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…

Influence of a spontaneous cover crop on the vineyard and soil erosion under Mediterranean climate

Sixty five % of the agricultural area of the Basque Country located in the DO Ca Rioja corresponds to vineyards. More than 40% of it has an average slope greater than 10%, which makes it sensitive to erosive processes. Furthermore, it is foreseeable that extreme weather events (storms, hail, extreme heat and cold, etc.) will be favored due to climate change. Cover cropping can mitigate this risk, and therefore the objective of this work is to evaluate the impact that a vegetable cover has on the agronomic behavior of the vineyard, the quality of the grape and soil erosion. For this, a trial has been carried out with a Graciano variety vineyard with a slope between 10% -20% during the years 2020 and 2021. Conventional tillage management in the area has been compared (4-6 passes per year of tillage machinery) versus spontaneous vegetation cover management in the vineyard. This implies not tilling and allowing the grass of the land to colonize the range between the lines of vines, controlling their height through 1-3 mowing passes per year, always trying to affect the surface of the land as little as possible. The vegetative growth, yield and quality of the grape and wine was measured. Furthermore, erosion has been measured using Gerlasch boxes. The yield was lower in the second year of the trial in the cover crop treatment, but erosion was significantly reduced.

Viticultural zoning applications at the detailed scale of a cooperative winery: terroirs in St­hilaire-d’Ozilhan (AOC Côtes-du-Rhône)

La maîtrise de la typicité du vin s’élabore au niveau local ou communal d’une exploitation viticole et/ou d’une cave, unité de vinification. La cave coopérative de Saint-Hilaire­-d’Ozilhan (AOC Côtes-du-Rhône), dont le territoire communal s’étend sur une superficie de 1 670 ha, couvre près de 310 ha cultivés en vigne. Elle réalise des vinifications «au terroir», en utilisant des regroupements d’unités de sol en 9 unités de terroir potentiellement viticoles, définies en s’appuyant sur la parenté des substrats. Diverses sélections d’une même unité peuvent aboutir aussi à des vins différents, ce qui suggère une hétérogénéité spatiale de certaines unités définies.

Matériel végétal et valorisation des terroirs viticoles

[lwp_divi_breadcrumbs home_text="IVES" use_before_icon="on" before_icon="||divi||400" module_id="publication-ariane" _builder_version="4.19.4" _module_preset="default" module_text_align="center" module_font_size="16px" text_orientation="center"...

Protein stabilization of white wines by stabilizing filtration: pilot studies

Protein stabilization is an important part of the winemaking process of white wines, and in this work we present the results of protein stabilization of different monovarietal wines (Xarel.lo, Chardonnay, and Muscat) by a continuous stabilizing filtration process using a column packed with zirconium oxide operating in a continuous regime in a closed loop at pilot scale.

Metabolomics comparison of non-Saccharomyces yeasts in Sauvignon blanc and Shiraz

Saccharomyces cerevisiae (SC) is the main driver of alcoholic fermentation however, in wine, non-Saccharomyces species can have a powerful effect on aroma and flavor formation. This study aimed to compare untargeted volatile compound profiles from SPME-GC×GC-TOF-MS of Sauvignon blanc and Shiraz wine inoculated with six different non-Saccharomyces yeasts followed by SC. Torulaspora delbrueckii (TD), Lachancea thermotolerans (LT), Pichia kluyveri (PK) and Metschnikowia pulcherrima (MP) were commercial starter strains, while Candida zemplinina (CZ) and Kazachstania aerobia (KA), were isolated from wine grape environments. Each fermentation produced a distinct chemical profile that was unique for both grape musts. The SC-monoculture and CZ-SC sequential fermentations were the most distinctly different in the Sauvignon blanc while the LT-SC sequential fermentations were the most different from the control in the Shiraz fermentations.