terclim by ICS banner
IVES 9 IVES Conference Series 9 Functional characterization of grapevine MLO genes to define their roles in Powdery mildew susceptibility by CRISPR/Cas9 genome editing

Functional characterization of grapevine MLO genes to define their roles in Powdery mildew susceptibility by CRISPR/Cas9 genome editing

Abstract

Successful powdery mildew (PM) infection in plants relies on Mildew Resistance Locus O (MLO) genes, which encode susceptibility factors essential for fungal penetration. In Arabidopsis, loss-of-function mutations in three clade-V MLOs, AtMLO2, 6, and 12 confer complete resistance to PM infection. Since then, efforts are on to discover MLO genes contributing to PM susceptibility in many species to introduce mlo-based PM-resistance. Earlier studies in tomato and grapevine, using the RNAi approach, attributed PM susceptibility to SlMLO1, 5, and 8 and VvMLO3, 13, and 17, respectively indicating likely functional redundancy among MLOs. Here, we disrupted the closest grapevine orthologues, VvMLO3, 4, 13, and 17 through CRISPR/Cas9-mediated mutagenesis in the microvine model with the goal of identifying the candidate MLO genes to introduce mlo-based PM resistance. Individual mutants mlo3, mlo4, mlo13 and mlo17 showed 8 to 50% less infection to E. necator, whereas double mutants, mlo3/4, mlo3/13 andmlo13/17 and triple mutant mlo3/13/17 showed 60 to 90% less infection. But the quadruple mlo3/4/13/17 mutant plants showed near complete PM resistance. Considerable differences were observed in the resistance level of clones among the triple and quadruple mutants due to the differences in editing efficiency of individual guide RNAs. Some mutants showed pleiotropic effects in the growth and development, ranging from early senescence and stunted growth to non-flowering phenotypes, which also seemed to depend on the percentage of gene-edited cells in the plant. The overarching goal is to excise the genome-integrated T-DNA cassette from the mutants using CRISPR Ribonucleoproteins for transgene-free PM resistance.

Download the abstract

DOI:

Publication date: June 13, 2024

Issue: Open GPB 2024

Type: Article

Authors

Satyanarayana Gouthu1*, Laurent Deluc2,3, Samuel Talbot1

1,2 Department of Horticulture, Oregon State University, Corvallis, OR, USA
3 Oregon Wine Research Institute, Oregon State University, Corvallis, OR, USA

Contact the author*

 

Keywords

Powdery mildew, Grapevine MLO, mildew-resistance, Gene Editing

Tags

IVES Conference Series | Open GPB | Open GPB 2024

Citation

Related articles…

Chemical profiling and sensory analysis of wines from resistant hybrid grape cultivars vs conventional wines

Recently, there has been a shift toward sustainable wine production, according to EU policy (F2F and Green Deal), to reduce pesticide usage, improve workplace health and safety, and prevent the impacts of climate change. These trends have gained the interest of consumers and winemakers. The cultivation of disease resistant hybrid grape cultivars (DRHGC), known as ‘PIWI’ grapes can help with these objectives [1]. This study aimed to profile white and red wines produced from DRHGC in South Tyrol (Italy). Wines produced from DRHGCs were compared with conventional wines produced by the same wineries. The measured parameters were residual sugars, organic acids, alcohol content, pigments and other phenolics by LC-QqQ/MS, colorimetric indexes (CIELab); and volatile profiles (HS-SPME-GCxGC-ToF/MS [2]).

Read More

Can plant shaking reduce the incidence of Botrytis?

Wine production is expanding in Scandinavia with a focus on organic growing, and Solaris becoming the signature grape of the region.

Read More

Terroir effects on wine aromatic metabolomics in the eastern foot of Helan Mountain, Ningxia, China

Aim: The eastern foot of Helan Mountain, Ningxia, China is one of the most important wine production regions in China and grape cultivation has spread in several sub-regions with different soils and cultivars. Large diversity in wine aromas have been observed at Ningxia region but which terroir factors drive those diversity in aromas remain to uncover. This study aims to investigate the impacts of grape varieties and soil chemical properties on wine aromas at Ningxia, in order to characterize the aromatic typicality of Ningxia wines and provide foundation for developing a ‘Protected Designation of Origin’ system. 

Read More

Maturità fenolica e cellulare come metodo di valutazione dell’interazione vitigno-ambiente: il caso del Cabernet-Sauvignon

ln the current work, phenolic and cellular maturation curves were used to assess the degree of adaptation of the cultivar Cabernet sauvignon to the sites under esamination. Five wine­-producing zones with different pedoclimatic characteristics and latitudes were considered (Marche, Toscana, Emilia, Friuli and Slovenia).

Read More

LC-MS based metabolomics and target analysis to study the chemical evolution of wines stored under different redox conditions

Oxygen is a key player in oenology, since its effects can be a blessing, benefiting wine quality, or a curse causing irreversible damage.

Read More