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Plant and Cell Physiology Advance Access published online on February 2, 2005

Plant and Cell Physiology, doi:10.1093/pcp/pci033
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Plant and Cell Physiology 2005 © The Japanese Society of Plant Physiologists (JSPP); all rights reserved.
Received September 29, 2004
Accepted December 12, 2004

Regular Paper

Combinatorial Microarray Analysis Revealing Arabidopsis Genes Implicated in Cytokinin Responses through the His-to-Asp Phosphorelay Circuitry

Takatoshi Kiba 1, Takahito Naitou 1, Nobuya Koizumi 1, Takafumi Yamashino 1, Hitoshi Sakakibara 2, and Takeshi Mizuno 1*

1 Laboratory of Molecular Microbiology, School of Agriculture, Nagoya University, Chikusa-ku, Nagoya 464-8601, Japan
2 Plant Science Center, RIKEN (Institute of Physical and Chemical Research), 1-7-22, Suehiro, Tsurumi, Yokohama 230-0045, Japan

* To whom correspondence should be addressed.
Takeshi Mizuno, E-mail: tmizuno{at}agr.nagoya-u.ac.jp


   Abstract

In Arabidopsis thaliana, the immediate early response of plants to cytokinin is formulated as the multistep AHK->AHP->ARR phosphorelay signaling circuitry, which is initiated by the cytokinin-receptor histidine protein kinases. In the hope of finding components (or genes) that function downstream of the cytokinin-mediated His->Asp phosphorelay signaling circuitry, we carried out genome-wide microarray analyses. To this end, we took a combinatorial microarray strategy by employing not only wild-type plants, but also certain transgenic lines in which the cytokinin-mediated His->Asp phosphorelay signaling circuitry has been genetically manipulated. These transgenic lines employed were ARR21-overexpressing and ARR22-overexpressing plants, each of which exhibits a characteristic phenotype with regard to the cytokinin-mediated His->Asp phosphorelay. The results of extensive microarray analyses with these plants allowed us to systematically identify a certain number of genes that were upregulated at the level of transcription in response to cytokinin directly or indirectly. Among them, some representatives were further examined in wild-type plants to support the idea that certain genes encoding transcription factors are rapidly and specifically induced at the level of transcription by cytokinin in a manner similar to that of the type-A ARR genes, which are the hallmarks of the His->Asp phosphorelay signaling circuitry. Several interesting transcription factors were thus identified as such cytokinin-responsive ones, including those belonging to AP2/EREBP-family, MYB-family, GATA-family, or bHLH-family. Including these, the presented list of cytokinin-upregulated (214) genes will provide us with valuable bases for understanding the His->Asp phosphorelay in Arabidopsis thaliana.

Keywords: Arabidopsis thaliana; Cytokinin; Microarray; Phosphorelay; Signal transduction.
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