title: Genome-wide direct target analysis reveals a role for SHORT-ROOT in root vascular patterning through cytokinin homeostasis.
abstract: SHORT-ROOT (SHR) is a key regulator of root growth and development in Arabidopsis (Arabidopsis thaliana). Made in the stele, the SHR protein moves into an adjacent cell layer, where it specifies endodermal cell fate; it is also essential for apical meristem maintenance, ground tissue patterning, vascular differentiation, and lateral root formation. Much has been learned about the mechanism by which SHR controls radial patterning, but how it regulates other aspects of root morphogenesis is still unclear. To dissect the SHR developmental pathway, we have determined the genome-wide locations of SHR direct targets using a chromatin immunoprecipitation followed by microarray analysis method. K-means clustering analysis not only identified additional quiescent center-specific SHR targets but also revealed a direct role for SHR in gene regulation in the pericycle and xylem. Using cell type-specific markers, we showed that in shr, the phloem and the phloem-associated pericycle expanded, whereas the xylem and xylem-associated pericycle diminished. Interestingly, we found that cytokinin level was elevated in shr and that exogenous cytokinin conferred a shr-like vascular patterning phenotype in wild-type root. By chromatin immunoprecipitation-polymerase chain reaction and reverse transcription-polymerase chain reaction assays, we showed that SHR regulates cytokinin homeostasis by directly controlling the transcription of cytokinin oxidase 3, a cytokinin catabolism enzyme preferentially expressed in the stele. Finally, overexpression of a cytokinin oxidase in shr alleviated its vascular patterning defect. On the basis of these results, we suggest that one mechanism by which SHR controls vascular patterning is the regulation of cytokinin homeostasis.
authors.affiliation: Department of Biological Science, Florida State University, Tallahassee, Florida 32306-4295, USA. hcui@bio.fsu.edu
authors.forename: Hongchang
authors.initials: H
authors.lastname: Cui
authors.forename: Yueling
authors.initials: Y
authors.lastname: Hao
authors.forename: Mikhail
authors.initials: M
authors.lastname: Kovtun
authors.forename: Viktor
authors.initials: V
authors.lastname: Stolc
authors.forename: Xing-Wang
authors.initials: XW
authors.lastname: Deng
authors.forename: Hitoshi
authors.initials: H
authors.lastname: Sakakibara
authors.forename: Mikiko
authors.initials: M
authors.lastname: Kojima
mesh_terms: D017360:Arabidopsis / Q000187:drug effects / Q000235:genetics* / Q000254:growth & development; D029681:Arabidopsis Proteins / Q000235:genetics / Q000378:metabolism*; D019521:Body Patterning / Q000187:drug effects / Q000235:genetics*; D047369:Chromatin Immunoprecipitation; D016000:Cluster Analysis; D003583:Cytokinins / Q000378:metabolism* / Q000494:pharmacology; D018506:Gene Expression Regulation, Plant / Q000187:drug effects; D018745:Genome, Plant / Q000235:genetics*; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006706:Homeostasis / Q000187:drug effects; D018613:Microscopy, Confocal; D009154:Mutation / Q000235:genetics; D020411:Oligonucleotide Array Sequence Analysis; D010641:Phenotype; D052585:Phloem / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D058526:Plant Vascular Bundle / Q000187:drug effects / Q000235:genetics / Q000254:growth & development*; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*
chemical_list: D029681:Arabidopsis Proteins; D003583:Cytokinins; C410510:SHORT ROOT protein, Arabidopsis; D014157:Transcription Factors; D049452:Green Fluorescent Proteins