Molecular Basis for a Cell Fate Switch in Response to Impaired Ribosome Biogenesis in the Arabidopsis Root Epidermis.

The Arabidopsis (Arabidopsis thaliana) root epidermis consists of a position-dependent pattern of root hair cells and non-hair cells. Underlying this cell type patterning is a network of transcription factors including a central MYB-basic helix-loop-helix-WD40 complex containing WEREWOLF (WER), GLABRA3 (GL3)/ENHANCER OF GLABRA3, and TRANSPARENT TESTA GLABRA1. In this study, we used a genetic enhancer screen to identify apum23-4, a mutant allele of the ribosome biogenesis factor (RBF) gene ARABIDOPSIS PUMILIO23 (APUM23), which caused prospective root hair cells to instead adopt the non-hair cell fate. We discovered that this cell fate switch relied on MYB23, a MYB protein encoded by a WER target gene and acting redundantly with WER. In the apum23-4 mutant, MYB23 exhibited ectopic expression that was WER independent and instead required ANAC082, a recently identified ribosomal stress response mediator. We examined additional RBF mutants that produced ectopic non-hair cells and determined that this cell fate switch is generally linked to defects in ribosome biogenesis. Furthermore, the flagellin peptide flg22 triggers the ANAC082-MYB23-GL2 pathway. Taken together, our study provides a molecular explanation for root epidermal cell fate switch in response to ribosomal defects and, more generally, it demonstrates a novel regulatory connection between stress conditions and cell fate control in plants.

Root Epidermal Cell Patterning Is Modulated by a Critical Residue in the WEREWOLF Transcription Factor.

The Arabidopsis (Arabidopsis thaliana) root epidermis exhibits a position-dependent pattern of root-hair and nonhair cell types. A highly orchestrated network of gene regulatory interactions, including the R2R3-type MYB transcription factor WEREWOLF (WER), is responsible for generating this cell pattern during root development. In this study, we identified a novel wer mutant from a genetic enhancer screen, designated wer-4, that exhibits an abnormal pattern of root-hair and nonhair cells. We established that wer-4 bears a single-residue substitution (D105N) in the DNA-binding R3 MYB repeat of WER, which differentially affects the transcription of WER target genes, including GLABRA2, CAPRICE, TRIPTYCHON, and ENHANCER OF TRY AND CPC1 This modulation of the gene regulatory network leads to altered levels and distributions of cell fate regulators in the differentiating epidermal cells that ultimately generate the abnormal cell-type pattern. We also created several WER variants with substitutions at the Asp-105 position, and these exhibited a variety of gene expression and cell-type pattern alterations, further supporting the critical role of this residue. These findings provide insight into WER protein function and its importance in generating the proper balance of downstream transcriptional factors in the gene regulatory network that establishes root epidermal cell fate.

Distinct relationships between GLABRA2 and single-repeat R3 MYB transcription factors in the regulation of trichome and root hair patterning in Arabidopsis.

*The patterning of epidermal cell types in Arabidopsis is an excellent model for studying the molecular basis of cell specification. Trichome and root hair formation is controlled by a transcriptional activator complex that induces the homeobox gene GLABRA2 (GL2) and some single-repeat R3 MYB genes (single MYB). However, it remains unclear how the actions of GL2 and single MYBs are coordinated to regulate epidermal patterning. *GL2 is thought to act downstream of single MYBs to regulate trichome and root hair development. In order to test this hypothesis genetically, double and higher order mutants between gl2 and single myb were generated. *In these mutants, the glabrous phenotypes observed in the gl2 single mutants were partially recovered, suggesting that single MYBs may not act solely through GL2 to regulate trichome development. On the other hand, double and higher order mutants between gl2 and single myb phenocopied the root hair phenotype of gl2 single mutants, suggesting that GL2 and single MYBs act in a common pathway to regulate root hair patterning. *These findings reveal distinct relationships between GL2 and single MYBs in the regulation of trichome vs root hair development, and provide new insights into the molecular mechanism of epidermal patterning.

The gene regulatory network for root epidermal cell-type pattern formation in Arabidopsis.

A fundamental aspect of multicellular development is the patterning of distinct cell types in appropriate locations. In this review, the molecular genetic control of cell-type pattern formation in the root epidermis of Arabidopsis thaliana is summarized. This developmental system represents a simple and genetically tractable example of plant cell patterning. The distribution of the two epidermal cell types, root-hair cells and non-hair cells, are generated by a combination of positional signalling and lateral inhibition mechanisms. In addition, recent evidence suggests that reinforcing mechanisms are used to ensure that the initial cell fate choice is adopted in a robust manner.

Cell identity mediates the response of Arabidopsis roots to abiotic stress.

Little is known about the way developmental cues affect how cells interpret their environment. We characterized the transcriptional response to high salinity of different cell layers and developmental stages of the Arabidopsis root and found that transcriptional responses are highly constrained by developmental parameters. These transcriptional changes lead to the differential regulation of specific biological functions in subsets of cell layers, several of which correspond to observable physiological changes. We showed that known stress pathways primarily control semiubiquitous responses and used mutants that disrupt epidermal patterning to reveal cell-layer-specific and inter-cell-layer effects. By performing a similar analysis using iron deprivation, we identified common cell-type-specific stress responses and revealed the crucial role the environment plays in defining the transcriptional outcome of cell-fate decisions.