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…

Anthocyanin accumulation and extractability during the maturation of the grapes of three varieties

Anthocyanin accumulation and extractability were studied in Tannat, Cabernet Sauvignon and Merlot grapes produced in the south of Uruguay in two consecutive seasons. Typical cultivation situations employed in the region for each variety were considered. A follow-up was carried out, considering 60 plants per vineyard, and the harvest was determined according to the technological indices of maturity. Samples of grapes were taken in duplicate in each vineyard periodically along grape maturation. The basic composition, polyphenolic potential and anthocyanin extractability were determined. Also, half of grapes were frozen and later peeled; skin extractions over 24 hs with a solution of 12% ethanol and pH 3.2 were carried out. The anthocyanin contents of the extracts obtained were determined by HPLC-DAD. The levels of anthocyanins reached the highest values before technological maturity. Anthocyanin extractability had a decrease during grape maturation.

Grapevine root system architecture: empirical insights and first steps towards in silico studies

Root System Architecture (RSA) is crucial for plant resilience and resource uptake, yet remains underexplored in viticulture.

MONOSACCHARIDE COMPOSITION AND POLYSACCHARIDE FAMILIES OF LYOPHILISED EXTRACTS OBTAINED FROM POMACES OF DIFFERENT WHITE GRAPE VARIETIES

The recovery of bioactive compounds from grape and wine by-products is currently an important and necessary objective for sustainability. Grape pomace is one of the main by-products and is a rich source of some bioactive compounds such as polyphenols, polysaccharides, fatty acids, minerals and seed oil. Polysaccharides contained in the grape cell wall can be rhamnogalacturonans type II (RG-II), polysaccharides rich in arabinose and galactose (PRAG), mannoproteins (MP), homogalacturonans (HG) and non pectic polysaccharides (NPP).

Pesticide removal in wine with a physical treatment by molecular sieving

All along the winemaking process, conditioning and aging, wine is susceptible to be contaminated by different molecules. Contaminations can have various origins, related to wine microorganisms or as a result of an exogenous contamination. The aforementioned contamination of the wine can be caused by the migration of molecules from the materials in contact with the wine or by a contamination from exogenous molecules present in the air. Regardless of the source of the contamination, mainly two types of consequences can be observed.

The impact of leaf canopy management on eco-physiology, wood chemical properties and microbial communities in root, trunk and cordon of Riesling grapevines (Vitis vinifera L.)

In the last decades, climate change required already adaptation of vineyard management. Increase in temperature and unexpected weather events cause changes in all phenological stages requiring new management tools. For example, defoliation can be a useful tool to reduce the sugar content in the berries creating differences in the wine profiles. In a ten-year field experiment using Riesling (Vitis vinifera L, planted 1986, Geisenheim, Germany), various mechanical defoliation strategies and different intensities were trialed until 2016 before the vineyard was uprooted. Wood was sampled from the plant compartments root, trunk, cordon and shoot for analyses of physicochemical properties (e.g. lignin and element content, pH, diameter), nonstructural carbohydrates and the microbial communities. The aim of the study was to investigate the influence of reduced canopy leaf area on the sink-source allocation into different compartments and potential changes of the fungal and prokaryotic wood-inhabiting community using a metabarcoding approach. Severe summer pruning (SSP) of the canopy and mechanical defoliation (MDC) above the bunch zone decreased the leaf area by 50% compared to control (C). SSP reduced the photosynthetic capacity, which resulted in an altered source-sink allocation and carbohydrate storage. With lower leaf area, less carbohydrates are allocated. This for example resulted in a decreased trunk diameter. Further, it affected the composition of the grapevine wood microbiota. SSP and MDC management changed significantly the prokaryotic community composition in wood of the root samples, but had no effect in other compartments. In general, this study found strong compartment and less management effects of the microbial community composition and associated physicochemical properties. The highest microbial diversities were identified in the wood of the trunk, and several species were recorded the first time in grapevine.