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…

Simulating climate change impact on viticultural systems in historical and emergent vineyards

Global climate change affects regional climates and hold implications for wine growing regions worldwide. Although winegrowers are constantly adapting to internal and external factors, it seems relevant to develop tools, which will allow them to better define actual and future agro-climatic potentials. Within this context, we develop a modelling approach, able to simulate the impact of environmental conditions and constraints on vine behaviour and to highlight potential adaptation strategies according to different climate change scenarios. Our modeling approach, named SEVE (Simulating Environmental impacts on Viticultural Ecosystems), provides a generic modeling framework for simulating grapevine growth and berry ripening under different conditions and constraints (slope, aspect, soil type, climate variability…) as well as production strategies and adaptation rules according to climate change scenarios. Each activity is represented by an autonomous agent able to react and adapt its reaction to the variability of environmental constraints. Using this model, we have recently analyzed the evolution of vineyards’ exposure to climatic risks (frost, pathogen risk, heat wave) and the adaptation strategies potentially implemented by the winegrowers. This approach, implemented for two climate change scenarios, has been initiated in France on traditional (Loire Valley) and emerging (Brittany) vineyards. The objective is to identify the time horizons of adaptations and new opportunities in these two regions. Carried out in collaboration with wine growers, this approach aims to better understand the variability of climate change impacts at local scale in the medium and long term.

Diversificazione e valorizzazione di produzioni tipiche sul territorio: I cesanesi

The zone in which the Cesanese vines are cultivated has a secular tradition of red wine­making. This zone is placed between the Simbruini mountains slopes and the surrounding hills and has pedologicai variability but a very homogeneous microclimate.

The impact of nutrition label formats on wine consumer preferences

Recent regulations concerning alcoholic beverages have prompted producers to revise their product labels to incorporate nutritional information. In this context, qr codes containing such information, known as e-labels, are now being employed on wine labels for the first time.

Influence of soil management and vine water regime on leaf gas exchange, berry composition and quality of Chasselas wines in Switzerland

A soil management and vine irrigation trial was carried out for 4 consecutive years from 2020 to 2023 at agroscope’s experimental vineyard in leytron (Valais, Switzerland) with the Chasselas grape variety (clone 14-33/4, grafted on 5bb). Two types of soil maintenance (bare soil with chemical weeding and sown grass) coupled with two water regimes (with and without drip irrigation from flowering to veraison) were compared in a randomized design with four replicates of 10 vines each.

Study of the aromatic oxidation markers of Tempranillo long aged wines

The aromatic quality of wines after a long aging period in bottle is one of key points for oenologists. The objective of this work is to determine the main representative aromatic compounds found in long aged wines from D.O.Ca. Rioja. This study was made by 32 wines from 1971 to 2010 vintages. Sotolon, acetaldehyde, phenylacetaldehyde, 1,1,6-trimethyl-1,2-dihydronaptalene (TDN), β-damascenone, Y-decalactone and Y-dodecalactone were determined as the most important oxidation markers by GC-MS analysis. Moreover, sensory analysis using triangular tests were performed from wines with and without the addition of the mentioned compounds. Four different concentrations of each odorant were added, as individual compounds and as mixtures. The additions were ranged from values close to the reference odour thresholds up to high level concentrations. The most identified aroma was sotolon, which is commonly associated to curry and coffee liqueur aromatic notes. Other oxidative compounds were easily detected by panellists, such as Y-decalactone (peach compote), Y-dodecalactone (ripe fruit). The mixtures of the odorants were most easily detected than the individual compounds. It should be noted that acetaldehyde and phenylacetaldehyde were rarely perceived and distinguished.