我国棉蚜田间种群对氧乐果的抗性监测及靶标抗性机制解析

doi:10.3864/j.issn.0578-1752.2026.18.007

Abstract

Abstract:

【Objective】The objective of this study is to determine the resistance level of Aphis gossypii populations from the main cotton planting areas of China to omethoate, to elucidate the predominant mutant types in acetylcholinesterase 1 (AChE1) (the target of omethoate) in these populations, and further analyze the impacts on the AChE1 structure by these major mutation types and their relationship with omethoate resistance.【Method】In 2019-2023, A. gossypii field populations were collected from the main cotton planting areas of China, and the resistance levels of these populations to omethoate were determined using the leaf-dipping method. Mutations in AChE1 were detected by single-aphid PCR amplification and sequencing, and the types of mutations carried by an individual aphid were analyzed. Monoclonal strains carrying different AChE1 mutations were established using single-female rearing protocols, and their contributions to omethoate resistance were subsequently assessed through toxicity bioassays and synergist experiments. The mechanisms of A. gossypii resistance to omethoate mediated by these mutations were revealed using homology modeling, molecular docking, and molecular dynamics simulations.【Result】In 2019-2023, 38 A. gossypii field populations in China developed moderate to high levels of resistance to omethoate (17.1-207.8-fold) and remained relatively stable over the years. G221A, A302S, A332V, and S431F mutations in AChE1 were prevalent in these populations, with S431F mutation frequency reaching 100%, predominantly in homozygous form. While the average frequencies of G221A, A302S, and A332V were 37.26%, 51.26%, and 74.64%, respectively. Detection of mutant types from a single aphid revealed that individuals carrying the A332V+S431F combination mutation had the highest frequency (35.24%), followed by G221A+A302S+A332V+S431F (26.80%) and A302S+A332V+S431F (12.12%). The average frequency of the S431F single mutation was 9.56%. The weighted frequencies of these major mutation types showed a certain linear correlation with omethoate resistance (R²=0.46, P<0.001). By constructing monoclonal mutant strains, it was confirmed that A. gossypii strains carrying the combination mutations A332V+S431F, G221A+A302S+S431F, A302S+A332V+S431F, and G221A+A302S+A332V+S431F developed a moderate level of resistance to omethoate, with resistance ratios of 46.1-77.8-fold, significantly higher than that of the S431F single-mutant strain (12.5-fold). Moreover, PBO, TPP, and DEM did not exhibit significant synergistic effects on omethoate toxicity in these mutant strains. Molecular docking and dynamics simulations confirmed that the S431F single mutation and A332V+S431F, G221A+A302S+S431F, A302S+A332V+S431F, and G221A+A302S+A332V+S431F combination mutations significantly increased the geometric center distance between the AChE1 catalytic triad and omethoate by 0.074-0.185 nm compared to the wild-type. These mutations also altered the hydrophobic area and radius of gyration of the AChE1 active center, thereby affecting the active center of A. gossypii AChE1 and its binding conformation with omethoate.【Conclusion】A. gossypii populations in major cotton-planting regions of China have developed moderate to high levels of resistance to omethoate. Four mutations were identified in AChE1 across these populations, and these site mutations altered the active center of AChE1 and its binding affinity for omethoate through different combination types, thereby mediating field resistance to this insecticide. In field control practices, it is advisable to reduce the use of AChE-targeting insecticides to prevent the development of cross-resistance mediated by target mutation and further escalation of resistance levels.

Key words: Aphis gossypii, omethoate, acetylcholinesterase, resistance, mutation

LI Ren, CHENG ShenHang, LIANG PingZhuo, ZHANG Lei, WANG ShanNing. Monitoring of Resistance to Omethoate in Field Populations of Aphis gossypii in China and Elucidation of Target Resistance Mechanisms[J].Scientia Agricultura Sinica, 2026, 59(18): 4033-4047.

Figures/Tables 11

Table 1

The collecting sites of A. gossypii field populations in 2019-2023"

年份 Year	种群 Population	位置 Location	经纬度 Longitude, latitude (E, N)	年份 Year	种群 Population	位置 Location	经纬度 Longitude, latitude (E, N)
2019	HeBCZ	河北沧州Cangzhou, Hebei	116.87°, 38.31°	2021	HNNY	河南南阳Nanyang, Henan	112.54°, 33.00°
	HeBHS	河北衡水Hengshui, Hebei	115.58°, 37.55°		SDBZ	山东滨州Binzhou, Shandong	118.02°, 37.43°
	SDDY	山东东营Dongying, Shandong	118.58°, 37.45°		SXYC	山西运城Yuncheng, Shanxi	111.00°, 35.02°
	SDHZ	山东菏泽Heze, Shandong	115.57°, 35.07°		XJCJ	新疆昌吉Changji, Xinjiang	87.31°, 44.01°
	SDJN	山东济宁Jining, Shandong	116.60°, 35.41°		XJKC	新疆库车Kuche, Xinjiang	86.17°, 42.03°
	XJKEL	新疆库尔勒Kuerle, Xinjiang	86.17°, 41.73°		XJKT	新疆奎屯Kuitun, Xinjiang	84.90°, 44.43°
	XJSHZ	新疆石河子Shihezi, Xinjiang	86.08°, 44.31°		XJTMSK	新疆图木舒克Tumushuke, Xinjiang	79.21°, 40.00°
	XJNWS	新疆农五师Nongwushi, Xinjiang	82.41°, 44.21°		XJTLF	新疆吐鲁番Tulufan, Xinjiang	89.19°, 42.94°
2020	SDDY	山东东营Dongying, Shandong	118.58°, 37.45°		XJYL	新疆伊犁Yili, Xinjiang	81.32°, 43.92°
	HeBHS	河北衡水Hengshui, Hebei	115.58°, 37.55°	2022	HuBJZ	湖北荆州Jingzhou, Hubei	112.02°, 30.05°
	SDXJ	山东夏津Xiajin, Shandong	116.00°, 36.95°		SDDY	山东东营Dongying, Shandong	118.41°, 37.05°
	SXYC	山西运城Yuncheng, Shanxi	111.00°, 35.02°		XJWJQ	新疆五家渠Wujiaqu, Xinjiang	87.54°, 44.17°
	XJBL	新疆博乐Bole, Xinjiang	82.05°, 44.85°		XJSW	新疆沙湾Shawan, Xinjiang	85.62°, 44.33°
	XJKEL	新疆库尔勒Kuerle, Xinjiang	86.17°, 41.73°	2023	SDTZ	山东滕州Tengzhou, Shandong	116.00°, 36.95°
	XJSHZ	新疆石河子Shihezi, Xinjiang	86.08°, 44.31°		SDDY	山东东营Dongying, Shandong	118.41°, 37.05°
	XJWS	新疆乌苏Wusu, Xinjiang	84.68°, 44.44°		HuBJZ	湖北荆州Jingzhou, Hubei	112.19°, 30.35°
	XJYL	新疆伊犁Yili, Xinjiang	81.32°, 43.92°		XJHYH	新疆胡杨河Huyanghe, Xinjiang	84.83°, 44.69°
2021	HeBHS	河北衡水Hengshui, Hebei	115.58°, 37.55°		XJKS	新疆喀什Kashi, Xinjiang	75.99°, 39.47°
2021	HNKF	河南开封Kaifeng, Henan	114.35°, 34.79°		XJHT	新疆和田Hetian, Xinjiang	79.92°, 37.11°

Table 1

Table 2

The resistance of A. gossypii field populations to omethoate in 2019-2023"

年份 Year	种群 Population	试虫数 Number of test insects	斜率±标准误Slope±SE	致死中浓度（95%置信限） LC₅₀(95% CL) (mg·L^-1)	抗性倍数Resistance ratio	卡方值（自由度） χ²(df)
2019	HeBCZ	434	2.39±0.23	247.79 (204.60-300.85)	41.2	12.47 (12)
	HeBHS	403	1.38±0.20	1022.21 (758.50-1567.85)	169.8	12.17 (13)
	SDDY	464	1.25±0.14	282.29 (210.85-359.73)	46.9	8.61 (14)
	SDHZ	457	1.47±0.14	400.68 (328.38-495.35)	66.6	11.25 (13)
	SDJN	463	1.41±0.15	372.81 (289.67-463.10)	61.9	12.48 (15)
	XJKEL	449	1.64±0.15	468.96 (380.87-570.35)	77.9	9.38 (14)
	XJNWS	525	1.49±0.17	102.90 (69.97-139.53)	17.1	8.29 (16)
	XJSHZ	445	2.08±0.20	383.24 (310.00-460.45)	63.7	10.71 (13)
2020	HeBHS	522	1.13±0.13	590.32 (453.63-807.69)	98.1	4.43 (11)
	SDDY	451	3.52±0.49	729.43 (573.32-864.53)	121.2	14.60 (12)
	SDXJ	484	2.29±0.48	508.70 (331.93-666.55)	84.5	14.61 (13)
	SXYC	544	1.63±0.18	1217.41 (944.02-1648.82)	202.2	16.35 (13)
	XJBL	536	1.19±0.14	636.75 (482.49-917.19)	105.8	4.77 (13)
	XJKEL	469	1.79±0.17	387.28 (308.77-475.30)	64.3	11.73 (13)
	XJSHZ	444	1.46±0.19	624.73 (444.95-980.10)	103.8	13.35 (10)
	XJWS	507	2.48±0.22	1251.02 (1015.26-1517.09)	207.8	18.63 (13)
	XJYL	505	2.30±0.18	389.62 (327.96-467.65)	64.7	13.58 (13)
2021	SDBZ	626	1.71±0.16	592.34 (476.68-720.05)	98.4	14.48 (16)
	XJCJ	597	1.75±0.17	343.13 (233.55-462.08)	57.0	18.72 (14)
	HeBHS	568	1.31±0.11	746.39 (577.30-980.07)	124.0	17.24 (16)
	HNKF	616	2.17±0.21	165.80 (117.15-217.33)	27.5	17.05 (14)
	XJKC	677	2.33±0.22	438.88 (349.80-527.30)	72.9	16.54 (16)
	XJKT	632	2.11±0.24	365.47 (263.43-465.18)	60.7	16.06 (15)
	HNNY	577	2.17±0.21	196.79 (127.87-274.43)	32.7	15.97 (10)
	XJTLF	610	1.70±0.14	674.63 (552.27-819.63)	112.1	12.80 (13)
	XJTMSK	597	1.92±0.18	1200.53 (1016.26-1434.19)	199.4	9.71 (14)
	XJYL	579	2.11±0.28	735.29 (512.96-955.97)	122.1	16.62 (13)
	SXYC	622	1.60±0.15	380.01 (274.01-504.93)	63.1	15.54 (13)
2022	SDDY	675	1.81±0.21	541.21 (409.16-669.77)	89.9	12.47 (16)
	HuBJZ	670	1.78±0.26	308.75 (147.39-465.93)	51.3	20.78 (12)
	XJSW	670	1.93±0.22	824.45 (605.06-1066.98)	137.0	19.38 (14)
	XJWJQ	582	1.82±0.23	1234.02 (940.00-1656.41)	205.0	19.01 (14)
2023	SDDY	492	2.39±0.34	372.95 (262.56-478.86)	62.0	12.67(12)
	XJHYH	470	2.41±0.43	645.37 (458.83-848.08)	107.2	18.56 (13)
	SDTZ	458	1.22±0.24	404.84 (238.08-602.15)	67.2	5.01 (13)
	HuBJZ	407	1.60±0.22	141.41 (72.98-218.58)	23.5	18.48 (13)
	XJKS	534	4.22±0.84	805.12 (630.18-942.79)	133.7	14.73 (10)
	XJHT	525	1.25±0.33	738.51 (452.45-1133.60)	122.7	8.40 (12)

Table 2

Fig. 1

Fig. 2

Fig. 3

Table 3

Fig. 4

Fig. 5

Fig. 6

Fig. 7

Table 4

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品系 Strain	药剂+增效剂 Insecticide+ synergist	试虫数 Number of test insects	斜率±标准误 Slope±SE	致死中浓度（95%置信限） LC₅₀(95% CL) (mg·L^-1)	P值 P value	抗性倍数 Resistance ratio	增效倍数 Synergism ratio
S431F (RR)	OME	534	3.70±0.38	75.0 (64.3-87.3)	0.889	12.5	—
	OME+TPP	539	2.18±0.19	81.0 (64.7-98.8)	0.897	13.5	0.93
	OME+DEM	533	2.23±0.21	86.5 (67.3-107.0)	0.909	14.4	0.87
	OME+PBO	523	2.05±0.19	78.5 (60.5-98.1)	0.843	13.0	0.96
A332V (RR)+S431F (RR)	OME	500	2.24±0.33	277.3 (195.3-352.8)	0.871	46.1	—
	OME+TPP	524	3.34±0.56	346.0 (234.4-427.2)	0.141	57.5	0.80
	OME+DEM	505	5.35±0.76	408.3 (338.8-471.1)	0.175	67.8	0.68
	OME+PBO	509	4.55±0.76	363.0 (297.2-414.7)	0.843	60.3	0.76
G221A (RS)+A302S (RR) +S431F (RR)	OME	500	4.59±0.72	410.8 (281.9-510.0)	0.124	68.2	—
	OME+TPP	561	2.93±0.39	379.3 (289.4-459.2)	0.230	63.0	1.08
	OME+DEM	515	2.71±0.35	468.4 (387.2-549.3)	0.529	77.8	0.88
	OME+PBO	520	2.58±0.57	405.6 (263.2-501.7)	0.773	67.4	1.01
A302S (RS)+A332V (RS) +S431F (RR)	OME	525	2.25±0.35	410.8 (311.8-499.8)	0.605	68.2	—
	OME+TPP	521	1.72±0.25	301.0 (208.5-391.3)	0.674	50.0	1.36
	OME+DEM	507	1.79±0.26	293.8 (192.2-390.1)	0.285	48.8	1.40
	OME+PBO	517	1.59±0.18	280.6 (192.3-381.9)	0.164	46.6	1.46
G221A (RS)+A302S (RS) +A332V (RS)+S431F (RR)	OME	536	2.39±0.55	468.6 (196.1-641.6)	0.074	77.8	—
	OME+TPP	525	2.16±0.23	264.4 (190.5-341.4)	0.195	43.9	1.77
	OME+DEM	522	2.63±0.33	393.3 (319.8-463.2)	0.915	65.3	1.19
	OME+PBO	528	2.08±0.33	287.6 (183.5-375.5)	0.388	47.8	1.63

突变类型 Mutation type	吉布斯自由能 Gibbs free energy	范德华能 Van der Waals energy	静电能 Electrostatic energy	溶剂化能极性部分 Polar solvation energy	溶剂化能非极性部分 Non-polar solvation energy	熵 Entropy
WT	-21.8±15.5	-126.4±10.8	-57.1±14.1	151.0±12.8	-15.9±0.5	-26.6
S431F	-15.9±16.3	-140.5±11.9	-74.0±11.9	183.7±17.3	-16.0±0.3	-30.8
A302S+A332V+S431F	-6.8±15.1	-137.7±10.6	-54.5±11.6	167.9±15.7	-16.2±0.4	-33.9
A332V+S431F	-7.5±18.3	-131.7±9.1	-47.6±8.8	166.0±18.5	-16.2±0.3	-22.1
G221A+A302S+S431F	-23.6±17.2	-129.7±11.3	-49.8±14.3	146.2±22.6	-16.2±0.5	-25.9
G221A+A302S+A332V+S431F	-20.8±15.0	-137.3±11.0	-61.1±19.3	146.2±15.7	-16.3±0.6	-47.6

Monitoring of Resistance to Omethoate in Field Populations of Aphis gossypii in China and Elucidation of Target Resistance Mechanisms

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