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目的记录上海崇明发现稻富库蚊(Culex inatomii)。方法本研究于2015-2016年的5-11月在上海市崇明区东滩采集稻富库蚊的幼虫和成蚊,进行形态学观察。提取成蚊基因组DNA,PCR扩增线粒体细胞色素氧化酶亚基Ⅰ(COⅠ)基因片段。Clustal W2软件进行COⅠ序列多重比对。采用MEGA v5.10软件计算稻富库蚊与其同属的凶小库蚊(Cx.modestus)、淡色库蚊(Cx.pipiens pallens)和三带喙库蚊(Cx.tritaeniorhynchus)的种内和种间遗传距离。采用邻接法、最大似然法和贝叶斯法构建4个蚊种的系统进化树。结果共采集稻富库蚊成蚊156只,均为雌蚊;捕捞幼虫36只,实验室羽化后获雌蚊17只,雄蚊19只。形态学观察可见,雄蚊尾器结构中,抱肢基节亚端叶毛分为两组,前组具有2根棒状毛,后组具有1根棒状毛和1根叶状毛,可与其近缘种凶小库蚊区分。线粒体COⅠ基因片段PCR扩增获得约650 bp大小的条带,测序后提交Gen Bank(登录号为KX555565~KX555570)。COⅠ序列多重比对结果显示,稻富库蚊和凶小库蚊COⅠ序列的相似度为96%。两者种内遗传距离分别为0.001和0.011,种间遗传距离达0.047。系统进化树结果可见,4个蚊种聚成单系群,每个种独立为1个分支;稻富库蚊与凶小库蚊的亲缘关系最接近,而与同属其他2个蚊种的关系较远。结论上海崇明发现稻富库蚊。
Objective To record the discovery of Culex inatomii in Chongming, Shanghai. Methods In this study, larvae and adult mosquitoes of Culex pipiens pallens were collected from May 2015 to May 2016 in Chongming District, Shanghai, and morphology was observed. Genomic DNA of adult mosquitoes was extracted and the mitochondrial cytochrome oxidase subunit Ⅰ (COⅠ) gene fragment was amplified by PCR. Clustal W2 software COI sequence multiple alignment. MEGA v5.10 software was used to calculate intraspecific and interspecific relationships between Culex pipiens pallens and its related genera Cx.modestus, Cx.pipiens pallens and Cx.tritaeniorhynchus Genetic distance. The phylogenetic tree of four mosquito species was constructed by using adjacency method, maximum likelihood method and Bayesian method. Results A total of 156 mosquitoes were collected from Culex pipiens pallens, all of which were female mosquitoes. Thirty-six larvae were caught and 17 female mosquitoes and 19 male mosquitoes were found after laboratory emergence. Morphological observation shows that in the structure of the male mosquito, the terminal leaf segments of the limbs of the limbs are divided into two groups, the former group has two ciliated hairs, the latter group has one ciliated hairs and one leafy hairs, Marginal killer Curse mosquito distinction. A 650 bp fragment was obtained by PCR amplification of the mitochondrial COI gene fragment and sequenced and submitted to Gen Bank (Accession No. KX555565 ~ KX555570). Multiple alignment of COⅠ sequences showed that the similarity of COⅠ sequences between Culex pipiens pallens and Culex pipiens pallens was 96%. The intraspecific genetic distance was 0.001 and 0.011, respectively, and the genetic distance between species was 0.047. The results of phylogenetic tree showed that 4 mosquito species clustered into monophyletic group, each species was independent of 1 branch. The relationship between Culex pipiens pallens and Culex pipiens pallens was the closest, but with the other 2 mosquito species Far. Conclusion Chongming in Fujian found Cufu mosquito.