登革热媒介白纹伊蚊mtDNA-COI和mtDNA-ND4基因多态性研究
发布时间：2019-07-03 21:01
【摘要】： 目的:研究不同地理种群白纹伊蚊(Aedes albopictus)的线粒体NADH脱氢酶亚基Ⅳ(mtDNA-ND4)和线粒体细胞色素氧化酶亚基Ⅰ(mtDNA-COI)基因序列特征,探索从分子水平对不同地理株进行鉴别,掌握不同地理种群白纹伊蚊遗传结构以及基因漂流,了解登革热媒介伊蚊遗传多态性,推测其与登革热流行的关系。评价mtDNA-ND4和mtDNA-COI作为分子标志物在白纹伊蚊种内鉴别上的应用价值。方法:采集福建省四个地区不同海拔高度、登革热流行区与非流行区的白纹伊蚊幼虫,实验室饲养至成蚊,形态鉴定并收集、制成标本,以国内其它地区(云南、上海、贵州)白纹伊蚊作为对照,参考相关文献设计的引物,研磨并提取线粒体DNA,采用PCR方法扩增mtDNA-ND4和mtDNA-COI基因,通过GenBank进行序列比对确认,运用基因分析软件与遗传分析软件进行遗传分析。结果:①获得mtDNA-ND4和mtDNA-COI基因片段,长度分别为324b和415bp,碱基A+T含量分别为73.2%和70.2%,碱基置换位点分别为17和8个,颠换率为62.5%和64.71%。mtDNA-ND4核苷酸多态性(pi)为0.012,大于mtDNA-COI(0.00277)。②基于mtDNA-ND4基因,发现福建19个个体存在5个单倍型,单倍型多态性(h)为0.442,单倍型家系网络图显示福建单倍型由两种单倍型分别进化而来。N-J法与UPGMA法构建分子系统树显示种间分类清楚,但不同地理种群分类杂乱。③基于mtDNA-COI基因,发现福建、云南、上海、贵州41个个体存在6种单倍型,其中福建16个个体存在4个单倍型,单倍型多态性为0.592,分子多样性指数显示福建高于其它地理种群。单倍型家系网络显示所有的单倍型都由一种单倍型进化而来,有三种单倍型在本次研究中未被发现。遗传距离结果显示不同地理种群出现较小程度的分化(FST=0.28136),很可能由遗传漂变引起的,AMOVA结果显示该遗传分化主要来自种群内(占71.86%)。N-J法与UPGMA法构建分子系统树显示,云南、东山、武夷山三个地理种群分别归为一类。结论:①种群遗传多态性分析结果显示不同地理种群的白纹伊蚊出现了小程度的遗传分化,很可能由遗传漂变引起,主要由种群内变异引起。福建种群的分子多态性高于其它地理种群。②在白纹伊蚊种内鉴别上,分子标志物mtDNA-COI基因序列优于mtDNA-ND4基因序列。③福建东山白纹伊蚊幼虫密度、线粒体遗传多态性高,与登革热流行地区毗邻,存在登革热暴发的危险,应引起足够的重视。
[Abstract]:Aim: to study the sequence characteristics of mitochondrial NADH dehydrogenase subunit IV (mtDNA-ND4) and mitochondrial cytochrome oxidase subunit I (mtDNA-COI) gene in different geographical populations of Aedes albopictus (Aedes albopictus), to explore the identification of different geographical strains at molecular level, to master the genetic structure and gene drifting of Aedes albopictus in different geographical populations, to understand the genetic polymorphism of Aedes albopictus in different geographical populations, and to speculate the relationship between the genetic polymorphism of Aedes albopictus and the epidemic of dengue fever. To evaluate the application value of mtDNA-ND4 and mtDNA-COI as molecular markers in intra-species identification of Aedes albopictus. Methods: the larvae of Aedes albopictus from different altitude, dengue fever epidemic area and non-epidemic area in Fujian Province were collected and raised to adult mosquitoes in laboratory. The morphology of Aedes albopictus was identified and collected. Aedes albopictus was used as control in other parts of China (Yunnan, Shanghai, Guizhou). The mtDNA-ND4 and mtDNA-COI genes were amplified by PCR with reference to the primers designed in the relevant literature. Sequence alignment was carried out by GenBank, and genetic analysis software were used for genetic analysis. Results: 1 the length of mtDNA-ND4 and mtDNA-COI gene fragments were 324b and 415bp, the content of base A T was 73.2% and 70.2%, the base replacement sites were 17 and 8, the transversion rate was 62.5%, and the (pi) of 64.71%.mtDNA-ND4 nucleotides polymorphism was 0.012, which was larger than that of mtDNA-COI (0.00277). 2 based on mtDNA-ND4 gene, it was found that there were 5 haplotypes in 19 individuals in Fujian. The haplotype polymorphism (h) was 0.442. The haplotype family network map showed that Fujian haplotype was evolved from two haplotypes respectively. N-J method and UPGMA method constructed molecular systematic tree to show clear interspecific classification, but different geographical population classification was disorderly. 3 based on mtDNA-COI gene, it was found that there were 6 haplotypes in 41 individuals in Fujian, Yunnan, Shanghai and Guizhou, among which 4 haplotypes existed in 16 individuals in Fujian. The haplotype polymorphism was 0.592, and the molecular diversity index showed that Fujian was higher than other geographical populations. Haplotype family network showed that all haplotypes evolved from one haplotype, and three haplotypes were not found in this study. The results of genetic distance showed that there was a small degree of differentiation (FST=0.28136) in different geographical populations, which was probably caused by genetic drift. AMOVA results showed that the genetic differentiation mainly came from within the population (41.86%). The construction of molecular systematic tree by N 鈮，
本文编号：2509664
[Abstract]:Aim: to study the sequence characteristics of mitochondrial NADH dehydrogenase subunit IV (mtDNA-ND4) and mitochondrial cytochrome oxidase subunit I (mtDNA-COI) gene in different geographical populations of Aedes albopictus (Aedes albopictus), to explore the identification of different geographical strains at molecular level, to master the genetic structure and gene drifting of Aedes albopictus in different geographical populations, to understand the genetic polymorphism of Aedes albopictus in different geographical populations, and to speculate the relationship between the genetic polymorphism of Aedes albopictus and the epidemic of dengue fever. To evaluate the application value of mtDNA-ND4 and mtDNA-COI as molecular markers in intra-species identification of Aedes albopictus. Methods: the larvae of Aedes albopictus from different altitude, dengue fever epidemic area and non-epidemic area in Fujian Province were collected and raised to adult mosquitoes in laboratory. The morphology of Aedes albopictus was identified and collected. Aedes albopictus was used as control in other parts of China (Yunnan, Shanghai, Guizhou). The mtDNA-ND4 and mtDNA-COI genes were amplified by PCR with reference to the primers designed in the relevant literature. Sequence alignment was carried out by GenBank, and genetic analysis software were used for genetic analysis. Results: 1 the length of mtDNA-ND4 and mtDNA-COI gene fragments were 324b and 415bp, the content of base A T was 73.2% and 70.2%, the base replacement sites were 17 and 8, the transversion rate was 62.5%, and the (pi) of 64.71%.mtDNA-ND4 nucleotides polymorphism was 0.012, which was larger than that of mtDNA-COI (0.00277). 2 based on mtDNA-ND4 gene, it was found that there were 5 haplotypes in 19 individuals in Fujian. The haplotype polymorphism (h) was 0.442. The haplotype family network map showed that Fujian haplotype was evolved from two haplotypes respectively. N-J method and UPGMA method constructed molecular systematic tree to show clear interspecific classification, but different geographical population classification was disorderly. 3 based on mtDNA-COI gene, it was found that there were 6 haplotypes in 41 individuals in Fujian, Yunnan, Shanghai and Guizhou, among which 4 haplotypes existed in 16 individuals in Fujian. The haplotype polymorphism was 0.592, and the molecular diversity index showed that Fujian was higher than other geographical populations. Haplotype family network showed that all haplotypes evolved from one haplotype, and three haplotypes were not found in this study. The results of genetic distance showed that there was a small degree of differentiation (FST=0.28136) in different geographical populations, which was probably caused by genetic drift. AMOVA results showed that the genetic differentiation mainly came from within the population (41.86%). The construction of molecular systematic tree by N 鈮，
本文编号：2509664
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