河南省家蝇种群对拟除虫菊酯类杀虫剂敏感性测定和击倒抗性基因突变研究

doi:10.11853/j.issn.1003.8280.2023.03.005

摘要/Abstract

摘要： 目的建立河南省家蝇对拟除虫菊酯类杀虫剂的敏感基线,分析部分野生家蝇种群的击倒抗性基因多态性。方法家蝇敏感品系为实验室饲养多年的家蝇种群,未接触过杀虫剂；野生种群为2019－2021年间采集自开封、安阳、南阳、驻马店和鹤壁市等地的野外家蝇。采用微量点滴法检测雌成蝇对氯菊酯、溴氰菊酯、高效氯氰菊酯的半数致死剂量（LD₅₀）。采用等位基因特异性PCR（AS-PCR）技术检测各家蝇种群击倒抗性基因的基因型及其突变率,并检验击倒抗性基因突变率与LD₅₀的相关性。采用DNA测序方法验证AS-PCR的结果。结果氯菊酯、溴氰菊酯、高效氯氰菊酯对家蝇敏感品系的LD₅₀分别为0.151 0、0.077 2和0.166 6 μg/♀,对开封种群的LD₅₀分别为0.321 6、0.130 6和2.235 4 μg/♀,对安阳种群的LD₅₀分别为0.867 7、0.459 2和1.591 6 μg/♀,对南阳种群的LD₅₀分别为5.173 7、1.037 2和0.416 1 μg/♀,对驻马店种群的LD₅₀分别为0.634 1、0.108 2和0.262 4 μg/♀,对鹤壁种群的LD₅₀分别为2.745 0、1.102 9和2.556 0 μg/♀。家蝇敏感品系仅检测到1种基因型,即敏感纯合子,野生种群共检测到5种基因型,分别为敏感纯合子（L/L）、敏感/1014F杂合子（L/F）、敏感/1014H杂合子（L/H）、1014F/1014H杂合子（F/H）和1014H纯合子（H/H）。野生种群的突变率分别为14.84%、17.97%、13.28%、10.16%和20.31%。相关性检验显示家蝇种群对高效氯氰菊酯的抗性与L1014H突变相关。对130个家蝇样本的5种基因型进行测序,AS-PCR检测结果的准确率为79.23%。结论建立了河南省家蝇敏感基线,可作为河南省家蝇抗药性监测的参考值。AS-PCR技术能够用于家蝇抗药性基因的日常监测。

关键词: 家蝇, 敏感基线, 击倒抗性, 等位基因特异性, 聚合酶链式反应

Abstract: Objective To establish the susceptibility baseline of Musca domestica to pyrethroid insecticides, and to analyze the knockdown resistance gene polymorphism in some wild populations of M. domestica in Henan province, China.Methods The susceptible strain of M. domestica was a laboratory population that had not been exposed to any insecticides. The wild populations were collected in Kaifeng, Anyang, Nanyang, Zhumadian, and Hebi cities in 2019-2021. Topical application method was used to determine the median lethal dose (LD₅₀) of permethrin, deltamethrin, and beta-cypermethrin in female adult flies. Allele-specific polymerase chain reaction (AS-PCR) was used to determine the genotypes and mutation rates of knockdown resistance genes. Spearman correlation analysis was used to test the correlation between mutation rate and LD₅₀. DNA sequencing was used to verified the results of AS-PCR.Results The LD₅₀ of permethrin, deltamethrin, and beta-cypermethrin was 0.151 0, 0.077 2, and 0.166 6 μg/♀ in the susceptible strain, 0.321 6, 0.130 6, and 2.235 4 μg/♀ in Kaifeng population, 0.867 7, 0.459 2, and 1.591 6 μg/♀ in Anyang population, 5.173 7, 1.037 2, and 0.416 1 μg/♀ in Nanyang population, 0.634 1, 0.108 2, and 0.262 4 μg/♀ in Zhumadian population, and 2.745 0, 1.102 9, and 2.556 0 μg/♀ in Hebi population, respectively. Only one genotype, i.e. sensitive homozygote, was detected in the susceptible strain. Five genotypes were detected in the wild populations, including sensitive homozygote (L/L), sensitive/1014F heterozygote (L/F), sensitive/1014H heterozygote (L/H), 1014F/1014H heterozygote (F/H), and 1014H homozygote (H/H). Mutation rates in the wild populations were 14.84%, 17.97%, 13.28%, 10.16%, and 20.31%, respectively. The correlation test showed that the resistance to beta-cypermethrin was related to L1014H mutation. Five genotypes from 130 M. domestica samples were sequenced, and the accuracy of AS-PCR results was 79.23%.Conclusions The susceptibility baseline of M. domestica to pyrethroid insecticides is established in Henan, and this baseline can be used as a reference value for insecticide resistance monitoring. AS-PCR can be used for routine monitoring of resistance genes in M. domestica.

Key words: Musca domestica, Susceptibility baseline, Knockdown resistance, Allele-specific, Polymerase chain reaction

中图分类号:

赵奇, 樊金星, 张叶, 岳思宁, 刘吉起. 河南省家蝇种群对拟除虫菊酯类杀虫剂敏感性测定和击倒抗性基因突变研究[J]. 中国媒介生物学及控制杂志, 2023, 34(3): 308-313.

ZHAO Qi, FAN Jin-xing, ZHANG Ye, YUE Si-ning, LIU Ji-qi. Susceptibility of Musca domestica to pyrethroid insecticides and knockdown resistance gene mutations in Henan, China[J]. Chinese Journal of Vector Biology and Control, 2023, 34(3): 308-313.

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