Polymorphism and resistance mutation frequency of the target genes of insecticides in Anopheles sinensis in Guangyuan of Sichuan province, China

doi:10.11853/j.issn.1003.8280.2021.05.006

Abstract

Abstract: Objective To investigate the mutations of insecticide resistance-related genes through a sequencing analysis of the target genes of insecticides in Anopheles sinensis collected in Guangyuan of Sichuan province, China, and to provide guidance for the chemical control of An. sinensis in this region. Methods A total of 35 female An. sinensis mosquitoes collected in Jianshe village of Longtan township in Guangyuan of Sichuan province in September 2018 were selected, and genomic DNA was extracted after the abdomen was removed. PCR was used to obtain the gene fragments of acetylcholinesterase (ace), voltage-gated sodium channel (VGSC), and gamma-amino butyric acid receptor Rdl subunit (rdl), and then a sequencing analysis was performed. Results In comparison to the gene sequence of reference species, 20.00% of the 35 An. sinensis mosquitoes had resistance mutations, with a frequency of 5.71% for ace resistance allele (119 S), 7.14% and 1.43% for VGSC resistance alleles (1014F and 1014C), and 14.29% for rdl resistance allele (296S). Although there was a low proportion of An. sinensis mosquitoes with resistance mutations (<20.00%), mutations of multiple target genes were observed in more than 70.00% of the mosquitoes with mutation. Conclusion There is a certain proportion of resistance alleles in this An. sinensis population, with a genetic basis of developing high-level resistance, and the presence of mosquitoes with multiple target gene mutations indicates that this An. sinensis population has the possibility of developing into a multidrug-resistant population.

Key words: Anopheles sinensis, Insecticide resistance, Acetylcholinesterase, Voltage-gated sodium channel, Gamma-amino butyric acid receptor

摘要： 目的对四川省广元市中华按蚊3种杀虫剂的靶标基因进行测序分析，检测抗药性相关的基因变异，为该地区的按蚊化学防治提供指导。方法对2018年9月采集于四川省广元市龙潭乡建设村的35只中华按蚊雌蚊去除腹部后提取基因组DNA，通过聚合酶链式反应（PCR）获得乙酰胆碱酯酶（ace）、电压门控钠离子通道（VGSC）和γ-氨基丁酸受体（rdl）基因片段序列后进行测序分析。结果对比参照品系基因序列，此次检测的35只中华按蚊个体中，20.00%的个体存在抗性突变，其中ace抗性等位基因（119S）频率为5.71%，VGSC抗性等位基因（1014F、1014C）频率分别为7.14%和1.43%，rdl抗性等位基因（296S）频率为14.29%。携带抗性突变的中华按蚊所占比例较低（<20.00%），但突变个体中有70.00%以上是多靶标基因突变。结论该中华按蚊种群存在一定比例的抗性等位基因，具有向高水平抗性发展的遗传基础，多靶标基因突变个体的存在预示该中华按蚊种群具有发展为多重抗药性种群的可能性。

关键词: 中华按蚊, 杀虫剂靶标抗性, 乙酰胆碱酯酶, 电压门控钠离子通道, γ-氨基丁酸受体

CLC Number:

ZHAO Qiong-yao, QIU Xing-hui, YANG Yan, LUO Xing, SONG Xiao-ming, JIA Yong-chao. Polymorphism and resistance mutation frequency of the target genes of insecticides in Anopheles sinensis in Guangyuan of Sichuan province, China[J]. Chinese Journal of Vector Biology and Control, 2021, 32(5): 541-545.

赵琼瑶, 邱星辉, 杨研, 罗兴, 宋晓明, 贾永朝. 四川省广元市中华按蚊杀虫剂靶标基因的多态性及抗性突变频率研究[J]. 中国媒介生物学及控制杂志, 2021, 32(5): 541-545.

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