The <i>Arabidopsis</i> basic–helix–loop–helix transcription factor LRL1 activates cell wall-related genes during root hair development

Lotus japonicus-ROOT HAIR LESS1-LIKE-1 (LRL1) of Arabidopsis thaliana encodes a basic helix-loop-helix (bHLH) transcription factor (TF) involved in root hair development. Root hair development is regulated by an elaborate transcriptional network, in which GLABRA2 (GL2), a key negative regulator, directly represses bHLH TF genes, including LRL1 and ROOT HAIR DEFECTIVE6 (RHD6). Although RHD6 and its paralogous TFs have been shown to connect downstream to genes involved in cell morphological events, such as endomembrane and cell wall modification, the downstream network of LRL1 remains elusive. We found that a mutation of LRL1 causes a short-root hair phenotype and that this phenotype can be partially rescued by a transgene encoding a glucocorticoid receptor (GR) domain-fused LRL1, LRL1-GR, in the presence of glucocorticoids. Using this conditional rescue system, we identified 46 genes that are activated downstream of LRL1. Among these, the cell wall-related genes were significantly enriched and many of them were found to be immediately downstream of LRL1 without de novo protein synthesis in between. We further analyzed three representative genes, PROLINE-RICH PROTEIN1 (PRP1), PRP3, and XYLOGLUCAN ENDOTRANSGLUCOSYLASE/HYDOLASE12 (XTH12). Reporter gene analyses showed that these genes are specifically transcribed in root hair cells including those in the root-hypocotyl junction, and that their proteins were localized to the cell wall of elongating root hairs, root hair bulges, and root hair bulge-expecting loci. A T-DNA insertion mutant of PRP3 showed a moderate short-root hair phenotype. Based on these results, LRL1 is likely to promote root hair development throughout the morphogenetic process by activating cell wall-related genes.