Yanhong Tang, Huifang Zhang, Huanqing Zhu, Siyan Bi, Xiaodi Wang, Shunxia Ji, Jianhang Ji, Dongfang Ma, Cong Huang, Guifen Zhang, Nianwan Yang, Fanghao Wan, Zhichuang Lü, Wanxue Liu
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Potential regulatory genes were screened via RNAi (RNA interference), and the DNA methylase in <i>Tuta absoluta</i> was cloned by RACE (Rapid amplification of cDNA ends). <i>TaDnmt1</i> was identified as a potential regulatory gene via bioinformatics; its expression was evaluated in response to temperature stress and during different development stages using real-time polymerase chain reaction. Results revealed that <i>TaDnmt1</i> participates in hot/cold tolerance, temperature preference and larval development. The full-length cDNA sequence of <i>TaDnmt1</i> is 3765 bp and encodes a 1254 kDa protein with typical Dnmt1 node-conserved structural features and six conserved DNA-binding active motifs. Moreover, <i>TaDnmt1</i> expression is significantly altered by temperature stress treatments and within different development stages. Hence, <i>TaDnmt1</i> likely contributes to temperature responses and organismal development. Furthermore, after treating with double-stranded RNA and exposing <i>Tuta absoluta</i> to 35°C heat shock or −12°C cold shock for 1 h, the survival rate significantly decreases; the preferred temperature is 2°C lower than that of the control group. In addition, the epidermal segments become enlarged and irregularly folded while the surface dries up. This results in a significant increase in larval mortality (57%) and a decrease in pupation (49.3%) and eclosion (50.9%) rates. Hence, <i>TaDnmt1</i> contributes to temperature stress responses and temperature perception, as well as organismal growth and development, via DNA methylation regulation. These findings suggest that the rapid geographic expansion of <i>T absoluta</i> has been closely associated with <i>TaDnmt1-</i>mediated temperature tolerance. 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The full-length cDNA sequence of <i>TaDnmt1</i> is 3765 bp and encodes a 1254 kDa protein with typical Dnmt1 node-conserved structural features and six conserved DNA-binding active motifs. Moreover, <i>TaDnmt1</i> expression is significantly altered by temperature stress treatments and within different development stages. Hence, <i>TaDnmt1</i> likely contributes to temperature responses and organismal development. Furthermore, after treating with double-stranded RNA and exposing <i>Tuta absoluta</i> to 35°C heat shock or −12°C cold shock for 1 h, the survival rate significantly decreases; the preferred temperature is 2°C lower than that of the control group. In addition, the epidermal segments become enlarged and irregularly folded while the surface dries up. This results in a significant increase in larval mortality (57%) and a decrease in pupation (49.3%) and eclosion (50.9%) rates. 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引用次数: 0
摘要
DNA 甲基化酶 1(Dnmt1)是与昆虫发育所需的生化信号有关的重要调节因子。它能对环境变化做出反应,并引发表型可塑性。与此同时,Tuta absoluta Meyrick(鳞翅目:Gelechiidae)--一种破坏性入侵害虫--能迅速入侵并适应不同的生境;然而,Dnmt1 在该生物中的作用尚未阐明。因此,本研究调查了 Tuta absoluta 快速适应温度胁迫的机制。通过RNAi(RNA干扰)筛选了潜在的调控基因,并通过RACE(cDNA末端快速扩增)克隆了Tuta absoluta中的DNA甲基化酶。通过生物信息学确定 TaDnmt1 为潜在的调控基因;利用实时聚合酶链式反应评估了其在温度胁迫和不同发育阶段的表达情况。结果表明,TaDnmt1 参与了冷热耐受、温度偏好和幼虫发育。TaDnmt1 的全长 cDNA 序列长达 3765 bp,编码 1254 kDa 蛋白,具有典型的 Dnmt1 节点保守结构特征和六个保守的 DNA 结合活性基序。此外,TaDnmt1 的表达受温度胁迫处理和不同发育阶段的影响而发生显著变化。因此,TaDnmt1 很可能对温度反应和生物体的发育做出了贡献。此外,用双链 RNA 处理 Tuta absoluta 并将其暴露于 35°C 热休克或 -12°C 冷休克 1 小时后,其存活率会显著下降;首选温度比对照组低 2°C。此外,表皮片段变大,呈不规则折叠,表面干枯。这导致幼虫死亡率显著上升(57%),化蛹率下降(49.3%),羽化率下降(50.9%)。因此,TaDnmt1 通过 DNA 甲基化调控,有助于温度胁迫反应和温度感知,以及生物体的生长和发育。这些研究结果表明,TaDnmt1 介导的温度耐受性与 T absoluta 的快速地理扩张密切相关。这项研究推动了对 "耐温 Dnmt "的研究,并为 RNAi- 驱动的 Tuta absoluta 调控提供了潜在靶标。
DNA methylase 1 influences temperature responses and development in the invasive pest Tuta absoluta
DNA methylase 1 (Dnmt1) is an important regulatory factor associated with biochemical signals required for insect development. It responds to changes in the environment and triggers phenotypic plasticity. Meanwhile, Tuta absoluta Meyrick (Lepidoptera: Gelechiidae)—a destructive invasive pest—can rapidly invade and adapt to different habitats; however, the role of Dnmt1 in this organism has not been elucidated. Accordingly, this study investigates the mechanism(s) underlying the rapid adaptation of Tuta absoluta to temperature stress. Potential regulatory genes were screened via RNAi (RNA interference), and the DNA methylase in Tuta absoluta was cloned by RACE (Rapid amplification of cDNA ends). TaDnmt1 was identified as a potential regulatory gene via bioinformatics; its expression was evaluated in response to temperature stress and during different development stages using real-time polymerase chain reaction. Results revealed that TaDnmt1 participates in hot/cold tolerance, temperature preference and larval development. The full-length cDNA sequence of TaDnmt1 is 3765 bp and encodes a 1254 kDa protein with typical Dnmt1 node-conserved structural features and six conserved DNA-binding active motifs. Moreover, TaDnmt1 expression is significantly altered by temperature stress treatments and within different development stages. Hence, TaDnmt1 likely contributes to temperature responses and organismal development. Furthermore, after treating with double-stranded RNA and exposing Tuta absoluta to 35°C heat shock or −12°C cold shock for 1 h, the survival rate significantly decreases; the preferred temperature is 2°C lower than that of the control group. In addition, the epidermal segments become enlarged and irregularly folded while the surface dries up. This results in a significant increase in larval mortality (57%) and a decrease in pupation (49.3%) and eclosion (50.9%) rates. Hence, TaDnmt1 contributes to temperature stress responses and temperature perception, as well as organismal growth and development, via DNA methylation regulation. These findings suggest that the rapid geographic expansion of T absoluta has been closely associated with TaDnmt1-mediated temperature tolerance. This study advances the research on ‘thermos Dnmt’ and provides a potential target for RNAi-driven regulation of Tuta absoluta.
期刊介绍:
Insect Molecular Biology has been dedicated to providing researchers with the opportunity to publish high quality original research on topics broadly related to insect molecular biology since 1992. IMB is particularly interested in publishing research in insect genomics/genes and proteomics/proteins.
This includes research related to:
• insect gene structure
• control of gene expression
• localisation and function/activity of proteins
• interactions of proteins and ligands/substrates
• effect of mutations on gene/protein function
• evolution of insect genes/genomes, especially where principles relevant to insects in general are established
• molecular population genetics where data are used to identify genes (or regions of genomes) involved in specific adaptations
• gene mapping using molecular tools
• molecular interactions of insects with microorganisms including Wolbachia, symbionts and viruses or other pathogens transmitted by insects
Papers can include large data sets e.g.from micro-array or proteomic experiments or analyses of genome sequences done in silico (subject to the data being placed in the context of hypothesis testing).