National Key Laboratory of Crop Genetic Improvement, National Engineering Research Center of Rapeseed, Hubei Hongshan Laboratory, Huazhong Agricultural University, Wuhan, China.
Shanghai Center for Plant Stress Biology, Center for Excellence in Molecular Plant Sciences, Chinese Academy of Sciences, Shanghai, China.
Nat Plants. 2024 Sep;10(9):1317-1329. doi: 10.1038/s41477-024-01743-7. Epub 2024 Aug 23.
Chromatin immunoprecipitation followed by sequencing (ChIP-seq) is crucial for profiling histone modifications and transcription factor binding throughout the genome. However, its application in economically important plant organs (EIPOs) such as seeds, fruits and flowers is challenging due to their sturdy cell walls and complex constituents. Here we present advanced ChIP (aChIP), an optimized method that efficiently isolates chromatin from plant tissues while simultaneously removing cell walls and cellular constituents. aChIP precisely profiles histone modifications in all 14 tested EIPOs and identifies transcription factor and chromatin-modifying enzyme binding sites. In addition, aChIP enhances ChIP efficiency, revealing numerous novel modified sites compared with previous methods in vegetative tissues. aChIP reveals the histone modification landscape for rapeseed dry seeds, highlighting the intricate roles of chromatin dynamics during seed dormancy and germination. Altogether, aChIP is a powerful, efficient and sensitive approach for comprehensive chromatin profiling in virtually all plant tissues, especially in EIPOs.
染色质免疫沉淀测序(ChIP-seq)对于在整个基因组中分析组蛋白修饰和转录因子结合至关重要。然而,由于其坚固的细胞壁和复杂的成分,其在经济上重要的植物器官(EIPOs)如种子、果实和花朵中的应用具有挑战性。在这里,我们提出了先进的 ChIP(aChIP),这是一种优化的方法,可以有效地从植物组织中分离染色质,同时去除细胞壁和细胞成分。aChIP 精确地分析了所有 14 种测试的 EIPOs 中的组蛋白修饰,并鉴定了转录因子和染色质修饰酶的结合位点。此外,与营养组织中的先前方法相比,aChIP 提高了 ChIP 的效率,揭示了许多新的修饰位点。aChIP 揭示了油菜干种子的组蛋白修饰图谱,突出了染色质动力学在种子休眠和萌发过程中的复杂作用。总之,aChIP 是一种强大、高效和敏感的方法,可用于几乎所有植物组织的全面染色质分析,特别是在 EIPOs 中。