Scientia Agricultura Sinica ›› 2022, Vol. 55 ›› Issue (20): 3948-3959.doi: 10.3864/j.issn.0578-1752.2022.20.007

• PLANT PROTECTION • Previous Articles     Next Articles

Mutation Analysis of Insecticide Target Genes in Populations of Spodoptera frugiperda in China

WANG ShuaiYu1(),ZHANG ZiTeng1,2,XIE AiTing1,DONG Jie1,YANG JianGuo1,ZHANG AiHuan2()   

  1. 1Beijing Plant Protection Station, Beijing 100029
    2College of Bioscience and Resource Environment, Beijing University of Agriculture/Key Laboratory of Urban Agriculture (North China), Ministry of Agriculture and Rural Affairs, Beijing 102206
  • Received:2022-06-28 Accepted:2022-07-20 Online:2022-10-16 Published:2022-10-24
  • Contact: ShuaiYu WANG,AiHuan ZHANG E-mail:shuaiyu3@126.com;zhangaihuan@126.com

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Abstract:

【Background】The fall armyworm (Spodoptera frugiperda) is a major migratory agricultural pest in the tropical and subtropical areas of America, it has rapidly expanded to more than 100 countries around the world since 2016, and has now invaded 27 provinces (Autonomous Region, Municipality) in China, posing a great threat to food security. Resistance may become an increasingly serious problem in many regions with the heavy reliance on insecticides for several years and lead to low control efficiency. S. frugiperda had spread to the north of the Great Wall rapidly. However, there are few studies on insecticide resistance and annual variation in China, especially the Great Wall protection zone in 2020 and 2021.【Objective】The objective of this study is to clarify the susceptibility and differences of S. frugiperda populations to insecticides in the Great Wall protection zone and other areas of China, and to provide guidance for scientific insecticide use for the prevention and control of S. frugiperda.【Method】Samples of S. frugiperda were collected from maize fields in 13 provinces (Autonomous Region, Municipality) in China, and the genomic DNA was extracted. The mutations of target genes of carbamate, pyrethroid and diamide insecticides in 362 individuals collected from 47 cities (districts) of 13 provinces (Autonomous Region, Municipality) in 2020 and 2021 were analyzed through direct sequencing of the PCR products individually.【Result】There were 6 genotypes of mutations in ace-1 to carbamate insecticides in all the samples. A201S and F290V mutations were detected in ace-1. The frequency of A201S heterozygous mutation was 8.4%, and the frequency of F290V homozygous and heterozygous mutations was 14.9% and 25.7%, respectively. No G227A mutation was detected. The mutation frequency at F290V in the key prevention region was higher than that in the migratory transition region and annual breeding region, while the mutation frequency at A201S was lower than that in the above two regions. The average frequency of ace-1 mutation was high in the Great Wall protection zone, Huang-Huai-Hai interception zone, Yangtze River Basin and Southwest and South China protection zone. There was no significant difference in the frequency of ace-1 mutation between the populations in 2021 and 2020. No mutations were detected in pyrethroid vgsc and diamide ryr.【Conclusion】A201S and F290V mutations with high frequency in ace-1 is one of the mechanisms conferring carbamate resistance in many populations in China. The use of carbamates should be restricted in the control of S. frugiperda. No mutations had been detected in the vgsc and ryr, suggesting susceptibility to pyrethroids and diamides for rational rotation, however, the low frequency of mutation in S. frugiperda and common mutation in Spodoptera exigua (relatives of S. frugiperda) to pyrethroids and diamides have been detected. In the future, the effect of high insecticide selection stress in the lab and field populations fed by different hosts on the target gene mutation should be evaluated, and the research of rapid diagnostics methods and products of insecticide resistance should be accelerated.

Key words: Spodoptera frugiperda, insecticide resistance, carbamate, pyrethroid, diamide, target gene, mutation frequency

Table 1

Collection information of S. frugiperda in 2020-2021"

省（自治区、直辖市）
Province (Autonomous Region, Municipality)		 市（区）
City (District)		 采集时间
Collection time (y-m-d)		 虫态
State		 数量（头）
Number
北京Beijing 顺义Shunyi 2020-09-01;2021-10-05 成虫Adult 4
丰台Fengtai 2020-09-01 成虫Adult 2
通州Tongzhou 2020-09-01;2021-09-14 成虫Adult 3
怀柔Huairou 2020-09-01 成虫Adult 2
朝阳Chaoyang 2021-10-04 成虫Adult 1
海淀Haidian 2020-09-01;2021-09-09 成虫Adult 3
昌平Changping 2020-09-01 成虫Adult 2
平谷Pinggu 2020-09-01;2021-09-03 成虫Adult 3
密云Miyun 2020-09-01 成虫Adult 2
延庆Yanqing 2021-09-03 成虫Adult 3
大兴Daxing 2021-09-03 成虫Adult 2
天津Tianjin 蓟州Jizhou 2020-08-31;2021-08-25 成虫Adult 5
东丽Dongli 2020-09-07 成虫Adult 8
滨海Binhai 2020-09-18 成虫Adult 1
静海Jinghai 2020-09-03 成虫Adult 3
宝坻Baodi 2020-09-21 成虫Adult 2
武清Wuqing 2020-09-27 成虫Adult 2
河北Hebei 石家庄Shijiazhuang 2020-10-14;2021-09-02 幼虫Larva 8
霸州Bazhou 2021-09-01 成虫Adult 3
唐山Tangshan 2021-10-08 成虫Adult 24
山东Shandong 济南Ji’nan 2020-10-13 幼虫Larva 3
河南Henan 周口Zhoukou 2020-10-20;2021-08-23 成虫Adult 6
开封Kaifeng 2020-10-11;2021-10-05 成虫Adult 33
驻马店Zhumadian 2021-10-08 成虫Adult 48
安徽Anhui 黄山Huangshan 2020-10-28 成虫Adult 2
池州Chizhou 2020-10-28 幼虫Larva 3
马鞍山Maanshan 2020-10-24 成虫Adult 6
芜湖Wuhu 2020-11-07;2021-10-06 成虫Adult 29
宿州Suzhou 2020-10-25 成虫Adult 5
安庆Anqing 2020-11-07;2021-10-09 成虫Adult 24
江苏Jiangsu 盐城Yancheng 2020-10-25 成虫Adult 3
大丰Dafeng 2020-10-25 成虫Adult 3
浙江Zhejiang 金华Jinhua 2020-10-16;2021-09-01 幼虫/成虫Larva/Adult 10
杭州Hangzhou 2020-10-18;2021-09-01 成虫Adult 19
温州Wenzhou 2020-10-11 成虫Adult 4
四川Sichuan 西昌Xichang 2020-11-16 成虫Adult 4
攀枝花Panzhihua 2021-10-03 幼虫Larva 3
云南Yunnan 红河Honghe 2020-09-20 幼虫Larva 3
西清Xiqing 2020-09-20 幼虫Larva 3
临沧Lincang 2020-09-20 成虫Adult 23
德宏Dehong 2020-09-20;2021-09-10 幼虫Larva 12
曲靖Qujing 2021-10-10 幼虫Larva 8
昆明Kunming 2021-10-10 成虫Adult 3
楚雄Chuxiong 2021-10-10 幼虫Larva 3
广东Guangdong 广州Guangzhou 2020-11-04 幼虫Larva 10
广西Guangxi 南宁Nanning 2020-11-01 幼虫Larva 3
海南Hainan 儋州Danzhou 2020-10-30 幼虫Larva 6

Table 2

Primers used for PCR amplification of ace-1, vgsc and ryr"

基因
Gene 		引物
<BOLD>P</BOLD>rimer		 引物序列
<BOLD>P</BOLD>rimer sequence (5′-3′)		 扩增长度
<BOLD>L</BOLD>ength (bp)		 位点
<BOLD>S</BOLD>ite		 来源
Source
ace-1 ACEF TAAGAACGCTGCTGTCATGC 760 A201S、F290V CARVALHO et al.[9]
ACER TGGCCTGTCTTCCAACATCA
ryr RYFG ACATAGACTGGCGTTACC 730 G4891E 本研究This study
RYRG CCTCGTCATCTTCCTCC
I4734F CGAGGACTTCTTCTACATGG 800 I4734M BOAVENTURA et al.[17]
I4734R AATTTACGGGCAATCTCC
vgsc VG9F CAAATTGGCAAAGTCATG 253 T929I、L932F 本研究This study
VG9R AGCAGCGATTGTATGTATCT
V10F CGTAGACCGTTTCCCG 360 L1014F 本研究This study
V10R CACCCAATCAATGAACCT

Table 3

Genotypes and mutation frequencies of ace-1 of different S. frugiperda populations in 2020-2021"

省（自治区、直辖市）Province (Autonomous Region, Municipality) 年份<BOLD>Y</BOLD>ear 数量（头）<BOLD>N</BOLD>umber A201S位点基因型Genotype at locus A201S (%) 突变频率<BOLD>M</BOLD>utation frequency (%) F290V位点基因型Genotype at locus F290V (%) 突变频率<BOLD>M</BOLD>utation frequency (%) G227A位点基因型Genotype at locus G227A (%) 突变频率<BOLD>M</BOLD>utation frequency (%)
SS RS RR SS RS RR SS RS RR
北京Beijing 2020 15 66.7 33.3 0 16.7 46.6 46.7 6.7 30 100 0 0 0
2021 12 100 0 0 0 58.4 33.3 8.3 25 100 0 0 0
天津Tianjin 2020 19 68.4 31.6 0 15.8 15.8 68.4 15.8 50 100 0 0 0
2021 2 100 0 0 0 0 50 50 75 100 0 0 0
河北Hebei 2020 3 100 0 0 0 100 0 0 0 100 0 0 0
2021 32 87.4 12.6 0 6.3 15.6 68.8 15.6 50 100 0 0 0
山东Shandong 2020 3 66.6 33.4 0 16.7 0 66.7 33.3 66.7 100 0 0 0
河南Henan 2020 33 91 9 0 4.5 33.3 45.5 21.2 69.4 100 0 0 0
2021 54 83.4 16.6 0 8.3 24 51.9 24.1 50 100 0 0 0
安徽Anhui 2020 45 88.8 11.2 0 5.6 42.2 42.2 15.6 36.7 100 0 0 0
2021 24 79.2 20.8 0 10.4 50 37.5 12.5 31.3 100 0 0 0
江苏Jiangsu 2020 6 83.4 16.6 0 8.3 16.6 66.7 16.7 50 100 0 0 0
浙江Zhejiang 2020 10 60 40 0 20 40 60 0 30 100 0 0 0
2021 23 87 13 0 6.5 52.2 47.8 0 23.9 100 0 0 0
四川Sichuan 2020 4 100 0 0 0 0 50 50 75 100 0 0 0
2021 3 66.7 33.3 0 16.7 66.7 33.3 0 16.7 100 0 0 0
云南Yunnan 2020 32 84.4 15.6 0 7.8 21.8 59.4 18.8 48.4 100 0 0 0
2021 23 82.6 17.4 0 8.7 47.9 47.8 4.3 28.3 100 0 0 0
广东Guangdong 2020 10 80 20 0 10 20 70 10 45 100 0 0 0
广西Guangxi 2020 3 100 0 0 0 66.7 33.3 0 16.7 100 0 0 0
海南Hainan 2020 6 50 50 0 25 16.6 66.7 16.7 50 100 0 0 0

Fig. 1

Frequency of A201S and F290V mutations in ace-1 of S. frugiperda in different provinces"

Fig. 2

Frequency of A201S and F290V mutations in ace-1 of S. frugiperda populations in three ecological regions and four protective zones"

Fig. 3

Difference in ace-1 A201S and F290V mutation frequencies of S. frugiperda populations in three ecological regions and four protective zones in 2020-2021"

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