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…

Vinhos de talha: to pitch or not to pitch

In Alentejo, south of Portugal there is a traditional way of fermenting wines in clay vessels, known as “Vinhos de Talha”. Clay vessels were traditionally impermeabilized using pine pitch, creating a barrier between the fermenting must and the clay. Due to this unusual production technology that uses of clay vessels, instead of inox or wood vessels, “Vinhos de Talha” present unique characteristics increasingly appreciated by national and international consumers when compared with wine obtained by the said traditional methods of winemaking. Although the positive consumers feedback, there is little literature about the physical-chemical characteristics of these wines (Martins et al, 2018; Cabrita et al, 2018). This work aims to characterize the volatile composition of white wines produced in clay vessels with different coatings and to contribute to the knowledge and preservation of these wines that are a unique cultural heritage. Wine samples were produced during 2019 vintage from white grapes, using the traditional technology associated to these wines.

IDENTIFICATION AND LEVELS OF PHENOLIC COMPOUNDS (TANINS, ANTHO-CYANS) IN RED VARIETAL WINES (PROKUPAC AND BLACK TAMJANIKA) FROM SERBIA

The phenolic compounds of red wines represent a source of numerous benefits for human health, which is why they are a constant subject of scientific research. Winemaking in Serbia has a growing economic significance, with particularly autochthonous varieties included [1]. This research identifies and quantifies phenolic compounds of Serbian red varietal wines of Prokupac and Black Tamjanika varieties. Quantification of the level of phenolics has been conducted, including molecular tannins [(+)-catechin, (-)-epicatechin, procyanidin dimers B1, B2, B3, B4], molecular anthocyanins, and the mean degree of polymerization of tannins by HPLC by UV detection, total antioxidant capacity via spectrophotometric methods and chromatic characteristics via CIELAB.

Impact of winemaking processes on wine polysaccharides, improving by qNMR

Today the knowledge in terms of molecular composition of the colloidal matrix is ​​not enough in order to control its stability, according to the number of winemaking and wine stabilization processes. The physico-chemical processes during the winemaking change the composition and quantity of wine macromolecules. The goal today is to determine which analytical techniques will allow to discriminate these winemaking processes in order to better understand their impact on colloidal matrix stability as well as which molecules are responsible for its instabilities. METHODS: Wines obtained after conventional winemaking were subjected to different fining and chemical stabilization treatments. Different methods were used to investigate the wine macromolecular composition and stability after chemical stabilization, including quantitative and qualitative analyzes of total soluble polysaccharides by extraction under acidified ethanol, and by size exclusion separation as well as qNMR metabolomics. RESULTS: Observation of a slight difference at the quantitative level using classical analysis between the winemaking processes was observed as well as a strong discrimination by qNMR metabolomics.

PHOTOCHEMICAL DEGRADATION OF TRYPTOPHAN IN MODEL WINE: IMPACT OF HEAVY METALS AND OXYGEN ON 2-AMINOACETOPHENONE FORMATION

The wine industry worldwide faces more and more challenges due to climate change, such as increased dryness in some areas, water stress, sunburn and early harvesting during hot summer temperatures¹. One of the resulting problems for the wine quality might be a higher prevalence of the untypical aging off-flavor (ATA)². A substance, which Rapp and Versini made responsible for ATA, is the 2-aminoace-tophenone (2-AAP)³. 2-AAP in wine causes a naphthalene, wet towels, wet wool, acacia flower or just a soapy note⁴.

CropManage online decision support tool for irrigation scheduling of vineyards

CropManage (CM) is an online decision support service (DSS) developed by the University of California, Division of Agriculture and Natural Resources for assisting farmers with efficiently managing water and nitrogen fertilizer to match the site-specific needs of their crops.