title: Comparative transcriptome analyses of deltamethrin-susceptible and -resistant Culex pipiens pallens by RNA-seq.
abstract: The widespread and improper use of pyrethroid insecticides, such as deltamethrin, has resulted in the evolution of resistance in many mosquito species, including Culex pipiens pallens. With the development of high-throughput sequencing, it is possible to massively screen pyrethroid resistance-associated gene. In this study, we used Illumina-Solexa transcriptome sequencing to identify genes that are expressed differently in deltamethrin-susceptible and -resistant strains of Culex pipiens pallens as a critical knowledge base for further studies. A total of 4,961,197,620 base pairs and 55,124,418 reads were sequenced, mapped to the Culex quinquefasciatus genome and assembled into 17,679 known genes. We recorded 1826 significantly differentially expressed genes (DEGs). Among them, 1078 genes were up-regulated and 748 genes were down-regulated in the deltamethrin-resistant strain compared to -susceptible strain. These DEGs contained cytochrome P450 s, cuticle proteins, UDP-glucuronosyltransferases, lipases, serine proteases, heat shock proteins, esterases and others. Among the 1826 DEGs, we found that the transcriptional levels of CYP6AA9 in the laboratory populations was elevated as the levels of deltamethrin resistance increased. Moreover, the expression levels of the CYP6AA9 were significantly higher in the resistant strains than the susceptible strains in three different field populations. We further confirmed the association between the CYP6AA9 gene and deltamethrin resistance in mosquitoes by RNA interfering (RNAi). Altogether, we explored massive potential pyrethroid resistance-associated genes and demonstrated that CYP6AA9 participated in the pyrethroid resistance in mosquitoes.
authors.affiliation: Department of Pathogen Biology, Nanjing Medical University, 210029, Nanjing, Jiangsu, China.|Department of Nuclear Medicine, Nanjing First Hospital, Nanjing Medical University, Nanjing, China.
authors.forename: Yuan
authors.initials: Y
authors.lastname: Lv
authors.affiliation: Department of Pathogen Biology, Nanjing Medical University, 210029, Nanjing, Jiangsu, China.
authors.forename: Weijie
authors.initials: W
authors.lastname: Wang
authors.affiliation: Department of Pathogen Biology, Nanjing Medical University, 210029, Nanjing, Jiangsu, China.
authors.forename: Shanchao
authors.initials: S
authors.lastname: Hong
authors.affiliation: Department of Pathogen Biology, Nanjing Medical University, 210029, Nanjing, Jiangsu, China.
authors.forename: Zhentao
authors.initials: Z
authors.lastname: Lei
authors.affiliation: Department of Pathogen Biology, Nanjing Medical University, 210029, Nanjing, Jiangsu, China.
authors.forename: Fujin
authors.initials: F
authors.lastname: Fang
authors.affiliation: Department of Pathogen Biology, Nanjing Medical University, 210029, Nanjing, Jiangsu, China.
authors.forename: Qin
authors.initials: Q
authors.lastname: Guo
authors.affiliation: Department of Pathogen Biology, Nanjing Medical University, 210029, Nanjing, Jiangsu, China.
authors.forename: Shengli
authors.initials: S
authors.lastname: Hu
authors.affiliation: Department of Pathogen Biology, Nanjing Medical University, 210029, Nanjing, Jiangsu, China.
authors.forename: Mengmeng
authors.initials: M
authors.lastname: Tian
authors.affiliation: Department of Pathogen Biology, Nanjing Medical University, 210029, Nanjing, Jiangsu, China.
authors.forename: Bingqian
authors.initials: B
authors.lastname: Liu
authors.affiliation: Department of Pathogen Biology, Nanjing Medical University, 210029, Nanjing, Jiangsu, China.
authors.forename: Donghui
authors.initials: D
authors.lastname: Zhang
authors.affiliation: Department of Pathogen Biology, Nanjing Medical University, 210029, Nanjing, Jiangsu, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Sun
authors.affiliation: Department of Pathogen Biology, Nanjing Medical University, 210029, Nanjing, Jiangsu, China.
authors.forename: Lei
authors.initials: L
authors.lastname: Ma
authors.affiliation: Department of Pathogen Biology, Nanjing Medical University, 210029, Nanjing, Jiangsu, China.
authors.forename: Bo
authors.initials: B
authors.lastname: Shen
authors.affiliation: Department of Pathogen Biology, Nanjing Medical University, 210029, Nanjing, Jiangsu, China. zdd1209love@163.com.
authors.forename: Dan
authors.initials: D
authors.lastname: Zhou
authors.affiliation: Department of Pathogen Biology, Nanjing Medical University, 210029, Nanjing, Jiangsu, China. clzhu@njmu.edu.cn.
authors.forename: Changliang
authors.initials: C
authors.lastname: Zhu
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular / Q000379:methods; D003465:Culex / Q000187:drug effects* / Q000235:genetics*; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics; D020869:Gene Expression Profiling / Q000379:methods; D019476:Insect Proteins / Q000235:genetics; D007305:Insecticide Resistance / Q000235:genetics*; D008969:Molecular Sequence Data; D009570:Nitriles / Q000494:pharmacology*; D011722:Pyrethrins / Q000494:pharmacology*; D012313:RNA / Q000235:genetics*; D016415:Sequence Alignment; D059467:Transcriptome / Q000235:genetics*
chemical_list: D019476:Insect Proteins; D009570:Nitriles; D011722:Pyrethrins; C017180:decamethrin; D012313:RNA; D003577:Cytochrome P-450 Enzyme System
keywords: CYP6AA9; Mosquitoes; Pyrethroids; RNA-seq; RNAi