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目的探讨不同地理区域伪威氏按蚊的群体遗传结构与差异,发现伪威氏按蚊的群体遗传与进化规律。方法对云南、西藏和缅甸伪威氏按蚊,采用mtDNA-Cytb基因进行群体遗传分析;测序结果以Chromas(Version 2.13)进行核对,序列比对采用ClustalX进行,MEGA软件包统计序列特征,运用ARLEQUIN(Version 3.0)统计和计算各群体的基因序列碱基分化参数;群体遗传结构由ARLEQUIN的AMOVA模块计算群体间的分化参数Fst(F-statistics)、基因流水平Nm(Nm=1-Fst/4Fst)等指标;TCS 1.21计算群体中的单倍型并构建单倍型间的家系网络图;MEGA软件构建单倍型之间的系统聚类树。结果伪威氏按蚊mtDNA-Cytb基因均各扩增出单一清晰的目的条带;mtDNA-Cytb基因分子标志的TCS单倍型家系网络图显示高水平平行演化;单倍型NJ聚类未显示出聚类关系与地理距离之间的相关性;AMOVA分析发现群体内差异远远大于群体间差异,伪威氏按蚊不同群体2分子标志的Fst和Nm值分别为0.01696,4.168。结论 mtDNA-Cytb基因可以作为研究按蚊群体遗传结构的理想分子标志;伪威氏按蚊群体间交流频繁,目前尚未发生群体分化。
Objective To investigate the population genetic structure and diversity of Anopheles Pseudoperis in different geographical regions and to find out the population genetic and evolutionary rules of Anopheles Pseudoperis. Methods Population genetic analysis was conducted using mtDNA-Cytb gene from Anopheles Pseudovirinae in Yunnan, Tibet and Myanmar. The sequence was verified by Chromas (Version 2.13). The sequence alignment was performed using ClustalX. The MEGA software package was used to characterize the sequence. ARLEQUIN (Version 3.0) Statistical analysis and calculation of base-line differentiation parameters of each group; Population genetic structure Fractional parameters Fst (F-statistics) and Nm (Nm = 1-Fst / 4Fst) were calculated by ARLEQUIN’s AMOVA module ) And other indicators; TCS 1.21 population haplotypes were calculated and the haplotype family network map constructed; MEGA software to build haplogroups between the system clustering tree. Results A single clear band was amplified from the mtDNA-Cytb gene of Anopheles pipiens; the TCS haplotype network of mtDNA-Cytb molecular markers showed high parallel evolution; the haplotype NJ clustering did not show The correlation between clustering relationship and geographical distance was found out. According to AMOVA analysis, the intra-population differences were far greater than those between populations. The Fst and Nm values of two molecular markers in different populations of Anopheles Pseudoperonodes were 0.01696 and 4.168, respectively. Conclusion The mtDNA-Cytb gene can be used as an ideal molecular marker for studying the genetic structure of Anopheles mosquitoes. The communication between populations of Anopheles pipiens is frequent, and no population differentiation has occurred yet.