北京市3个公园白纹伊蚊种群Rdl抗性基因突变分析

Frequency analysis of Rdl resistance gene mutations in Aedes albopictus from 3 parks in Beijing

目的 对北京市3个大型森林公园白纹伊蚊(Aedes albopictus)野外种群的Rdl基因抗性突变进行检测,评估其对γ-氨基丁酸(γ-aminobutyric acid,GABA)受体类杀虫剂的抗性风险。方法 利用序列比对分析获得白纹伊蚊编码GABA受体的Rdl基因序列,通过PCR扩增Rdl基因片段并进行测序,分析与抗性有关位点(第A296和V327位点)的突变情况。结果 白纹伊蚊Rdl基因由10个外显子(Exon)和9个内含子(Intron)组成,编码区(CDS)全长1 608 bp,编码535个氨基酸;白纹伊蚊Rdl基因编码氨基酸序列与埃及伊蚊(Aedes aegypti)和淡色库蚊(Culex pipiens pallens)相应序列的相似性分别为99.63%,与黑腹果蝇(Drosophila melanogaster)相似性为91.55%;在3个公园采集野外白纹伊蚊种群中(90只标本)均未检测到上述Rdl靶标抗性突变,均为敏感型(GCA和GTA)。结论 北京市3个大型的城市森林公园环境中白纹伊蚊种群对环戊二烯类有机氯杀虫剂和苯基吡唑类杀虫剂氟虫腈的靶标(即GABA受体)Rdl基因与其他昆虫表现出不同程度的同源性和高度保守的基因结构特征,3个受试白纹伊蚊野外种群未检测到Rdl基因抗性相关突变,提示以靶标敏感为导向反向研发杀虫剂。

Objective To analyze Rdl resistance mutations in three park-collected populations of Aedes albopictus in Beijing,and assess their risk of resistance to cyclopentadienyl insecticides.Methods Rdl gene sequence encoding the γ-aminobutyric acid(GABA) receptor in A.albopictus was obtained by sequence retrieve and alignment analysis,then amplified by PCR and sequenced to analyze mutations at resistance-related loci(A296 and V327).Results Rdl gene of A.albopictus consists of 10 exons and 9 introns,with a coding region(CDS) of 1 608 bp in length,it codes for 535 amino acids;The amino acid sequence of the Rdl gene of A.albopictus showed 99.63% similarity to the corresponding sequences of A.aegypti,Culex pipiens pallens,and 91.55% similarity to Drosophila melanogaster.No mutation was detected in the above loci in three field populations of A.albopictus(90 specimens),all were sensitive genotype(GCA and GTA).Conclusions The study was conducted in three large populations of A.albopictus in urban forest park environments in Beijing,and investigated the gene characteristics and resistance mutations of the targets(GABA receptors) of cyclopentadiene organochlorine insecticides and the phenyl pyrazole insecticide fipronil,and found that the Rdl gene of A.albopictus showed different degrees of homology and highly conserved gene structure characteristics with other insects,and no Rdl resistance-related mutation was detected in three field populations of A.albopictus,suggesting that target-sensitive-oriented reverse development of insecticides.