简介:Akirin作为一种高度保守的转录因子,对增长产生了深远的影响,发展,免疫反应,和动物的生殖过程。褐飞虱(BPH),Nilaparvatalugens,亚洲水稻生产中的主要害虫,具有很高的生殖能力,导致水稻产量下降的关键因素。本研究的目的是证明Akirin在BPH繁殖中的调节作用。方法:在本研究中,定量PCR(qPCR)检测基因的mRNA表达。RNA干扰(RNAi)用于下调Akirin基因的表达,和RNA测序(RNA-seq)用于筛选由Akirin下调引起的差异表达基因。用酶联免疫吸附试验(ELISA)测定激素含量,用二辛可宁酸(BCA)方法评估蛋白质含量。结果:使用BPH基因组数据,我们筛选了Akirin基因(NlAkirin)。对组织特异性表达的分析表明,NlAkirin在雌性BPH的所有测试组织中均表达,但其在卵巢中的表达水平最高。在抑制BPH女性中NlAkirin的mRNA表达后,产卵的数量,孵化率,卵巢数量减少。进行转录组测序,在NlAkirin双链RNA处理之后。与对照的基因相比,用GFP双链RNA注射,有438个上调基因和1012个下调基因;卵黄蛋白原(Vg)和卵黄蛋白原受体(VgR)基因的表达以及与雷帕霉素靶蛋白(TOR)相关基因的mRNA表达,幼体激素(JH),参与Vg合成的胰岛素途径显著下调。由于NlAkirin击倒,JHIII和蜕皮激素(Ecd)的滴度在未交配的雌性中下调,但在交配的雌性中恢复到正常水平。未交配和交配雌性的卵巢蛋白质含量均下调。讨论与结论:我们的结果表明,NlAkirin通过调节与Vg相关基因的mRNA表达来影响雌性BPH的繁殖,VgR,TOR,JH,和胰岛素信号通路,除了JHIII和Ecd的滴度。这项研究的发现为Akirin在昆虫繁殖能力中的调节作用提供了新的见解。
Introduction: Akirin as a highly conserved transcription factor, exerts a profound influence on the growth, development, immune response, and reproductive processes in animals. The brown planthopper (BPH), Nilaparvata lugens, a major pest in rice production in Asia, possesses high reproductive capacity, a critical factor contributing to reduced rice yields. The aims of this study were to demonstrate the regulatory role of Akirin in the reproduction of BPH. Methods: In this study, quantitative PCR (qPCR) was used to detect the mRNA expression of genes. RNA interference (RNAi) was used to downregulate the expression of Akirin gene, and RNA sequencing (RNA-seq) was used to screen for differentially expressed genes caused by Akirin downregulation. Hormone contents were measured with the enzyme linked immunosorbent assay (ELISA), and protein content was evaluated with the bicinchoninic acid (BCA) method. Results: Using BPH genome data, we screened for an Akirin gene (NlAkirin). An analysis of tissue-specific expressions showed that NlAkirin was expressed in all tissues tested in female BPH, but its expression level was highest in the ovary. After inhibiting the mRNA expression of NlAkirin in BPH females, the number of eggs laid, hatching rate, and number of ovarioles decreased. Transcriptome sequencing was performed, following a NlAkirin double-stranded RNA treatment. Compared with the genes of the control, which was injected with GFP double-stranded RNA, there were 438 upregulated genes and 1012 downregulated genes; the expression of
vitellogenin (Vg) and
vitellogenin receptor (VgR) genes as well as the mRNA expression of genes related to the target of rapamycin (TOR), juvenile hormone (JH), and insulin pathways involved in Vg synthesis was significantly downregulated. As a result of NlAkirin knockdown, the titers of JH III and Ecdysone (Ecd) were downregulated in unmated females but returned to normal levels in mated females. The ovarian protein contents in both unmated and mated females were downregulated. Discussion and conclusion: Our results suggest that NlAkirin affects female BPH reproduction by regulating the mRNA expression of genes related to the Vg, VgR, TOR, JH, and insulin signaling pathways, in addition to the titers of JH III and Ecd. The findings of this research provide novel insights into the regulatory role of Akirin in insect reproductive capacity.