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…

Phenology, thermal requirements and maturation of the SR 0.501-17 wine grape hybrid cultivated in contrasting climate

The use of hybrids in viticulture is one of the alternatives for sustainable production in hot and rainy regions during grapevine maturation. This sustainable production concerns the reduction of pesticide use, adaptation to climate and control of vine decline. The SR 0.501-17 wine grape hybrid, developed in the grapevine program of the Agronomic Institute of Campinas (IAC), is characterized by producing white grapes with small spherical berries with seeds. The agronomic characterization of this hybrid, especially in different climatic conditions, as well as the evaluation of its performance in winemaking are necessary. The objective of this work was to characterize the duration and thermal requirements of the different phenological stages and the influence of rainfall on the physicochemical characteristics of the must in two contrasting climate regions of the State of São Paulo.

Resilience analysis in viticulture: an approach based on expert knowledge elicitation

The study aims to address the issue of resilience to climate change in viticulture through the adoption of the expert knowledge elicitation (EKE) approach.

Correlation between grape and wine quality, landscape diversity, on-field biodiversity, in doc gioia del colle, italy

Analysis of aerial photos by using GIS tools and on-field surveys of flora are used to characterize territories from an agro-ecological point of view and to assess the level of diversity of given agro-ecosystems. More and more correlations between landscape characteristics, sustainability and quality of agriculture production were speculated. In last three years a study was carried out in the area of DOC “Gioia del Colle” in Apulia, South Italy, in order to characterize and investigate different vineyards and sites and find out possible interactions and correlations between the landscape diversity, the biodiversity of fields and the quality of grapes and wines.

Use of a new, miniaturized, low-cost spectral sensor to estimate and map the vineyard water status from a mobile 

Optimizing the use of water and improving irrigation strategies has become increasingly important in most winegrowing countries due to the consequences of climate change, which are leading to more frequent droughts, heat waves, or alteration of precipitation patterns. Optimized irrigation scheduling can only be based on a reliable knowledge of the vineyard water status.

In this context, this work aims at the development of a novel methodology, using a contactless, miniaturized, low-cost NIR spectral tool to monitor (on-the-go) the vineyard water status variability. On-the-go spectral measurements were acquired in the vineyard using a NIR micro spectrometer, operating in the 900–1900 nm spectral range, from a ground vehicle moving at 3 km/h. Spectral measurements were collected on the northeast side of the canopy across four different dates (July 8th, 14th, 21st and August 12th) during 2021 season in a commercial vineyard (3 ha). Grapevines of Vitis vinifera L. Graciano planted on a VSP trellis were monitored at solar noon using stem water potential (Ψs) as reference indicators of plant water status. In total, 108 measurements of Ψs were taken (27 vines per date).

Calibration and prediction models were performed using Partial Least Squares (PLS) regression. The best prediction models for grapevine water status yielded a determination coefficient of cross-validation (r2cv) of 0.67 and a root mean square error of cross-validation (RMSEcv) of 0.131 MPa. This predictive model was employed to map the spatial variability of the vineyard water status and provided useful, practical information towards the implementation of appropriate irrigation strategies. The outcomes presented in this work show the great potential of this low-cost methodology to assess the vineyard stem water potential and its spatial variability in a commercial vineyard.

Mobile device to induce heat-stress on grapevine berries

Studying heat stress response of grapevine berries in the field often relies on weather conditions during the growing season. We constructed a mobile heating device, able to induce controlled heat stress on grapes in vineyards. The heater consisted of six 150 W infrared lamps mounted in a profile frame. Heating power of the lamps could be controlled individually by a control unit consisting of a single board computer and six temperature sensors to reach a pre-set temperature. The heat energy applied to individual berries within a cluster decreases by the squared distance to the heat source, enabling the establishment of temperature profiles within individual clusters. These profiles can be measured by infrared thermography once a steady state has been reached. Radiant flux density received by a berry depending on the distance was calculated based on a view factor and measured lamp surface temperature and resulted to 665 Wm-2 at 7cm. Infrared thermography of the fruit surface was in good agreement with measurements conducted with a thermocouple inserted at epidermis level. In combination with infrared thermography, the presented device offers possibilities for a wide range of applications like phenotyping for heat tolerance in the field to proceed in the understanding of the complex response of plants to heat stress. Sunburn necrosis symptoms were artificially induced with the aid of the device for cv. Bacchus and cv. Sylvaner in the 2020 and 2021 growing season. Threshold temperatures for sunburn induction (LT5030min) were derived from temperature data of single berries and visual sunburn assessment, applying logistic regression. A comparison of threshold temperatures for the occurrence of sunburn necrosis confirmed the higher susceptibility of cv. Bacchus. The lower susceptibility of cv. Sylvaner did not seem to be related to its phenolic composition, rendering a thermoprotective role of berry phenolic compounds unlikely.