中国血吸虫病防治杂志 ›› 2023, Vol. 35 ›› Issue (1): 22-.

• 论著 • 上一篇    下一篇

基于细胞色素b和内转录间隔区基因序列的4个棕脊足螨地理种群遗传变异分析

陶香林,马飞,李政,阚心蕊,叶长江,孙恩涛*   

  1. 皖南医学院检验学院(安徽 芜湖 241002)
  • 出版日期:2023-03-14 发布日期:2023-03-14
  • 作者简介:陶香林,女,硕士,讲师。研究方向:微生物学
  • 基金资助:
    国家自然科学基金(31870352);皖南医学院校级重点科研基金项目 (WK2019Z02)

Genetic variations in four geographical isolates of Gohieria fusca based on cytochrome b and internal transcribed spacer genes

TAO Xianglin, MA Fei, LI Zheng, KAN Xinrui, YE Changjiang, SUN Entao*   

  1. School of Laboratory Medicine, Wannan Medical College, Wuhu, Anhui 241002, China
  • Online:2023-03-14 Published:2023-03-14

摘要: 目的 分析不同地理种群棕脊足螨(Gohieria fusca)遗传多样性和遗传分化。方法 自安徽省芜湖市(WH)、蚌埠市(BB)、亳州市(BZ)和浙江省嘉兴市(JX)等4个地区采集螨虫标本并筛选出棕脊足螨,采用PCR技术扩增棕脊足螨线粒体细胞色素b(cytochrome b, Cytb)和核糖体DNA内转录间隔区(internal transcribed spacer, ITS)基因并对扩增产物测序。采用Chromas 2和DNASTAR 1.00软件进行序列校对和拼接,采用DnaSP 5.10.00软件计算各种群单倍型多样性(Hd)、核苷酸多态性(Pi),应用MEGA 10.2软件包计算遗传分化指数(Fst)和基因流(Nm)。采用Arlequin 3.1软件计算Tajima’s D和Fu’s FS值,并进行中性检验和分子方差分析;采用Network 10.2软件基于Median⁃joining法构建单倍型网络图。结果 棕脊足螨样本Cytb基因序列长度为372 bp、ITS基因序列长度为1 301 ~ 1 320 bp。基于线粒体Cytb基因和核糖体ITS基因序列分析发现,4个棕脊足螨地理种群均表现出较高遗传多样性(Hd = 0.804,Pi = 0.006 91)。分子方差分析发现,4个棕脊足螨地理种群间存在遗传分化(Fst = 0.202 40, P < 0.05),且遗传变异主要来自种群内(79.76%)。基因流分析发现,4个棕脊足螨地理种群间局部存在高水平基因流(Nm > 1)。基于Cytb基因的中性检验发现,棕脊足螨WH种群Tajima’s D值和Fu’s FS值分别为-1.796 31和-3.293 98(P均< 0.05),表明WH种群可能发生过历史扩张。单倍型网络图和系统发育树分析结果一致,均表明不同棕脊足螨地理种群无明显地理分布格局。基于ITS基因序列分析发现,4个棕脊足螨地理种群单倍型多样性(Hd)与核苷酸多态性(Pi)均较高,整体值分别为0.985和0.011 97,具有较高遗传多样性。分子方差分析发现,棕脊足螨Fst值为0.104 62(P < 0.05),种群间存在中等程度遗传分化。基于ITS基因的中性检验发现,棕脊足螨整体Tajima’s D值和Fu’s FS值分别为-6.088 20和-1.935 99(P均> 0.05),无明显历史扩张。结论 4个棕脊足螨地理种群遗传多样性较高,存在显著遗传分化。由于不同棕脊足螨地理种群间局部发生高水平基因流,未发现其明显地理分布格局。  

关键词: 棕脊足螨, 细胞色素b, 内转录间隔区, 种群多样性, 遗传分化, 基因流

Abstract: Objective To investigate the genetic diversity and genetic differentiation of different geographical isolates of Gohieria fusca. Methods G. fusca isolates were sampled from Wuhu (WH), Bengbu (BB) and Bozhou cities (BZ) of Anhui Province and Jiaxing City of Zhejiang Province (JX). Mitochondrial cytochrome b (Cytb) and ribosomal internal transcribed spacer (ITS) genes were amplified in WH, BB, BZ and JX isolates of G. fusca using PCR assay. The gene sequences were edited and aligned using the software Chromas 2 and DNASTAR 1.00, and the haplotype, haplotype diversity (Hd) and nucleotide polymorphism (Pi) of each isolate were calculated using the software DnaSP 5.10.00. The genetic differentiation among isolates (Fst) and gene flow value (Nm) were estimated using the software MEGA 10.2, and a phylogenetic tree was built. Tests of neutrality and analysis of molecular variance (AMOVA) were performed using the software Arlequin 3.1 and a haplotype network was built based on the Median⁃Joining network using the software Network 10.2. Results PCR assay showed that the sizes of the Cytb and ITS genes were 372 bp and 1 301 to 1 320 bp, respectively. All four isolates of G. fusca presented high genetic diversity based on mitochondrial Cytb and ITS genes (Hd = 0.804,Pi = 0.006 91). AMOVA showed genetic differentiation among geographical isolates of G. fusca (Fst = 0.202 40, P < 0.05), and the genetic variation was mainly caused by intra⁃population variations (79.76%). Gene flow analysis showed a high level of gene flow among G. fusca isolates (Nm > 1). Tests of neutrality based on Cytb gene measured a Tajima’s D value of -1.796 31 (P < 0.05) and a Fu’s FS value of -3.293 98 (P < 0.05) in WH isolate of G. fusca, indicating population expansion in WH isolate of G. fusca. Haplotype network analysis and phylogenetic analysis revealed no remarkable geographical distribution pattern among different geographical isolates of G. fusca. All four isolates of G. fusca presented high genetic diversity (Hd = 0.985,Pi = 0.011 97). AMOVA showed moderate level of genetic differentiation between four isolates (Fst = 0.104 62, P < 0.05). The tests of neutrality based on ITS genes measured a Tajima’s D value of -6.088 20 and a Fu’s FS value of -1.935 99 (both P > 0.05) in the whole isolate of G. fusca, indicating no obviously population expansion. Conclusions  The four geographical isolates of G. fusca have high genetic diversity and remarkable genetic differentiation. Since a high level of gene flow is detected among different geographical isolates of G. fusca, no obvious geographical distribution pattern of G. fusca is found.

Key words: Gohieria fusca, Cytochrome b, Internal transcribed spacer, Genetic diversity, Genetic differentiation, Gene flow

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