Department of Tropical Infectious Diseases, Second Military Medical University, Shanghai, 200433, China.
Team ten Cadet Brigade, Second Military Medical University, Shanghai, 200433, China.
Infect Dis Poverty. 2017 Oct 2;6(1):149. doi: 10.1186/s40249-017-0362-7.
Anopheles sinensis (Diptera: Culicidae) is a primary vector of Plasmodium vivax and Brugia malayi in most regions of China. In addition, its phylogenetic relationship with the cryptic species of the Hyrcanus Group is complex and remains unresolved. Mitochondrial genome sequences are widely used as molecular markers for phylogenetic studies of mosquito species complexes, of which mitochondrial genome data of An. sinensis is not available.
An. sinensis samples was collected from Shandong, China, and identified by molecular marker. Genomic DNA was extracted, followed by the Illumina sequencing. Two complete mitochondrial genomes were assembled and annotated using the mitochondrial genome of An. gambiae as reference. The mitochondrial genomes sequences of the 28 known Anopheles species were aligned and reconstructed phylogenetic tree by Maximum Likelihood (ML) method.
The length of complete mitochondrial genomes of An. sinensis was 15,076 bp and 15,138 bp, consisting of 13 protein-coding genes, 22 transfer RNA (tRNA) genes, 2 ribosomal RNA (rRNA) genes, and an AT-rich control region. As in other insects, most mitochondrial genes are encoded on the J strand, except for ND5, ND4, ND4L, ND1, two rRNA and eight tRNA genes, which are encoded on the N strand. The bootstrap value was set as 1000 in ML analyses. The topologies restored phylogenetic affinity within subfamily Anophelinae. The ML tree showed four major clades, corresponding to the subgenera Cellia, Anopheles, Nyssorhynchus and Kerteszia of the genus Anopheles.
The complete mitochondrial genomes of An. sinensis were obtained. The number, order and transcription direction of An. sinensis mitochondrial genes were the same as in other species of family Culicidae.
中华按蚊（双翅目：蚊科）是中国大部分地区间日疟原虫和马来丝虫的主要传播媒介。此外，它与 Hyrcanus 组隐种的系统发育关系复杂，尚未解决。线粒体基因组序列被广泛用作蚊科种复合体的系统发育研究的分子标记，而中华按蚊的线粒体基因组数据尚不可用。
从中国山东采集中华按蚊样本，并用分子标记物进行鉴定。提取基因组 DNA，然后进行 Illumina 测序。使用冈比亚按蚊的线粒体基因组作为参考，组装和注释了两个完整的线粒体基因组。对 28 种已知的按蚊属物种的线粒体基因组序列进行了比对，并采用最大似然法（ML）重建了系统发育树。
中华按蚊的完整线粒体基因组长度分别为 15076bp 和 15138bp，由 13 个蛋白质编码基因、22 个转移 RNA（tRNA）基因、2 个核糖体 RNA（rRNA）基因和一个富含 AT 的控制区组成。与其他昆虫一样，大多数线粒体基因都在 J 链上编码，除了 ND5、ND4、ND4L、ND1、两个 rRNA 和八个 tRNA 基因在 N 链上编码。ML 分析中的自举值设置为 1000。系统发育分析恢复了亚科按蚊属内的亲缘关系。ML 树显示了四个主要分支，对应于属按蚊的 Cellia、Anopheles、Nyssorhynchus 和 Kerteszia 亚属。
获得了中华按蚊的完整线粒体基因组。中华按蚊线粒体基因的数量、顺序和转录方向与其他蚊科物种相同。
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