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.

Download the abstract

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…

An exploration of South Tyrolean Pinot blanc wines and their quality potential in vineyard sites across a range of altitudes

Aim: Pinot Blanc is the third most planted white wine grape in northern Italy’s region of South Tyrol, where small-scale viticultural production permits the examination of the wine’s diverse expressive potential in a small area across a wide range of climatic variables. This study aimed to explore the qualitative potential of Pinot Blanc across a range of climatic variation leading to site-specific terroir expression in a cool climate region.

Read More

Nutrients and heavy metals in a vineyard soil under organic, biodynamic and conventional management

Promoting sustainable agricultural practices is one of the challenges of the last decades. Organic and biodynamic viticulture can be an alternative to intensive viticulture, furthermore contributing to reduction of impact on environment and human health and guaranteeing soil preservation and quality products1. The aim of this experimentation was to evaluate the medium and long-term effects of different agronomic practices in viticulture on nutrient availability and heavy metal accumulation in soil.

Read More

Influence of ‘pinotage’ defoliation on fruit and wine quality

Among the different management techniques in Viticulture, which have
been developed with the purpose of optimizing the interception of sunlight, the photosynthetic capacity of
the plant and the microclimate of the clusters, especially in varieties that show excess vigor, the management of defoliation presents great importance. The defoliation consists of the removal of leaves that cover or that are in direct contact with the curls, which can cause physical damages in the berries, and aims to balance the relation between part area and number of fruits, providing the aeration and insolation in the interior of the vineyard, as well as reduce the incidence of rot in order to achieve greater efficiency in phytosanitary treatments and quality musts.

Read More

Effect of microwave maceration and SO2 free vinification on volatile composition of red wines

This study evaluates the effect of microwave treatment in grape maceration on the content of free and glycosidically bound varietal compounds) of must and wine and on the overall aroma of wines produced in the presence and absence of SO2.

Read More

Old but gold: the interest for minor grapevine varieties – a case study in the Veneto region (Italy)

The biodiversity of cultivated vines has been significantly reduced due to a series of factors that have favoured the cultivation of a limited number of varieties and clones over time. In veneto, since 1980, a series of important actions have been implemented to counter this process. These actions have focused on the conservation of germplasm identified in the territory and the recovery of varieties historically present in the region, which were in danger of being abandoned and disappearing.

Read More