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.

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…

OPTIMISATION OF THE AROMATIC PROFILE OF UGNI BLANC WINE DISTILLATE THROUGH THE CONTROL OF ALCOHOLIC FERMENTATION

The online monitoring of fermentative aromas provides a better understanding of the effect of temperature on the synthesis and the loss of these molecules. During fermentation, gas and liquid phase concentrations as well as losses and total productions of volatile compounds can be followed with an unprecedented acquisition frequency of about one measurement per hour. Access to instantaneous production rates and total production balances for the various volatile compounds makes it possible to distinguish the impact of temperature on yeast production (biological effect) from the loss of aromatic molecules due to a physical effect³.

Agronomic and qualitative behaviour of cv. Tempranillo according to three vine spacing on two different hydric-edaphic situations in the Duero river valley

The knowledge of the influence of soil conditions on the effects that different plant densities provoke in the agronomic grapevine behaviour becomes very interesting since it allows to focus the vineyard management on the optimization of the natural, hydric and human resources.

Integrated approach to grape stalks valorization: sustainable recovery of bioactive compounds and biofuel production

Grape stalks are a byproduct of the winemaking process and represent a valuable and inexpensive source of bioactive compounds. While their direct use in whole bunch fermentation is known, the majority of grape stalks are discarded, posing environmental and economic challenges.

Terroir et marché : exemples de stratégie pour les vins d’une petite région (Muscadet – Anjou – Touraine)

The designations of origin of the Loire Valley wine have been recognized according to customs and notoriety established over the centuries since the Middle Ages. There are four main production basins going up the Loire, from Nantes to the Sancerrois region: Nantes, Anjou-Saumur, Touraine and the vineyards of the Centre. In each of these basins, there is a wide range of appellations of origin which has been established according to a logic which may not seem obvious to the uninformed.

Bioclimatic shifts and land use options for Viticulture in Portugal

Land use, plays a relevant role in the climatic system. It endows means for agriculture practices thus contributing to the food supply. Since climate and land are closely intertwined through multiple interface processes, climate change may lead to significant impacts in land use. In this study, 1-km observational gridded datasets are used to assess changes in the Köppen–Geiger and Worldwide Bioclimatic (WBCS)