Roles of DEMETER in regulating DNA methylation in vegetative tissues and pathogen resistance,Journal of Integrative Plant Biology

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Roles of DEMETER in regulating DNA methylation in vegetative tissues and pathogen resistance
Journal of Integrative Plant Biology ( IF 9.3 ) Pub Date : 2020-11-25 , DOI: 10.1111/jipb.13037
Wenjie Zeng _{1,

2} , Huan Huang ₁ , Xueqiang Lin _{1,

2} , Chen Zhu ₁ , Ken-Ichi Kosami _{1,

3} , Chaofeng Huang ₁ , Huiming Zhang ₁ , Cheng-Guo Duan ₁ , Jian-Kang Zhu _{1,

4} , Daisuke Miki ₁

Affiliation

DNA methylation is an epigenetic mark important for genome stability and gene expression. In Arabidopsis thaliana, the 5‐methylcytosine DNA glycosylase/demethylase DEMETER (DME) controls active DNA demethylation during the reproductive stage; however, the lethality of loss‐of‐function dme mutations has made it difficult to assess DME function in vegetative tissues. Here, we edited DME using clustered regularly interspaced short palindromic repeats (CRISPR) /CRISPR‐associated protein 9 and created three weak dme mutants that produced a few viable seeds. We also performed central cell‐specific complementation in a strong dme mutant and combined this line with mutations in the other three Arabidopsis demethylase genes to generate the dme ros1 dml2 dml3 (drdd) quadruple mutant. A DNA methylome analysis showed that DME is required for DNA demethylation at hundreds of genomic regions in vegetative tissues. A transcriptome analysis of the drdd mutant revealed that DME and the other three demethylases are important for plant responses to biotic and abiotic stresses in vegetative tissues. Despite the limited role of DME in regulating DNA methylation in vegetative tissues, the dme mutants showed increased susceptibility to bacterial and fungal pathogens. Our study highlights the important functions of DME in vegetative tissues and provides valuable genetic tools for future investigations of DNA demethylation in plants.

中文翻译：

DEMETER在调节营养组织DNA甲基化和病原体抗性中的作用

DNA 甲基化是一种对基因组稳定性和基因表达很重要的表观遗传标记。在拟南芥中，5-甲基胞嘧啶 DNA 糖基化酶/去甲基化酶 DEMETER (DME) 在生殖阶段控制活跃的 DNA 去甲基化；然而，功能丧失的dme突变的致死性使得评估植物组织中的 DME 功能变得困难。在这里，我们使用成簇的规则间隔短回文重复序列 (CRISPR)/CRISPR 相关蛋白 9编辑DME，并创建了三个弱dme突变体，这些突变体产生了一些可行的种子。我们还在一个强dme突变体中进行了中心细胞特异性互补，并将这条线与其他三个突变体结合起来。拟南芥脱甲基酶基因产生dme ros1 dml2 dml3 ( drdd ) 四重突变体。DNA 甲基化组分析表明，植物组织中数百个基因组区域的 DNA 去甲基化需要 DME。drdd突变体的转录组分析表明，DME 和其他三种脱甲基酶对于植物对营养组织中生物和非生物胁迫的反应很重要。尽管DME的在调节营养组织DNA甲基化的作用有限，在DME突变体对细菌和真菌病原体的易感性增加。我们的研究突出了 DME 在营养组织中的重要功能，并为未来植物 DNA 去甲基化的研究提供了宝贵的遗传工具。

更新日期：2020-11-25

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