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…

A multivariate clustering approach for a gis based territorial characterization of the montepulciano d’abruzzo DOCG “Colline Teramane”

The aim of the project was to characterize the Premium Denomination of Guaranteed Origin (DOCG) “Colline Teramane” wine-growing region and to delineate and define homogeneous zones (terroir units) within it, by applying a multivariate clustering approach combined with geomatics.

Impact of crop load management on terpene content in gewürztraminer grapes

Context and purpose of the study ‐ Crop load management by cluster thinning can improve ripening and the concentration of key metabolites for grape and wine quality. However, little work has been done on testing the impact of crop load management on terpene content of white grapes. The goal of the study was to assess if by reducing crop load via cluster thinning growers can increase terpene concentration of grapes, as well as to test if the timing of thinning application affects terpene concentration.

Étude des relations sol-vigne sur le vignoble de Côte Rôtie

La topographie du vignoble de Côte Rôtie, la prédominance de la non culture ainsi que la structure très légère des sols amènent les vignerons à s’interroger sur l’entretien du sol, la conduite de la fertilisation de leurs parcelles ainsi que sur le développement racinaire de la vigne.

Effect of different plant fibers on the elimination of undesirable compounds in red wine. Correlation with its polysaccharide composition

The presence of undesirable compounds in wines, such as OTA, biogenic amines and pesticides residues, affects wine quality and can cause health problems for the consumer. The main tool that a winemaker has to reduce their content in the wine is fining. However, some of the fining agents commonly used in the winery can cause allergies or even increase the protein content in the wine, increasing the turbidity. To avoid these problems, the use of plant fibers may be an alternative, such as those from grape pomace[1] or other plant origins.

Machines and fire: developing a rapid detection system for grapevine smoke contamination using NIR spectroscopy and machine learning modelling

Bushfires are a common occurrence throughout Australia and their incidence is predicted to both rise and increase in severity due to climate change. Many of these bushfires occur in areas close to wine regions, which receive different levels of exposure to smoke. Wine produced from smoke-affected grapes are characterised by unpalatable smoky aromas such as “burning rubber”, “smoked meats” and “burnt wood”. These smoke tainted wines are unprofitable and result in significant financial losses for winegrowers.