Establishment of a CRISPR-based system for rapidly detecting the target-site resistance of American sloughgrass (Beckmannia syzigachne) to Pinoxaden.

BACKGROUND Weeds resistant to herbicides pose significant challenges in crop production, making early resistance monitoring crucial for timely control of resistant weeds. Prolonged use of ACCase-inhibiting herbicides, like pinoxaden, has led to the evolution of high-level resistance in weed populations over time. American sloughgrass (Beckmannia syzigachne), is a noxious grass weed, that severely impacts the yield and quality of wheat and rapeseed crops. RESULTS To accurately and rapidly detect the mutations in the target gene of B. syzigachne, we developed a novel rapid detection method based on the CRISPR-Cas12b/sgRNA system to evaluate the target mutation at amino acid position 1781 of the ACCase gene. By optimizing various reaction conditions, the novel detection system can assess target-site resistance of B. syzigachne to pinoxaden within 40 min at a constant temperature of 54 °C. This novel system exhibits excellent specificity, high sensitivity, simplicity in procedure, also with time-efficient and high throughput. CONCLUSION This study presents an efficient method based on the CRISPR-Cas12b system for rapidly detecting the target-site resistance, which will facilitate the precise management of resistant weeds. © 2025 Society of Chemical Industry.