海南省不同地形白纹伊蚊线粒体细胞色素C氧化酶亚基Ⅰ基因遗传多样性

Genetic mtDNA-COⅠdiversity of Aedes albopictus populations in different terrains of Hainan Province

目的研究海南省平原、丘陵、山地地区白纹伊种群遗传多样性,分析海南省不同地形区白纹伊蚊种群遗传结构。方法于海南省三亚和海口的平原地区,儋州和屯昌的丘陵,白沙的山区采集白纹伊蚊,单只蚊子提取DNA,储存于-80℃冰箱中备用,用聚合酶链反应(polymerase chain reaction,PCR)扩增线粒体细胞色素C氧化酶亚基Ⅰ基因(mitochondrial cytochrome C oxidase subunitⅠgene,mtDNA-COⅠ)并测序,在美国国立生物技术信息中心(National Center for Biotechnology Information,NCBI)上进行比对,DnaSP v6计算遗传多样性参数、PopART 1.7绘制单倍型网络图,Arlequin 3.5计算分子方差分析(analysis of molecular variance,AMOVA)值,分析种群间的差异。结果5个地区白纹伊蚊种群mtDNA-COⅠ测序共获得414条序列,长度均为663 bp,全部序列有5个突变位点,G+C含量为32.86%,A+T含量为67.14%,符合线粒体DNA的特征,与其他种群相比,平原地区的三亚和丘陵地区的儋州、屯昌种群核苷酸多样性较高,平均核苷酸差异数较高,单倍型分析共获得6个单倍型,单倍型H02为优势单倍型,三亚和屯昌种群单倍型数量最多,平原地区海口和三亚种群有独享的单倍型,丘陵和山地白纹伊蚊种群单倍型数量较少,中性结果和错配分布图显示所有地区白纹伊蚊种群近期均有种群扩张。AMOVA值显示种群间差异大于种群内差异。结论mtDNA-COⅠ基因可以作为研究白纹伊蚊种群遗传多样性的分子标志,平原地区的三亚种群显示出较高的遗传多样性,丘陵地区的2个种群遗传多样性也较高,但单倍型数量只有平原地区的三亚和丘陵地区的儋州种群较高。

Objective To explore the genetic diversity of Aedes albopictus populations in plain,hilly,and mountainous areas of Hainan Province,and to analyze the genetic structure of Aedes albopictus populations in different terrain regions of Hainan Province.Methods Aedes albopictus were collected from the plain areas of Sanya and Haikou City,the hills of Danzhou and Tunchang City,and the mountainous areas of Baisha City in Hainan Province.DNA was extracted from a single mosquito and stored in a-80℃refrigerator for use.Mitochondrial cytochrome C oxidase subunitⅠ(mtDNA-COⅠ)was amplified by PCR and sequenced.The results were compared on the National Center for Biotechnology Information(NCBI)website,sequence peaks were observed using BioEdit 7.0,genetic diversity parameters were calculated using DnaSP v6,haplotype networks were constructed using PopART 1.7,and analysis of molecular variance(AMOVA)values were calculated using Arlequin to analyze differences between populations.Results A total of 414 mtDNA-COⅠsequences were obtained from Aedes albopictus populations in 5 regions,with lengths of 663 bp.All sequences had five mutation sites,with a G+C content of 32.86%and an A+T content of 67.14%,consistent with mitochondrial DNA characteristics.Compared with other populations,the nucleotide diversity of the Danzhou and Tunchang populations in the plain and hilly areas was higher.The average nucleotide differences were higher in the Sanya,Tunchang,and Danzhou populations.Haploid analysis revealed 6 haplotypes,with H02 being the dominant haplotype.The Sanya and Tunchang populations had the highest number of haplotypes,while the Haikou and Sanya populations had exclusive haplotypes.Only the plains populations had unique haplotypes,while the hill and mountainous populations had relatively fewer haplotypes.The neutral results and mismatch distribution map indicated that the population of Aedes albopictus has recently expanded in all regions.The AMOVA value showed that the intra-population differences were greater than the inter-population differences.Conclusions The mtDNA-COⅠgene can serve as a molecular marker for studying the genetic diversity of Aedes albopictus populations.The Sanya population in the plain region shows higher genetic diversity,and the two populations in the hilly region also have higher genetic diversity.However,the number of haplotypes is only higher in the Sanya population in the plains and the Danzhou population in the hills.