海南热带臭虫分子鉴定及内共生菌Wolbachia检测  

Molecular identification of Cimex hemipterus and detection of the endosymbiont Wolbachia in Hainan Province

作　　者：芦亚君[1,2] 陈子杰 赵秋宇 解芸芸 夏琪 程诗 赵九莹 夏乾峰 LU Ya-jun;CHEN Zi-jie;ZHAO Qiu-yu;XIE Yun-yun;XIA Qi;CHENG Shi;ZHAO Jiu-Ying;XIA Qian-feng(School of Tropical Medicine,Hainan Medical University,Haikou 571199,China;Department of Pathogen Biology and Immunology,School of Basic Medical Sciences,Xi’an Jiaotong University Health Science Center)

机构地区：[1]海南医学院热带医学院,海南海口571199 [2]西安交通大学医学部基础医学院病原生物学与免疫学系

出　　处：《中国病原生物学杂志》2022年第11期1288-1292,共5页Journal of Pathogen Biology

基　　金：海南省重大科技计划项目(No.ZDKJ202003,ZDKJ2021036);海南省自然科学基金项目(No.2019RC223)。

摘　　要：目的了解海南臭虫种类及内共生菌Wolbachia的感染情况,并探讨海南臭虫COI基因和Wolbachia的wsp基因的序列特征及系统发育关系。方法2019年5月采集海南臭虫92只,提取基因组DNA,PCR扩增臭虫COI基因和Wolbachia表面蛋白基因wsp序列并测序。BLAST多序列比对确定物种;MEGA 11软件对序列进行碱基含量、保守位点、可变位点的计算和系统发育分析;SDT软件对序列两两比对,进行聚类分析。结果海南热带臭虫COI基因序列长度637 bp,G+C含量为37.21%,A+T含量为62.79%。与GenBank中热带臭虫COI序列比对显示,保守位点占99.84%。从92只海南热带臭虫中,检测出26只热带臭虫感染Wolbachia,感染率为28.26%。26条Wolbachia wsp序列长度约为610 bp,G+C含量为39%~40%,A+T含量为60%~61%,保守位点占99.03%。系统发育关系显示海南热带臭虫Wolbachia与GenBank中其他地理株的热带臭虫Wolbachia聚为一支,与梳毛臭虫、温带臭虫的Wolbachia系统发育关系较远。结论热带臭虫为海南臭虫的优势虫种,且自然感染内共生菌Wolbachia,为热带臭虫防控技术提供了理论依据。Objective To understand the bedbug species and the infection of endosymbiont Wolbachia in Hainan Province,and to explore the sequence characteristics and phylogenetic relationship based on the cytochrome oxidase(COI)gene of bedbugs and the Wolbachia surface protein(wsp)gene in Hainan Province.Methods In May 2019,92 bedbugs were collected in Hainan Province,genomic DNA was extracted,and the COI gene and the Wolbachia wsp sequences were amplified by PCR and sequenced.The multiple sequence alignment was carried out to determine the bedbug species via BLAST.The nucleotide base content,the conserved sites,and the variable sites were calculated and the phylogenetic relationship was analyzed using the MEGA 11 software.Pairwise comparisons between the sequences were performed and the cluster analysis was conducted with SDT software.Results The COI gene sequence of Cimex hemipterus in Hainan Province was 637 bp in length with a G+C content of 37.21%and an A+T content of 62.79%.Alignment with the COI sequences of Cimex hemipterus in GenBank revealed that the conserved sites accounted for 99.84%.The Wolbachia infection was detected in 26 Cimex hemipterus of 92 Cimex hemipterus in Hainan Province,with an infection rate of 28.26%.The 26 Wolbachia wsp sequences were about 610 bp in length,with a G+C content of 39%-40%,an A+T content of 60%-61%,and the conserved sites accounting for 99.03%.The phylogenetic relationships demonstrated that the Wolbachia of Cimex hemipterus in Hainan Province clustered into one clade with the Wolbachia of Cimex hemipterus at the other geographical location retrieved in GenBank,and had a remote phylogenetic relationship with Wolbachia of Cimex lectularius and Cimex pilosellus.Conclusion Cimex hemipterus was the dominant species of bedbugs in Hainan Province,and was naturally infected with the endosymbiont Wolbachia,which provided a theoretical basis for the prevention and control of Cimex hemipterus.

关 键 词：热带臭虫 COI基因 WOLBACHIA 分子鉴定 系统发育分析 

分 类 号：R384.3[医药卫生—医学寄生虫学]