稗草是中国东北稻田的一种恶性杂草，目前其抗药性问题日益严重。我们的研究以 HJHL-715 E. phyllopogon 种群为中心，通过全植物生物测定，该种群对五氟磺草胺表现出较高的抗性。用 P450 抑制剂马拉硫磷预处理可显着提高抗性植物对五氟磺草胺的敏感性。为了确定抗性群体的抗性机制，我们从单株植物中纯化了抗性群体，并分离了靶点抗性（TSR）和非靶点抗性（NTSR）材料。耐药群体的乙酰乳酸合酶 (ALS) 1 和 ALS2 中的 Pro-197-Thr 和 Trp-574-Leu 突变导致五氟磺草胺对靶位点 ALS 的敏感性降低，这是主要的耐药机制。为了充分了解 NTSR 机制，通过使用 RNA 测序 (RNA-seq) 结合参考基因组对 NTSR 材料进行了研究。高效液相色谱-串联质谱 (HPLC-MS/MS) 分析进一步支持 NTSR 材料中五氟磺草胺代谢增强。基因表达数据和定量逆转录聚合酶链反应 (qRT-PCR) 验证证实，在五氟磺草胺处理下有 29 个基因过表达，其中 16 个基因在二氯喹啉酸和恶唑酰草胺处理下同时上调。总体而言，我们的研究证实了 E. phyllopogon 对 ALS 抑制剂的抗性中并存的 TSR 和 NTSR 机制。

Echinochloa phyllopogon, a malignant weed in Northeast China's paddy fields, is currently presenting escalating resistance concerns. Our study centered on the HJHL-715 E. phyllopogon population, which showed heightened resistance to penoxsulam, through a whole-plant bioassay. Pretreatment with a P450 inhibitor malathion significantly increased penoxsulam sensitivity in resistant plants. In order to determine the resistance mechanism of the resistant population, we purified the resistant population from individual plants and isolated target-site resistance (TSR) and nontarget-site resistance (NTSR) materials. Pro-197-Thr and Trp-574-Leu mutations in acetolactate synthase (ALS) 1 and ALS2 of the resistant population drove reduced sensitivity of penoxsulam to the target-site ALS, the primary resistance mechanisms. To fully understand the NTSR mechanism, NTSR materials were investigated by using RNA-sequencing (RNA-seq) combined with a reference genome. High-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) analysis further supported the enhanced penoxsulam metabolism in NTSR materials. Gene expression data and quantitative reverse transcription polymerase chain reaction (qRT-PCR) validation confirmed 29 overexpressed genes under penoxsulam treatment, with 16 genes concurrently upregulated with quinclorac and metamifop treatment. Overall, our study confirmed coexisting TSR and NTSR mechanisms in E. phyllopogon's resistance to ALS inhibitors.