one hand the behavior of the vine (which is the cause), and on the other hand, its effects on the grapes and finally on the wine.

Saccharomyces cerevisiae intraspecies differentiation by metabolomic signature and sensory patterns in wine

AIM: The composition and quality of wine are directly linked to microorganisms involved in the alcoholic fermentation. Several studies have been conducted on the impact of Saccharomyces cerevisiae on volatile compounds composition after fermentation. However, if different studies have dealt with combined sensory and volatiles analyses, few works have compared so far the impact of distinct yeast strains on the global metabolome of the wine.

Rootstock regulation of scion phenotypes: the relationship between rootstock parentage and petiole mineral concentration

Grapevine is grown grafted in most of the world largely because of Phylloxera. Rootstocks not only provide tolerance to Phylloxera, but also ensure the supply of water and mineral nutrients to the scion. Rootstocks are an important means of adaptation to environmental conditions if we want to conserve the typical features of the currently used scion genotypes. To aid this adaptation, we can exploit the large diversity of rootstocks used worldwide. To fully explore this existing rootstock diversity, this work benefits from the unique GreffAdapt vineyard, in which four scion genotypes were studied onto 55 commercial rootstocks in three blocks. The aim of this study was to characterise rootstock regulation of scion mineral status and how it relates to scion development.

Flor yeast diversity and dynamics in biologically aged wines

Wine biological aging is characterized by the development of yeast strains that form a biofilm on the wine surface after alcoholic fermentation. These yeasts, known as flor yeasts, form a velum that protects the wine from oxidation during aging. Thirty-nine velums aged from 1 to 6 years were sampled from “Vin jaune” from two different cellars. We show for the first time that these velums possess various aspects in term of color and surface aspects. Surprisingly, the heterogeneous velums are mostly composed of one species, S. cerevisiae. Scanning electron microscope observations of these velums revealed unprecedented biofilm structures and various yeast morphologies formed by the sole S. cerevisiae species.