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Plant and Cell Physiology Advance Access originally published online on October 20, 2006
Plant and Cell Physiology 2006 47(11):1443-1456; doi:10.1093/pcp/pcl023
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© The Author 2006. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oxfordjournals.org

Rapid Paper

The ENHANCER OF SHOOT REGENERATION 2 gene in Arabidopsis Regulates CUP-SHAPED COTYLEDON 1 at the Transcriptional Level and Controls Cotyledon Development

Yoshihisa Ikeda1,3, Hiroharu Banno1,4, Qi-Wen Niu1, Stephen H. Howell2 and Nam-Hai Chua1,*

1 Laboratory of Plant Molecular Biology, The Rockefeller University, 1230 York Avenue, New York, NY 10021-6399, USA
2 Plant Sciences Institute, Iowa State University, Ames, IA 50011, USA

* Corresponding author: E-mail, chua{at}mail.rockefeller.edu; Fax, +1-212-327-8327.


  Abstract

Cytokinins stimulate shoot regeneration in tissue culture but the genes required for this developmental process are not well understood. Here we show that the Arabidopsis gene, ENHANCER OF SHOOT REGENERATION2 (ESR2), encoding an AP2-domain transcriptional factor, functions in the regeneration of shoots in tissue culture. ESR2 overexpression conferred cytokinin-independent shoot regeneration from cre1/ahk4 mutant explants, suggesting that CRE1 is not required for ESR2-mediated shoot regeneration. ESR2 directly targeted CYCLIN D1;1, ARABIDOPSIS PHOSPHOTRANSMITTER 6 (AHP6) and CUP-SHAPED COTYLEDON 1 (CUC1) in assays involving the translocation of ESR2–ER (estradiol receptor) fusions to the nucleus. Knock-down of ESR2 expression by RNA interference (RNAi) reduced CUC1 expression and resulted in altered cotyledon phenotypes at a low frequency, including partially fused cotyledons and triple cotyledons. Phenotypes of induced ESR2 expression in a cuc1-1 mutant background were suppressed. Our results suggest that ESR2 plays a role in shoot regeneration through transcriptional regulation of CUC1.

Keywords: Cell cycle - Cotyledon - Cytokinin - Gene expression - Shoot regeneration

Abbreviations: AHK, ARABIDOPSIS HISTIDINE KINASE; AHP, ARABIDOPSIS HISTIDINE PHOSPHOTRANSMITTER; ANT, AINTEGUMENTA; ARR, ARABIDOPSIS RESPONSE REGULATOR; BP, BREVIPEDICELLUS; CHX, cycloheximide; CIM, callus-inducing medium; CUC, CUP-SHAPED COTYLEDON; CKI1, CYTOKININ-INDEPENDENT 1; CRE1, CYTOKININ RESPONSE 1; CYC, CYCLIN; DRN, DORNRÖSCHEN; EAR, ERF-associated amphiphilic repression; ERF, ETHYLENE RESPONSE FACTOR; ER, estrogen receptor; ESR, ENHANCER OF SHOOT REGENERATION; GFP, green fluorescent protein; GUS, ß-glucuronidase; HLS, HOOKLESS; IPT, isopentenyl transferase; KNAT, KNOTTED-like from Arabidopsis thaliana; PIN, PIN FORMED; RIM, root induction medium; RNAi, RNA interference; RT–PCR, reverse transcription–PCR; SAM, shoot apical meristem; SIM, shoot induction medium; STM, SHOOT MERISTEMLESS; WOL, WOODEN LEG; WUS, WUSCHEL

3 Present address: Umeå Plant Science Centre, Department of Forest Genetics and Plant Physiology, Swedish University of Agricultural Sciences, SE-901 83 Umeå, Sweden.

4 Present address: Department of Environmental Biology, School of Bioscience and Biotechnology, Chubu University, 1200 Matsumoto-cho, Kasugai, Aichi, 487-8501 Japan.

The nucleotide sequence data reported herein have been deposited in the EMBL/GenBank databases under the following accession numbers: ESR1, AF353577 (At1g12980); ESR2, AAN27907 (At1g24590).

(Received September 7, 2006; Accepted October 8, 2006)
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