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

  • Na Xu
  • Xiao-Hong Sun
  • Zhi-Han Liu
  • Yang Xu
  • Yan Sun
  • Dan Zhou
  • Bo ShenEmail author
  • Chang-Liang ZhuEmail author
Original Article


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.


Culex pipiens pallens Deltamethrin resistance eQTL P450s Regulatory network 



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

Author 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.


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.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflicts of interest.

Research involving human participants and/or animals

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

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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Xuzhou Central HospitalXuzhouPeople’s Republic of China
  2. 2.Department of Pathogen BiologyNanjing Medical UniversityNanjingPeople’s Republic of China
  3. 3.Key Laboratory of Pathogen Biology of Jiangsu ProvinceNanjingPeople’s Republic of China

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