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…

Validation of the viticulture zoning methodology applied to determine the homogenous soil units present on D.O. Ribera de Duero region

The methodology to viticulture zoning developed and proposed by Gómez-Miguel and Sotés (1992) has been studied in order to validate it. This was the main aim of this work

THE EFFECT OF BENTONITE FINING ON THE VOLATILE AND NON-VOLATILE PROFILE OF ITALIAN WHITE WINES

Marselan wines have an unusual high proportion of seed derived tannins from grapes having high proportions of skins, which are rich in tannins. But the causes behind this characteristic have not yet been identified. In vintage 2023 wines were made at experimental scale (9 kg by experimental unit) from Arinarnoa, Marselan and Tannat Vitis vinifera grape cultivars by traditional maceration, and by techniques aimed to increase the wine content in skin derived tannin: addition of extraction enzymes, addition at vatting of grape-skin enological tannins, or by extended maceration, known to increase the seed derived tannin contents of wines. Macerations were of 7 days, except in the extended macerations that were of 15 days.

Energy optimization of the Charmat-Martinotti refermentation process

The european union has estimated that energy consumption for wine production is about 1,750 million kwh per year, of which 500 million kwh is attributable to italy. In recent years, Italy has emerged as the world’s leading wine producer with about 50 million hectoliters per year. About 20 percent (9.8 million hectoliters) of Italian wine is marketed after refermentation according to the Charmat-Martinotti method.

Partial dealcoholisation of red wine by reverse osmosis-evaporative perstraction: impact on wine composition

Around the world, the alcohol content of wine has been steadily increasing; partly as a consequence of climate change, but also due to improvements in viticultural management practices and winemaking techniques [1,2]. Concurrently, market demand for wines with lower alcohol levels has increased as consumers seek to reduce alcohol intake for social and/or health reasons [3]. As such, there is increasing demand for both innovative methods that allow winemakers to produce ‘reduced alcohol wines’ (RAW) and a better understanding of the impact of such methods on the composition of RAW. This study therefore aimed to investigate compositional changes in two red wines resulting from partial alcohol removal following treatment by one such method, involving a combination of reverse osmosis and evaporative perstraction (RO-EP).

Vino e paesaggio: materiali per il governo del territorio vitivinicolo. Il piano regolatore delle città del vino

S’intende per Piano Regolatore delle Città del Vino la metodologia per redigere la parte strutturale dello strumento comunale di governo del territorio. Parliamo, infatti, del principale strumento comunale di governo del territorio, così come è venuto maturando nella riflessione delle Città del Vino, strumento che si misura con la sfida di governare tutto il territorio in modo coerente e sostenibile, a partire dal riconoscimento del valore del “sistema vigneto” e della sua intrinseca fragilità.