【摘要】： 目的:研究不同地理種群白紋伊蚊(Aedes albopictus)的線粒體NADH脫氫酶亞基Ⅳ(mtDNA-ND4)和線粒體細胞色素氧化酶亞基Ⅰ(mtDNA-COI)基因序列特征,探索從分子水平對不同地理株進行鑒別,掌握不同地理種群白紋伊蚊遺傳結(jié)構(gòu)以及基因漂流,了解登革熱媒介伊蚊遺傳多態(tài)性,推測其與登革熱流行的關(guān)系。評價mtDNA-ND4和mtDNA-COI作為分子標志物在白紋伊蚊種內(nèi)鑒別上的應(yīng)用價值。方法:采集福建省四個地區(qū)不同海拔高度、登革熱流行區(qū)與非流行區(qū)的白紋伊蚊幼蟲,實驗室飼養(yǎng)至成蚊,形態(tài)鑒定并收集、制成標本,以國內(nèi)其它地區(qū)(云南、上海、貴州)白紋伊蚊作為對照,參考相關(guān)文獻設(shè)計的引物,研磨并提取線粒體DNA,采用PCR方法擴增mtDNA-ND4和mtDNA-COI基因,通過GenBank進行序列比對確認,運用基因分析軟件與遺傳分析軟件進行遺傳分析。結(jié)果:①獲得mtDNA-ND4和mtDNA-COI基因片段,長度分別為324b和415bp,堿基A+T含量分別為73.2%和70.2%,堿基置換位點分別為17和8個,顛換率為62.5%和64.71%。mtDNA-ND4核苷酸多態(tài)性(pi)為0.012,大于mtDNA-COI(0.00277)。②基于mtDNA-ND4基因,發(fā)現(xiàn)福建19個個體存在5個單倍型,單倍型多態(tài)性(h)為0.442,單倍型家系網(wǎng)絡(luò)圖顯示福建單倍型由兩種單倍型分別進化而來。N-J法與UPGMA法構(gòu)建分子系統(tǒng)樹顯示種間分類清楚,但不同地理種群分類雜亂。③基于mtDNA-COI基因,發(fā)現(xiàn)福建、云南、上海、貴州41個個體存在6種單倍型,其中福建16個個體存在4個單倍型,單倍型多態(tài)性為0.592,分子多樣性指數(shù)顯示福建高于其它地理種群。單倍型家系網(wǎng)絡(luò)顯示所有的單倍型都由一種單倍型進化而來,有三種單倍型在本次研究中未被發(fā)現(xiàn)。遺傳距離結(jié)果顯示不同地理種群出現(xiàn)較小程度的分化(FST=0.28136),很可能由遺傳漂變引起的,AMOVA結(jié)果顯示該遺傳分化主要來自種群內(nèi)(占71.86%)。N-J法與UPGMA法構(gòu)建分子系統(tǒng)樹顯示,云南、東山、武夷山三個地理種群分別歸為一類。結(jié)論:①種群遺傳多態(tài)性分析結(jié)果顯示不同地理種群的白紋伊蚊出現(xiàn)了小程度的遺傳分化,很可能由遺傳漂變引起,主要由種群內(nèi)變異引起。福建種群的分子多態(tài)性高于其它地理種群。②在白紋伊蚊種內(nèi)鑒別上,分子標志物mtDNA-COI基因序列優(yōu)于mtDNA-ND4基因序列。③福建東山白紋伊蚊幼蟲密度、線粒體遺傳多態(tài)性高,與登革熱流行地區(qū)毗鄰,存在登革熱暴發(fā)的危險,應(yīng)引起足夠的重視。
[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，

本文編號：2509664