Wang Shengnan, Li Yu, Li Zijuan, Li Chao, Xu Lin, Zhai Ning
School of Life Sciences, East China Normal University, Shanghai, 200241, China.
Key Laboratory of Plant Carbon Capture, CAS Center for Excellence in Molecular Plant Sciences, Shanghai Institute of Plant Physiology and Ecology, Chinese Academy of Sciences, Shanghai, 200032, China.
Plant Methods. 2025 Mar 14;21(1):36. doi: 10.1186/s13007-025-01352-1.
The root apical meristem (RAM) and the shoot apical meristem (SAM), which contain stem cells and their lineages, are the centers for underground and aboveground organ formation in vascular plants. Accurately tracing stem cell lineage is important to understand how the RAM and SAM are formed and how they give rise to new organs. Here, we introduce the all-in-one CRE/LOX system, which is assembled in a single plasmid and can be used to efficiently trace cell lineage during de novo formation of meristems and cell differentiation within meristems in Arabidopsis thaliana. Using the CRE/LOX-GUS system driven by the WUSCHEL-RELATED HOMEOBOX5 (WOX5) promoter (i.e., WOX5-CRE/LOX-GUS), we show that cell division of the quiescent center (QC) within the stem cell niche of the RAM might replenish initial/stem cells in plants grown on mannitol-containing medium or in soil. The results obtained using WOX5-CRE/LOX-GUS also show that a group of shoot progenitor cells acts together to initiate the SAM during de novo shoot regeneration from callus in tissue culture. We also demonstrate the use of the CRE/LOX-RUBY system for real-time in vivo tracing of cell lineages in live organs. Overall, this paper not only introduces the all-in-one CRE/LOX tool, but also reveals the cell lineages during meristem development and regeneration.
根顶端分生组织（RAM）和茎顶端分生组织（SAM）包含干细胞及其谱系，是维管植物地下和地上器官形成的中心。准确追踪干细胞谱系对于理解RAM和SAM如何形成以及它们如何产生新器官至关重要。在这里，我们介绍了一种一体化的CRE/LOX系统，该系统组装在单个质粒中，可用于在拟南芥分生组织从头形成和分生组织内细胞分化过程中高效追踪细胞谱系。使用由WUSCHEL相关同源盒5（WOX5）启动子驱动的CRE/LOX-GUS系统（即WOX5-CRE/LOX-GUS），我们发现，在含甘露醇的培养基上生长的植物或土壤中的植物中，RAM干细胞龛内静止中心（QC）的细胞分裂可能会补充初始/干细胞。使用WOX5-CRE/LOX-GUS获得的结果还表明，一组茎祖细胞共同作用，在组织培养中从愈伤组织进行从头茎再生期间启动SAM。我们还展示了CRE/LOX-RUBY系统在实时体内追踪活器官中细胞谱系的应用。总体而言，本文不仅介绍了一体化的CRE/LOX工具，还揭示了分生组织发育和再生过程中的细胞谱系。
