Root stem cell niche (SCN) consists of a quiescent center (QC) and surrounding stem cells. Disrupted symplastic communication leads to loss of stemness in the whole SCN. Several SCN regulators were reported to move between cells for SCN main-tenance. However, single mutant of these regulators is insu?cient to abolish QC stemness despite the high differentiation rate in surrounding stem cells. To dis-sect the mechanism behind such distinct stemness in SCN, we combined the mis-expression strategy with pWOX5:icals3m system in which QC is symplastically isolated. We found the starch accumulation in QC could be synergistically repressed by WUSCHEL-RE-LATED HOMEOBOX 5 (WOX5), SHORT-ROOT (SHR), SCARCROW (SCR), and PLETHORA (PLT). Like PLTs, other core regulators also exhibited dimorphicfunctions by inhibiting differentiation at a higher dose while promoting cell division at a low protein level. Being located in the center of the intersected ex-pression zones, QC cells receive the highest level of core regulators, forming the most robust stemness within SCN. WUSCHEL-RELATED HOMEOBOX 5 was su?cient to activate PLT1/2 expression, contributing to the QC-enriched PLTs. Our results provide ex-perimental evidence supporting the long-standing hypothesis that the combination of spatial ex-pression, synergistic function and dosage effect of core regulators result in spatially distinct stemness in SCN.