DNA methylation and histone modifications regulate de novo shoot regeneration in Arabidopsis by modulating WUSCHEL expression and auxin signaling.
DNA methylation and histone modifications regulate de novo shoot regeneration in Arabidopsis by modulating WUSCHEL expression and auxin signaling.
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DNA 甲基化和组蛋白修饰通过调节 WUSCHEL 表达和生长素信号传导来调节拟南芥芽再生
DOI：
10.1371/journal.pgen.1002243
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发表时间：
2011-08
期刊：
影响因子：
4.5
通讯作者：
Zhang XS
中科院分区：
文献类型：
--
作者：
Li W;Liu H;Cheng ZJ;Su YH;Han HN;Zhang Y;Zhang XS
植物具有从分化的体细胞组织再生器官的深刻能力，基于此，离体繁殖植物成为可能。离体芽再生技术除了在生物技术中的应用外，也是研究器官发生的重要系统。植物激素和转录因子WUSCHEL（WUS）在这一过程中起着关键作用，但表观遗传修饰是否以及如何参与尚不清楚。在这里，我们报告了DNA甲基化和组蛋白修饰的表观遗传标记通过调节WUS表达和生长素信号传导来调节拟南芥从头芽再生。首先，关键表观遗传基因的功能丧失-包括编码DNA甲基转移酶的甲基转移酶1（MET 1），编码组蛋白3赖氨酸9（H3 K9）甲基转移酶的KRYPTONITE（KYP），编码组蛋白3赖氨酸4（H3 K4）脱甲基酶的JMJ 14和编码组蛋白乙酰转移酶的HAC 1-导致体外再生芽的WUS表达和发育速率改变。其次，我们通过亚硫酸氢盐测序和染色质免疫沉淀显示，WUS的调控区域在发育过程中受到DNA甲基化和组蛋白修饰的调控。第三，WUS调控区的DNA甲基化在met 1突变体中丢失，从而导致WUS表达及其定位增加。第四，我们进行了全基因组转录分析，发现野生型和met 1之间的一些差异表达基因参与了植物激素生长素的信号转导。我们证实了生长素反应因子3（ARF 3）在met 1中的表达增加确实是由于DNA去甲基化，这表明DNA甲基化通过调节生长素信号传导来调节从头芽再生。我们认为DNA甲基化和组蛋白修饰通过调节WUS表达和生长素信号传导来调节从头芽再生。这项研究表明，尽管参与器官发生的分子组分在植物和动物中进化不同，但表观遗传修饰在这一过程中发挥着进化趋同的作用。
Plants have a profound capacity to regenerate organs from differentiated somatic tissues, based on which propagating plants in vitro was made possible. Beside its use in biotechnology, in vitro shoot regeneration is also an important system to study de novo organogenesis. Phytohormones and transcription factor WUSCHEL (WUS) play critical roles in this process but whether and how epigenetic modifications are involved is unknown. Here, we report that epigenetic marks of DNA methylation and histone modifications regulate de novo shoot regeneration of Arabidopsis through modulating WUS expression and auxin signaling. First, functional loss of key epigenetic genes—including METHYLTRANSFERASE1 (MET1) encoding for DNA methyltransferase, KRYPTONITE (KYP) for the histone 3 lysine 9 (H3K9) methyltransferase, JMJ14 for the histone 3 lysine 4 (H3K4) demethylase, and HAC1 for the histone acetyltransferase—resulted in altered WUS expression and developmental rates of regenerated shoots in vitro. Second, we showed that regulatory regions of WUS were developmentally regulated by both DNA methylation and histone modifications through bisulfite sequencing and chromatin immunoprecipitation. Third, DNA methylation in the regulatory regions of WUS was lost in the met1 mutant, thus leading to increased WUS expression and its localization. Fourth, we did a genome-wide transcriptional analysis and found out that some of differentially expressed genes between wild type and met1 were involved in signal transduction of the phytohormone auxin. We verified that the increased expression of AUXIN RESPONSE FACTOR3 (ARF3) in met1 indeed was due to DNA demethylation, suggesting DNA methylation regulates de novo shoot regeneration by modulating auxin signaling. We propose that DNA methylation and histone modifications regulate de novo shoot regeneration by modulating WUS expression and auxin signaling. The study demonstrates that, although molecular components involved in organogenesis are divergently evolved in plants and animals, epigenetic modifications play an evolutionarily convergent role in this process.
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