terclim by ICS banner
IVES 9 IVES Conference Series 9 DNA-Free genome editing confers disease resistance in grapevine

DNA-Free genome editing confers disease resistance in grapevine

Abstract

Grapevine (Vitis Vinifera L.), one of the most important cultivated fruit crops, is facing significant challenges due to climate change. Specifically, increasing temperatures negatively impact the physiological traits and disrupt plant phenology. Additionally, increased virulence in pathogen attacks and pests leads to significant yield loss, requiring widespread application of plant protection products. Traditional agronomic practices offer only partial mitigation, requiring the development of precise and effective intervention strategies. The economic worth of viticulture has prompted continuous efforts in grapevine genetic improvement programs, traditionally involving conventional breeding and clonal selection that, however, are complex and time-consuming approaches. Instead, the advent of New Breeding Techniques, especially genome editing via the CRISPR/Cas9 system, presents a promising avenue for the development of tools suitable to mitigate the current viticulture challenges, including fungal diseases. We report the application of a DNA-free genome editing approach to induce targeted mutations in the VviMLO17, a gene associated with powdery mildew susceptibility in grapevine. CRISPR/Cas9 ribonucleoparticles were introduced into protoplasts isolated from embryogenic calli. Through protoplast regeneration, a homozygous edited grapevine plant mutated in the VviMLO17 gene was obtained. This mutation confers resistance to Erysiphe necator, as evidenced by phenotypic analyses that demonstrated a reduced susceptibility to pathogen attack. The success of DNA-free CRISPR/Cas9 application for the improvement of target traits establishes a foundation for promoting viticulture sustainability yet preserving the identity of the grapevine cultivars. This advancement aligns with market and legislative demands, paving the way for a resilient and environmentally conscious winegrowing system.

DOI:

Publication date: June 13, 2024

Issue: Open GPB 2024

Type: Article

Authors

Marianna Fasoli1, Edoardo Bertini2, Erica D’Incà2, Luca Cattaneo1, Stefania Zattoni1, Sara Lissandrini1, Clarissa Ciffolillo1, Annalisa Polverari1, Giovanni Battista Tornielli1,3, Sara Zenoni1*

1 Department of Biotechnology, University of Verona, Verona, Italy
2 EdiVite S.r.l., San Pietro Viminario, Padua, Italy
3 Current address: Department of Agronomy, Food, Natural Resources, Animals and the Environment., University of Padua, Padua, Italy

Contact the author*

Keywords

genome editing DNA-free, CRISPR/Cas9 system, protoplast regeneration, powdery mildew resistance, sustainable viticulture

Tags

IVES Conference Series | Open GPB | Open GPB 2024

Citation

Related articles…

Methodological advances in relating deep root activity to whole vine physiology

Full understanding of grapevine responses to variable soil resources requires
assessing the grapevine root system. Grapevine root systems are expansive and examining deep roots (i.e., >40 cm)
is particularly important in conditions where grapevines increase reliance on deep soil resources, such as drought
or plant competition. Traditional methods of assessing roots rely on morphological traits associated specific
functions (e.g., root color, diameter, length), while recent methodological advances allow for estimating root
function more directly (e.g., omics). Yet, the potential of applying refined methods remains underexplored for roots
at deep depths.

THE EFFECT OF PRE-FERMENTATIVE GLYPHOSATE ADDITION ON THE METABOLITE PROFILE OF WINE

The synthetic herbicide glyphosate has been used extensively in viticulture over many decades to combat weeds. Despite this, the possible influence of residual glyphosate on both the alcoholic fermentation of grape juice and the subsequent metabolite profile of wines has not been investigated. In this study, Pinot noir juice supplemented with different concentrations of glyphosate (0 µg L-1, 10 µg L-1 and 1000 µg L-1) was fermented with commercial Saccharomyces cerevisiae yeast strains. Using a combination of analytical methods, 80 metabolites were quantified in the resulting wines.

Traditional agroforestry vineyards, sources of inspiration for the agroecological transition of viticulture

A unique “terroir” can be found in southern Bolivia, which combines the specific features of climate, topography and altitude of high valleys, with the management of grapevines staked on trees. It is one of the rare remnants of agroforestry viticulture. A survey was carried out among 29 grapegrowers in three valleys, to characterize the structure and management of these vineyards, and identify the services they expect from trees. Farms were small (2.2 ha on average) and 85% of vineyards were less than 1 ha. Viticulture was associated with vegetable, fruit and fodder production, sometimes in the same fields. Molle trees were found in all plots, together with one or two other native tree species. Traditional grapevine varieties such as Negra Criolla, Moscatel de Alejandría and Vicchoqueña were grown with a large range of densities from 1550 to 9500 vines ha-1. From 18 to 30% of them were staked on trees, with 1.2 to 4.9 vines per tree. The management of these vineyards (irrigation, fertilization and grapevine protection) was described, the most particular technical operation being the coordinated pruning of trees and grapevines. Three types of management could be identified in the three valleys. Grapegrowers had a clear idea of the ecosystem services they expected from trees in their vineyards. The main one was protection against climate hazards (hail, frost, flood). Then they expected benefits in terms of pest and disease control, improvement of soil fertility and resulting yield. At last, some producers claimed that tree-staking was quicker and cheaper than conventional trellising. It can be hypothesized then that agroforestry is a promising technique for the agroecological transition of viticulture. Its contribution to the “terroir” of the high valleys of southern Bolivia and its link with the specificities of the wines and spirits produced there remain to be explored.

Novel insights into Passito wines aroma typicality. Rationalizing the markers of varietal and geographical origin of Amarone DOCG

Valpolicella is a famous Italian wine-producing region (Paronetto & Dellaglio, 2011), whose main characteristic is the extensive use of the post-harvest withering technique, which takes place in naturally ventilated rooms called ‘fruttai’ (Bellincontro et al., 2016).

On quality assurance of winemaking components

This report examines product quality assurance issues arising when technological aids and food additives are utilized in winemaking.