Fang Fujin, Wang Weijie, Zhang Donghui, Lv Yuan, Zhou Dan, Ma Lei, Shen Bo, Sun Yan, Zhu Changliang
Department of Pathogen Biology, Nanjing Medical University, 140 Hanzhong Road, Nanjing, Jiangsu, 210029, China.
Jiangsu Province Key Laboratory of Modern Pathogen Biology, 140 Hanzhong Road, Nanjing, Jiangsu, 210029, China.
Parasitol Res. 2015 Dec;114(12):4421-9. doi: 10.1007/s00436-015-4683-9. Epub 2015 Sep 4.
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.
杀虫剂抗性一直是一项重大的公共卫生挑战。研究杀虫剂抗性机制迫在眉睫。在我们之前的研究中，基于淡色库蚊溴氰菊酯抗性和敏感蛋白质组及转录组，发现了14种差异积累的昆虫表皮蛋白（ICPs）。为了研究这些ICPs是否与溴氰菊酯抗性相关，通过定量实时聚合酶链反应（qRT-PCR）检测了实验室和野外种群的敏感（DS）和抗性（DR）品系中14种ICPs的不同转录水平。在将DS品系短期暴露于溴氰菊酯后，也测定了这14种ICPs的表达水平。从淡色库蚊中克隆了编码14种ICPs之一的CpCPLCG5基因的全长互补DNA（cDNA）。与其他一些昆虫进行了同源性分析和系统发育分析。此外，使用小干扰RNA（siRNA）降低CpCPLCG5基因的表达水平，以表征其对溴氰菊酯抗性的作用。结果表明，在实验室和野外种群中，DR品系中CpCPLCG5基因的表达水平均高于DS品系，而其他13种ICPs的表达下调。CpCPLCG5基因的全长cDNA为732 bp，开放阅读框为390 bp，推导的氨基酸有129个（GenBank/KF723314,2013）。敲低CpCPLCG5基因增加了DR品系的敏感性，而其他13种ICPs的表达水平升高。我们的研究结果表明，表皮蛋白与淡色库蚊对溴氰菊酯的抗性有关。
