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感染植原体的泡桐全基因组 DNA 甲基化分析。

Genome-wide DNA methylation analysis of paulownia with phytoplasma infection.

机构信息

Institute of Paulownia, Henan Agricultural University, 95 Wenhua Road, Jinshui District, 450002 Zhengzhou, Henan, PR China; College of Forestry, Henan Agricultural University, 95 Wenhua Road, Jinshui District, 450002 Zhengzhou, PR China.

Forestry Academy of Henan, Zhengzhou, PR China.

出版信息

Gene. 2020 Sep 10;755:144905. doi: 10.1016/j.gene.2020.144905. Epub 2020 Jun 12.

Abstract

DNA methylation, an important epigenetic modification, regulates a wide range of biological processes. Previous MSAP results showed that the occurrence of PaWB related to changes of DNA methylation level; however, the relationship between DNA methylation and gene expression remains obscure in paulownia. Therefore, in the present study, we applied WGBS and RNA-seq techniques to investigate the DNA methylation and gene expression changes between healthy Paulownia fortunei seedlings and the phytoplasma-infected ones. A map of methylated cytosines at the single base pair resolution of paulownia was constructed. Compared to the healthy seedlings, the DNA methylation level increased after phytoplasma infection, and the change of mCHH was the main methylation pattern. DMR analysis showed that 422,662 DMRs in the genome were identified, in which, 27,871 DMR-associated genes were differentially expressed. Finally, 436 genes with significant differences in their methylation levels and mRNA expression profiles were identified through integrated analysis of the DNA methylomic and transcriptomic. KEGG pathway analysis revealed that these genes are mainly involved in plant hormone signal transduction, carbon metabolism, and starch and sucrose metabolism pathways. Two of DMR-associated genes were verified by BS- PCR. Finally, we selected TRP 1 and R2R3-MYB protein were closely related to the occurrence of PaWB. Our findings provide valuable insight into the mechanism of PaWB at the epigenetic level.

摘要

DNA 甲基化是一种重要的表观遗传修饰,调节着广泛的生物学过程。先前的 MSAP 结果表明,PaWB 的发生与 DNA 甲基化水平的变化有关;然而,在泡桐中,DNA 甲基化与基因表达之间的关系仍不清楚。因此,在本研究中,我们应用 WGBS 和 RNA-seq 技术来研究健康泡桐幼苗和受植原体感染的泡桐幼苗之间的 DNA 甲基化和基因表达变化。构建了泡桐单碱基分辨率的甲基化胞嘧啶图谱。与健康幼苗相比,植原体感染后 DNA 甲基化水平增加,mCHH 的变化是主要的甲基化模式。DMR 分析表明,在基因组中鉴定出 422662 个 DMR,其中 27871 个 DMR 相关基因差异表达。最后,通过对 DNA 甲基组学和转录组学的综合分析,鉴定出 436 个在甲基化水平和 mRNA 表达谱上有显著差异的基因。KEGG 通路分析表明,这些基因主要参与植物激素信号转导、碳代谢以及淀粉和蔗糖代谢途径。通过 BS-PCR 验证了两个与 DMR 相关的基因。最后,我们选择了与 PaWB 发生密切相关的 TRP1 和 R2R3-MYB 蛋白。我们的研究结果为 PaWB 在表观遗传水平的机制提供了有价值的见解。

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