山东省小麦田播娘蒿对双氟磺草胺抗性水平及靶标抗性机理

doi:10.3864/j.issn.0578-1752.2020.12.008

Abstract

Abstract:

【Background】 Descurainia sophia is the most widely distributed and harmful broad-leaved weed in winter wheat field, and florasulam is the most widely used ALS inhibitor herbicide for controlling broad-leaved weeds. After several years of application, the control effect of florasulam on D. sophia has been found to decline in some wheat fields, which may be related to the resistance. 【Objective】 The objective of this study is to clarify the resistance level and mechanism of D. sophia to florasulam, and to provide theoretical basis for the establishment of precise regional control of broad-leaved weeds in wheat field. 【Method】 A total of 40 populations of D. sophia were collected from winter wheat fields. Whole-plant dose response experiments were conducted to determine the resistance level of 40 populations to florasulam, tribenuron-methyl and MCPA in the greenhouse. At the same time, according to the ALS gene sequence of D. sophia, the genomic DNA of a single plant with high resistance to florasulam was extracted. The ALS gene sequences were obtained and compared with that of sensitive type Arabidopsis thaliana to find out the mutation site and the resistance mechanism. 【Result】 The results of resistance level determination showed that 32 of the 40 D. sophia populations were sensitive to florasulam, accounting for 80.00%. There were 3, 3 and 2 populations of low resistance, medium resistance and high resistance, respectively. JN-1, JNI-2 and LY-2 belonged to low resistance populations. Three populations LC-3, LY-4 and YT-1 belonged to medium resistance with resistance index (RI) of 49.00, 26.44 and 21.09, respectively. BZ-1 and DZ-3 belonged to high resistance populations with RI of 52.00 and 194.00, respectively. The ALS sequence analysis showed that there were mutations from CCT (pro) to TCT (Ser) or CTT (Leu) in amino acid 197 of ALS gene in BZ-1, while TGG (Trp) to TTG (Leu) in amino acid 574 of ALS gene in DZ-3. In addition, 19 of the 40 D. sophia populations were resistant to tribenuron-methyl, accounting for 47.50%, among them, 11 populations were low resistance, 6 populations were medium resistance with RI of 38.05, 13.55, 11.54, 10.45, 11.50, 11.02, and DZ-3 and LY-4 belonged to high resistance populations with RI of 244.75 and 68.50, respectively. All 40 populations were sensitive to MCPA. 【Conclusion】 Among the 40 populations of D. sophia collected from wheat fields in Shandong Province, 20.00% of them are resistance to florasulam, and the substitutions of amino acids occur in different positions in the populations with high resistance to florasulam. The resistance of D. sophia to tribenuron-methyl is still very serious, but all the populations are not resistant to MCPA. In view of the resistance area of D. sophia, the alternative and mixed use of herbicides with multiple mechanisms should be promoted, which can not only delay and control the development of resistance in weeds, but also expand the weed control spectrum and reduce the use of herbicides.

Key words: florasulam, Descurainia sophia, resistance level, resistance mechanism

GAO XingXiang,ZHANG YueLi,LI Mei,LI Jian,FANG Feng. Resistance Level and Mechanism of Descurainia sophia to Florasulam in Wheat Field of Shandong Province[J].Scientia Agricultura Sinica, 2020, 53(12): 2399-2409.

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Figures/Tables 5

Table 1

Collection sites information of 40 D. sophia populations"

序号 Number	采集地点 Collection site	序号 Number	采集地点 Collection site
BZ-1	滨州市邹平县台子镇 Taizi Town, Zouping County, Binzhou City	LY-2	临沂市郯城县胜利镇 Shengli Town, Tancheng County, Linyi City
BZ-2	滨州市惠民县胡集镇 Huji Town, Huimin County, Binzhou City	LY-3	临沂市费县探义镇 Tanyi Town, Fei County, Linyi City
DY-1	东营市利津县北宋镇 Beisong Town, Lijin County, Dongying City	LY-4	临沂市沂南县青驼镇 Qingtuo Town, Yi’nan County, Linyi City
DZ-1	德州市庆云县常家镇 Changjia Town, Qingyun County, Dezhou City	QD-1	青岛市莱西区河头店镇 Hetoudian Town, Laixi District, Qingdao City
DZ-2	德州市平原县恩城镇 Encheng Town, Pingyuan County, Dezhou City	QD-2	青岛市平度市云山镇 Yunshan Town, Pingdu County, Qingdao City
DZ-3	德州市武城县武城镇 Wucheng Town, Wucheng County, Dezhou City	RZ-1	日照市莒县小店镇 Xiaodian Town, Ju County, Rizhao City
HZ-1	菏泽市成武县张楼镇 Zhanglou Town, Chengwu County, Heze City	RZ-2	日照市五莲县西湖镇 Xihu Town, Wulian County, Rizhao City
HZ-2	菏泽市巨野县田桥镇 Tianqiao Town, Juye County, Heze City	TA-1	泰安市东平县大洋镇 Dayang Town, Dongping County, Taian City
JN-1	济南市长清区张夏镇 Zhangxia Town, Changqing District, Ji’nan City	TA-2	泰安市肥城县老城街道 Laocheng Street, Feicheng County, Taian City
JN-2	济南市历城区王舍人镇 Wangsheren Town, Licheng District, Ji’nan City	TA-3	泰安市宁阳县磁窑镇 Ciyao Town, Ningyang County, Taian City
JN-3	济南市章丘区刁镇 Diao Town, Zhangqiu District, Ji’nan City	WF-1	潍坊市潍城区于河街道 Yuhe Street, Weicheng District, Weifang City
JNI-1	济宁市汶上县郭仓镇 Guocang Town, Wenshang County, Jining City	WF-2	潍坊市昌邑市卜庄镇 Bozhuang Town, Changyi County, Weifang City
JNI-2	济宁市梁山县韩岗镇 Hangang Town, Liangshan County, Jining City	WF-3	潍坊市临朐县沂山镇 Yishan Town, Linqu County, Weifang City
JNI-3	济宁市嘉祥县疃里镇 Tuanli Town, Jiaxiang County, Jining City	WH-1	威海市乳山市夏村镇 Xiacun Town, Rushan County, Weihai City
JNI-4	济宁市任城区南张镇 Nanzhuang Town, Rencheng District, Jining City	YT-1	烟台市莱州市沙河镇 Shahe Town, Laizhou County, Yantai City
LC-1	聊城市莘县古城镇 Gucheng Town, Shen County, Liaocheng City	YT-2	烟台市莱阳市河头店镇 Hetoudian Town, Laiyang County, Yantai City
LC-2	聊城市阳谷县西湖镇 Xihu Town, Yanggu County, Liaocheng City	ZB-1	淄博市周村区南郊镇 Nanjiao Town, Zhoucun District, Zibo City
LC-3	聊城市东昌府区侯营镇 Houying Town, Dongchangfu District, Liaocheng City	ZB-2	淄博市临淄区齐陵街道 Qiling Street, Linzi District, Zibo City
LW-1	莱芜市莱城区口镇 Kou Town, Laicheng District, Laiwu City	ZZ-1	枣庄市滕州市鲍沟镇 Baogou Town, Tengzhou County, Zaozhuang City
LY-1	临沂市兰陵县卢柞镇 Luzuo Town, Lanling County, Linyi City	ZZ-2	枣庄市薛城区周营镇 Zhouying Town, Xuecheng District, Zaozhuang City

Table 1

Table 2

Resistance level of D. sophia populations to florasulam"

序号 Number	回归方程 Regression equation (y=)	相关系数 Correlation coefficient	GR₅₀(g·hm^-2) (95% CL)	相对抗性指数 RI
BZ-1	3.0871+2.5252x	0.9220	5.72 (3.89-10.98)	52.00
BZ-2	6.7548+2.0988x	0.8926	0.15 (0.01-0.39)	1.36
DY-1	6.1746+1.2088x	0.9405	0.11 (0.02-0.26)	1.00
DZ-1	6.1210+1.8228x	0.8825	0.24 (0.08-0.45)	2.18
DZ-2	6.1178+1.5045x	0.8717	0.18 (0.001-0.62)	1.64
DZ-3	3.7060+0.9736x	0.8519	21.34 (13.81-39.62)	194.00
HZ-1	5.6433+1.2505x	0.9203	0.31 (0.14-0.51)	2.82
HZ-2	6.2837+2.3134x	0.8914	0.28 (0.10-0.50)	2.55
JN-1	5.4469+1.6999x	0.9475	0.55 (0.33-0.78)	5.00
JN-2	5.5627+1.0454x	0.8902	0.29 (0.002-0.97)	2.64
JN-3	6.1607+1.3459x	0.9459	0.14 (0.03-0.31)	1.27
JNI-1	5.6161+2.1229x	0.9679	0.51 (0.30-0.74)	4.64
JNI-2	5.3038+1.5719x	0.9333	0.64 (0.18-1.20)	5.82
JNI-3	6.4349+1.5358x	0.8804	0.12 (0.01-0.30)	1.09
JNI-4	6.5989+1.6727x	0.9630	0.11 (0.01-0.31)	1.00
LC-1	6.6323+1.9998x	0.8911	0.15 (0.02-0.38)	1.36
LC-2	6.2404+1.7165x	0.9655	0.19 (0.05-0.39)	1.73
LC-3	3.3624+2.2387x	0.9631	5.39 (3.89-8.22)	49.00
LW-1	5.6180+1.4674x	0.9352	0.38 (0.19-0.59)	3.45
LY-1	6.4307+2.3490x	0.8925	0.25 (0.07-0.48)	2.27
LY-2	5.3477+1.7628x	0.9479	0.64 (0.41-0.87)	5.82
LY-3	6.0852+1.7614x	0.9592	0.24 (0.08-0.45)	2.18
LY-4	4.0274+2.0848x	0.9589	2.93 (2.48-3.44)	26.64
QD-1	5.9423+1.3467x	0.9401	0.20 (0.07-0.38)	1.82
QD-2	6.0594+1.2737x	0.9363	0.15 (0.04-0.32)	1.27
RZ-1	5.4556+1.6946x	0.9474	0.54 (0.15-1.01)	4.91
RZ-2	6.0120+1.6168x	0.8734	0.24 (0.004-0.70)	2.18
TA-1	5.9847+1.6603x	0.8744	0.26 (0.01-0.72)	2.36
TA-2	6.0133+1.9703x	0.8839	0.31 (0.13-0.52)	2.82
TA-3	6.1886+1.3238x	0.9432	0.13 (0.03-0.29)	1.18
WF-1	6.4373+1.8136x	0.8868	0.16 (0.03-0.37)	1.45
WF-2	6.4515+2.7972x	0.8939	0.30 (0.10-0.54)	2.73
WF-3	6.1546+2.1186x	0.8883	0.29 (0.11-0.50)	2.64
WH-1	5.7762+1.1081x	0.9022	0.20 (0.07-0.38)	1.82
YT-1	4.5119+1.3342x	0.8726	2.32 (1.26-3.85)	21.09
YT-2	6.2982+1.7011x	0.9642	0.17 (0.04-0.37)	1.55
ZB-1	5.4225+1.4153x	0.9245	0.50 (0.13-0.98)	4.55
ZB-2	6.2430+1.5091x	0.8754	0.15 (0.03-0.33)	1.36
ZZ-1	6.5223+2.1754x	0.8918	0.20 (0.04-0.43)	1.82
ZZ-2	6.0763+1.4651x	0.9454	0.18 (0.05-0.37)	1.64

Table 2

Table 3

Resistance level of D. sophia populations to tribenuron-methyl"

序号 Number	回归方程 Regression equation (y=)	相关系数 Correlation coefficient	GR₅₀(g·hm^-2) (95% CL)	相对抗性指数 RI
BZ-1	4.4592+0.4070x	0.9305	21.31 (7.80-41.91)	38.05
BZ-2	3.4995+2.1764x	0.8938	4.89 (1.73-9.05)	8.73
DY-1	4.5408+1.5010x	0.8883	2.02 (0.27-5.44)	3.61
DZ-1	4.5400+1.3410x	0.8811	2.20 (0.42-5.40)	3.93
DZ-2	3.7377+1.7915x	0.8886	5.07 (1.96-9.11)	9.05
DZ-3	2.7435+1.0560x	0.9480	137.06 (58.29-448.11)	244.75
HZ-1	4.0109+1.4039x	0.9423	0.56 (0.03-11.73)	1.00
HZ-2	4.0697+1.2949x	0.9481	5.23 (2.24-9.17)	9.34
JN-1	4.1869+0.9237x	0.9913	7.59 (3.73-12.60)	13.55
JN-2	4.7999+0.9194x	0.9239	1.65 (0.34-4.16)	2.95
JN-3	4.2133+1.3990x	0.9533	3.65 (1.18-7.25)	6.52
JNI-1	4.5819+1.1696x	0.9575	2.28 (0.53-5.29)	4.07
JNI-2	3.4317+1.9362x	0.8894	6.46 (2.96-10.70)	11.54
JNI-3	4.4136+1.5958x	0.8898	2.33 (0.37-5.92)	4.16
JNI-4	4.1771+1.2621x	0.9501	4.49 (1.76-8.23)	8.02
LC-1	3.7939+1.5722x	0.9610	5.85 (2.59-9.96)	10.45
LC-2	4.4694+1.8515x	0.8941	1.93 (0.12-6.05)	3.45
LC-3	4.2146+0.9707x	0.9104	6.44 (3.02-10.96)	11.50
LW-1	4.8448+0.7822x	0.8931	1.58 (0.33-4.02)	2.82
LY-1	4.3215+1.7573x	0.8925	2.43 (0.35-6.26)	4.34
LY-2	4.7095+1.5411x	0.8909	1.54 (0.10-5.04)	2.75
LY-3	4.4512+1.5314x	0.8883	2.28 (0.37-5.77)	4.07
LY-4	3.3832+1.0208x	0.8737	38.36 (18.32-67.62)	68.50
QD-1	4.0803+1.2919x	0.9503	5.15 (2.19-9.07)	9.20
QD-2	3.7652+1.5620x	0.9589	6.17 (2.83-10.34)	11.02
RZ-1	4.3648+1.5478x	0.8879	2.57 (0.50-6.12)	4.59
RZ-2	4.5092+1.3450x	0.8808	2.32 (0.47-5.55)	4.14
TA-1	3.9601+1.6906x	0.8879	4.12 (1.35-8.00)	7.36
TA-2	3.2026+2.4882x	0.8944	5.28 (1.95-9.52)	9.43
TA-3	4.2321+1.4666x	0.9509	3.34 (0.96-6.92)	5.96
WF-1	4.8405+1.4052x	0.8883	1.30 (0.07-4.49)	2.32
WF-2	4.7478+1.2172x	0.8773	1.61 (0.23-4.46)	2.88
WF-3	4.4687+1.5162x	0.9677	2.24 (0.36-5.70)	4.00
WH-1	4.7373+1.4737x	0.8894	1.51 (0.10-4.85)	2.70
YT-1	4.3582+0.9489x	0.9037	4.75 (1.94-8.67)	8.48
YT-2	4.6091+1.5464x	0.8902	1.79 (0.17-5.28)	3.20
ZB-1	4.4016+1.2996x	0.8750	2.89 (0.03-10.85)	5.16
ZB-2	4.3259+1.8489x	0.8936	2.32 (0.26-6.31)	4.14
ZZ-1	4.8315+1.4640x	0.8900	1.30 (0.06-4.64)	2.32
ZZ-2	4.3491+1.7430x	0.8924	2.36 (0.32-6.18)	4.21

Table 3

Table 4

Resistance level of D. sophia populations to MCPA"

序号 Number	回归方程 Regression equation (y=)	相关系数 Correlation coefficient	GR₅₀(g·hm^-2) (95% CL)	相对抗性指数 RI
BZ-1	2.0945+2.5738x	0.8928	13.45 (2.43-29.10)	3.61
BZ-2	3.3409+1.6892x	0.8819	9.60 (1.80-21.89)	2.57
DY-1	3.9377+1.3693x	0.8715	5.97 (0.69-16.27)	1.60
DZ-1	4.3261+1.0307x	0.9189	4.51 (0.60-12.56)	1.21
DZ-2	3.7099+1.5489x	0.8802	6.91 (0.73-18.38)	1.85
DZ-3	3.4617+1.7590x	0.8866	7.49 (0.66-20.53)	2.01
HZ-1	4.2287+0.9926x	0.9061	5.98 (1.17-14.62)	1.60
HZ-2	4.9736+0.7651x	0.8886	3.88 (0.40-12.68)	1.04
JN-1	3.2623+1.7257x	0.8826	10.16 (2.04-22.62)	2.72
JN-2	3.7622+1.4396x	0.9516	7.24 (1.10-18.09)	1.94
JN-3	3.4954+1.7896x	0.8880	6.93 (0.41-20.41)	1.86
JNI-2	3.3834+1.6609x	0.8811	9.40 (1.75-21.55)	2.52
JNI-2	3.6806+1.4785x	0.9582	7.81 (1.29-18.91)	2.09
JNI-3	3.4141+1.6799x	0.8826	8.79 (1.39-21.08)	2.36
JNI-4	4.3825+1.0806x	0.9246	3.73 (0.32-11.68)	1.00
LC-2	3.7705+1.5307x	0.8802	6.36 (0.59-17.83)	1.71
LC-2	3.5698+1.4654x	0.8705	9.46 (0.03-34.75)	2.54
LC-3	3.2963+1.5965x	0.8749	11.67 (3.24-23.58)	3.13
LW-1	3.0031+2.1808x	0.8924	8.24 (0.33-24.44)	2.21
LY-1	3.4446+1.8365x	0.8889	7.03 (0.38-20.88)	1.88
LY-2	3.4498+1.6801x	0.8831	8.37 (1.18-20.70)	2.24
LY-3	4.0256+1.3975x	0.8761	4.98 (0.33-15.47)	1.34
LY-4	3.0374+1.8191x	0.8837	11.99 (2.97-24.79)	3.21
QD-1	3.1072+1.9785x	0.8897	9.05 (0.95-23.20)	2.43
QD-2	3.1567+1.9338x	0.8890	8.98 (1.00-22.84)	2.41
RZ-1	3.5592+1.7137x	0.8860	6.93 (0.52-19.76)	1.86
RZ-2	4.1233+1.4701x	0.8825	3.95 (0.07-15.40)	1.06
TA-1	3.2322+1.6788x	0.8794	11.30 (2.83-23.52)	3.03
TA-2	3.7666+1.4893x	0.8774	6.73 (0.80-17.88)	1.80
TA-3	3.0389+2.0898x	0.8914	8.68 (0.61-23.81)	2.33
WF-1	4.2364+1.2589x	0.9485	4.04 (0.24-13.34)	1.08
WF-2	3.5311+1.8064x	0.8888	6.50 (0.27-20.34)	1.74
WF-3	3.5460+1.6892x	0.8849	7.26 (0.68-19.82)	1.95
WH-1	3.2738+1.8467x	0.8876	8.60 (0.98-21.99)	2.31
YT-1	3.4549+1.8182x	0.8884	7.08 (0.42-20.75)	1.90
YT-2	3.7438+1.3591x	0.9397	8.40 (1.82-18.96)	2.25
ZB-1	3.0505+1.6576x	0.9504	15.00 (5.39-27.35)	4.02
ZB-2	3.6460+1.7483x	0.8881	5.95 (0.20-19.50)	1.60
ZZ-1	3.3559+1.8120x	0.8873	8.08 (0.81-21.35)	2.17
ZZ-2	4.2640+1.1111x	0.9285	4.60 (0.55-13.04)	1.23

Table 4

Table 5

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序号Number	相对抗性指数RI	ALS基因突变ALS gene mutation	检出比例Detection rate
BZ-1	52.00	Pro-197-Leu;Pro-197-Ser	Pro-197-Leu,40%;Pro-197-Ser,60%
DZ-3	194.00	Trp-574-Leu	100%
DY-1	1.00	Wild type	—

Resistance Level and Mechanism of Descurainia sophia to Florasulam in Wheat Field of Shandong Province

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