Graduate School of Human Development and Environment, Kobe University, Kobe, Japan.
Research Center for Genetic Engineering, National Research and Innovation Agency, Bogor, Indonesia.
Methods Mol Biol. 2023;2672:233-244. doi: 10.1007/978-1-0716-3226-0_14.
Recent evidence has demonstrated that specific epigenetic changes are also related to plant growth and development. Immunostaining enables the detection and characterization of chromatin modification, e.g., histone H4 acetylation (H4K5ac), histone H3 methylation (H3K4me2 and H3K9me2), and DNA methylation (5mC) with unique and specific patterns in plant tissues. Here we describe experimental procedures to determine the histone H3 methylation (H3K4me2 and H3K9me2) patterns in 3D-chromatin in whole roots tissue and 2D-chromatin in single nuclei in rice. To analyze both iron and salinity treatments, we show how to test for changes to the epigenetic chromatin landscape using heterochromatin (H3K9me2) and euchromatin (H3K4me) markers for chromatin immunostaining, especially in the proximal meristem region. To elucidate the epigenetic impact of environmental stress and external plant growth regulators, we demonstrate how to apply a combination of salinity, auxin, and abscisic acid treatments. The results of these experiments provide insights into the epigenetic landscape during rice root growth and development.
最近的证据表明，特定的表观遗传变化也与植物的生长和发育有关。免疫染色可以检测和描述染色质修饰，例如组蛋白 H4 乙酰化 (H4K5ac)、组蛋白 H3 甲基化 (H3K4me2 和 H3K9me2) 和 DNA 甲基化 (5mC)，这些修饰在植物组织中具有独特和特定的模式。在这里，我们描述了确定水稻整个根部组织中的三维染色质和单个核中的二维染色质中组蛋白 H3 甲基化 (H3K4me2 和 H3K9me2) 模式的实验程序。为了分析铁和盐处理，我们展示了如何使用异染色质 (H3K9me2) 和常染色质 (H3K4me) 标记物进行染色质免疫染色来检测表观遗传染色质景观的变化，特别是在近基部长。为了阐明环境胁迫和外部植物生长调节剂的表观遗传影响，我们展示了如何结合盐度、生长素和脱落酸处理。这些实验的结果提供了在水稻根生长和发育过程中表观遗传景观的见解。
