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: 13/06/2024

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…

The moment of preharvest elicitor application influence its final effect on winegrapes quality

Phenolic compounds are secondary metabolites of grapes. Plants produce a wide variety of this type of metabolites through diverse biosynthesis pathways and their production is sometimes a response to external stimuli, either environmental or biotic stresses. Some of them may act as chemical defenses against pathogens or herbivores and their synthesis is increased when the attack exists. However, it is remarkable that the synthesis of these interesting compounds can be activated even when the stimulus is not present, with the use of elicitors. These are substances that when applied exogenously trigger the biosynthetic pathways conducting to the synthesis of these defense compounds.

A viticultural perspective of Meso-scale atmospheric modelling in the Stellenbosch wine growing area, South Africa

La brise de mer et les facteurs climatiques qu’elle entraîne (accélération de la vitesse du vent au cours de l’après midi, augmentation de l’humidité et baisse de la temperature) sont d’un intérêt particulier pour la viticulture.

Using ΔC13 to assess viticultural and oenological suitability for Sangiovese of different pedoclimatic conditions in Chianti

A two years trial was carried out in Chianti (Central Italy) to assess at the detailed scale the viticultural and oenological suitability for Sangiovese of different pedoclimatic conditions, by means of the ΔC13 measured in the must sugars

Vitamins in musts : an unexplored field

Vitamins are major compounds, involved in several prime yeast metabolic pathways. Yet, their significance in oenology has remained mostly unexplored for several decades and our current knowledge on the matter still remaining obscure to this day. While the vitaminic contents of grape musts have been approached in these ancient investigation

IMPACT OF CLIMATIC ZONES ON THE AROMATIC PROFILE OF CORVINA WINES IN THE VALPOLICELLA REGION

In Italy, in the past two decades, the rate of temperature increases (0.0369 °C per year) was slightly higher compared to the world average (0.0313 °C per year). It has also been indicated that the number and intensity of heat waves have increased considerably in the last decades. (IEA, 2022). Viticultural zones can be classified with climatic indexes. Huglin’s index (HI) considers the temperature in a definite area and has been considered as reliable to evaluate the thermal suitability for winegrape production (Zhang et al., 2023).