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

Abstract

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

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

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

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