terclim by ICS banner
IVES 9 IVES Conference Series 9 Novel approaches and promising perspectives for enhancing grapevine editing and regeneration

Novel approaches and promising perspectives for enhancing grapevine editing and regeneration

Abstract

Grapevine (Vitis vinifera L.) is a challenging plant species to transform and regenerate due to its complex genome and biological characteristics. This limits the development of cisgenic and gene-edited varieties. One hurdle is selecting the best starting tissue for the transformation process, much like isolating suitable tissue for protoplasts. One promising method involves delivering CRISPR/Cas components to protoplasts isolated from embryogenic calli, which are then induced to regenerate. However, this process is inefficient, time-consuming, and only applicable to a few genotypes. To enhance grapevine regeneration efficiency, the expression of developmental and plant growth regulators shows promise in escaping the recalcitrance encountered in traditional tissue culture methods. A strategy based on the Bean yellow dwarf virus (BeYDV) allows for the temporary expression of regulators while minimizing the risk of obtaining transgenic plants. Additionally, the high copy number replication of BeYDV allows for high CRISPR/CAS levels, thereby improving editing. The goal of this study is to evaluate the effects of individual or combined developmental regulators’ expression on grapevine embryogenic tissues. We conducted preliminary assays using BeYDV-derived vectors for luciferase reporter gene expression to optimize delivery efficiencies. Assays were performed on ‘Chardonnay’ calli and protoplasts using both agrobacterium-mediated transformation and protoplast transfection approaches. The present study seeks to enhance the transformation protocols and regeneration processes, with the ultimate aim of realizing the full potential of editing technologies in grapevine.

DOI:

Publication date: June 13, 2024

Issue: Open GPB 2024

Type: Poster

Authors

Flavia Angela Maria Maggiolini1*, Margherita D’Amico1, Lucia Rosaria Forleo1, Annalisa Prencipe2, Bruna Suriano1, Mario Ventura2, Maria Francesca Cardone1, Riccardo Velasco1, Carlo Bergamini1

1 Council for Agricultural Research and Economics – Research Center Viticulture and Enology (CREA-VE), Via Casamassima 148-70010 Turi (Ba), Italy
2 Department of Biosciences, Biotechnology and Environment, University of Bari “Aldo Moro”, 70125 Bari, Italy.

Contact the author*

Keywords

Vitis vinifera, genome editing, protoplasts, developmental regulators, BeYDV

Tags

IVES Conference Series | Open GPB | Open GPB 2024

Citation

Related articles…

What happens with the glutathione during winemaking and the storage of the wine?

We tried to give a part of the answer to this question by monitoring glutathione during winemaking and storage. The novelty of our approach is to quantify simultaneously the three known forms of glutathione: free glutathione (GSH), oxidized form (GSSG) and glutathione-S-sulfonate (GSSO3H).

A DNA-free editing approach to help viticulture sustainability: dual editing of DMR6-1 and DMR6-2 enhances resistance to downy mildew 

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.

A sensometabolomic approach to understand wine mouthfeel percepts

Targeted analytical methods can overlook compounds that are a priori unknown to play a role in the mouthfeel sensations. This limitation can be overcome with the information provided by untargeted metabolomic analysis using UPLC‐QTOF-MS. To this end, an untargeted metabolomic approach applied to 42 red wines has allowed development of a model with predictive capacity by cross-validation for the “dry”, “oily” and “unctuous” sensations perceived by a sensory panel. The optimal PLS model for “dry” retained compounds with positive regression coefficients (≥ 0.17) including a trimer procyanidin, a peptide, and four anthocyanins.

Leaf removal to regulate fruit ripening in Cabernet-Sauvignon

Aim: Under the effects of climate change it is becoming increasingly common to observe excessively fast sugar accumulation while anthocyanin and flavour development are lagging behind. Understanding the impact of different leaf removal techniques on ripening will provide vineyard managers with a canopy management strategy suitable for

Impact of long term agroecological and conventional practices on subsurface soil microbiota in Macabeu and Xarel·lo vineyards

There is a growing trend on the transition from conventional to agroecological management of vineyards. However, the impact of practices, such as reduced-tillage, organic fertilization and cover crops, is not well-understood regarding the soil microbial diversity, and its relationship with the soil physicochemical properties in the subsurface depth near the rooting zone. Soil bacterial diversity is an important contributor towards plant health, productivity and response to environmental stresses. A field experiment was conducted by sampling subsurface soil bacterial community (NGS and qPCR) near to the root zone of Macabeu and Xarel·lo vineyards, located at the Penedes. 3 organic (ECO) and 3 conventional (CON) vineyards, with more than 10 years of respective management were sampled (n=5 each plot). ECO practices did not affect bacterial and fungal abundance but increased significantly the ammonium oxidizing bacteria and alpha-diversity (Inv.Simpson). Interestingly beta-diversity was significantly affected by the management strategy. ANOSIM-tests revealed a significative effect of the management (ecological vs conventional) and plot, on the soil microbial structure (ASV abundance). Main phyla depicted were Proteobacteria, Actinobacteria and Acidobacteria, whose relative abundances were not affected by the management. EdgeR assay revealed a significant increase of Cyanobacteria and decrease of Gemmatimonadetes and Firmicutes phyla in ECO. Interestingly, the grapevine variety was not correlated with the soil microbial community structure. Mantel-test revealed an important correlation (Spearman) of some physicochemical parameters with the soil microbiota structure, in order of importance: texture, EC, pH Ca/Mg, Mg/P, K+, Mg2+, Ca2+, SO42-, and OM. N-NH4 and NTK, which were higher in the ECO managed soils, did not correlated significantly with the soil microbiome population. The results revealed the importance of combining a deep physicochemical characterization of each replicate with the microbial diversity assessment to gain better insights on the relationship between soil microbiome and vineyard management.