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.

Download the abstract

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…

Sensory and chemical profiles of Cabernet Sauvignon wines exposed to different irrigation regimes during heatwaves

Heatwaves, defined as three or more consecutive days above average historical maximum temperatures, are having a significant impact on agricultural crop yields and quality, especially in arid or semi-arid regions with reduced water availability during the growing season.

Read More

Potentiel des sols viticoles et qualité des vins

La qualité des vins dépend de différents facteurs et procédés, notamment de la nature des terrains viticoles. Dans ce travail, nous avons cherché à établir les liens entre descripteurs pédologiques des parcelles et descripteurs sensoriels des vins. Sur la base de Classifications Ascendantes Hiérarchiques (CAH) et d’Analyses en Composante Principale (ACP), il a été possible d’établir des liens entre la nature des parcelles (sableuse, argileuse, sablo-graveuleuse) et certains descripteurs sensoriels des vins (chaleur, astringence, fruit noir) et plus globalement avec le type de vins élaborés.

Read More

Understanding colloidal instability in white wine model solutions: A study focused on the effect of polysaccharides and salts onto bentonite efficiency

A white wine model solution (12% v/v ethanol, 4 g/L tartaric acid, pH 3.2) was used to assess wine colloidal instability as well as the influence of several wine components on bentonite performance in protein removal.

Read More

The effects of perennial cover crop management on soil temperature and vine water status

The implications of perennial cover crop management on vine vigor and yield have been well documented. However, whereas multiple studies show that cover crop management affects grapevine dry matter production, water, and nutrient status, the specific effects of a new hybrid perennial cover crop on soil temperature and its relationship to vine water status in vineyards has not been explored. This study will compare 3 different perennial cover crop combinations and tillage practices with a no-till seeding of a new hybrid perennial, Poa bulbosa (Pb).

Read More

Integrated approaches for the functional characterization of miRNAs in grapevine

Micro(mi)RNAs are small non-coding RNAs that regulate several pathways and are widely recognised as key players in plant development, tissue differentiation, and many other important physiological processes, including plant adaptation to biotic and abiotic stresses. The release of plant genomes and the application of high throughput sequencing have considerably extended miRNA discovery across many species, including grapevine (Vitis spp.). Despite their relevance in plant development, functional studies in grapevine to clarify the function of miRNAs are not yet available. Through the grapevine genetic improvement platform IMPROVIT at CNR-IPSP (http://www.ipsp.cnr.it/en/thematics/turin-headquarter-thematics/improvit/), we developed integrated approaches to discover miRNA function in grapevine.

Read More