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…

The importance of rural extension and advisory services to achieve a sustainable viticulture in a climate change scenario

A healthy and dynamic agricultural sector is an important foundation of rural development, generating strong bonds to other economic sectors.

VineAI: artificial intelligence for fungal disease

Early and accurate grapevine disease detection and surveillance are crucial for optimizing vineyard management practices.

Under-vine management effects on grapevine production, soil properties and plant communities in South Australia

Under-vine (UV) management has traditionally consisted of synthetic herbicide use to limit competition between weeds and grapevines. With growing global interest towards non-synthetic chemical use, this study aimed to capture the effects of alternative UV management at two commercial Shiraz vineyards in South Australia, where the sole management variables were UV management since 2016. In adjacent treatment blocks, cultivation (CU) was compared to spontaneous vegetation (SV) in McLaren Vale (MV), and herbicide was compared to SV in Eden Valley (EV). Soil water infiltration rates were slower and grapevine stem water potential was lower in CU compared to SV in MV, with the latter having a plant community dominated by soursob (Oxalis pes-caprae) during winter; while in EV, there was little separation between the treatments. Yields were affected at both sites, with SV being higher in MV and HE being higher in EV. In MV, the only effect on grape must was a lower 13C:12C isotope ratio in CU, indicating greater grapevine water stress. In the grape must at EV, SV had higher total soluble solids, total phenolics, anthocyanins, and yeast available nitrogen; and lower pH and titratable acidity. Pruning weights were not affected by the treatments in MV, while they were higher in HE at EV. Assessments revealed that the differing soil types at the two sites were likely the main determinants of the opposing production outcomes associated with UV management. In the silty loam soil of MV, the higher yields in SV were likely due to more plant-available water, as a potential result of the continuous soil bio-pores formed by winter UV vegetation. Conversely, in the loamy sand soils of EV with a lower cation exchange capacity, the lower yields and pruning weights in SV suggest the UV vegetation competed significantly with the grapevines for available water and nutrients.

Bilan de l’impact des pratiques viticoles sur la qualité biologique des sols

Dans le cadre de TerclimPro 2025, Laure Gontier a présenté un article IVES Technical Reviews. Retrouvez la présentation ci-dessous ainsi que l’article associé : https://ives-technicalreviews.eu/article/view/8481

NEW TREATMENTS FOR TEMPRANILLO WINES BY USING CABERNET SAUVIGNON VINE-SHOOTS AND MICRO-OXYGENATION

Toasted vine-shoots as enological additive represents a promising topic due to their significant effect on wine profile. However, the use of this new enological tool with SEGs varieties different than wine and combined with others winemaking technologies, such as micro-oxygenation (MOX), has not been studied so far, despite this combination could result in wine with high chemical and organoleptic quality.