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Quiescent centre (QC) cells represent an integral part of the root stem cell niche. They typically display a low division frequency that has been reported to be controlled by hormone signaling and different regulators, including the ETHYLENE RESPONSE FACTOR 115 (ERF115) transcription factor and D-type cyclins. Here, we applied a three-dimensional (3D) imaging to visualize the Arabidopsis QC cell number, volume and division patterns, including visualization of anticlinal divisions that cannot be deduced from longitudinal 2D imaging. We found that 5-day-old seedlings possess on average eight QC cells which are organized in a monolayered disc. In a period of 7 d, half of the QC cells undergo anticlinal division in a largely invariant space. Ectopic expression of ERF115 and CYCLIN D1;1 (CYCD1;1) promote both anticlinal and periclinal QC cell divisions, the latter resulting in a dual-layered QC zone holding up to 2-fold more QC cells compared with the wild type. In contrast, application of cytokinin or ethylene results in an increase in the number of periclinal, but a decrease in anticlinal QC divisions, suggesting that they control the orientation of QC cell division. Our data illustrate the power of 3D visualization in revealing unexpected QC characteristics. [ABSTRACT FROM AUTHOR]