江苏省淮安市蚊虫感染沃尔巴克氏体的调查
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摘要
目的了解淮安市蚊种自然感染沃尔巴克氏体Wolbachia的特点及Wolbachia种群遗传多样性研究,为蚊媒及蚊媒传染病的生物防治提供科学依据。方法用诱蚊灯法采集蚊虫,对采集的蚊种进行DNA提取,采用PCR扩增细胞色素C氧化酶亚基Ⅰ(mt COI)基因及wsp基因、测序及系统进化分析。结果在淮安市捕获蚊种5种共计1 640只,Wolbachia在蚊虫样本组的总感染率为28. 05%(23/82)。淮安市蚊种携带的Wolbachia的核苷酸序列相似性为71. 5%~99. 9%,氨基酸序列相似性为70. 7%~99. 9%。系统进化分析显示淮安蚊种携带的Wolbachia被划分为Mors、Dei和Pip组。淮安蚊种携带的Wolbachia种群有4个单倍型,Wolbachia的单倍型多样性Hd=0. 597,核苷酸多样性π=0. 120,多态性位点S=151。结论淮安市蚊虫普遍存在Wolbachia的感染,其基因型为Mors、Dei和Pip。
Objective To determine the infection state of Wolbachia from mosquitoes population and population genetic diversity of Wolbachia in Huai'an,and lay the foundation for mosquito and mosquito-borne disease control.Methods Light traps were applied to collect mosquitoes,genomic DNA was extracted for PCR amplification of the mitochondrial cytochrome c oxidase subunitⅠ( mt COI) gene of mosquitoes and wsp gene of Wolbachia,followed by sequencing and phylogenetic analysis. Results A total of 1 640 mosquitoes,belonging to five species,were captured in Huai'an,twenty three groups out of eighty two groups were infected Wolbachia with the group infection rate of28. 05%. Nucleotide sequence similarity of Wolbachia strains in Huai'an was 71. 5%-99. 9%,and the amino acid sequence similarity was 70. 7%-99. 9%. The phylogenetic analysis indicated the Wolbachia belonged to group Mors,Dei and Pip. Four haplotyes in Wolbachia populations in Huai 'an were identified with the genetic diversity of populations( Hd = 0. 597,π = 0. 120,S = 151). Conclusion Wolbachia widely spread in mosquitoes in Huai'an,and its genotypes belong to group Mors,Dei and Pip.
Objective To determine the infection state of Wolbachia from mosquitoes population and population genetic diversity of Wolbachia in Huai'an,and lay the foundation for mosquito and mosquito-borne disease control.Methods Light traps were applied to collect mosquitoes,genomic DNA was extracted for PCR amplification of the mitochondrial cytochrome c oxidase subunitⅠ( mt COI) gene of mosquitoes and wsp gene of Wolbachia,followed by sequencing and phylogenetic analysis. Results A total of 1 640 mosquitoes,belonging to five species,were captured in Huai'an,twenty three groups out of eighty two groups were infected Wolbachia with the group infection rate of28. 05%. Nucleotide sequence similarity of Wolbachia strains in Huai'an was 71. 5%-99. 9%,and the amino acid sequence similarity was 70. 7%-99. 9%. The phylogenetic analysis indicated the Wolbachia belonged to group Mors,Dei and Pip. Four haplotyes in Wolbachia populations in Huai 'an were identified with the genetic diversity of populations( Hd = 0. 597,π = 0. 120,S = 151). Conclusion Wolbachia widely spread in mosquitoes in Huai'an,and its genotypes belong to group Mors,Dei and Pip.
引文
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[11]刘倩倩,吴炳耀,何江,等.江苏口岸常见蚊虫沃尔巴克氏体感染调查研究[J].中国媒介生物学及控制杂志,2018,29(2):138-142.
[12] Nugapola N,De WS,Karunaratne S. Distribution and phylogeny of Wolbachiastrains in wild mosquito populations in Sri Lanka:[J].Parasit Vectors,2017,10(1):230.
[13] Kittayapong P,Baisley KJ,Baimai V,et al. Distribution and diversity of Wolbachia infections in Southeast Asian mosquitoes(Diptera:Culicidae)[J]. J Med Entomol,2000,37(3):340-345.
[14]杨鹏飞,燕清丽,苏琦,等.基于mt COI基因序列的DNA条形码技术在江苏沿江长角血蜱鉴定中的应用[J].江苏预防医学,2018(2):135-139.
[15]胡双双,吴炳耀,何江,等.江苏口岸小家鼠细胞色素氧化酶亚基COI基因的多态性研究[J].中华卫生杀虫药械,2017(1):65-69.
[2] Kittayapong P,Baisley KJ,Baimai V,et al. Distribution and diversity of Wolbachia infections in Southeast Asian mosquitoes(Diptera:Culicidae)[J]. J Med Entomol,2000,37(3):340-345.
[3]林立丰,吴德,张欢,等.广东省蚊虫感染沃尔巴克氏体初步调查研究[J].中国媒介生物学及控制杂志,2014,25(2):113-115.
[4] Van Meer MM,Witteveldt J,Stouthamer R. Phylogeny of the arthropod endosymbiont Wolbachia based on the wsp gene[J].Insect Molecular Biology,2010,8(3):399-408.
[5] Moreira LA. Wolbachia,in Aedes,mosquitoes:towards biological control of vector-borne diseases[M]. Holand:Wageningen Academic Publishers,2013:155-165.
[6] Reumer BM,van Alphen JJ,Kraaijeveld K. Population genetics of Wolbachia-infected,parthenogenetic and uninfected,sexual populations of Tetrastichus coeruleus(Hymenoptera:Eulophidae)[J]. Molecular Ecol,2013,22(17):4433-4444.
[7] Ben Longdon. Male-killing Wolbachia do not protect Drosophila bifasciata against viral infection[J]. BMC Microbiol,2012,12(Suppl 1):1-6.
[8] Kondo N,Shimada M,Fukatsu T. Infection density of Wolbachia endosymbiont affected by co-infection and host genotype[J]. Biol Letters,2005,1(4):488-491.
[9] Zhou W,Rousset F,O'Neil S. Phylogeny and PCR-based classification of Wolbachia strains using wsp gene sequences.[J].Proc Biol Sci,1998,265(1395):509-515.
[10] Díaz-Nieto L M,MaciáA,Parisi G,et al. Distribution of Mosquitoes in the South East of Argentina and First Report on the Analysis Based on 18S r DNA and COI Sequences[J]. Plos One,2013,8(9):e75516.
[11]刘倩倩,吴炳耀,何江,等.江苏口岸常见蚊虫沃尔巴克氏体感染调查研究[J].中国媒介生物学及控制杂志,2018,29(2):138-142.
[12] Nugapola N,De WS,Karunaratne S. Distribution and phylogeny of Wolbachiastrains in wild mosquito populations in Sri Lanka:[J].Parasit Vectors,2017,10(1):230.
[13] Kittayapong P,Baisley KJ,Baimai V,et al. Distribution and diversity of Wolbachia infections in Southeast Asian mosquitoes(Diptera:Culicidae)[J]. J Med Entomol,2000,37(3):340-345.
[14]杨鹏飞,燕清丽,苏琦,等.基于mt COI基因序列的DNA条形码技术在江苏沿江长角血蜱鉴定中的应用[J].江苏预防医学,2018(2):135-139.
[15]胡双双,吴炳耀,何江,等.江苏口岸小家鼠细胞色素氧化酶亚基COI基因的多态性研究[J].中华卫生杀虫药械,2017(1):65-69.