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Sequencing and analysis of the complete mitochondrial genome of Culex tritaeniorhynchus (Dipera: Culicidae) in China.

ABSTRACT: The complete sequence of the mitochondrial genome of the Culex tritaeniorhynchus (Diptera: Culicidae) is presented using traditional Sanger sequencing. Its mitogenome are 15,123?bp in length, consisting of 13 protein-coding genes, 2 ribosomal RNA genes (rRNA) and 22 transfer RNA (tRNA) genes. The percent of A?+?T of the mitogenome is 77.73%. Most mitochondrial genes are encoded on the heavy strand, except for NAD5, NaD4, NAD4L, NAD1, two rRNA and nine tRNA genes, which are encoded on the light strand. The results of phylogenetic analyzes showed that the Cx. tritaeniorhynchus has close relationship with the species of genus Culex.

SUBMITTER: Chu H

PROVIDER: S-EPMC7800139 | biostudies-literature | 2016 Jun

REPOSITORIES: biostudies-literature

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Sequencing and analysis of the complete mitochondrial genome of Culex tritaeniorhynchus (Dipera: Culicidae) in China.

Chu Hongliang H Wu Zhiming Z Zhang Hengduan H Li Chunxiao C Wang Gang G Zhou Minghao M Zhao Tongyan T

Mitochondrial DNA. Part B, Resources 20160620 1

The complete sequence of the mitochondrial genome of the Culex tritaeniorhynchus (Diptera: Culicidae) is presented using traditional Sanger sequencing. Its mitogenome are 15,123 bp in length, consisting of 13 protein-coding genes, 2 ribosomal RNA genes (rRNA) and 22 transfer RNA (tRNA) genes. The percent of A + T of the mitogenome is 77.73%. Most mitochondrial genes are encoded on the heavy strand, except for NAD5, NaD4, NAD4L, NAD1, two rRNA and nine tRNA gene ...[more]

PMID: 33473485

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Project description:BackgroundCulex tritaeniorhynchus and Culex pipiens pallens are the major vectors of the Japanese encephalitis virus and Wuchereria bancrofti, the causative agent of filariasis. The knowledge of mitochondrial genomes has been widely useful for the studies on molecular evolution, phylogenetics and population genetics.MethodsIn this study, we sequenced and annotated the mitochondrial (mt) genomes of Cx. tritaeniorhynchus and Cx. p. pallens, and performed a comparative analysis including four known mt genomes of species of the subgenus Culex (Culex). The phylogenetic relationships of Cx. tritaeniorhynchus, Cx. p. pallens and four known Culex mt genome sequences were reconstructed by maximum likelihood based on concatenated protein-coding gene sequences.ResultsCulex tritaeniorhynchus and Cx. p. pallens mt genomes are 14,844 bp and 15,617 bp long, both consists of 13 PCGs, 22 tRNAs, 2 rRNAs and 1 CR (not sequenced for Cx. tritaeniorhynchus). The initiation and termination codons of PCGs are ATN and TAA, respectively, except for COI starting with TCG, and COI and COII terminated with T. tRNAs have the typical clover-leaf secondary structures except for trnS ((AGN)) that is lacking the DHU stem. 16S rRNA and 12S rRNA secondary structures were drawn for the first time for mosquito mt genomes. The control region of Cx. p. pallens mt genome is 747 bp long and with four tandem repeat structures. Phylogenetic analyses demonstrated that the mt genome of Cx. tritaeniorhynchus was significantly separated from the remaining five mt genomes of Culex spp. Culex p. pipiens, Cx. p. pallens and Cx. p. quinquefasciatus formed a monophyletic clade with Cx. p. quinquefasciatus linked in the middle of the clade, and Cx. p. pallens should have the same taxonomic level as Culex p. pipiens and Cx. p. quinquefasciatus.ConclusionsThe mt genomes of Cx. tritaeniorhynchus and Cx. p. pallens share the same gene composition and order with those of two other Culex species. Culex p. pallens of the Pipiens complex should have the same taxonomic level as Culex p. pipiens and Cx. p. quinquefasciatus investigated. We enriched the Culex mt genome data and provided a reference basis for further Culex mt genome sequencing and analyses.

Project description:BackgroundMosquitoes carry a variety of viruses that can cause disease in humans, animals and livestock. Surveys for viruses carried by wild mosquitoes can significantly contribute to surveillance efforts and early detection systems. In addition to mosquito-borne viruses, mosquitoes harbor many insect-specific viruses (ISVs). Quang Binh virus (QBV) is one such example, categorized as an ISV within the Flavivirus genus (family Flaviviridae). QBV has been specifically documented in Vietnam and China, with reports limited to several mosquito species.MethodsThe homogenate obtained from female mosquitoes was cultured on C6/36 (Aedes albopictus) and BHK-21 (baby hamster kidney) cell lines. Positive cultures were identified by reverse transcription-polymerase chain reaction (RT‒PCR) with taxon- or species-specific primers. Next-generation sequencing was employed to sequence the complete genomes of the identified positive samples. Subsequently, phylogenetic, gene homology, molecular evolutionary and genetic variation analyses were conducted.ResultIn 2021, a total of 32,177 adult female mosquitoes were collected from 15 counties in Guizhou Province, China. The predominant mosquito species identified were Culex tritaeniorhynchus, Armigeres subalbatus and Anopheles sinensis. Among the collected mosquitoes, three positive cultures were obtained from Cx. tritaeniorhynchus pools, revealing the presence of Quang Binh virus (QBV) RNA sequences. Phylogenetic analysis indicated that the three Guizhou isolates, along with the prototype isolate from Vietnam, formed distinct branches. These branches were primarily closely related to other QBV isolates reported in China. Comparative analysis revealed a high degree of nucleotide and amino acid homology between the Guizhou isolates and both Vietnamese and other indigenous Chinese isolates. Additionally, nonsynonymous single-nucleotide variants (SNVs) were observed in these strains compared to the QBV prototype strain.ConclusionThis study represents the first report of QBV presences in Cx. tritaeniorhynchus mosquitoes in Guizhou Province, China. Phylogenetic tree analysis showed that the three Guizhou isolates were most closely related to the QBV genes found in China. In addition, the study of the genetic characteristics and variation of this virus provided a deeper understanding of QBV and enriched the baseline data of these insect-specific flaviviruses (ISFVs).

Dataset Information

Sequencing and analysis of the complete mitochondrial genome of Culex tritaeniorhynchus (Dipera: Culicidae) in China.

Publications

Sequencing and analysis of the complete mitochondrial genome of <i>Culex tritaeniorhynchus</i> (Dipera: Culicidae) in China.

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