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.

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…

Exploring relationships among grapevine chemical and physiological parameters and mycobiome composition under drought stress

Improving our knowledge on biotic and abiotic factors that influence the composition of the grapevine mycobiome is of great agricultural significance, due to potential effects on plant health, productivity, and wine characteristics. Among the various environmental factors affecting the morphological, physiological, biochemical and molecular attributes of grapevine, drought stress is one of the most severe, becoming increasingly an issue worldwide.

AROMA ASSESSMENT OF COMMERCIAL SFORZATO DI VALTELLINA WINES BYINSTRUMENTAL AND SENSORY METHODOLOGIES

Sforzato di Valtellina DOCG is a special dry red wine produced from partially dehydrated Nebbiolo wine-grapes growing in the Rhaetian Alps valley of Valtellina (Lombardy, Italy). Valtellina terraced vineyards are located at an altitude of 350–800 m according to ‘heroic’ viticulture on steep slopes. The harvested grape bunches are naturally dehydrated indoors, where a slow and continuous withering occurs (about 20% w/w of weight loss), until at least 1st December when the grapes reach the desired sugar content and can be processed following a normal winemaking with maceration.

Late pruning, an alternative for rainfed vine varieties facing new climatic conditions

In Chile there is a dry farming area known as a traditional wine region, where varieties brought by the Spanish conquerors still persist. These varieties, in general, are cultivated under traditional systems, with low use of technical and economic resources, and low profitability for their grapes and wines. In this region, as in other wine grape growing areas, climatic conditions have changed significantly in recent decades. In particular, the occurrence of spring frosts, when bud break has already begun, have generated significant losses for these growers.

Improving grapevine cloning material of Welshriesling by comprehensive analysis

The important grape variety Welschriesling for Austrian and Southeast European viticulture has been selectively bred over the years for improving some quantitative traits. Collected genotypes as well as the local clones were examined from agricultural, analytical, sensory, and genetic perspectives.

Impact of dried stems in winemaking on Veneto Passito wines

The use of stems during fermentation is generally avoided due to the herbaceous off-odors they can impart to the wine. [1].