Arndell Taj, Chen Jian, Sperschneider Jana, Upadhyaya Narayana M, Blundell Cheryl, Niesner Nathalie, Outram Megan A, Wang Aihua, Swain Steve, Luo Ming, Ayliffe Michael A, Figueroa Melania, Vanhercke Thomas, Dodds Peter N
CSIRO Agriculture and Food, Canberra, Australian Capital Territory, Australia.
Centre for Agriculture and the Bioeconomy, Queensland University of Technology, Brisbane, Queensland, Australia.
Nat Plants. 2024 Apr;10(4):572-580. doi: 10.1038/s41477-024-01641-y. Epub 2024 Feb 26.
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 years, 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.
由于病原体毒力的快速进化，培育具有持久抗病性的作物品种具有挑战性。尽管近年来抗性（R）基因克隆和累加方面取得了进展，但许多病原体中相应无毒（Avr）基因的鉴定因缺乏高通量筛选方法而受到阻碍。为了填补这一技术空白，我们开发了一个用于在植物原生质体中进行混合文库筛选的平台，以便快速鉴定相互作用的R-Avr对。通过针对单个R基因筛选设计的假定效应子文库后，从小麦条锈菌（Puccinia graminis f. sp. tritici）中分离出已知和新的Avr基因，我们验证了该平台。快速鉴定Avr基因提供了分子工具，可通过基因型监测来理解和追踪病原体毒力进化，这反过来又将导致优化的R基因累加和部署策略。该平台应广泛适用于许多作物病原体，并有可能适用于筛选参与其他原生质体可选择性状的基因。
