CAJ | 학술논문

目的：探讨我国中华按蚊种群遗传多样性、遗传分化及种群系统发育关系。方法本实验采用2010-2012年间在我国山东，安徽，江苏，贵州，广西和云南等6个不同地理环境采集的中华按蚊样品，进行线粒体COI基因扩增并测序。采用Bioedit 7.0软件对测序结果进行比对分析；运用DnaSP 5.0软件确定种群遗传结构特点；应用Arlequin 3.1软件计算种群的遗传距离及遗传分化系数；通过IBDWS在线软件来明确遗传距离与地理距离的相关性；使用PHYLIP 3.6软件构建种群系统发育进化树。结果六个中华按蚊种群共123个雌蚊个体的PCR产物成功扩增和测序。PCR扩增中华按蚊线粒体COI基因序列大小为814 bp，平均A+T含量（71.2%）>平均G+C含量（28.8%）。线粒体COI序列分析结果显示，种群具有丰富的遗传多样性；分子变异等级分析表明中华按蚊遗传分化主要来自于种群内部；种群间存在一定的地理隔离现象。中性检验，错配分析及聚类进化树结果显示：中华按蚊种群发展经历扩张状态，而云南种群相对其他种群较独立，在系统树中为孤立一支。结论线粒体COI基因可以作为研究中华按蚊种群遗传结构和系统进化的理想分子标志。云南种群遗传发育较其他地理种群具有一定特殊性。
목적：탐토아국중화안문충군유전다양성、유전분화급충군계통발육관계。방법본실험채용2010-2012년간재아국산동，안휘，강소，귀주，엄서화운남등6개불동지리배경채집적중화안문양품，진행선립체COI기인확증병측서。채용Bioedit 7.0연건대측서결과진행비대분석；운용DnaSP 5.0연건학정충군유전결구특점；응용Arlequin 3.1연건계산충군적유전거리급유전분화계수；통과IBDWS재선연건래명학유전거리여지리거리적상관성；사용PHYLIP 3.6연건구건충군계통발육진화수。결과륙개중화안문충군공123개자문개체적PCR산물성공확증화측서。PCR확증중화안문선립체COI기인서렬대소위814 bp，평균A+T함량（71.2%）>평균G+C함량（28.8%）。선립체COI서렬분석결과현시，충군구유봉부적유전다양성；분자변이등급분석표명중화안문유전분화주요래자우충군내부；충군간존재일정적지리격리현상。중성검험，착배분석급취류진화수결과현시：중화안문충군발전경력확장상태，이운남충군상대기타충군교독립，재계통수중위고립일지。결론선립체COI기인가이작위연구중화안문충군유전결구화계통진화적이상분자표지。운남충군유전발육교기타지리충군구유일정특수성。
Objective To study the population genetic variation, genetic diversity and phylogenesis of Anopheles sinensis in China. Methods Anopheles sinensis samples collected from Shandong, Anhui, Jiangsu, Guizhou, and Yunnan Provinces and Guangxi Zhuang Autonomous Region with different geographical conditions between 2010 and 2012 were analyzed by mitochondrial DNA cytochrome oxidase subunit I (mtDNA-COI) gene amplification and sequencing. Bioedit 7.0 and DnaSP 5.0 software was used to compare the gene sequences and analyze the population genetic structure, respectively. Arlequin 3.1 was used to calculate the genetic distance and parameters of population differentiation. The relationship between the geographic and genetic distances was analyzed using IBD Web Service. PHYLIP 3.6 was used to construct the phylogenetic tree. Results PCR amplification and sequencing was performed successfully for 6 Anopheles sinensis populations containing 123 female mosquitoes. The length of mtDNA-COI gene fragment was 841 bp with an average A+T content of 71.2% and G+C content of 28.8%. High nucleotide diversity and genetic differentiation were observed among the Anopheles sinensis populations based on mtDNA-COI gene. Analysis of the molecular variance revealed a greater variation between populations than that within populations with isolation by distance between the populations. The Anopheles sinensis populations appeared to have undergone expansion, but the Yunnan population constituted an isolated branch in the phylogenetic tree. Conclusion mtDNA-COI can serve as the molecular marker to analyze population genetic variation and phylogenesis of Anopheles sinensis. The Yunnan population shows a phylogenetic difference from the other populations analyzed in this study.