College of Animal Science, Shanxi Agricultural University, Jinzhong, Shanxi, China.
College of Life Sciences, Shanxi Agricultural University, Jinzhong, Shanxi, China.
BMC Genomics. 2024 May 22;25(1):506. doi: 10.1186/s12864-024-10399-8.
Long non-coding RNAs (lncRNAs) are crucial modulators of post-transcriptional gene expression regulation, cell fate determination, and disease development. However, lncRNA functions during short-term heat stress in adult worker bees are poorly understood. Here, we performed deep sequencing and bioinformatic analyses of honeybee lncRNAs. RNA interference was performed by using siRNA targeting the most highly expressed lncRNA. The silencing effect on lncRNA and the relative expression levels of seven heat shock protein (HSP) genes, were subsequently examined. Overall, 7,842 lncRNAs and 115 differentially expressed lncRNAs (DELs) were identified in adult worker bees following heat stress exposure. Structural analysis revealed that the overall expression abundance, length of transcripts, exon number, and open reading frames of lncRNAs were lower than those of mRNAs. GO analysis revealed that the target genes were mainly involved in "metabolism," "protein folding," "response to stress," and "signal transduction" pathways. KEGG analysis indicated that the "protein processing in endoplasmic reticulum" and "longevity regulating pathway-multiple species" pathways were most enriched. Quantitative real-time polymerase chain reaction (qRT-PCR) detection of the selected DELs confirmed the reliability of the sequencing data. Moreover, the siRNA experiment indicated that feeding siRNA yielded a silencing efficiency of 77.51% for lncRNA MSTRG.9645.5. Upon silencing this lncRNA, the expression levels of three HSP genes were significantly downregulated (p < 0.05), whereas those of three other HSP genes were significantly upregulated (p < 0.05). Our results provide a new perspective for understanding the regulatory mechanisms of lncRNAs in adult worker bees under short-term heat stress.
长链非编码 RNA（lncRNA）是转录后基因表达调控、细胞命运决定和疾病发展的关键调节剂。然而，在成年工蜂的短期热应激中，lncRNA 的功能仍知之甚少。在这里，我们对蜜蜂 lncRNA 进行了深度测序和生物信息学分析。使用针对表达最丰富的 lncRNA 的 siRNA 进行 RNA 干扰。随后检查了 lncRNA 的沉默效果以及 7 个热休克蛋白（HSP）基因的相对表达水平。总体而言，在成年工蜂暴露于热应激后，鉴定出了 7842 个 lncRNA 和 115 个差异表达的 lncRNA（DEL）。结构分析表明，lncRNA 的整体表达丰度、转录本长度、外显子数量和开放阅读框均低于 mRNA。GO 分析表明，靶基因主要参与“代谢”、“蛋白质折叠”、“应激反应”和“信号转导”途径。KEGG 分析表明，“内质网中的蛋白质加工”和“长寿调节途径-多种物种”途径最为丰富。对选定的 DEL 进行定量实时聚合酶链反应（qRT-PCR）检测证实了测序数据的可靠性。此外，siRNA 实验表明，喂食 siRNA 可使 lncRNA MSTRG.9645.5 的沉默效率达到 77.51%。沉默该 lncRNA 后，三个 HSP 基因的表达水平显著下调（p<0.05），而其他三个 HSP 基因的表达水平显著上调（p<0.05）。我们的研究结果为理解短期热应激下成年工蜂中 lncRNA 的调控机制提供了新视角。
