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…

Swiss program for the creation of fungal disease resistant grape varieties in Switzerland

Grapevine breeding is part of the research program of Agroscope in Switzerland since 1965. From 1965 to 1995, the aim of the Vitis vinifera crosses was to obtain a high resistance to grey rot (Botrytis cinerea), one of the most virulent fungal pathogens in the Swiss vineyard. In 2021, the grape varieties released from this first breeding program covered 936 ha of the 15’000 ha of the Swiss vineyard.
In 1996, a second breeding program aimed at obtaining, by classical interspecific hybridization, grape varieties resistant to downy mildew (Plasmopara viticola) and powdery mildew (Erisyphe necator) and less sensitive to grey rot (Botrytis cinerea). In order to accelerate and make the selection process more reliable, an early biochemical test was developed based on the natural defense mechanisms of the vine against downy mildew (stilbene phytoalexins). The synthesis of stilbenes (i.e., resveratrol and its oxidized dimers - and -viniférine) and pterostilbenes (methylated derivative) is among the most efficient induced defense mechanisms of grapevine against fungal pathogens on both the leaves and the clusters.

Read More

The influence of native flora on Argentine white terroir cv. Torrontes Riojano

The main objective of this paper is to establish considerable differences between wines from three wine areas or terroir, made with cv Torrontes Riojano.

Read More

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.

Read More

Closing the carbon loop: evaluating the potential of grapevine-derived biochar as a soil conditioner in warm climate vineyards

Significant increases in anthropogenic carbon dioxide (CO2) emissions due to combustion of fossil fuels and intensive land management practices that release CO2 into the atmosphere have resulted in higher air temperatures due to the greenhouse effect.

Read More

Insights from selected ion flow tube mass spectrometry (SIFT-MS) and chemometrics applied to the quick discrimination of grapevine varieties

Selected Ion Flow Tube Mass Spectrometry (SIFT-MS) is an innovative analytical method based on soft chemical ionization to analyze thecomposition in volatile compounds of a gas phase

Read More