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…

Winemaking techniques and wine tasting methods at the end of the Middle Ages

Les pratiques de vinification et de dégustation du vin sont souvent perçues, à travers un discours marketing très puissant, sous l’angle d’une tradition millénaire qui perdure depuis le Moyen Âge. En Bourgogne, il est courant de rattacher les racines de ces pratiques à l’activité des institutions ecclésiastiques qui possédaient d

The dynamics of δ13C and δ18O in musts during berries development

Aim: Many processes or reactions that occur in plants involved isotopic discrimination. Water availability, for example, affects the isotopic ratio of carbon (δ13C) and oxygen (δ18O). In viticulture, δ13C is used in experiments related to water relations and irrigation in vineyards. δ18O is used much less but it could be a good complement to δ13C. The aim of this study was to generate knowledge on how these isotopic ratios, measured in musts, could help to better understand the water behavior of grape varieties. 

Exploring grapevine water relations in the context of fruit growth at pre- and post-veraison

Climate change is increasing the frequency of water deficit in many grape-growing regions. Grapevine varieties differ in their stomatal behavior during water deficit, and their ability to regulate water potential under dry soil conditions is commonly differentiated using the concept of isohydricity. It remains unclear whether stomatal behavior, water potential regulation, and the resulting degree of isohydricity has a relationship with changes to fruit growth during water deficit. This study was conducted on four varieties (`Cabernet Franc`, `Semillon`, `Grenache`, and `Riesling`) subjected to both short-term, severe water deficit and long-term, moderate water deficit applied at both pre- and post-veraison.

Effect of different plant fibers on the elimination of undesirable compounds in red wine 

The presence of undesirable compounds in wines, such as ota, biogenic amines and pesticides residues, affects wine quality and can cause health problems for the consumer. Additionally, an excess of tannins can produce an undesirable increase in the astringency and bitterness of the wine, so tannins are also a target for reduction. The main tool that a winemaker has to reduce their content in the wine is fining. However, some of the fining agents commonly used in the winery can cause allergies or even increase the protein content in the wine, increasing the turbidity.

Effet terroir et arômes des muscats

L’étude porte sur trois terroirs du Roussillon, classés dans l’A.O.C. Muscat de Rivesaltes et concerne les 2 cépages de cette appellation : le muscat à petits grains et le muscat d’Alexandrie. Elle a pour objectif de connaître pour un terroir donné le meilleur choix de cépage.