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…

Effects of post-fermentative cold maceration on chemical and sensory characteristics of Syrah, Cabernet Franc and Montepulciano wines

Astringency sensation decreases slowly during the aging of red wine. Complex reactions of condensation and precipitation of wine polyphenols are involved in this phenomenon. Wine composition and conditions of aging, such as temperature and oxygen availability, strongly influence evolution of the phenol matrix. Recently, a Post-Fermentative cold Maceration (PFM) technique was tested with the aim of accelerating reactions leading to the reduction of astringency and exploiting chemical compounds not extracted from the solid parts of grapes during the previous traditional maceration phase. To this purpose, an innovative maceration system was engineered and used to perform PFM trials on marc derived from vinification of different varieties of red grapes.

Read More

Use of mathematical modelling and multivariate statistical process control during alcoholic fermentation of red wine

Cyberphysical systems can be seen in the wine industry in the form of precision oenology. Currently, limitations exist with established infrared chemometric models and first principle mathematical models in that they require a high degree of sample preparation, making it inappropriate for use in-line,

Read More

Unleashing the power of artificial intelligence for viticulture and oenology on earth and space

Implementing artificial intelligence (AI) in viticulture and enology is a rapidly growing field of research with an essential number of potential practical applications.

Read More

HOW OXYGEN CONSUMPTION INFLUENCES RED WINES VOLTAMMETRIC PROFILE

Phenolic compounds play a central role in sensory characteristics of wine, such as colour, mouthfeel, flavour and determine its shelf life. Furthermore, the major non-enzymatic wine oxidation process is due to the catalytic oxidation of phenols in quinones. Due their importance, during the years have been developed different analytical methods to monitor the concentration of phenols in wine, such as Folin-Ciocalteu method, spectrophotometric techniques and HPLC. These methods can also be used to follow some oxidation-related chemical transformations.

Read More

WHAT’S FUTURE FOR SANTORINI’S VITICULTURE IN THE CONTEXT OF CLIMATE CHANGE

The own-rooted vineyard of Santorini is a unique case of vineyard worldwide that is been cultivated for thousands of years. On the island’s volcanic soil, the vines are still cultivated with traditional techniques, which are adapted to the specific and extreme weather conditions that prevail on it. While climate change is a reality in the Mediterranean region, will Santorini vineyard endure its impact? The study of the traditional training systems, techniques and vine density, as well as the application of sustainable solutions (cover crops and use of kaolin etc.) revealed sustainable methods for the adaptation of the local viticulture to new climatic phenomena that tend to be more and more frequent in the region due to climate change.

Read More