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…

Berry weight loss in Vitis vinifera (L.) cultivars during ripening

Berry shriveling (BS) in vineyards are caused by numerous factors such as sunburn, dehydration, stem necrosis. Climate change results in an increase in day and night temperatures, rainfall throughout the year, changes in the timing and quantities, long dry summers and a combination of climatic variability such as floods, droughts and heatwaves). Grape development and its composition at harvest is influenced by the latter as grape metabolites are sensitive to the environmental conditions. The grape berry experiences water loss and an increase in flavour development as a result of the BS. An increased sugar content in grapes will result in higher alcohol wines and concentration of grape aromas which may be detrimental to the final wine quality.

Spontaneous fermentation dynamics of indigenous yeast populations and their effect on the sensory properties of Riesling

Varietal Riesling aroma relies strongly on the formation and liberation of bound aroma compounds. Floral monoterpenes, green C6-alcohols, fruity C13-norisoprenoids and spicy volatile phenols are predominantly bound to disaccharides, which are produced and stored in the grape berry during berry maturation. Grape processing aims to extract maximum amount of the precursors from the berry skin to increase the potential for a strong varietal aroma in the wine. Subsequent yeast selection plays an important part in this process.

Variability of Tempranillo phenology within the toro do (Spain) and its relationship to climatic characteristics

Aims: The objective of this research was to analyse the spatial and temporal variability of vine phenology of the Tempranillo variety in the Toro Designation of Origen (DO) related to climatic conditions at present and under future climate change scenarios.

Effect of quercus alba oak barrels from different forest on the volatile composition of Tempranillo wines

The species and origin used for red wine oak aging determines the physiological composition of the wood and thus the finished wines. In America, oak is grown primarily

Iso-/anisohydric behavior in wine grapes may be a matter of soil moisture

There are claims that wine grape cultivars are either isohydric or anisohydric; the former maintaining, and the latter decreasing, their plant water status as soil moisture declines. However, available information is inconsistent. There are those that show an existence of a continuum in cultivar response to soil moisture rather than a distinct categorization. Others even show both behaviors in the same cultivar grown in different environments. In this study we investigated the behavior of 30 own rooted Vitis vinifera cultivars during successive drydown and rewatering cycles over two growing seasons in arid eastern Washington (<200 mm annual precipitation).