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…

Climate change impacts on European grapevine yields through a dynamic crop modelling approach

Climate has a predominant role on growth and development of grapevines. Therefore, climate change represents an important challenge to the winemaking sector.

Viticultural zoning applications at the detailed scale of a cooperative winery: terroirs in St­hilaire-d’Ozilhan (AOC Côtes-du-Rhône)

La maîtrise de la typicité du vin s’élabore au niveau local ou communal d’une exploitation viticole et/ou d’une cave, unité de vinification. La cave coopérative de Saint-Hilaire­-d’Ozilhan (AOC Côtes-du-Rhône), dont le territoire communal s’étend sur une superficie de 1 670 ha, couvre près de 310 ha cultivés en vigne. Elle réalise des vinifications «au terroir», en utilisant des regroupements d’unités de sol en 9 unités de terroir potentiellement viticoles, définies en s’appuyant sur la parenté des substrats. Diverses sélections d’une même unité peuvent aboutir aussi à des vins différents, ce qui suggère une hétérogénéité spatiale de certaines unités définies.

Présentation d’une méthodologie de caractérisation des terroirs et valorisation par l’étude de l’effet terroir sur la typicité et l’originalité du produit vin dans la région des Côtes du Rhône

In the global economic context, an Appellation d’Origine Contrôlée must now more than ever control the typicity and originality of the wines it produces. It is in this spirit that the Côtes du Rhône have decided to acquire the means necessary for this control.

Utility of leaf removal timing and irrigation amounts on grape berry flavonoids under climate change

Context and purpose of the study – The dormant and growing season temperatures in California USA have been increasing with more clear sky days. A consequence increasing temperatures and clear sky days is water deficit conditions. Viticulturists must determine appropriate balances of canopy management and irrigation budgeting to produce suitable yields without compromising berry chemistry. In response, a study designed to test the interactive effects of leaf removal timing and applied water amounts on Cabernet Sauvignon/110R in Napa Valley, CA.

On-farm monitoring of grapevine water and nitrogen status in relation to different soil management practices in Valais, Switzerland

In response to increasing societal demands for environmentally-friendly viticulture, winegrowers are adapting their cultivation techniques, particularly by reducing the use of herbicides.