Key Laboratory of Integrated Pest Management in Crops in Eastern China (Ministry of Agriculture of China), College of Plant Protection, Nanjing Agricultural University, Nanjing, 210095, People's Republic of China.
Present address: Institute of Entomology, Jiangxi Agricultural University, Nanchang, 330045, China.
BMC Genomics. 2020 Feb 3;21(1):120. doi: 10.1186/s12864-020-6533-0.
Fluralaner is a novel isoxazoline insecticide with a unique action site on the γ-aminobutyric acid receptor (GABAR), shows excellent activity on agricultural pests including the common cutworm Spodoptera litura, and significantly influences the development and fecundity of S. litura at either lethal or sublethal doses. Herein, Illumina HiSeq Xten (IHX) platform was used to explore the transcriptome of S. litura and to identify genes responding to fluralaner exposure.
A total of 16,572 genes, including 451 newly identified genes, were observed in the S. litura transcriptome and annotated according to the COG, GO, KEGG and NR databases. These genes included 156 detoxification enzyme genes [107 cytochrome P450 enzymes (P450s), 30 glutathione S-transferases (GSTs) and 19 carboxylesterases (CarEs)] and 24 insecticide-targeted genes [5 ionotropic GABARs, 1 glutamate-gated chloride channel (GluCl), 2 voltage-gated sodium channels (VGSCs), 13 nicotinic acetylcholine receptors (nAChRs), 2 acetylcholinesterases (AChEs) and 1 ryanodine receptor (RyR)]. There were 3275 and 2491 differentially expressed genes (DEGs) in S. litura treated with LC or LC concentrations of fluralaner, respectively. Among the DEGs, 20 related to detoxification [16 P450s, 1 GST and 3 CarEs] and 5 were growth-related genes (1 chitin and 4 juvenile hormone synthesis genes). For 26 randomly selected DEGs, real-time quantitative PCR (RT-qPCR) results showed that the relative expression levels of genes encoding several P450s, GSTs, heat shock protein (HSP) 68, vacuolar protein sorting-associated protein 13 (VPSAP13), sodium-coupled monocarboxylate transporter 1 (SCMT1), pupal cuticle protein (PCP), protein takeout (PT) and low density lipoprotein receptor adapter protein 1-B (LDLRAP1-B) were significantly up-regulated. Conversely, genes encoding esterase, sulfotransferase 1C4, proton-coupled folate transporter, chitinase 10, gelsolin-related protein of 125 kDa (GRP), fibroin heavy chain (FHC), fatty acid synthase and some P450s were significantly down-regulated in response to fluralaner.
The transcriptome in this study provides more effective resources for the further study of S. litura whilst the DEGs identified sheds further light on the molecular response to fluralaner.
氟虫腈是一种新型的异噁唑啉类杀虫剂,其作用靶标为γ-氨基丁酸受体(GABAR),对鳞翅目农业害虫具有优异的活性,包括斜纹夜蛾(Spodoptera litura),并以致死或亚致死剂量显著影响斜纹夜蛾的发育和繁殖。本研究采用 Illumina HiSeq Xten(IHX)平台,对斜纹夜蛾的转录组进行了研究,并鉴定了对氟虫腈暴露有反应的基因。
共观察到斜纹夜蛾转录组中的 16572 个基因,包括 451 个新鉴定的基因,根据 COG、GO、KEGG 和 NR 数据库进行了注释。这些基因包括 156 个解毒酶基因[107 个细胞色素 P450 酶(P450s)、30 个谷胱甘肽 S-转移酶(GSTs)和 19 个羧酸酯酶(CarEs)]和 24 个杀虫剂靶标基因[5 个离子型 GABARs、1 个谷氨酸门控氯离子通道(GluCl)、2 个电压门控钠离子通道(VGSCs)、13 个烟碱型乙酰胆碱受体(nAChRs)、2 个乙酰胆碱酯酶(AChEs)和 1 个肌醇 1,4,5-三磷酸受体(RyR)]。用 LC 或 LC 浓度的氟虫腈处理斜纹夜蛾后,分别有 3275 个和 2491 个差异表达基因(DEGs)。在 DEGs 中,有 20 个与解毒相关[16 个 P450s、1 个 GST 和 3 个 CarEs],5 个与生长相关(1 个几丁质和 4 个保幼激素合成基因)。对 26 个随机选择的 DEGs 进行实时定量 PCR(RT-qPCR)验证,结果显示,编码几个 P450s、GSTs、热休克蛋白 68(HSP68)、液泡蛋白分选相关蛋白 13(VPSAP13)、钠偶联单羧酸转运蛋白 1(SCMT1)、蛹皮蛋白(PCP)、蛋白提取(PT)和低密度脂蛋白受体衔接蛋白 1-B(LDLRAP1-B)的基因的相对表达水平显著上调。相反,编码酯酶、磺基转移酶 1C4、质子偶联叶酸转运蛋白、几丁质酶 10、125kDa 凝胶蛋白相关蛋白(GRP)、丝氨酸重链(FHC)、脂肪酸合成酶和一些 P450s 的基因在响应氟虫腈时表达显著下调。
本研究的转录组为进一步研究斜纹夜蛾提供了更有效的资源,而鉴定的 DEGs 进一步阐明了斜纹夜蛾对氟虫腈的分子反应。