Tomasiak Alicja, Piński Artur, Milewska-Hendel Anna, Andreu Godall Ignasi, Borowska-Żuchowska Natalia, Morończyk Joanna, Moreno-Romero Jordi, Betekhtin Alexander
Institute of Biology, Biotechnology and Environmental Protection, Faculty of Natural Sciences, University of Silesia in Katowice, Katowice, Poland.
Centre for Research in Agricultural Genomics (CRAG), CSIC-IRTA-UAB-UB, Barcelona, Spain.
Front Plant Sci. 2024 Sep 25;15:1465514. doi: 10.3389/fpls.2024.1465514. eCollection 2024.
Epigenetic changes accompany the dynamic changes in the cell wall composition during the development of callus cells. H3K4me3 is responsible for active gene expression and reaction to environmental cues. Chromatin immunoprecipitation (ChIP) is a powerful technique for studying the interplay between epigenetic modifications and the DNA regions of interest. In combination with sequencing, it can provide the genome-wide enrichment of the specific epigenetic mark, providing vital information on its involvement in the plethora of cellular processes. Here, we describe the genome-wide distribution of H3K4me3 in morphogenic and non-morphogenic callus of . Levels of H3K4me3 were higher around the transcription start site, in agreement with the role of this mark in transcriptional activation. The global levels of methylation were higher in the non-morphogenic callus, which indicated increased gene activation compared to the morphogenic callus. We also employed ChIP to analyse the changes in the enrichment of this epigenetic mark on the cell wall-related genes in both calli types during the course of the passage. Enrichment of H3K4me3 on cell wall genes was specific for callus type, suggesting that the role of this mark in cell-wall remodelling is complex and involved in many processes related to dedifferentiation and redifferentiation. This intricacy of the cell wall composition was supported by the immunohistochemical analysis of the cell wall epitopes' distribution of pectins and extensins. Together, these data give a novel insight into the involvement of H3K4me3 in the regeneration processes in callus tissue culture.
在愈伤组织细胞发育过程中，表观遗传变化伴随着细胞壁成分的动态变化。H3K4me3负责活跃基因表达以及对环境信号的反应。染色质免疫沉淀（ChIP）是研究表观遗传修饰与感兴趣的DNA区域之间相互作用的强大技术。与测序相结合，它可以提供全基因组范围内特定表观遗传标记的富集情况，为其参与众多细胞过程提供重要信息。在此，我们描述了H3K4me3在[具体植物名称]的形态发生和非形态发生愈伤组织中的全基因组分布。H3K4me3水平在转录起始位点周围较高，这与该标记在转录激活中的作用一致。非形态发生愈伤组织中的整体甲基化水平较高，这表明与形态发生愈伤组织相比，基因激活增加。我们还采用ChIP分析了传代过程中两种愈伤组织类型中该表观遗传标记在细胞壁相关基因上的富集变化。H3K4me3在细胞壁基因上的富集对愈伤组织类型具有特异性，这表明该标记在细胞壁重塑中的作用是复杂的，并且参与了许多与去分化和再分化相关的过程。细胞壁表位中果胶和伸展蛋白分布的免疫组织化学分析支持了细胞壁组成的这种复杂性。总之，这些数据为H3K4me3参与愈伤组织组织培养中的再生过程提供了新的见解。
