DNA-free editing to improve stress resilience of wine grape genotypes recalcitrant-to-regeneration

The need to mitigate the yield impact of Spring frosts in vineyards remains a significant challenge around the world.

Conventional molecular analyses provide bulk genomic/transcriptomic data that are unable to reveal the cellular heterogeneity and to precisely define how gene networks orchestrate organ development. We will profile gene expression and identify open chromatin regions at the individual cells level, allowing to define cell-type specific regulatory elements, developmental trajectories and transcriptional networks orchestrating organ development and function. We will perform scRNA-seq and snATAC-seq on leaf/berry protoplasts and nuclei and combine them with the leaf/berry bulk tissues obtained results, where the analysis of transcripts, chromatin accessibility, histone modification and transcription factor binding sites showed that a large fraction of phenotypic variation appears to be determined by regulatory rather than coding variation and that many variants have an organ-specific effect.

In grapevine, phenology from bud break to berry maturation, depends on temperature and water availability. Increases in average temperatures accelerates initiation of bud break, exposing newly formed shoots to detrimental environmental stresses. It is therefore essential to identify genotypes that could delay phenology in order to adapt to the environment. The use of different rootstocks has been applied to change scion’s characteristics, to adapt and resist to abiotic and biotic stresses[1].

Proseccco is a famous italian Protected Designation of Origin (PDO) produced in two regions: Veneto e Friuli Venezia Giulia, however, the production is mainly concentrated in the province of Treviso. These territories are characterized by plains with some hilly areas and temperate climate. Its Production regulation provides a minimum utilization of 85% of Glera grapes, a local white grape variety, and up to a maximum of 15% of other local and international varieties. Prosecco second fermentation takes place, according to the Charmat method, in autoclaves.

Several Eurasian wild grapevine populations were found along Extremadura region (southwestern Spain). For conservation and study, one individual from four different populations (named L1, L2, L5 and L6) was vegetatively propagated and planted at Instituto de Investigaciones Agrarias Finca La Orden (CICYTEX), Badajoz. The aim of the present work was to characterize those conserved individuals from four different populations based on both an ampelographic description and a molecular analysis. Three vines per individual were studied.