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Identification and classification of differentially expressed genes in pyrethroid-resistant Culex pipiens pallens

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Abstract

Culex pipiens pallens is an important vector that transmits Bancroftian filariasis, Japanese encephalitis and other diseases that pose a serious threat to human health. Extensive and improper use of insecticides has caused insecticide resistance in mosquitoes, which has become an important obstacle to the control of mosquito-borne diseases. It is crucial to investigate the underlying mechanism of insecticide resistance. The aims of this study were to identify genes involved in insecticide resistance based on the resistance phenotype, gene expression profile and single-nucleotide polymorphisms (SNPs) and to screen for major genes controlling insecticide resistance. Using a combination of SNP and transcriptome data, gene expression quantitative trait loci (eQTLs) were studied in deltamethrin-resistant mosquitoes. The most differentially expressed pathway in the resistant group was identified, and a regulatory network was built using these SNPs and the differentially expressed genes (DEGs) in this pathway. The major candidate genes involved in the control of insecticide resistance were analyzed by qPCR, siRNA microinjection and CDC bottle bioassays. A total of 85 DEGs that encoded putative detoxification enzymes (including 61 P450s) were identified in this pathway. The resistance regulatory network was built using SNPs, and these metabolic genes, and a major gene, CYP9AL1, were identified. The functional role of CYP9AL1 in insecticide resistance was confirmed by siRNA microinjection and CDC bottle bioassays. Using the eQTL approach, we identified important genes in pyrethroid resistance that may aid in understanding the mechanism underlying insecticide resistance and in targeting new measures for resistance monitoring and management.

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Data Availability

The datasets generated and/or analyzed in the current study are available in the NCBI repository, (http://www.ncbi.nlm.nih.gov/ and https://www.ncbi.nlm.nih.gov/sra/SRP129856). The data supporting the conclusions of this article are included in the article and additional files.

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Acknowledgements

We thank Meng-Xue Hu for her assistance with mosquito collection.

Funding

This study was funded by the National Natural Science Foundation of China (Grant numbers 81471984, 81672056, 81672058 and 81772227), the National Institutes of Health (NIH) of the USA (Grant number 2R01AI075746) and the Priority Academic Program Development of Jiangsu Higher Education Institutions.

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Authors and Affiliations

  1. Xuzhou Central Hospital, 29 Taihang Road, Yunlong District, Xuzhou, 221111, People’s Republic of China

    Na Xu

  2. Department of Pathogen Biology, Nanjing Medical University, 101 Longmian Avenue, Jiangning District, Nanjing, 211166, People’s Republic of China

    Xiao-Hong Sun, Zhi-Han Liu, Yang Xu, Yan Sun, Dan Zhou, Bo Shen & Chang-Liang Zhu

  3. Key Laboratory of Pathogen Biology of Jiangsu Province, 101 Longmian Avenue, Jiangning District, Nanjing, 211166, People’s Republic of China

    Yan Sun, Dan Zhou, Bo Shen & Chang-Liang Zhu

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  1. Na Xu
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  4. Yang Xu
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  5. Yan Sun
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Contributions

BS, C-LZ and NX conceived the study. NX collected the data and performed the experiment. X-HS performed the data analysis. Z-HL performed the supplementary experiment duiring revision. YX, DZ and YS supported the development of the statistical methodology. NX and BS drafted the manuscript. All authors critically reviewed, read and approved the final manuscript.

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Correspondence to Bo Shen or Chang-Liang Zhu.

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This article does not contain any studies with human participants or animals performed by any of the authors.

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Xu, N., Sun, XH., Liu, ZH. et al. Identification and classification of differentially expressed genes in pyrethroid-resistant Culex pipiens pallens. Mol Genet Genomics 294, 861–873 (2019). https://doi.org/10.1007/s00438-018-1521-7

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  • DOI: https://doi.org/10.1007/s00438-018-1521-7

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