目的 了解淡色库蚊与骚扰库蚊触角中差异性表达的嗅觉基因,为揭示二者遗传分化特征提供参考依据。方法 分别设置淡色库蚊吸血前、后,骚扰库蚊吸血前、后等4个实验组。每组共设4个生物学重复样品,每个样品取20只蚊虫触角。采用TRIzol法提取各组样品总RNA,采用NEBNext Ultra^TM RNA^® 文库构建试剂盒构建文库。对转录组基因进行测序,通过FPKM值差异性分析,筛选淡色库蚊与骚扰库蚊分别在吸血前、后表达差异显著的嗅觉基因,并进行实时荧光定量PCR（qRT-PCR）验证。结果 淡色库蚊吸血前、后触角转录组共注释了50个气味结合蛋白（OBP）和31个气味受体（OR）基因,骚扰库蚊吸血前、后触角转录组共注释了36个OBP和12个OR。淡色库蚊吸血后较吸血前显著上调的OBP有4个,无显著下调的OBP。骚扰库蚊吸血后较吸血前显著上调和下调的OBP分别有3个。淡色库蚊吸血后较吸血前无显著上调的OR,显著下调的OR有3个。骚扰库蚊吸血后较吸血前无显著上调的OR,显著下调的OR有1个。qRT-PCR验证结果显示,淡色库蚊吸血后OBP17显著上调;骚扰库蚊吸血后OBP31、OBP44表达量显著上调,其中OBP44与测序结果一致,OBP31与测序结果相反。吸血前,骚扰库蚊OBP17显著高于淡色库蚊;吸血后,骚扰库蚊OBP31、OBP44显著高于淡色库蚊,OBP99a显著低于淡色库蚊。骚扰库蚊吸血后OR31、OR65、CPIJ001069表达量均显著上调,其中OR65和CPIJ001069表达量显著高于淡色库蚊(P < 0.05)。淡色库蚊吸血后无显著变化的OR基因。结论 淡色库蚊与骚扰库蚊的嗅觉基因OBP和OR表现出了进化分化,OBP17、OBP44可能参与调控骚扰库蚊吸血后行为。OR31、OR65、CPIJ001069可能参与调控骚扰库蚊吸血后行为。

Objective To analyze the difference in antennal olfactory gene expression between Culex pipiens pallens and Cx. pipiens molistus to provide a reference for revealing the characteristics of their genetic differentiation. Methods Total RNAs were extracted from antennas of Cx. pipiens pallens and Cx. pipiens molistus before and after blood feeding, and the RNA libraries were constructed using NEBNext Ultra^TM RNA ^® RNA Library Prep Kit. The transcriptome were obtained by RNAseq and their difference in different species of Cx. was analyzed by FPKM. Those olfactory genes with significant difference before and after blood feeding were identified by qRT-PCR. Results RNA-Seq results identified 50 odorant binding protein (OBP) genes and 31 odor receptor(OR) genes expressed in Cx. pipiens pallens, 36 OBPs and 12 ORs in Cx. pipiens molestus. The regulation trend of OBP44 was, in consistence, the same with the RNAseq results while OBP31 was contrary to that. The expression of OBP17 was significantly higher in Cx. pipiens molestus than in Cx. pipiens pallens before blood-sucking, and the expressions of OBP31 and OBP44 displayed the same trend after blood-sucking while the expression of OBP99a was significantly lower than Cx. pipiens pallens(P < 0.05). Expression of OR31, OR65 and CPIJ001069 were significantly up-regulated in Cx. pipiens molestus post blood feeding, but not in Cx. pipiens pallens, consistent with RNAseq results. Expression of OR65 and CPIJ001069 was significantly higher in Cx. pipiens molestus than that in Cx. pipiens pallens post blood feeding(P < 0.05). There was no significant change of OR gene expression in Cx. pipens pallents post blood feeding. Conclusion OBPs and ORs of Cx. pipiens pallens and Cx. pipiens molestus showed genetic differentiation. OBP17 and OBP44 is possibly involved in the regulation of behavior post blood feeding of Cx. pipiens molestus. OR31, OR65, and CPIJ001069 could be involved in regulating the behavior of Cx. pipiens molestus post blood feeding.