Stem Cell Activity And Regeneration In Roots Require Non-Cell Autonomous Regulation From The Ground Tissue

Stem cells are central to plant development. During root postembryonic development stem cells generating tissues are patterned in layers around a stem cell organizer or quiescent center (QC). How stem cell lineages are initially patterned is unclear. Here, we describe a role for BLUEJAY (BLJ), JACKDAW (JKD) and SCARECROW (SCR) transcription factors in patterning of cell lineages during growth and in patterning reestablishment during regeneration through regulation of number of QC cells and their regenerative capacities. In blj jkd scr mutants, QC cells are progressively lost which results in loss of tissue layers. Upon laser ablation blj jkd scr is impaired in QC division and specification resulting in severe impairment in pattern regeneration. Beyond direct regulation of QC activity by these transcription factors, reduced levels of SHORT-ROOT (SHR) and of PIN auxin transporters were observed in the vasculature of blj jkd scr, leading to strong reduction in the auxin response in the QC. We narrowed down non-cell-autonomous regulation of vascularly expressed genes in blj jkd scr to C-REPEAT BINDING FACTOR 3 (CBF3). cbf3 mutant shows reduced levels of SHR in the vasculature, and in addition QC disorganization and downregulation of the QC regulator WUSCHEL-RELATED HOMEODOMAIN 5 (WOX5). CBF3 gene is primarily expressed in the ground tissue downstream of BLJ, JKD and SCR, while CBF3 protein may move. Targeted-expression of CBF3 to the ground tissue of blj jkd scr recovers radial patterning and regeneration. We propose that BLJ, JKD and SCR regulate QC-mediated patterning, and that part of this regulation involves CBF3.