油菜5N对ALS抑制剂类除草剂的交互抗性机理

doi:10.3864/j.issn.0578-1752.2026.09.005

中国农业科学 ›› 2026, Vol. 59 ›› Issue (9): 1887-1902.doi: 10.3864/j.issn.0578-1752.2026.09.005

油菜5N对ALS抑制剂类除草剂的交互抗性机理

郭月¹^,²(), 张云龙¹, 刘德才¹, 王俊禹¹, 刘湟捷¹^,⁴, 邵晓琪¹^,⁴, 彭琦¹^,², 于仕程¹^,⁴, 祁余容¹^,⁴, 高建芹¹, 张洁夫¹, 胡茂龙¹^,³^,⁴()

¹ 江苏省农业科学院经济作物研究所/国家油料作物改良中心南京分中心/农业农村部长江下游棉花与油菜重点实验室，南京 210014
² 江苏大学食品与生物工程学院，江苏镇江 212013
³ 江苏大学生命科学学院，江苏镇江 212013
⁴ 福建农林大学农学院，福州 350002

收稿日期:2025-12-22 接受日期:2026-01-27 出版日期:2026-05-01 发布日期:2026-05-06
通信作者:
胡茂龙，E-mail：humolon@163.com
联系方式: 郭月，E-mail：guoguo23.25@163.com。
基金资助:
江苏省基础研究计划(BK20251970); 科技创新2030—重大项目(2023ZD0404203); 国家自然科学基金青年科学基金(31901503); 江苏省农业科技自主创新资金(CX(22)3084); 国家农业现代产业技术体系建设专项(CARS-12); 江苏省农业科学院科研基金(KYJJ(25)0503)

The Cross-Resistance Mechanism of Rapeseed 5N to ALS Inhibitor Herbicides

GUO Yue¹^,²(), ZHANG YunLong¹, LIU DeCai¹, WANG JunYu¹, LIU HuangJie¹^,⁴, SHAO XiaoQi¹^,⁴, PENG Qi¹^,², YU ShiCheng¹^,⁴, QI YuRong¹^,⁴, GAO JianQin¹, ZHANG JieFu¹, HU MaoLong¹^,³^,⁴()

¹ Institute of Industrial Crops, Jiangsu Academy of Agricultural Sciences/Nanjing Sub-Center, National Center of Oil Crops Improvement/Key Laboratory of Cotton and Rapeseed (Nanjing), Ministry of Agriculture and Rural Affairs, Nanjing 210014
² School of Food and Biological Engineering, Jiangsu University, Zhenjiang 212023, Jiangsu
³ School of Life Sciences, Jiangsu University, Zhenjiang 212023, Jiangsu
⁴ College of Agriculture, Fujian Agriculture and Forestry University, Fuzhou 350002

Received:2025-12-22 Accepted:2026-01-27 Published:2026-05-01 Online:2026-05-06

摘要/Abstract

摘要：

【目的】高抗磺酰脲（SU）类除草剂油菜5N已广泛参与抗除草剂油菜育种利用，但其对其他4类ALS抑制剂类除草剂的交互抗性谱系及机制尚不清晰。本研究旨在系统评价5N对5大类ALS抑制剂的交互抗性水平，并阐明其抗性机制，为该种质的科学利用提供理论依据。【方法】通过苗期外源喷施5类ALS抑制剂类除草剂（磺酰脲-SU、磺酰胺羧基三唑啉酮-SCT、咪唑啉酮-IMI、嘧啶水杨酸-PB和三唑嘧啶-TP），鉴定其抗性水平和生物量的毒理学响应；通过体外酶活性抑制试验与实时荧光定量PCR（qRT-PCR）技术分析靶标酶敏感性及两个靶基因的表达模式；同时测定丙二醛含量及多种抗氧化酶活性以评估氧化应激响应。【结果】5N对SU和SCT类除草剂表现出16倍田间推荐浓度（16×RC）的高抗性，对IMI类为4×RC的中等抗性，对PB和TP类抗性较低；生物量毒理学响应结果与上述抗性谱趋势一致。机理研究表明：（1）靶标机制：5N的ALS酶对各类除草剂（20—300 µmol·L^-1）的敏感性均显著低于N131，且其ALS的表达上调幅度与持续时间显著高于N131；（2）非靶标机制：SU和SCT类除草剂处理后，5N中丙二醛含量、超氧化物歧化酶、过氧化氢酶、过氧化物酶和谷胱甘肽还原酶活性以及解毒基因BnCYP86A2、BnCYP71B3、BnGST39和BnGST78的表达量均显著高于N131，而在PB、IMI和TP类除草剂处理组较为复杂。【结论】油菜5N对ALS抑制剂类除草剂存在广谱但差异性的交互抗性，该抗性主要由靶标酶的低敏感性与靶基因的突变共同介导，而增强的解毒和抗氧化能力等非靶标抗性机制对抗性表型的产生也起到了辅助作用。研究明确了SCT类除草剂可作为苗期SU类的安全替代选择，并为在抗除草剂育种中合理利用5N的交互抗性性状提供了技术支撑。

关键词: 油菜, 5N, ALS抑制剂类除草剂, 交互抗性, 抗性机制

Abstract:

【Objective】The high sulfonylurea (SU) resistance germplasm 5N has been widely used in the breeding of rapeseed resistant to herbicides. However, the cross-resistance spectrum and mechanisms of 5N to the other four classes of ALS-inhibitor herbicides remain unclear. The study aims to systematically evaluate the cross-resistance level of 5N to the five classes of ALS inhibitors and elucidate its resistance mechanisms, which will provide a theoretical basis for the scientific utilization of 5N.【Method】The resistance levels and toxicological responses in biomass were detected by exogenous spraying of five kinds of ALS inhibitor herbicides (sulfonylurea-SU, sulfonlyaminocarbonyl-triazolinone-SCT, imidazolinone-IMI, pyrimidyl-benzoate-PB and triazolopyrimidine-TP) at the seedling stage. The sensitivity of the target enzyme and the expression patterns of two target genes were analyzed through in vitro enzyme activity inhibition assays and qRT-PCR. Simultaneously, malondialdehyde content and the activities of multiple antioxidant enzymes were measured to evaluate the oxidative stress response.【Result】5N exhibited the cross-resistance of high level of 16 times the recommended concentration (16×RC) to SU and SCT herbicides, mid-level of 4×RC to IMI herbicides, low level to PB and TP herbicides, which was consistent with the trend in the phytotoxic response of biomass. The mechanism revealed that: (1) Target-site mechanism: The sensitivity of the ALS enzyme in 5N to various herbicides (20-300 µmol·L^-1) was significantly lower than that in N131, and the upregulated magnitude and duration of its ALS expression were notably higher than those in N131; (2) Non-target-site mechanism: After treatment with SU and SCT herbicides, the malondialdehyde content, the activities of superoxide dismutase, catalase, peroxidase, and glutathione reductase, as well as the expression levels of detoxification genes such as BnCYP86A2, BnCYP71B3, BnGST39, and BnGST78 in 5N were significantly higher than those in N131. In contrast, the responses under PB, IMI, and TP treatments were more complex and diverse.【Conclusion】5N exhibits broad-spectrum but differential cross-resistance to ALS inhibitors. This resistance is primarily mediated by both the low sensitivity of the target enzyme and mutations in the target gene, while non-target-site resistance mechanisms such as enhanced detoxification and antioxidant capacity, also have an assistant effect on the resistant phenotype. This study shows that SCT herbicides can serve as a safe alternative to SU herbicides at the seedling stage, and provides technical support for the rational utilization of the cross-resistance trait of 5N in herbicide-resistant breeding.

Key words: rapeseed (Brassica napus), 5N, ALS inhibitor herbicide, cross-resistance, resistance mechanism

郭月, 张云龙, 刘德才, 王俊禹, 刘湟捷, 邵晓琪, 彭琦, 于仕程, 祁余容, 高建芹, 张洁夫, 胡茂龙. 油菜5N对ALS抑制剂类除草剂的交互抗性机理[J]. 中国农业科学, 2026, 59(9): 1887-1902.

GUO Yue, ZHANG YunLong, LIU DeCai, WANG JunYu, LIU HuangJie, SHAO XiaoQi, PENG Qi, YU ShiCheng, QI YuRong, GAO JianQin, ZHANG JieFu, HU MaoLong. The Cross-Resistance Mechanism of Rapeseed 5N to ALS Inhibitor Herbicides[J]. Scientia Agricultura Sinica, 2026, 59(9): 1887-1902.

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除草剂类别 Herbicide category	除草剂及有效成分含量 Herbicides and active ingredient content	田间推荐使用浓度1倍（1×RC） Recommended concentration for field use	对应国标单位浓度 Corresponding to the national standard unit concentration (mmol·L^-1)
SU	25%噻吩磺隆Thifensulfuron-methyl	8 g/667 m²	0.17
SU	10%苄嘧磺隆Bensulfuron-methyl	20 g/667 m²	0.16
TP	25 g·L^-1五氟磺草胺Penoxsulam	80 mL/667 m²	0.14
TP	50 g·L^-1双氟磺草胺Florasulam	6 mL/667 m²	0.028
PB	10%双草醚Bispyribac-sodium	20 mL/667 m²	0.15
PB	5%嘧啶肟草醚Pyribenzoxim	50 mL/667 m²	0.14
SCT	70%氟唑磺隆Flucarbazone-sodium	3.5 g/667 m²	0.21
SCT	99.5%噻酮磺隆Thiencarbazone-methyl	4 g/667 m²	0.34
IMI	4%甲氧咪草烟Imazamox	80 mL/667 m²	0.35
IMI	5%咪唑乙烟酸Imazethapyr	133 mL/667 m²	0.72

名称Name	序列Sequence (5′-3′)	大小Size (bp)	名称Name	序列Sequence (5′-3′)	大小Size (bp)
BnALS1-F2	TTCTCCTTAACCCCACAGAAAGA	23	CYP86A2-F	AAGGACGGAGAGAAACGACG	20
BnALS1-R2	GGGAGCGTAGCGGGAGAC	18	CYP86A2-R	AGGAGACGCCTTCACCAATG	20
BnALS3-F2	CTCCTTAACCCCACAGAAACC	21	CYP71B3-F	CTTGAGTGTTGCACAAGACCG	21
BnALS3-R2	GGGAGCGTAGCGGGAGAT	18	CYP71B3-R	CACCATATGGCGCAAAGCAA	20
EIF4B-F	GTGAGCTGGACGACAAAGTC	20	BnGST39-F	GGCTAGCTACATCCAGTTACAT	22
EIF4B-R	CTCTGATCATCGGCAGGTCT	20	BnGST39-R	TCACAATGTCAACTAGGCTCAT	22
BnALS1-P	Fam+CCGTCAATGTCGCACCTCCTTCC+Tamra	23	BnGST78-F	CCTCAAAGAGTCTTGCTTTGTC	22
BnALS3-P	Fam+CAACTCACCCGTCAATGTCGCACC+Tamra	24	BnGST78-R	ACGAAGAAGATGTTCAGGTCTT	22
EIF4B-P	Fam+TGCCTGCACCAGACCCGGGT+Tamra	20

种类Category	名称 Name	植株 Plant	喷施剂量Spray dose (RC)
种类Category	名称 Name	植株 Plant	0×	1/4×	1/2×	1×	2×	4×	8×	16×	20×
SU	噻吩磺隆Thifensulfuron-methyl	5N/N131	R/R	R/S	R/S	R/S	R/S	R/S	R/S	R/S	M/S
SU	苄嘧磺隆Bensulfuron-methyl	5N/N131	R/R	R/S	R/S	R/S	R/S	R/S	R/S	R/S	M/S
TP	五氟磺草胺Penoxsulam	5N/N131	R/R	R/S	R/S	R/S	R/S	S/S	S/S	S/S	S/S
TP	双氟磺草胺Florasulam	5N/N131	R/R	R/S	R/S	R/S	R/S	M/S	S/S	S/S	S/S
PB	双草醚Bispyribac-sodium	5N/N131	R/R	R/S	R/S	R/S	R/S	M/S	S/S	S/S	S/S
PB	嘧啶肟草醚Pyribenzoxim	5N/N131	R/R	R/S	R/S	R/S	R/S	M/S	S/S	S/S	S/S
SCT	氟唑磺隆Flucarbazone-sodium	5N/N131	R/R	R/S	R/S	R/S	R/S	R/S	R/S	R/S	M/S
SCT	噻酮磺隆Thiencarbazone-methyl	5N/N131	R/R	R/S	R/S	R/S	R/S	R/S	R/S	R/S	M/S
IMI	甲氧咪草烟Imazamox	5N/N131	R/R	R/S	R/S	R/S	R/S	R/S	S/S	S/S	S/S
IMI	咪唑乙烟酸Imazethapyr	5N/N131	R/R	R/S	R/S	R/S	R/S	R/S	S/S	S/S	S/S

油菜 Rapeseed	噻吩磺隆 Thifensulfuron-methyl (mmol·L^-1)	双氟磺草胺 Florasulam (μmol·L^-1)	双草醚 Bispyribac-sodium (μmol·L^-1)	噻酮磺隆 Thiencarbazone-methyl (mmol·L^-1)	咪唑乙烟酸 Imazethapyr (mmol·L^-1)
N131	0.019±0.004	0.154±0.033	7.300±2.311	0.039±0.007	0.050±0.012
5N	2.771±0.862	6.116±1.046	295.701±39.020	5.462±0.637	3.097±0.486

油菜 Rapeseed	噻吩磺隆 Thifensulfuron-methyl	双氟磺草胺 Florasulam	双草醚 Bispyribac-sodium	噻酮磺隆 Thiencarbazone-methyl	咪唑乙烟酸 Imazethapyr
N131	29.510±1.654	25.240±2.444	14.180±1.756	19.980±4.847	23.850±1.482
5N	934.700±24.640	76.020±9.142	35.130±5.421	608.300±47.060	249.700±61.860

油菜5N对ALS抑制剂类除草剂的交互抗性机理

The Cross-Resistance Mechanism of Rapeseed 5N to ALS Inhibitor Herbicides

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