Yin Hanqi, Ge Zhixun, Yao Sai, Chen Guoqi, Yuan Shuzhong, Yang Qian, Deng Wei
College of Plant Protection, Yangzhou University, Yangzhou, China.
Jiangsu Key Laboratory of Crop Genetics and Physiology/Jiangsu Key Laboratory of Crop Cultivation and Physiology, Agricultural College/Research Institute of Rice Industrial Engineering Technology of Yangzhou University, Jiangsu Co-Innovation Center for Modern Production Technology of Grain Crops, Yangzhou University, Yangzhou, China.
Pest Manag Sci. 2025 Sep;81(9):5581-5588. doi: 10.1002/ps.8911. Epub 2025 May 21.
Leptochloa fusca is a new dominant grass weed in paddy ecosystem of local region in China. The extensive herbicide application has selected the cyhalofop-butyl-resistant L. fusca populations. Here, we aimed to (i) characterize the target-site resistance mechanisms to ACCase inhibitors, and (ii) establish an efficient and rapid method for detecting mutations.
The L. fusca population (SFC-R) was resistant to cyhalofop-butyl (RF = 5.4), metamifop (RF = 5.5), and fenoxaprop-P-ethyl (RF = 6.8). Gene sequencing revealed that the Ile-1781-Val mutation was the molecular basis of resistance. A recombinase-aided amplification (RAA) combined with CRISPR/Cas12a method was successfully developed, which could rapidly and visually detect the single nucleotide variation underling the Ile-1781-Val substitution in L. fusca.
This study confirmed that target-site mutation (Ile-1781-Val) endowed resistance to ACCase inhibitors in L. fusca, and represented the first report of applying the RAA-CRISPR/Cas12a assay for mutation detection in resistant weeds. Considering its simplicity and specificity, this technique may be used in monitoring and early warning of resistant weeds. © 2025 Society of Chemical Industry.
细叶千金子是中国当地稻田生态系统中一种新的优势禾本科杂草。广泛使用除草剂已筛选出对氰氟草酯具有抗性的细叶千金子种群。在此，我们旨在：（i）表征对乙酰辅酶A羧化酶（ACCase）抑制剂的靶标位点抗性机制，以及（ii）建立一种高效快速的突变检测方法。
细叶千金子种群（SFC-R）对氰氟草酯（抗性倍数RF = 5.4）、噁唑酰草胺（RF = 5.5）和精噁唑禾草灵（RF = 6.8）具有抗性。基因测序表明，Ile-1781-Val突变是抗性的分子基础。成功开发了一种重组酶辅助扩增（RAA）与CRISPR/Cas12a相结合的方法，该方法可快速、直观地检测细叶千金子中Ile-1781-Val替换的单核苷酸变异。
本研究证实靶标位点突变（Ile-1781-Val）赋予细叶千金子对ACCase抑制剂的抗性，这是首次报道将RAA-CRISPR/Cas12a检测方法应用于抗性杂草的突变检测。鉴于其简单性和特异性，该技术可用于抗性杂草的监测和预警。© 2025化学工业协会。
