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…

Closing the carbon loop: evaluating the potential of grapevine-derived biochar as a soil conditioner in warm climate vineyards

Significant increases in anthropogenic carbon dioxide (CO2) emissions due to combustion of fossil fuels and intensive land management practices that release CO2 into the atmosphere have resulted in higher air temperatures due to the greenhouse effect.

Spatial Analysis of Climate in Winegrape Growing Regions in Portugal

Spatial climate data at a 1 km resolution has allowed for a comprehensive mapping and assessment of viticulture DOs regions in Portugal. Overall the 50 regions and sub-regions in Portugal range

Tropical fruit aroma in white wines: the role of fermentation esters and volatile thiols

Volatile thiols are impact aroma compounds, well-known in the literature for imparting tropical fruit aromas such as passion fruit, guava, grapefruit, and citrus in white wines [1]. More recent evidence suggests that tropical fruit aromas are also caused by other aroma compounds besides thiols, such as fermentation esters, or the interaction between these volatile families. Therefore, the objective of this study was to investigate the effects of combining esters and/or thiols to determine their impact on the fruitiness aroma perception of white wines. Pinot gris wine was produced at the OSU research winery and was dearomatized using Lichrolut® EN. Combinations of fermentation volatile compounds were added to the wine, forming the aroma base. Treatment wines were composed of additions of different concentrations and combinations of thiols and/or esters. Samples were subjected to sensory analysis where forty-six white wine consumers evaluated the orthonasal aroma of the wines and participated in Check-All-That-Apply (CATA).

Zoning like base instrument for the agronomist’s work in vineyard

Ad una prima analisi l’interesse dimostrato dal settore produttivo nei confronti della zonazione vitivinicola è da ricondursi al fatto che dopo i primi approcci puramente accademici

Trials with machine harvested sauvignon blanc: the importance of grape transport time and temperature

It is well known that free varietal thiols, in particular 3-mercaptohexanol (3MH) and 3-mercaptohexyl ace-tate (3MHA), are important constituents to the aroma of New Zealand Sauvignon blanc wines.