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…

Identification of key-odorants in Sauternes Wines

The aim of the present work was to investigate Sauternes wines aromas. The flavor profiles of two wines (vintages 2002 and 2003) were investigated. Key-odorants have been determined by AEDA applied to Amberlite XAD-2 resin extracts. Various complementary techniques were used to identify the compounds (pHMB extraction, chemical synthesis of non-commercial standards, co-injections on two capillary columns, odor description at the sniffing port, GC-MS and GC-PFPD).

Read More

Caractérisation de la dynamique d’accumulation des sucres à l’échelle d’un territoire

Dans le cadre de TerclimPro 2025, Laure de Rességuier a présenté un article IVES Technical Reviews. Retrouvez la présentation ci-dessous ainsi que l’article associé : https://ives-technicalreviews.eu/article/view/8495

Read More

Use of fumaric acid on must or during alcoholic fermentation

Fumaric acid has been approved by the OIV in 2021 for its application on wine to control the growth and activity of lactic acid bacteria. Fumaric acid is currently being evaluated by the OIV as an acidifier of must and wine. Investigations during the 2023 vintage provided further information on its use on must or during AF, thus completing information provided during the previous vintage.

Read More

Comprehensive exploration of wine aroma-related compounds as promoted by alternative vinification procedures in case of Zelen (Vitis vinifera L.) grapes processing

Not only vintner’s decisions in the vineyard, but also winemaker’s choices of technology approaches in the cellar play a significant role in the final wine style and quality. Whereas traditional technologies within chosen terroir are quite well explored and thus somehow predictable, there is no proper knowledge available on possible outcomes in case of implementing novel, alternative winemaking strategies. To reveal their effects on wine aroma compounds and sensory characteristics, two alternative strategies
(cryoextraction or addition of whole grape berries during last stages of fermentation) were compared to classical Vipava valley winemaking approach as normally used for an autochthonous variety Zelen. After separate vinification and bottling, all the experimental wines were subjected to semiquantitative metabolic profiling of volatile compounds (VOCs) by means of GC/MS and were then also sensorialy evaluated by pre-trained panel.

Read More

The effect of pedoclimatic conditions on the yeast assimilable nitrogen concentration on white cv. Doral in Switzerland

Aims: Agroscope investigated the efficiency of nitrogen fertilization via foliar urea application at veraison with the aim of raising the YAN (yeast assimilable nitrogen) content in the musts. The observations were conducted on the white grapevine cultivar Doral (Chasselas x Chardonnay) in several pedoclimatic conditions of the Leman wine region, Switzerland, in the years 2012 and 2013. Knowing that the YAN in must plays a key role in wine quality, the aim was finding the main parameters affecting the final YAN level in order to better control them.

Read More