Mathematical modelling of WOX5- and CLE40-mediated columella stem cell homeostasis in Arabidopsis.

The root meristem of Arabidopsis thaliana harbours a pool of stem cells, which divide to give rise to the differentiated cells of the various root tissues. Regulatory networks of inter-cellular signalling molecules control the homeostasis of stem cell number and position so that both stem and differentiated cells are consistently available. This work focuses on the transcription factor WUSCHEL-RELATED HOMEOBOX 5 (WOX5), the signalling peptide CLAVATA3/EMBRYO-SURROUNDING REGION 40 (CLE40) and the feedback loops involving them, which maintain the columella stem cells (CSCs). WOX5 signals from the quiescent centre (QC) to promote stem cell fate, while CLE40 is secreted from the differentiated columella cells (CCs) to promote differentiation. Our analyses of mathematical models of this network show that, when cell fate is controlled primarily by antagonistic factors, homeostasis requires a spatial component and inter-cellular signalling. We have also found that WOX5 contributes to, but is not absolutely necessary for, CSC maintenance. Furthermore, our modelling led us to postulate an additional signalling molecule that promotes CSC maintenance. We propose that this WOX5-independent signal originates in the QC, is targeted by CLE40 signalling and is capable of maintaining CSCs.

Moderation of Arabidopsis root stemness by CLAVATA1 and ARABIDOPSIS CRINKLY4 receptor kinase complexes.

BACKGROUND: The root system of higher plants originates from the activity of a root meristem, which comprises a group of highly specialized and long-lasting stem cells. Their maintenance and number is controlled by the quiescent center (QC) cells and by feedback signaling from differentiated cells. Root meristems may have evolved from structurally distinct shoot meristems; however, no common player acting in stemness control has been found so far. RESULTS: We show that CLAVATA1 (CLV1), a key receptor kinase in shoot stemness maintenance, performs a similar but distinct role in root meristems. We report that CLV1 is signaling, activated by the peptide ligand CLAVATA3/EMBRYO SURROUNDING REGION40 (CLE40), together with the receptor kinase ARABIDOPSIS CRINKLY4 (ACR4) to restrict root stemness. Both CLV1 and ACR4 overlap in their expression domains in the distal root meristem and localize to the plasma membrane (PM) and plasmodesmata (PDs), where ACR4 preferentially accumulates. Using multiparameter fluorescence image spectroscopy (MFIS), we show that CLV1 and ACR4 can form homo- and heteromeric complexes that differ in their composition depending on their subcellular localization. CONCLUSIONS: We hypothesize that these homo- and heteromeric complexes may differentially regulate distal root meristem maintenance. We conclude that essential components of the ancestral shoot stemness regulatory system also act in the root and that the specific interaction of CLV1 with ACR4 serves to moderate and control stemness homeostasis in the root meristem. The structural differences between these two meristem types may have necessitated this recruitment of ACR4 for signaling by CLV1.

A signaling module controlling the stem cell niche in Arabidopsis root meristems.

The niches of the Arabidopsis shoot and root meristems, the organizing center (OC) and the quiescent center (QC), orchestrate the fine balance of stem cell maintenance and the provision of differentiating descendants. They express the functionally related homeobox genes WUSCHEL (WUS) and WOX5, respectively, that promote stem cell fate in adjacent cells. Shoot stem cells signal back to the OC by secreting the CLAVATA3 (CLV3) dodecapeptide, which represses WUS expression. However, the signals controlling homeostasis of the root stem cell system are not identified to date. Here we show that the differentiating descendants of distal root stem cells express CLE40, a peptide closely related to CLV3. Reducing CLE40 levels delays differentiation and allows stem cell proliferation. Conversely, increased CLE40 levels drastically alter the expression domain of WOX5 and promote stem cell differentiation. We report that the receptor kinase ACR4, previously shown to control cell proliferation, is an essential component, and also a target, of CLE40 signaling. Our results reveal how, in contrast to the shoot system, signals originating from differentiated cells, but not the stem cells, determine the size and position of the root niche.