Ban Yaxin, Zhang Zhanzhan, Wei Jianguo, Xu Gaofeng, Ma Youmei, Yin Shaojing, Dong Liyao, Feng Zhike
State Key Laboratory of Agricultural and Forestry Biosecurity, College of Plant Protection, Nanjing Agricultural University, Nanjing, China.
Institute of Agricultural Environment and Resources Research, Yunnan Academy of Agricultural Sciences, Kunming, China.
Pest Manag Sci. 2025 Sep;81(9):5649-5658. doi: 10.1002/ps.8919. Epub 2025 May 26.
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.
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.
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.
抗除草剂杂草给作物生产带来重大挑战，因此早期抗性监测对于及时控制抗性杂草至关重要。随着时间的推移，像炔草酯这样的乙酰辅酶A羧化酶（ACCase）抑制性除草剂的长期使用导致杂草种群产生了高水平抗性。美国碱茅（Beckmannia syzigachne）是一种有害杂草，严重影响小麦和油菜作物的产量和质量。
为了准确快速地检测美国碱茅靶基因中的突变，我们基于CRISPR-Cas12b/sgRNA系统开发了一种新型快速检测方法，以评估ACCase基因第1781位氨基酸处的靶突变。通过优化各种反应条件，该新型检测系统能够在54℃恒温下40分钟内评估美国碱茅对炔草酯的靶位点抗性。这种新型系统具有出色的特异性、高灵敏度、操作简单，且省时、高通量。
本研究提出了一种基于CRISPR-Cas12b系统的高效快速检测靶位点抗性的方法，这将有助于抗性杂草的精准治理。© 2025化学工业协会。
