淮安市常见蚊种感染沃尔巴克氏体的分子鉴定及遗传多样性研究

倪文宇; 燕清丽; 何芳; 吕蒙蒙; 行岳真; 殷德辉; 彭海燕; 杨鹏飞

doi:10.3784/jbjc.202505210382

淮安市常见蚊种感染沃尔巴克氏体的分子鉴定及遗传多样性研究

Molecular Identification and Genetic Diversity of Wolbachia Infection in Common Mosquito Species in Huai'an

摘要

摘要:
目的初步研究江苏省淮安市常见蚊种中沃尔巴克氏体（Wolbachia）的感染情况、遗传多样性、种群动态及传播模式，为蚊传疾病的科学防治提供参考。
方法 2022年7-10月，使用灯诱法在淮安市的机场、货站、绿化带以及牛棚、猪圈、民房等生境中采集蚊虫，利用形态学和DNA条形码（mt COI）鉴定种类。利用Wolbachia表面蛋白基因WSP进行分子鉴定。使用IQ-TREE 2.2.0软件构建系统发育树，R 4.4.2软件进行单倍型分析，PopART 1.7软件构建单倍型网络图。使用R 4.4.2软件计算遗传分化（AMOVA，FST），DnaSP 6.1软件进行中性检验（Tajima's D、Fu's Fs）。使用R 4.4.2软件探究协同进化信号（PACo，ParaFit），Jane 4软件探究Wolbachia和宿主间的进化关系。
结果在淮安市各监测点采集到8种总计2252只蚊虫，Wolbachia总感染率为14.03%，其中淡色库蚊的感染率最高（22.01%）。共鉴定出ωPip型单倍型41个，发现跨宿主共享单倍型。中性检验（D=-2.476，P<0.010；Fs=-24.130，P<0.001）显示ωPip型单倍型Wolbachia显著偏离中性进化假设。不同蚊种宿主间种群分化差异无统计学意义（P=0.795；ΦST=-0.062，P=0.944）。宿主与Wolbachia之间缺乏显著的协同进化模式（Global ss=0.894，P=0.725；ParaFitGlobal=0.046，P=0.607）以及发现23次宿主转换事件。
结论淮安市蚊虫携带Wolbachia以超级B组ωPip型为主且具有极高的遗传多样性，水平转移是该菌株的重要传播方式。本地区生物防治需优先引入母系遗传稳定的异型菌株。

Abstract:
Objective To preliminarily study the infection status, genetic diversity, population dynamics and transmission mode of Wolbachia in common mosquito species in Huai'an City, Jiangsu Province, and to provide reference for scientific prevention and control of mosquito-borne diseases.
Methods From July to October 2022, using lamp trapping method in Huai'an City airport, freight station, green belt as well as cattle sheds, pigpens, civilian houses and other habitats collected mosquitoes, using morphology and DNA barcode (mt COI) identified species. Using Wolbachia surface protein gene WSP carried out molecular identification. Using IQ-TREE 2.2.0 software constructed phylogenetic tree, R 4.4.2 software carried out haplotype analysis, PopART 1.7 software constructed haplotype network diagram. Using R 4.4.2 software calculated genetic differentiation (AMOVA, FST), DnaSP 6.1 software carried out neutral test (Tajima's D, Fu's Fs). Using R 4.4.2 software explored co-evolution signal (PACo, ParaFit), Jane 4 software explored Wolbachia and host between evolutionary relationship.
Results In Huai'an City various monitoring points collected 8 species total 2252 mosquitoes, Wolbachia total infection rate was 14.03%, among them Culex pipiens pallens infection rate was highest (22.01%). Total identified ωPip type haplotypes 41, discovered cross-host shared haplotypes. Neutral test (D=-2.476, P<0.010; Fs=-24.130, P<0.001) showed ωPip type haplotype Wolbachia significantly deviated from neutral evolution hypothesis. Different mosquito species hosts between population differentiation difference no statistical significance (P=0.795; ΦST=-0.062, P=0.944). Host and Wolbachia between lacked significant co-evolution pattern (Global ss=0.894, P=0.725; ParaFitGlobal=0.046, P=0.607) and discovered 23 times host switch events.
Conclusion Huai'an City mosquitoes carried Wolbachia with supergroup B ωPip type as main and has extremely high genetic diversity, horizontal transfer is this strain's important transmission mode. This region biological control need priority introduce maternal inheritance stable heterotypic strain.

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