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…

Evaluating the suitability of hyper- and multispectral imaging to detect endogenic diseases in grapevine

Endogenic diseases often arise from pathogens that exist within the plant tissue, including fungi, bacteria, and viruses, which can remain latent and then emerge under stress conditions or favorable environmental conditions, causing symptoms that weaken vines or can lead to plant death.

Investigation on the potentiality of a biostimulant by Fabaceae tissues and rich in triacontanol to enhance grapevine resilience under drought stress

The primary objective of this research was to investigate the potential benefits of a Fabaceae-based product rich in triacontanol (a long-chain alcohol) applied to Vitis vinifera cv. Merlot, on key physiological and productive parameters of grapevines under controlled water stress conditions.

Non-Saccharomyces yeast nitrogen consumption and metabolite production during wine fermentation

Over the last decade, the use of non-Saccharomyces yeasts in the winemaking process has been re-assessed and accepted by winemakers. These yeasts can be used to achieve specific objectives such as lowering the ethanol content, preventing wine spoilage and increasing the production of specific aroma compounds. Since these species are unable to complete alcoholic fermentation, strategies of co- and sequential inoculation of non-Saccharomyces and Saccharomyces cerevisiae have been developed. However, when mixed starter cultures are used, several parameters (e.g. strain yeast, inoculation timing and nutrient competitions) impact the growth of the individual yeasts, the fermentation kinetics and the metabolites/aroma production. In particular, competition for nitrogen compounds could have a major impact, potentially leading to sluggish fermentation when the yeast assimilable nitrogen (YAN) availability is low. Moreover, many aroma compounds produced by the yeasts are directly produced and influenced by nitrogen metabolism such as higher alcohols, acetate esters and ethyl esters which participate in the organoleptic complexity of wine.

How to improve the success of dead vine replacement: insights into the impacts of young plant‘s environment 

Grapevine faces multiple biotic and/or abiotic stresses, which are interrelated. Depending on their incidence, they can have a negative impact on the development and production of the plant, but also on its longevity, leading to vine dieback. One of the consequences of vine dieback on production is the increased replacement rate of dead or missing vines within a parcel.

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.