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…

A blueprint for managing vine physiological balance at different spatial and temporal scales in Champagne

In Champagne, the vine adaptation to different climatic and technical changes during these last 20 years can be seen through physiological balance disruptions. These disruptions emphasize the general grapevine decline. Since the 2000s, among other nitrogen stress indicators, the must nitrogen has been decreasing. The combination of restricted mineral fertilizers and herbicide use, the growing variability of spring rainfall, the increasing thermal stress as well as the soil type heterogeneity are only a few underlying factors that trigger loss of physiological balance in the vineyards. It is important to weigh and quantify the impact of these factors on the vine. In order to do so, the Comité Champagne uses two key-tools: networking and modelization. The use of quantitative and harmonized ecophysiological indicators is necessary, especially in large spatial scales such as the Champagne appellation. A working group with different professional structures of Champagne has been launched by the Comité Champagne in order to create a common ecophysiology protocol and thus monitor the vine physiology, yearly, around 100 plots, with various cultural practices and types of soil. The use of crop modelling to follow the vine physiological balance within different pedoclimatic conditions enables to understand the present balance but also predict the possible disruptions to come in future climatic scenarios. The physiological references created each year through the working group, benefit the calibration of the STICS model used in Champagne. In return, the model delivers ecophysiology indicators, on a daily scale and can be used on very different types of soils. This study will present the bottom-up method used to give accurate information on the impacts of soil, climate and cultural practices on vine physiology.

Veraison as determinant for wine quality and its potential for climate adapted breeding

The evaluation of new grapevine genotypes regarding their potential to produce high quality wines is the time limiting factor in the process of grapevine breeding. Hence, the development of quality-related markers useable in marker-assisted selection (MAS) as well as in prediction models for this bottleneck trait will tremendously enhance breeding efficiency. In extensive studies a training set of a segregating white wine F1 population (150 F1 genotypes = POP150; `Calardis Musqué´ x `Villard Blanc´) was deeply phenotyped and genotyped for model development and QTL analysis.

Montpellier vine & wine sciences (M-WineS)

The Occitanie Region is the first vine-growing area in France: 270 000 hectares of vineyard and an annual production of 15 million hectoliters. Its annual income reaches 1 900 million euros, of which 900 million euros in export.The vine and wine sector is facing many issues: inputs reduction, adaptation to climate change, maintaining the production competitiveness, digital tools integration in production and transformation processes, and the production of quality wines meeting the consumer demand.

Soil clay mineralogy and potassium buffer capacity as potential wine quality determining factors in Western Cape vineyards

The potassium (K) supply characteristics and clay mineralogies of a population of Western Cape soils were investigated to determine their potential effects on vine K uptake and wine quality. The total K contents of granite-, shale- and sandstone-derived soils varied, averaging 33.7, 26.1 and 4.5 cmol(+)/kg, respectively. Corresponding M NH4Cl exchangeable soil K levels were: 0.172, 0.042 and 0.035 cmol/kg.

Genetic traceability of the varietal origin of wines: a robust application for must and wines during alcoholic fermentation

Industry and regulatory agencies have developed regulations to ensure authenticity and compliance with wine composition limits. However, this can be truncated by the absence of simple and robust analytical methodologies, uninfluenced by the environment, different oenological techniques and cultural practices. Genetic fingerprinting is the most powerful tool for unequivocal varietal identification; it is not affected by the environment or agronomic practices; however, its usefulness in musts and wines has been controversial and there is currently no routine certification of varietal origin based on DNA analysis.