terclim by ICS banner
IVES 9 IVES Conference Series 9 Open-GPB 9 Open-GPB-2024 9 Flash - New biotechnological tools 9 Enhancing hydric stress tolerance by editing the VviMYB60 promoter with CRISPR/Cas9 

Enhancing hydric stress tolerance by editing the VviMYB60 promoter with CRISPR/Cas9 

Abstract

Climate change presents increasing challenges to viticulture, particularly with rising water stress contributing significantly to yield losses and damages. The identification of the MYB60 transcription factor, which regulates stomatal opening and closing in Arabidopsis thaliana and Vitis vinifera, offers potential solutions. Notably, knockout studies in Arabidopsis have shown reduced stomatal opening and increased drought tolerance in myb60 mutants. Additionally, the grapevine ortholog, VviMYB60, can restore the wild-type phenotype of Arabidopsis myb60 mutants. Further investigation of the Arabidopsis promoter region has revealed that mutations in DOF motifs lead to reduced expression of AtMYB60.

Utilizing the advancements in CRISPR/Cas9 genome editing, this project aims to modify the VviMYB60 promoter region to lower gene expression, thereby reducing stomatal opening in grapevines. Binary vectors for genome editing were constructed to target two specific regions of the VviMYB60 promoter. Agrobacterium-mediated transformation was performed on Chardonnay embryogenic calli, resulting in the successful regeneration of plants under selection conditions. Sanger sequencing analysis of the targeted region confirmed the occurrence of genetic edits in four of the six lines analyzed so far.

To further characterize the edited lines, next-generation sequencing will be utilized, providing a more comprehensive understanding of the mutations, as well as gene expression of VviMYB60 will be evaluated to confirm that the editing reduces its expression. Morphological and physiological parameters will be measured after acclimatation in greenhouse and finally these edited lines will undergo drought tolerance testing to assess their performance.

DOI:

Publication date: June 13, 2024

Issue: Open GPB 2024

Type: Article

Authors

Manuela Campa1*, Loredana Moffa2, Luca Nerva2, Walter Chitarra2, Johan Burger1

1 Genetics Department, Faculty of Agrisciences, Stellenbosch University, 7600 Stellenbosch, South Africa
2 Research Centre for Viticulture and Enology, Council for Agricultural Research and Economics (CREA-VE), Via XXVIII Aprile 26, 31015 Conegliano (Italy)

Contact the author*

Keywords

CRISPR/Cas9, VviMYB60, promoter, drought tolerance, stomatal regulation

Tags

IVES Conference Series | Open GPB | Open GPB 2024

Citation

Related articles…

Ellagitannins and flavano-ellagitannins: concentration ranges in different areas and sensory evaluation

C-Glucosidic ellagitannins, which are the main polyphenolic compounds in oak heartwood, are extracted by wine during aging in oak barrels. Although such maturing of alcoholic beverages in oak barrels is a multi-centennial practice, very little is known on the impact of these ellagitannins on the organoleptic properties of red wine. The objectives of the present investigation were (i) to isolate oak ellagitannins and to hemisynthesize some made-in-wine flavano-ellagitannins, such as acutissimin A; (ii) to analyse their concentration ranges depending on the cultivar area and (iii) to evaluate their sensory impact on the basis of their human threshold concentrations and dose/response relationships in different types of solutions.

Application to the wine sector of European Convention on the landscapes

The landscape is defined by the European convention of the landscape (Florence, October 20, 2000) like part of the territory as perceived by the populations, whose character results from the action of natural and/or human factors and their interrelationships. This convention is based on the contribution cultural, ecological, environmental, social of the landscapes and aims at a reinforcement of the tools of protection and valorization in particular in the agricultural policies, of regional planning and town planning. Moreover, it encourages a step of identification and qualification of the landscapes and underlines the need for developing the sensitizing and the training of the actors concerned.

Impact of acidification by fumaric acid at vatting on Cabernet-Sauvignon wine during winemaking

Acidity of grape berries is lowered due to climate changes (1), resulting in musts and wines with higher pHs. These higher pHs induce microbiological instability

Synergistic effect of fumaric acid and chitosan on the inhibition of malolactic fermentation

During wine storage and aging, microorganisms capable of degrading malic acid in an undesirable manner can proliferate.

The impact of acetaldehyde on phenolic evolution of a free-SO2 red wine

Some wine producers, in good years, can produce free-SO2 red wines and decide to add the minimum amount of sulphur dioxide only at bottling. To manage this addition