›› 2018, Vol. 61 ›› Issue (4): 488-497.doi: 10.16380/j.kcxb.2018.04.011

• 研究论文 • 上一篇    下一篇

基于线粒体COI基因的青海省44种蚤类的分子鉴定及系统发育分析

马英1, 李海龙1, 何建1, 赵延梅1, 杨汉青1, 鲁亮2,*, 刘起勇2   

  1.  (1. 青海省地方病预防控制所, 西宁 811602; 2. 中国疾病预防控制中心传染病预防控制所, 北京 102200)
  • 出版日期:2018-04-20 发布日期:2018-04-20

Molecular identification and phylogenetic analysis of 44 species of fleas in Qinghai Province, western China based on mtDNA COI gene sequence

MA Ying1, LI Hai-Long1, HE Jian1, ZHAO Yan-Mei1, YANG Han-Qing1, LU Liang2,*, LIU Qi-Yong2   

  1. (1. Qinghai Institute for Endemic Disease Prevention and Control, Xining 811602, China; 2. National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102200, China)
  • Online:2018-04-20 Published:2018-04-20

摘要:  【目的】本研究应用DNA条形码技术对青海省部分蚤种进行鉴定,旨在弥补蚤类传统形态分类方法的不足。【方法】通过PCR扩增3总科6科22属44种共182头蚤类标本的线粒体COI基因片段(约600 bp)并进行测序和比对;用K2P模型计算种内及种间遗传距离;以邻接法(neighbor-joining, NJ)构建系统发育树。【结果】共测得182条COI基因序列片段(GenBank登录号: MG138154-MG138335);分析可知蚤种内遗传距离0.01%~2.90%,种间遗传距离4%~12%,种间遗传距离显著大于种内遗传距离。系统发育树显示,同一物种的不同个体均形成高支持率的单系,种间分支很明显。【结论】本研究结果表明COI基因可以用于蚤类的快速鉴定。

关键词: COI基因, 蚤类, DNA条形码技术, 分子鉴定, 系统发育, 青海

Abstract: 【Aim】 DNA barcoding method was used to identify flea samples from Qinghai Province, western China in order to make up the deficiency of the traditional morphological classification method in this study. 【Methods】 The partial fragments (about 600 bp) of the mtDNA COI gene of 182 flea individuals that belong to 3 superfamilies, 6 families, 22 genera and 44 species were amplified by PCR, sequenced and aligned. The intra-and inter-species genetic distances were calculated with K2P model, and a phylogenetic tree was constructed with the neighbor-joining (NJ) method. 【Results】 Totally 182 COI gene sequences (GenBank accession no.: MG138154-MG138335) were sequenced successfully. The inter-species genetic distance (4%-12%) was significantly greater than the intra-species genetic distance (0.01%-2.90%). The NJ tree obtained showed that samples of the same species formed monophyletic groups with high support value, and inter-species branches were clear. 【Conclusion】 The results confirm that COI gene can be used in molecular identification of fleas.

Key words: COI gene, flea, DNA barcoding, molecular identification, phylogeny, Qinghai