terclim by ICS banner
IVES 9 IVES Conference Series 9 New breeding frontiers: application of the CRISPR-cas9 system in grapevine (V. vinifera L.) and improvements in plant regeneration

New breeding frontiers: application of the CRISPR-cas9 system in grapevine (V. vinifera L.) and improvements in plant regeneration

Abstract

Nowadays, many policies are being adopted for direct agriculture towards more sustainable approaches. To continue to maintain a high production using fewer fertilizers, pesticides and water resources, agronomic techniques must be combined with biotechnological approaches. In grapevine, the breeding programs are restricted by the fact that it has a highly heterozygous genome, therefore, if on the one hand, we try to improve the characteristics, on the other hand it is necessary to preserve the original genome of the varieties. CRISPR-cas9 system is one of the smartest tools to carry out highly precise genetic modifications leaving the genetic background unchanged. To produce edited DNA-free grapevine plant is necessary to dispose of an efficient delivery system to introduce the preassembled ribonucleoproteins (RNP) and to ensure the subsequent step of regeneration of the edited plant. Protoplasts are the best system for this purpose: they represent a highly regenerative platform accessible to most of transformation techniques. The regeneration is possible through somatic embryogenesis. Grapevine is recalcitrant to the regeneration process leading to a low rate of plant recovery. The aim of this study is to optimise the in vitroregeneration process of Cabernet Sauvignon and Glera varieties to apply a DNA-free genome editing approach to improve agronomical and oenological traits. Here, we also provide preliminary studies on the enhancing effect of the current in the regeneration process through the application of an external electric field and evaluating the expression of fluorescent transcriptional reporters of transcription factors involved in shoot regeneration in A.thaliana.

DOI:

Publication date: June 13, 2024

Issue: Open GPB 2024

Type: Poster

Authors

Stefania Zattoni 1*, Edoardo Bertini 2, Erica D’Incà 2, Maddalena Salvalaio 3, Giovanni Sena 3, Sara Zenoni 1

1 Department of Biotechnology, University of Verona, Strada Le Grazie 15, 37134, Verona, Italy
2 Edivite s.r.l, San Pietro Viminario, Quartiere San Mauro 30, 35020 Padova
3 Laboratory of plant morphogenesis, Imperial College London, South Kensington Campus, London SW7 2AZ, UK

Contact the author*

Keywords

Protoplast, Somatic embryogenesis, CRISPR-cas9, in vitro regeneration, Electric field

Tags

IVES Conference Series | Open GPB | Open GPB 2024

Citation

Related articles…

Characterization of bunch compactness and identification of associated genes in a diverse collection of cultivars of Vitis vinifera L.

Compactness is a complex trait of V. vinifera L. and is defined ultimately by the portion of free space within the bunch which is not occupied by the berries. A high degree of compactness results in poor ventilation and consequently a higher susceptibility to fungal diseases, diminishing the quality of the fruit. The easiness to conceptualize the trait and its importance arguably contrasts with the difficulty to measure and quantify it. However, recent technical advancements have allowed to study this attribute more accurately over the last decade. Our main objective was to explore the underlying genetics determining bunch compactness by applying updated phenotyping methods in a collection of V. vinifera L. cultivars with a wide genetic diversity.

AROMA ASSESSMENT OF COMMERCIAL SFORZATO DI VALTELLINA WINES BYINSTRUMENTAL AND SENSORY METHODOLOGIES

Sforzato di Valtellina DOCG is a special dry red wine produced from partially dehydrated Nebbiolo wine-grapes growing in the Rhaetian Alps valley of Valtellina (Lombardy, Italy). Valtellina terraced vineyards are located at an altitude of 350–800 m according to ‘heroic’ viticulture on steep slopes. The harvested grape bunches are naturally dehydrated indoors, where a slow and continuous withering occurs (about 20% w/w of weight loss), until at least 1st December when the grapes reach the desired sugar content and can be processed following a normal winemaking with maceration.

Effect of different winemaking practices on chemical composition, aroma profile and sensory perception of ribolla gialla sparkling wines

This study aims at evaluating the effects of different refermentation methods (Martinotti/Charmat vs. Classic) on the chemical composition, aroma profile and sensory characteristics of Ribolla Gialla sparkling wines; furthermore, certain winemaking practices (skin contact and use of pectolytic enzymes) were investigated considering the extraction of varietal aromas and aroma precursors. METHODS: Sparkling wines were produced at pilot-plant scale. Concerning refermentation methods, traditional Martinotti (MB – 30 days length), extended Martinotti (ML) with 4 months of aging on lees and Classic method (CL) with 11 months of aging on lees were compared; in a second trial, skin contact (MM), enzyme addition on must also subjected to maceration (ME), and enzyme addition on base wine (VE) were evaluated. All experimental trials were performed in triplicate. Basic chemical composition, varietal (terpenes and C13-norisoprenoids in free and bound form) and non-varietal aroma compounds were evaluated by LLE-GCMS analysis; finally, sensory analysis was also performed, by descriptive testing.

Use of fumaric acid on must or during alcoholic fermentation

Fumaric acid has been approved by the OIV in 2021 for its application on wine to control the growth and activity of lactic acid bacteria. Fumaric acid is currently being evaluated by the OIV as an acidifier of must and wine. Investigations during the 2023 vintage provided further information on its use on must or during AF, thus completing information provided during the previous vintage.

Effects of using cationic exchange for reducing pH on the composition and quality of sparkling wine

In this video recording of the IVES science meeting 2023, Fernando Zamora (Department of biochemistry and biotechnology, Faculty of oenology, Universitat Rovira i Virgili, Spain) speaks about the effects of using cationic exchange for reducing pH on the composition and quality of sparkling wine. This presentation is based on an original article accessible for free on OENO One.