iGReD, CNRS, Inserm, Université Clermont Auvergne, Clermont-Ferrand, France.
Methods Mol Biol. 2025;2873:247-262. doi: 10.1007/978-1-0716-4228-3_14.
Embryo development and seed germination are characterized by important alterations in nuclear and chromatin organization. It is however challenging to analyze these dynamic changes because the Arabidopsis embryo is embedded in the seed coat and a layer of endosperm tissue and is therefore difficult to access. Here, we describe a protocol to analyze nuclear organization in whole-mount dry seed embryos from Arabidopsis thaliana and to investigate the distribution of histone variants and modifications using immunostaining on isolated embryo nuclei. This protocol should be useful to elucidate the dry seed-specific chromatin organization and to follow dynamic changes in nuclear organization during seed germination.
胚胎发育和种子萌发的特点是核和染色质组织的重要改变。然而，由于拟南芥胚胎嵌入在种皮和一层胚乳组织中，因此很难进行分析，所以这些动态变化的分析具有一定的挑战性。在这里，我们描述了一种从拟南芥中分析整个干燥种子胚胎核组织的方案，并使用分离胚胎核的免疫染色来研究组蛋白变体和修饰的分布。该方案应该有助于阐明干燥种子特有的染色质组织，并在种子萌发过程中跟踪核组织的动态变化。
