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  

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

作　　者：陶香林 马飞 李政 阚心蕊 叶长江 孙恩涛 TAO Xianglin;MA Fei;LI Zheng;KAN Xinrui;YE Changjiang;SUN Entao(School of Laboratory Medicine,Wannan Medical College,Wuhu,Anhui 241002,China)

机构地区：[1]皖南医学院检验学院,安徽芜湖241002

出　　处：《中国血吸虫病防治杂志》2023年第1期22-28,共7页Chinese Journal of Schistosomiasis Control

基　　金：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)。

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(F_(st))和基因流(N_(m))。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1301~1320 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=0.00691)。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(F_(st)=0.20240,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(N_(m)>1)。基于Cytb基因的中性检验发现,棕脊足螨WH种群Tajima’sD值和Fu’s FS值分别为-1.79631和-3.29398(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Hd与Pi均较高,整体值分别为0.985和0.01197,具有较高遗传多样性。分子方差分析发现,棕脊足螨Fst值为0.10462(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-6.08820和-1.93599(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,�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(F_(st))and gene flow value(N_(m))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 1301 to 1320 bp,respectively.All four isolates of G.fusca presented high genetic diversity based on mitochondrial Cytb and ITS genes(Hd=0.804,Pi=0.00691).AMOVA showed genetic differentiation among geographical isolates of G.fusca(F_(st)=0.20240,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(N_(m)>1).Tests of neutrality based on Cytb gene measured a Tajima’s D value of-1.79631(P<0.05)and a Fu’s FS value of-3.29398(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=0.01197).AMOVA showed moderate level of genetic differentiation between four isolates(F_(

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 

分 类 号：R384.4[医药卫生—医学寄生虫学]