A Gene Regulatory Network for Root Epidermis Cell Differentiation in Arabidopsis

Abstract

The root epidermis of Arabidopsis provides an exceptional model for studying the molecular basis of cell fate and differentiation. To obtain a systems-level view of root epidermal cell differentiation, we used a genome-wide transcriptome approach to define and organize a large set of genes into a transcriptional regulatory network. Using cell fate mutants that produce only one of the two epidermal cell types, together with fluorescence-activated cell-sorting to preferentially analyze the root epidermis transcriptome, we identified 1,582 genes differentially expressed in the root-hair or non-hair cell types, including a set of 208 “core” root epidermal genes. The organization of the core genes into a network was accomplished by using 17 distinct root epidermis mutants and 2 hormone treatments to perturb the system and assess the effects on each gene's transcript accumulation. In addition, temporal gene expression information from a developmental time series dataset and predicted gene associations derived from a Bayesian modeling approach were used to aid the positioning of genes within the network. Further, a detailed functional analysis of likely bHLH regulatory genes within the network, including MYC1, bHLH54, bHLH66, and bHLH82, showed that three distinct subfamilies of bHLH proteins participate in root epidermis development in a stage-specific manner. The integration of genetic, genomic, and computational analyses provides a new view of the composition, architecture, and logic of the root epidermal transcriptional network, and it demonstrates the utility of a comprehensive systems approach for dissecting a complex regulatory network.

Author Summary

A current challenge in the field of developmental biology is to define the composition and organization of gene networks that direct the pattern and differentiation of cells, tissues, and organs. In this study, we address this problem using Arabidopsis root epidermis development, a relatively simple model for studies of cell pattern formation and differentiation in plants. We used a tissue-specific cell sorting approach to define more than 1,500 genes whose transcripts differentially accumulate in the developing root epidermis. A series of transcriptome analyses were performed with 17 root epidermal mutants and 2 plant hormone treatments to dissect the regulatory relationships between 208 core genes. In addition, gene expression information from a developmental time series dataset was used to organize genes temporally. The results provide insight into the composition, organization, and logic of a developmental gene regulatory network. Furthermore, this work demonstrates the utility of an integrated analysis in gene regulatory network construction using genetic, genomic, and computational approaches.