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…

AGING PATTERNS OF VARIETAL VOLATILE PROFILES OF WHITE WINES: A CASE STUDY ON 18 ITALIAN VARIETAL WHITE WINES

During wine aging many compositional changes take place. In particular, aroma undergoes dramatic modifications through a wide range of reactions that to date are only partly understood. Italy owns one of the largest ampelographic heritages worldwide, with over three-hundred different varieties. Among these, many white grapes are employed for the production of dry still white wines. Some of these wines are consumed young while others are more prone to aging. For many of these wines, the aging patterns related to volatile composition are still unknown.

Read More

Usefulness and limits of the crop water stress index obtained from leaf temperature for vine water status monitoring

Aims: This work aimed i) to calibrate the accuracy of estimating vineyard water status by crop water stress index (CWSI) compared to stem water potential; ii) to determine the time interval during the day that best correlates to stem water potential and iii) to understand the its usefulness.

Read More

Budburst delay and berry ripening after vegetal oil application in Austria

Occurrence of freezing temperatures in early spring when grapevine shoots are developing is termed late frost in viticulture. Young green tissues are very sensible to temperatures below zero and damages often lead to important yield and quality losses such as the case in Europe in 2017. An indirect method to avoid late frost damage in vineyards consist in delaying the budburst. Previous research reported similar effects by applying vegetal oil on dormant buds. Here, we tested the application of rapeseed vegetal oil during late winter to delay the budburst on two V.vinifera cultivars of interest in Austria, Grüner Veltliner (GV) and Zweigelt (ZW).

Read More

Rootstocks and climate change: adding up means learning faster

In this video recording of the IVES science meeting 2025, Gonzaga Santesteban (Public University of Navarra, Pamplona, Spain) speaks about rootstocks, climate change and meta-analysis. This presentation is based on an original article accessible for free on OENO One.

Read More

Assessing bunch architecture for grapevine yield forecasting by image analysis

It is fundamental for wineries to know the potential yield of their vineyards as soon as possible for future planning of winery logistics. As such, non-invasive image-based methods are being investigated for early yield prediction. Many of these techniques have limitations that make it difficult to implement for practical use commercially. The aim of this study was to assess whether yield can be estimated using images taken in-field with a smartphone at different phenological stages. The accuracy of the method for predicting bunch weight at different phenological stages was assessed for seven different varieties.

Read More