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…

Comparison of plant nutrients in the soil solution and bleeding sap of grapevine cvs

In this study macro and micro nutrients of plants (N = NH4 + NO3 , P, K, Ca, Na, Zn, Mn, Fe and Cu) were determined both in soil solution and bleeding sap and compared each other. Bleeding sap was collected from the nine varieties of grapevine Cvs. grafted on 5BB rootstock and grown in different soil conditions. For all varieties, plant nutrients content in bleeding sap as higher than in soil solution except for Ca and Na. While in soil solution Ca content was found at 10209 ppm, this value in bleeding sap was 49.20 ppm (Kozak Beyazy), 55.38 ppm (Trakya Ylkeren), 50.37 (Cardinal) and 74.27 ppm (Tekirdaô Çekirdeksizi) respectively. For the same varieties the Na values were as follows : 7.16 ppm (in soil solution) : 4.8, 3.23, 4.21,4.58 ppm (in bleeding sap) respectively. K content in bleeding sap was higher than in soil solution for a few varieties, and lower in some varieties. Traces of Fe and Cu were found in both media.

Read More

Artificial intelligence (AI)-based protein modeling for the interpretation of grapevine genetic variants

Genetic variants known to produce single residue missense mutations have been associated with phenotypic traits of commercial interest in grapevine. This is the case of the K284N substitution in VviDXS1 associated with muscat aroma, or the R197L in VviAGL11 causing stenospermocarpic seedless grapes. The impact of such mutations on protein structure, stability, dynamics, interactions, or functional mechanism can be studied by computational methods, including our pyDock scoring, previously developed. For this, knowledge on the 3D structure of the protein and its complexes with other proteins and biomolecules is required, but such knowledge is not available for virtually none of the proteins and complexes in grapevine.

Read More

Determination of quality related polyphenols in chilean wines by absorbance-transmission and fluorescence excitation emission matrix (a-teem) analyses

Phenolic composition is essential to wine quality (Cleary et al., 2015; Bindon et al., 2020; Niimi et al., 2020) and its assessment is a strong industrial need to quality management.

Read More

Cultivation forms and viticulture models adapting to adverse “environmental” conditions

One of the main problems in viticultural production in Istria (Croatia) is a labour shortage in periods of intensive works, mainly during summer, respectively during tourist season.

Read More

Changes in white wine composition after treatment with cationic exchange resin: impact on wine oxidation after 8 years of bottle storage

Samples from 3 wine types were treated with a cationic exchange resin (7 lots) and stored for 8 years (47 samples). Forty-seven parameters were determined, including (1) important substrates with impact in white wine oxidation and (2) markers of oxidation. From group 1, sugars, elements, phenolic compounds, α-dicarbonyls and SO2 and from group 2, browning (A420), acetaldehyde, alkanals, furanic compounds were quantified.

Read More