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The callus formation capacity of strawberry leaf explant is modulated by DNA methylation.
Horticulture Research ( IF 8.7 ) Pub Date : 2022-01-19 , DOI: 10.1093/hr/uhab073
Decai Liu 1 , Qin Mu 1 , Xianyang Li 1 , Sheng Xu 1 , Yi Li 2 , Tingting Gu 1
Affiliation  

Shoot regeneration from leaf tissue requires de-differentiation of cells from a highly differentiated state into an active dividing state, but how this physiological transition occurs and is regulated especially at epigenetic level remains obscure. Here we have characterized the DNA methylome represented by 5-methylcytosine (5mC) in leaf and the callus tissue derived from the leaf explant of woodland strawberry Fragaria vesca. We detected an overall increase of DNA methylation and distinct 5mC enrichment patterns in the CG, CHG and CHH sequence contexts in genetic and transposable elements. Our analyses revealed an intricate relation between DNA methylation and gene expression levels in leaf or leaf-derived callus. However, when considering the genes involved in callus formation and shoot regeneration, e.g. FvePLT3/7, FveWIND3, FveWIND4, FveLOG4 and FveIAA14, their dynamic transcription levels were associated with the differentially methylated regions located in the promoters or gene bodies, indicating a regulatory role of DNA methylation in the transcriptional regulation of pluripotency acquisition in strawberry. Furthermore, application of a DNA methyltransferase inhibitor 5'-azacytidine (5'-Aza) hampered both callus formation and shoot regeneration from the leaf explant. We further showed that 5'-Aza down-regulated the genes involved in cell wall integrity, such as expansin, pectin lyase and pectin methylesterase genes, suggesting an essential role of cell wall metabolism during callus formation. This study reveals the contribution of DNA methylation in callus formation capacity and will provide a basis for developing a strategy to improve shoot regeneration for basic and applied research applications.

中文翻译:

草莓叶外植体的愈伤组织形成能力受 DNA 甲基化的调节。

叶组织的枝条再生需要将细胞从高度分化状态去分化为活跃的分裂状态,但这种生理转变如何发生并特别是在表观遗传水平上受到调节仍然不清楚。在这里,我们已经表征了叶子中以 5-甲基胞嘧啶 (5mC) 为代表的 DNA 甲基化组,以及来自林地草莓草莓叶外植体的愈伤组织组织。我们在遗传和转座因子的 CG、CHG 和 CHH 序列环境中检测到 DNA 甲基化的总体增加和不同的 5mC 富集模式。我们的分析揭示了叶或叶衍生愈伤组织中 DNA 甲基化与基因表达水平之间的复杂关系。然而,当考虑与愈伤组织形成和枝条再生有关的基因时,例如 FvePLT3/7、FveWIND3、FveWIND4、FveLOG4和FveIAA14,它们的动态转录水平与位于启动子或基因体的差异甲基化区域相关,表明DNA甲基化在草莓多能性获得的转录调控中的调节作用。此外,DNA 甲基转移酶抑制剂 5'-氮杂胞苷 (5'-Aza) 的应用阻碍了叶外植体的愈伤组织形成和枝条再生。我们进一步表明,5'-Aza 下调了参与细胞壁完整性的基因,如扩展蛋白、果胶裂解酶和果胶甲基酯酶基因,表明细胞壁代谢在愈伤组织形成过程中的重要作用。
更新日期:2022-01-19
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