热带病与寄生虫学

热带病与寄生虫学 ›› 2021, Vol. 19 ›› Issue (5): 254-.

• 实验研究 • 上一篇    下一篇

安徽省不同地区日本血吸虫群体线粒体基因遗传变异研究

马晓荷,汪敏,朱磊,郭见多,李清越,刘婷,翟杜娟,孙成松,张世清,汪天平
  

  1. 安徽省血吸虫病防治研究所,安徽 合肥 230601
  • 出版日期:2021-10-20 发布日期:2021-11-03
  • 通信作者: 汪天平,E-mail:tpwang906@ 163. com
  • 作者简介:马晓荷,女,硕士,检验师,研究方向:微生物学。 E-mail:xiaohema1987@ hotmail. com
  • 基金资助:
    安徽省血吸虫病防治研究所青年科研基金项目( XF2018004);安徽省自然科学基金项目(2008085QH430);安徽省重点研究与开发
    计划项目    (202104j07020001)

Investigation on the genetic variation of mitochondrial genes in Schistosoma japonicum species in different areas of Anhui Province

MA Xiao-he,WANG Min,ZHU Lei,GUO Jian-duo,LI Qing-yue,LIU Ting,ZHAI Du-juan,SUN Cheng-song,ZHANG Shi-qing,WANG Tian-ping#br#   

  1. Anhui Provincial Institute of Schistosomiasis Control,Hefei 230601,Anhui Province,China
  • Online:2021-10-20 Published:2021-11-03

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摘要： 目的 探讨安徽省不同地区日本血吸虫种群的遗传进化关系。 方法 分别从安徽省山丘型流行区(池州市石台县)和湖沼型流行区(安庆市大观区、铜陵市枞阳县)采集钉螺,逸出尾蚴后感染小鼠,每只小鼠感染(50±3)条尾蚴。 35 d 后解剖小鼠,门静脉灌注法收集成虫,提取成虫 DNA,PCR 扩增、克隆和测序成虫的线粒体 NADH(还原型烟酰胺腺嘌呤二核苷酸)脱氢酶 1(ND1)和细胞色素 C 氧化酶亚基Ⅰ( COⅠ)基因,利用生物信息学软件 Dnasp6. 0 和 Mega X 分析所得序列基本特征并构建系统进化树作遗传进化分析。 结果 安徽省三个地区日本血吸虫样本的 ND1 基因序列存在 15 个变异位点,COⅠ序列存在 74 个变异位点;ND1 基因和 COⅠ基因的核苷酸多样性(Pi)分别为 0. 004 7、0. 017 3,样本间 COⅠ基因序列差异高于 ND1 基因;来自湖沼型流行区的样本 ND1 基因和 COⅠ 基因的变异位点数、核苷酸多样性(Pi)均高于来自山丘型流行区样本。 来自山丘型流行区的样本在 ND1 系统进化树中与来自湖沼型流行区的样本聚集在一起,而在 COⅠ基因系统进化树中单独成簇。 结论 来自湖沼型流行区的日本血吸虫样本群体的基因多样性高于来自山丘型流行区的样本。 安徽省三个地区日本血吸虫群体的 COⅠ基因存在遗传分化,而 ND1 基因尚未产生较明显的遗传分化。

关键词: 日本血吸虫, 遗传分化, NADH 脱氢酶 1 基因, 细胞色素 C 氧化酶亚基Ⅰ基因, 安徽省

Abstract: Objective To investigate the genetic evolutionary relationship among Schistosoma japonicum populations in different areas within Anhui Province. Methods Naturally Oncomelania hupensis were collected form the hilly / mountainous regions(Shitai County in Chizhou City) and swamp / lake areas( Daguan district in Anqing City,Zongyang County in Tongling City). Cercariae escaped from the infected snails obtained in diverse locality were used to infect laboratory-raised female BALB / c mice by dose of (50±3) cercariae in individual mouse. Then the mice were dissected 35 days after infection to collect the adult worms via portal vein perfusion technique. DNA was extracted from the adults to undergo PCR amplification,cloning and sequencing of the mitochondrial dehydrogenase 1( ND1) and cytochrome C oxidase subunit Ⅰ( COⅠ) gene fragments. The sequences were analyzed by bioinformatics software Dnasp 6. 0 and Mega X. Evolutionary trees were constructed using the neighbor-joining method with MEGA software ( version 10. 2. 2) to observe the evolution clues. Results Fifteen nucleotide variation sites in ND1 gene and 74 in COⅠ gene were identified in the samples obtained in three districts. The nucleotide diversity (Pi) of ND1 and COⅠ gene was 0. 004 7 and 0. 017 3,respectively. The variation of CO Ⅰ between these samples was a little higher than that of ND1 gene. Variable sites of ND1 and COⅠ genes and Pi were higher in the samples from swamp / lake areas than in those from hilly / mountainous regions. Samples from Shitai County were independent in the tree based on COⅠ gene,yet all of the samples were gathered together in the tree at the foundation of ND1 gene. Conclusion The genetic diversity is higher in S. japonicum from marshy regions than in that from hilly regions. The results indicated that there was some differences between the COⅠ gene of S. japonicum from hilly region and marshy region in Anhui province, and meanwhile there was no significant differences between the ND1 gene of these S. japonicum worms by now.

Key words: Schistosoma japonicum, Genetic variation, NADH dehydrogenase subunit 1 gene, Cytochrome C oxidase subunit Ⅰ gene, Anhui Province

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