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The cuticle proteins: a putative role for deltamethrin resistance in Culex pipiens pallens

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Abstract

Insecticide resistance has been a major public health challenge. It is impendent to study the mechanism on insecticide resistance. In our previous study, 14 differentially accumulated insect cuticle proteins (ICPs) based on insecticide resistance proteomes and transcriptomes were found in the deltamethrin-resistant (DR) and -susceptible (DS) strains of Culex pipiens pallens. To investigate if these ICPs are associated with deltamethrin resistance, different transcriptional levels of the 14 ICPs were detected in the DS and DR strains from laboratory and field populations by using quantitative real-time polymerase chain reaction (qRT-PCR). The expression levels of the 14 ICPs were also measured after short-term exposure of the DS strain to deltamethrin. The full-length complementary DNA (cDNA) of CpCPLCG5 gene, which encodes one of the 14 ICPs, was cloned from Cx. pipiens pallens. Homology analysis and phylogenetic analysis were carried out with some other insects. Furthermore, small interfering RNA (siRNA) was used to knockdown the expression level of CpCPLCG5 gene for characterizing its contribution to deltamethrin resistance. The results showed that the expression level of CpCPLCG5 gene was higher in DR strain than in DS strain both in laboratory and field populations while the other 13 ICPs were downregulated. The full-length cDNA of CpCPLCG5 gene was 732 bp, with the ORF of 390 bp and deduced 129 amino acids (GenBank/KF723314,2013). Knockdown of CpCPLCG5 gene increased the susceptibility of the DR strain while the expression level of the other 13 ICPs elevated. Our findings indicate that the cuticle proteins are associated with deltamethrin resistance in Cx. pipiens pallens.

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Acknowledgments

This work was supported by the National Institutes of Health of US (NIH) (Grant No. 2R01AI075746), the National Natural Science Foundation of China (Grant No. 30901244, 81171610, and 81301458), the National S & T Major Program (Grant No. 2012ZX10004-219 and 2012ZX10004-220) and Priority Academic Program Development of Jiangsu Higher Education Institutions.

We thank Maoqing Gong from Shandong Institute of Parasitic Diseases and Xuelian Chang from BengBu Medical College for their help with mosquito collection and Dr. Rongfeng Li and Qiang Liu for their expert technical assistance in gene injection.

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Authors and Affiliations

  1. Department of Pathogen Biology, Nanjing Medical University, 140 Hanzhong Road, Nanjing, Jiangsu, 210029, China

    Fujin Fang, Weijie Wang, Donghui Zhang, Yuan Lv, Dan Zhou, Lei Ma, Bo Shen, Yan Sun & Changliang Zhu

  2. Jiangsu Province Key Laboratory of Modern Pathogen Biology, 140 Hanzhong Road, Nanjing, Jiangsu, 210029, China

    Fujin Fang, Weijie Wang, Donghui Zhang, Yuan Lv, Dan Zhou, Lei Ma, Bo Shen, Yan Sun & Changliang Zhu

  3. Clinical Laboratory, The Third People’s Hospital of Bengbu, 38 Middle Shengli Road, Bengbu, Anhui, 233000, China

    Fujin Fang

  4. Department of Pathogen Biology, Hebei Medical University, 361 East Zhongshan Road, Shijiazhuang, Hebei, 050017, China

    Weijie Wang

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  1. Fujin Fang
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  2. Weijie Wang
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  3. Donghui Zhang
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  9. Changliang Zhu
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Correspondence to Yan Sun.

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Table S1

Summary of the 14 ICPs identified based on insecticide-resistance proteomes and transcriptomes in the deltamethrin-resistant (DS) and –susceptible (DR) strains of Cx. pipiens pallens. (DOC 42 kb)

Table S2

Sequences of primers used in this study. (DOC 46 kb)

Table S3

Relative expression ratios of the 14 ICPs in DS strain after exposure to a sublethal dosage of deltamethrin. (DOC 42 kb)

Table S4

Mortality of the gene-injected mosquitoes in WHO insecticide susceptibility bioassay. (DOC 36 kb)

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Fang, F., Wang, W., Zhang, D. et al. The cuticle proteins: a putative role for deltamethrin resistance in Culex pipiens pallens . Parasitol Res 114, 4421–4429 (2015). https://doi.org/10.1007/s00436-015-4683-9

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