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…

Long term influence of a cover crop in the agronomic and oenological performance of CV. Chardonnay

Cover crops are acknowledged to be an interesting tool to produce
higher quality grapes in red varieties, as they generally reduce vine vigour and yield. However, their incidence in white wine quality is not clear, since higher nitrogen availability can play an important positive
role, and cover crops may compete for this nutrient. The possible reduction in available nitrogen can also modify the fermentation processes, as well as the synthesis of aromas in the wine. The aim of this work was to evaluate the long-term effect of a grass cover crop on grape and wine quality.

Mapping of canopy features in commercial vineyards using machine vision

Vineyard canopy features such canopy porosity and fruit exposure influenced microclimate, fungal disease incidence and grape composition. An objective, rapid and non-invasive method to assess and map the canopy status is needed to apply in precision viticulture. A new method for canopy status assessment and mapping based on non-invasive machine vision was applied in commercial vineyards in this work.

Soil or geology? And what’s the difference? Some observations from the New World

Observational historical geology seeks to establish the evolutionary history of the surface of Earth. This approach is applicable not only to bedrock, but to the soft material that lies at the surface, the stuff called soil by most people. The geologic perspective provides a view of this material that is quite different from that of soil science, at least as practiced by many in America.

Typologie des paysages de vigne: un outil de planification

La culture de la vigne dessine un paysage rural original. En effet, de par ses qualités physiologiques, ses exigences agronomiques et les techniques qu’elle requiert, elle est à l’origine d’un portrait de nature sculpté, architecturé, parfois même comparé à l’art des jardins. A ce que l’on pourrait le cas échéant qualifier d’« art involontaire »

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.