Plant resistance (R) and pathogen avirulence (Avr) gene interactions play key roles in crop resistance to pathogens. Efficient molecular screening tools for crop species are required to define interactions that trigger host resistance and to understand the virulence mechanisms of pathogenic fungi. Avr identification supports genome-based pathogen surveillance to enhance pre-emptive breeding of disease resistant cereal crops. To this end, we have developed a novel wheat protoplast assay that enables efficient screening of Avr/R interactions at scale. Our assay allows access to the extensive gene pool of phenotypically described R genes because it does not require the overexpression of cloned R genes and is applicable to all wheat cultivars tested so far. It is suitable for multiplexed Avr screening, with interactions tested in pools of up to ten Avr candidates. Our assay is based on newly identified Avr/R-induced marker genes which we used to create promoter-luciferase reporter constructs. We combined these reporter constructs with a dual-color ratiometric reporter system that normalizes read-outs accounting for experimental variability and Avr/R-induced cell death. Our new assay increases the throughput of Avr candidate screening, accelerating the study of cellular defense signaling and resistance gene identification in wheat. We anticipate that the uptake of our assay by the community will significantly accelerate Avr identification for many wheat pathogens, leading to improved genomic pathogen surveillance and breeding of disease resistant crops.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.