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…

Physiological responses of crimson seedless (Vitis vinifera) grapevines to altered micro climatic conditions and different water treatments in the Breede River Valley of South Africa

Challenging conditions created by limited water supply and changes in the climate require an understanding of the physiological status of table grapes along the whole value chain. This is critical to develop tools for regulatory management of growth balances and grape quality. This study aimed to determine the impact of different amounts of water and an altered micro-climate (complete covering of vineyards with plastic) on the physiological reaction of the grapevine during the growth season.

Vineyard floor management intensity impacts soil health indicators and biodiversity across South Australian viticultural landscapes

Vineyard floors in warm, dry landscapes including those in South Australia, have traditionally been managed using intensive practices such as tillage and herbicides to control weeds and vegetation, thereby limiting competition with grapevines for water and nutrients in order to not compromise yields.

Reconfiguring wine prescription : from traditional critics to digital social networks

The integration of digital social networks (DSN) has profoundly transformed communication practices within the wine industry, reorganizing the dynamics of prescription and marketing.

REMEDIATION OF SMOKE TAINTED WINE USING MOLECULARLY IMPRINTED POLYMERS

In recent years, vineyards in Australia, the US, Canada, Chile, South Africa and Europe have been exposed to smoke from wildfires. Wines made from smoke-affected grapes often exhibit unpleasant smoky, ashy characters, attributed to the presence of smoke-derived volatile compounds, including volatile phenols (which occur in free and glycosylated forms). Various strategies for remediation of smoke tainted wine have been evaluated. The most effective strategies involve the removal of smoke taint compounds via the addition of adsorbent materials such as activated carbon, which can either be added directly or used in combination with nanofiltration. However, these treatments often simultaneously remove wine constituents responsible for desirable aroma, flavour and colour attributes.

Exploring multisensory interactions through the study of astringency diversity of mono-varietal Italian red wines

According to the OIV Focus 2017 estimating the vine varieties distribution in the world, Italy is the richest grape producing country in terms of varieties.