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…

Stability of 3-mercaptohexanol during white wine storage in relationship to must pre-fermentative fining

3-Mercaptohexanol (3MH) is a volatile thiol occurring in several white and red wines, where it can contribute to fruity attributes. Its content is typically high in wines from certain grape varieties, in particular Sauvignon blanc, where it is considered a varietal marker. The strong nucleophilic character of thiols makes 3MH rather unstable during wine storage, due to the presence of several strong electrophilic species. Among these electrophilics, those arising from the oxidation of flavan3-ols such as catechin and epi-catechin have been indicated as critical for 3MH stability. Accordingly, there is a generalized interest towards the ability of vinification practices to reduce 3MH loss during aging through the management of wine flavan-3-ols content.

Determination of target compounds in cava quality using liquid chromatography. Application of chemometric tools in data analysis

According to the Protected Designation of Origin (PDO), Cava is protected in the quality sparkling wines made by the traditional Champenoise method were the wine realize a second fermentation inside the own bottle1. Geographical and human peculiarities of each bottle are the main way for the final quality2. The aim of this study is to find correlations and which target compounds are the most representative of the quality of two different grape varieties, Pinot Noir and Xarel·lo. The quality of these two types of grapes is being studied for each variety by a previous classification of the vineyard made by the company who provided the samples (qualities A,B,C,D, being A the better one and D the worst one). The target compounds studied are organic acids and polyphenols. The methodology for the determination of organic acids is HPLC-UV/vis and for some of them the enzymatic methodology.

Enhancing grapevine transformation and regeneration: A novel approach using developmental regulators and BeYDV-mediated expression

Grapevine (Vitis vinifera L.) is a challenging plant species to transform and regenerate due to its complex genome and biological characteristics. This limits the development of cisgenic and gene-edited varieties. One hurdle is selecting the best starting tissue for the transformation process, much like isolating suitable tissue for protoplasts. One promising method involves delivering crispr/cas components to protoplasts isolated from embryogenic calli, which are then induced to regenerate.

Les micro-zones et les technologies traditionnelles de la viniculture en Géorgie

La Géorgie est un pays d’une tradition très ancienne de viticulture et de viniculture. Là, dans les micro zones spécifiques, en précisant le lieu on produit de différents types du vin d’une

The effects of calcite silicon-mediated particle film application on leaf temperature and grape composition of Merlot (Vitis vinifera L.) vines under different irrigation conditions

This study examined whether the application of calcite-silicon mediated particle film (CaPF) at veraison can mitigate a drought-induced increase in leaf temperature on grapevine, thus contributing to improved leaf functionality, yield and grape composition traits. A total of 48 five-year-old Merlot (Vitis vinifera L.)