中国农业科学 ›› 2022, Vol. 55 ›› Issue (20): 3948-3959.doi: 10.3864/j.issn.0578-1752.2022.20.007
王帅宇1(),张子腾1,2,谢爱婷1,董杰1,杨建国1,张爱环2(
)
收稿日期:
2022-06-28
接受日期:
2022-07-20
出版日期:
2022-10-16
发布日期:
2022-10-24
通讯作者:
王帅宇,张爱环
基金资助:
WANG ShuaiYu1(),ZHANG ZiTeng1,2,XIE AiTing1,DONG Jie1,YANG JianGuo1,ZHANG AiHuan2(
)
Received:
2022-06-28
Accepted:
2022-07-20
Online:
2022-10-16
Published:
2022-10-24
Contact:
ShuaiYu WANG,AiHuan ZHANG
摘要:
【背景】 草地贪夜蛾(Spodoptera frugiperda)原产于美洲热带和亚热带地区,是重大迁飞性农业害虫。2016年以来迅速扩散至全球100多个国家,目前已入侵我国27省(自治区、直辖市),对粮食安全构成极大威胁。化学防治是草地贪夜蛾的重要应急防控措施,经过几年的药剂防治,草地贪夜蛾的抗药性可能会发生变化,进而影响防治策略的有效性。其发生范围持续北扩,而目前2020—2021年国内不同种群及长城防线种群抗药性现状以及年度间变化性研究较少。【目的】 明确我国不同区域种群(包括长城防线)的抗药性现状与差异,为草地贪夜蛾防控的科学用药提供指导。【方法】 采集13省(自治区、直辖市)的草地贪夜蛾田间种群样本,通过对单头样本杀虫剂靶标基因的PCR扩增产物直接测序,分析2020—2021年采自13省(自治区、直辖市)47市(区)草地贪夜蛾种群的362头个体氨基甲酸酯类、拟除虫菊酯类和双酰胺类杀虫剂靶标基因的突变情况。【结果】 草地贪夜蛾部分个体在氨基甲酸酯类药剂靶标乙酰胆碱酯酶(acetylcholine esterase)基因ace-1上存在突变,种群有6种ace-1基因型。ace-1基因检测到A201S和F290V位点突变;其中A201S均为抗性杂合突变,突变频率为8.4%;F290V为14.9%的抗性纯合突变和25.7%的杂合突变,未检测到G227A位点突变。重点防范区F290V位点突变频率高于迁飞过渡区和周年繁殖区,而在A201S位点突变频率均低于两区。长城防线、黄淮海阻截攻坚带、长江流域以及西南华南监测防控带ace-1基因突变频率均较高。A201S和F290V突变频率在2021年均较2020年种群略有下降,但无显著性差异。拟除虫菊酯类药剂靶标电压门控钠通道(voltage-gated sodium channel)基因vgsc和双酰胺类药剂靶标鱼尼丁受体(ryanodine receptor)基因ryr均未检测到靶标基因突变。【结论】 草地贪夜蛾种群氨基甲酸酯类靶标基因已发生较高频率的突变,说明对氨基甲酸酯类杀虫剂已产生较高水平的抗药性,在草地贪夜蛾的防控中应尽量减少使用。本研究显示草地贪夜蛾拟除虫菊酯类和双酰胺类杀虫剂靶标基因尚未检测到突变,这两类杀虫剂可与其他作用机制药剂科学轮换用于草地贪夜蛾的防控,鉴于目前已检测到这两类药剂的靶标基因突变个体,以及其国内近缘种甜菜夜蛾对这两类药剂存在较为普遍的杀虫剂靶标基因突变等情况,今后应开展草地贪夜蛾在室内外药剂高选择压力下以及寄主变化对主要防控药剂的靶标基因突变的影响,并加快抗药性快速检测技术和产品研究。
王帅宇,张子腾,谢爱婷,董杰,杨建国,张爱环. 我国草地贪夜蛾种群杀虫剂靶标基因突变分析[J]. 中国农业科学, 2022, 55(20): 3948-3959.
WANG ShuaiYu,ZHANG ZiTeng,XIE AiTing,DONG Jie,YANG JianGuo,ZHANG AiHuan. Mutation Analysis of Insecticide Target Genes in Populations of Spodoptera frugiperda in China[J]. Scientia Agricultura Sinica, 2022, 55(20): 3948-3959.
表1
2020—2021年草地贪夜蛾采集信息"
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 |
表2
PCR扩增ace-1、vgsc和ryr所用引物"
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.[ |
ACER | TGGCCTGTCTTCCAACATCA | ||||
ryr | RYFG | ACATAGACTGGCGTTACC | 730 | G4891E | 本研究This study |
RYRG | CCTCGTCATCTTCCTCC | ||||
I4734F | CGAGGACTTCTTCTACATGG | 800 | I4734M | BOAVENTURA et al.[ | |
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 |
表3
2020—2021年草地贪夜蛾不同种群ace-1基因型及突变频率"
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 |
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