terclim by ICS banner
IVES 9 IVES Conference Series 9 A DNA-free editing approach to help viticulture sustainability: dual editing of DMR6-1 and DMR6-2 enhances resistance to downy mildew 

A DNA-free editing approach to help viticulture sustainability: dual editing of DMR6-1 and DMR6-2 enhances resistance to downy mildew 

Abstract

The sustainability of viticulture hinges on maintaining quality and yield while reducing pesticide use. Promising strides in this direction involve the development of clones with enhanced disease tolerance, particularly through the knockout of plant susceptibility genes. Knocking out of Downy Mildew Resistant 6 (DMR6) led to increased levels of endogenous salicylic acid (SA), a regulator of immunity, resulting in enhanced tolerance to Downy Mildew (DM) and other diseases in various crops.

Mutations in both DMR6-1 and DMR6-2 genes were introduced into two grapevine cultivars using CRISPR-Cas9 using two methods. In the first case, transgene delivery mediated by A. tumefaciens was employed, while in the second case, we developed a ‘single-cell technology’ for gene editing, creating non-transgenic grapevine mutants through the regeneration of protoplasts previously edited with the CRISPR/Cas9 ribonucleoprotein.

We tested the susceptibility of single and double mutants to DM through artificial inoculation assays on detached leaves and whole plants. Our findings indicate that a simultaneous mutation in both DMR6-1 and DMR6-2 is needed to significantly enhance resistance to DM, with the double mutant (dmr6-1-dmr6-2) outperforming either single mutant in both cultivars. Elevated levels of endogenous SA were only observed in the double mutant, while single mutation in DMR6-1 or DMR6-2 proved ineffective. Collectively, our data highlight the need for a double knockout to achieve appreciable results against DM-susceptibility.

Currenlty, we are adapting the ‘single-cell technology’ to generate edited vines from various agronomically relevant cultivars. In parallel, we are assessing the performance of plants edited in different susceptibility genes.

Download the abstract

DOI:

Publication date: June 13, 2024

Issue: Open GPB 2024

Type: Poster

Authors

Lisa Giacomelli1*, Tieme Zeilmaker2, Oscar Giovannini1, Umberto Salvagnin3, Domenico Masuero1, Pietro Franceschi1, Urska Vrohvsek1, Simone Scintilla4, Jeroen Rouppe van der Voort2, Claudio Moser1

1 Research and Innovation Centre, Fondazione Edmund Mach, San Michele all’Adige, Italy
2 Enza ZadenEnza Zaden Research & Development B.V., Enkhuizen, The Netherlands  
3 C.I.VIT. Consorzio Innovazione Vite, Trento, Italy
4 Hudson River Biotechnology, Wageningen, The Netherlands

Contact the author*

Keywords

DMR6, grapevine, DNA-free, gene editing, downy mildew, susceptibility gene

Tags

IVES Conference Series | Open GPB | Open GPB 2024

Citation

Related articles…

Usefulness and limits of the crop water stress index obtained from leaf temperature for vine water status monitoring

Aims: This work aimed i) to calibrate the accuracy of estimating vineyard water status by crop water stress index (CWSI) compared to stem water potential; ii) to determine the time interval during the day that best correlates to stem water potential and iii) to understand the its usefulness.

Read More

Sustainability as system innovation: sustainability as system innovation: a returnable system for glass wine bottles

Introduction increasing sustainability is essential and a societal challenge, requiring fundamental changes in behaviour and attitudes. This applies to both producers and consumers. For the wine industry in particular, such a change is a major challenge. An eip-agri research project is evaluating the introduction of a returnable glass system in the german wine industry as a key solution for increasing sustainability. Given the need for change associated with a returnable system, the project is theoretically grounded in systems innovation, as this approach provides solutions for complex, transformative change.

Read More

Quantitative assessment of must composition using benchtop NMR spectroscopy: comparative evaluation with FTIR and validation by reference

The foundation of wine production lies in the use of high-quality grapes. To produce wines that meet the highest standards, a fast and reliable analytical assessment of grape quality is essential. Many wineries currently employ Fourier-Transform Middle-Infrared Spectroscopy (FTIR) for this purpose.

Read More

Antifungal and Laccase-Suppressing Activity of Phenolic Compounds and Their Oxidation Products on Grey Mold-Fungus Botrytis cinerea

Botrytis cinerea causes grey mold that results in severe problems for wine makers worldwide. Infected grapes lead to quality deterioration including formation of off-flavors or browning. The latter is caused by the enzyme laccase which is capable of oxidizing a wide range of phenolic compounds. Since the use of conventional pesticides is associated with many concerns of consumers and authorities regarding environmental and health related issues and may result in fungicide resistance, the development of green alternatives is gaining more attention.

Read More

Hormonal and associated metabolic changes in susceptible harvest-ripe grapes under asymptomatic and symptomatic Esca disease

Esca complex is a disease affecting grapevine trunks, characterized by the colonization of the wood by xylem-residing fungi (Phaeomoniella chlamydospora, Phaeoacremonium minimum and Fomitiporia mediterranea), and posing significant risks to vineyard longevity since no efficient treatment is available. Despite its prevalence, the mechanisms beyond symptomatic manifestations like interveinal chlorosis and leaf necrosis remain unclear. Preliminary findings indicated a more pronounced metabolic reprogramming in fruits compared to vegetative organs and a putative impact on wine quality by using fruits from symptomatic grapevines.

Read More