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…

The effect of cropload on the volatile aroma characteristics of ‘Beihong’ and ‘Beimei’ red wine

Beihong and Beimei were bred as winemaking cultivars released by Institute of Botany, the Chinese Academy of Sciences in 2008. The cultivars are selected from the population of ‘Muscat Hamburg’ (Vitis vinifera) ×V. amurensis. They are extended to most provinces in North of China because they have strong resistance to cold and disease and need not be buried in soil in winter. To better understand the effect of cropload on volatile compounds during wine-making, we surveyed volatiles composition and content of different cropload level in 3-years-old ‘Beihong’ and ‘Beimei’ vines which planted in east foot of Helan mountain of Ningxia (EHN).

Oenological compatibility of biocontrol yeasts applied to wine grapes 

Antagonistic yeasts applied to wine grapes must be compatible with the thereafter winemaking process, avoiding competition with the fermentative Saccharomyces cerevisiae or affecting wine flavour. Therefore, fifteen epiphytic yeasts (6 Metschnikowia sp., 6 Hanseniaspora uvarum, 3 Starmerella bacillaris) previously selected for its biocontrol ability against Alternaria on wine grapes were evaluate for possible competition with S. cerevisiae by the Niche Overlap Index (NOI) employing YNB agar media with 10 mM of 17 different carbonate sources present in wine grapes (proline, asparagine, alanine, glutamic acid, tirosine, arginine, lisine, methionine, glicine, malic acid, tartaric acid, fructose, melibiose, raffinose, rhamnose, sucrose, glucose).

Enological potential of autochtonous grape cultivars from Castilla y León (Spain) to elaborate sparkling wines: polyphenolic and biogenic amines and amino acid composition of base wines

In white wines, Verdejo wine stands out because of its high content in total amino acids. The total content in biogenic amines was low in all wines analyzed and putrescine was the predominant biogenic amine.

Utilization of remote sensing technology to detect riesling vineyard variability

ineyard blocks can vary spatially with respect to several viticulturally significant qualities such as soil variables, vine vigor, vine physiology

Postveraison shoot trimming in Tannat and Merlot: preliminary results on yield components, plant balance and berry composition

There is currently a trend towards the production of wines with low alcohol content. To achieve this, grapes with low sugar content must be used. There are techniques at the vineyard level that can delay ripening and avoid excessive sugar accumulation without, a priori, affecting the final polyphenol content. Postveraison shoot trimming (PVST) is experimentally evaluated for these purposes, but its impact under Uruguayan climatic conditions with high interannual variability is not known. The aim of this work is to assess the PVST in Tannat and Merlot cultivars and their impact on yield components, plant balance and berry primary composition. In this study, two commercial vineyards of 10 years old Tannat and Merlot (grafted on SO4) at Canelones Department were selected. During the 2020-201 growing season, grapevines were submitted to PVST when grapes reached 15º Brix. In a randomized block, trimmed (T) and control (C) plants were evaluated with three repetitions each cultivar. Evaluation of the evolution of primary berry composition during ripening, measurement of yield components and plant balance were performed. For both cultivars, PVST did not affect yield components. Merlot reached 5.4 kg per plant and Tannat 7.1 kg, with not statistical significance between treatments. However, statistical differences were observed in terms of plant balance. In Merlot Ravaz Index reached a difference of 5.3 (12.0 in T and 6.7 in C) meanwhile Tannat reached 3.5 of statistical difference (13.7 in T and 10.2 in C). The tendency to imbalance for the treated plants had an impact on the final grape composition. Merlot grapes showed statistical difference in final total acidity (0.3 g of difference between treatments) while treatments impact final sugar content on Tannat grapes (10.0 g of difference between treatments). Further studies are needed to assess the impact of different canopy management techniques in our conditions.