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…

Sustainable management of grapevine trunk diseases

Grapevine trunk diseases (GTD) occur wherever grapes are grown and are considered the main biotic factor reducing yields and shortening vineyards’ lifespan. Currently, no product is available to eradicate GTD once grapevines are infected. Therefore, prophylactic strategies based on pruning wound protection and ‘remedial surgery’, the only eradication method based on the elimination of infected wood and renewal of the vine by means of new canes or suckers, are the only effective strategies available. The Canadian grape and wine industry focusses on a sustainable production and thus, looking for alternatives to chemicals for disease management is a top priority.

Study of the volatil profile of minority white varieties

The genetic material preservation is a priority issue in winemaking research. The recovery of minority grape varieties can control the genetic erosion, contributing also to preserve wine typical characteristics. In D.O.Ca. Rioja (Spain) the number of grown white varieties has been very limited, representing Viura the 91% of the cultivated white grape area in 2005, while the others, Garnacha Blanca and Malvasía riojana, hardly were grown. For this reason, a recovery and characterization study of plant material was carried out in this region. In 2008, the results obtained allowed the authorization of three minority white varieties: Tempranillo Blanco, Maturana Blanca and Turruntés.

Directed Evolution of Oenococcus oeni: optimising yeast-bacteria interactions for improved malolactic fermentation

Malolactic fermentation (MLF) is a secondary step in the vinification process and it follows alcoholic fermentation (AF) which is predominantly carried out by Saccharomyces cerevisiae. These two processes result in the degradation of metabolites to produce secondary metabolites which also contribute to the final wine flavour and quality. AF results in the production of ethanol and carbon dioxide from sugars and MLF stems from the degradation of L-malic acid (a dicarboxylic acid) to L-lactic acid (a monocarboxylic acid). The latter process results in a smoother texture as the acidity of the wine is reduced by the process, it also adds to the flavour complexity of the wine.

applicazione dei metodi isotopici e dell’analisi sensoriale negli studi sull’origine dei vini

Traceability of agro-alimentary products is very important to certify their origin. This work aimed to characterize wines obtained by the same cultivar (Nero d’Avola and Fiano) – grown in regions with different soil and climate conditions during three vintages (2003-2005) – employing isotopic analyses (NMR and IRMS) and sensory analyses.

PIWIs’ variation in drought response under semi-controlled conditions 

Grapevine interspecific hybrids (PIWIs, from German “pilzwiderstandsfähige Rebsorten” meaning fungus tolerant grapevine cultivars), offer a promising alternative to traditional cultivars in many wine regions due to their tolerance to certain fungal diseases. This makes them a potential solution for sustainable wine production, particularly under organic systems. Despite extensive research on certain agronomic traits and susceptibility to biotic diseases, such as powdery and downy mildews, the response of these cultivars to abiotic stressors, such as drought, remains unclear. Our study aims to investigate the eco-physiological traits of two commercial PIWI cultivars, Muscaris and Souvignier gris, at the leaf level to evaluate their response to drought stress.