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…

Sustainable wine industry: supercritical fluid extraction as key technology for biorefinery enhancement

Supercritical carbon dioxide (sc-CO2) extraction is an environmentally friendly technology employed for bioactive compounds recovery from various natural sources and biomasses. The advantages of sc-co2 extraction include its selectivity, relatively mild operating conditions, which minimize the degradation of sensitive compounds, and the absence of potentially harmful organic solvents.

Arbuscular mycorrhizal fungi as biomarkers of vineyard yield in Champagne

The vine is colonized by a multitude of micro-organisms (fungi, bacteria, oomycetes) mainly coming from the microbial reservoir constituted by the soil. These microorganisms have positive or negative effects on the vine (protection against pathogens, resistance to abiotic stress, nutrition, but also triggering of diseases) (Fournier, Pellan et al. 2022). In addition to these functional roles, they respond quickly to environmental changes (climate, cultural practices) which could make them good bioindicators of the functioning of the wine ecosystem.

Predictive Breeding for Wine Quality: From Sensory Traits to Grapevine Genome

New pathogen resistant varieties allow an efficient and greatly reduced use of fungicides. These new varieties promise, therefore, an enormous potential to reach the European Green Deal aim of a 50% reduction of pesticides in EU agriculture by 2030.

Identification and biological properties of new resveratrol derivatives formed in red wine

Resveratrol is a well-known wine constituent with a wide range of activities. In wines, resveratrol can be oxidized to form various derivatives including oligomers [1]. In this study, resveratrol derivative transformation in hydroalcoholic solution was investigated by oxidative coupling using metals. De novo resveratrol derivatives were synthetized and analysed by NMR and MS experiments

Biotic and abiotic factors affecting physiological aspects underlying vegetative vigour in two commercial grapevine varieties

Grapevine vigour, defined as the propensity to assimilate, store and/or use non-structural sugars for allowing fast growth of shoots and producing large canopies[1], is crucial to optimize vineyard management. Recently, a model has been proposed for predicting the vigor of young grapevines through the measurement of the vegetative growth and physiological parameters, such as water status and gas exchange[2]. Our objectives were (1) to explore the influence of the association of two grapevine varieties (Tempranillo and Cabernet Sauvignon, grafted onto R110 rootstocks) with arbuscular mycorrhizal fungi (AMF) on the vegetative vigour of young plants; and (2) to assess the effect of environmental factors linked to climate change on the vegetative vigour of Cabernet Sauvignon.