Tracing meristem cell lineages during plant development and regeneration by the all-in-one CRE/LOX system

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_pro-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_pro-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.