DNA-free genome editing confers disease resistance in grapevine varieties

Abstract

The grapevine is facing significant challenges due to climate change, as rising temperatures impact its 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. The New Genomic Techniques that exploit genome editing via CRISPR/Cas9 present an avenue for developing tools to mitigate the challenges of viticulture, including fungal diseases.

We report the application of a DNA-free genome editing approach to induce targeted mutations in grapevine susceptibility genes. CRISPR/Cas9 ribonucleoparticles were introduced into protoplasts isolated from embryogenic calli. Through protoplast regeneration, edited grapevine plants of different commercial varieties were obtained. These mutations reduced susceptibility to pathogen attack and confer resistance to pests.

The success of this 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.