학술논문

Pooled effector library screening in protoplasts rapidly identifies novel Avr genes
Document Type
Original Paper
Source
Nature Plants. 10(4):572-580
Subject
Language
English
ISSN
2055-0278
Abstract
Crop breeding for durable disease resistance is challenging due to the rapid evolution of pathogen virulence. While progress in resistance (R) gene cloning and stacking has accelerated in recent years1–3, the identification of corresponding avirulence (Avr) genes in many pathogens is hampered by the lack of high-throughput screening options. To address this technology gap, we developed a platform for pooled library screening in plant protoplasts to allow rapid identification of interacting R–Avr pairs. We validated this platform by isolating known and novel Avr genes from wheat stem rust (Puccinia graminis f. sp. tritici) after screening a designed library of putative effectors against individual R genes. Rapid Avr gene identification provides molecular tools to understand and track pathogen virulence evolution via genotype surveillance, which in turn will lead to optimized R gene stacking and deployment strategies. This platform should be broadly applicable to many crop pathogens and could potentially be adapted for screening genes involved in other protoplast-selectable traits.
The authors developed a platform for rapid identification of interacting plant immune receptors and pathogen avirulence proteins by library screening in protoplasts, then used it to identify new wheat stem rust Avr genes recognized by known wheat resistance genes.