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Plant and Cell Physiology Advance Access originally published online on February 5, 2007
Plant and Cell Physiology 2007 48(3):441-450; doi:10.1093/pcp/pcm014
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© The Author 2007. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oxfordjournals.org

Phosphoenolpyruvate Carboxykinase in Arabidopsis: Changes in Gene Expression, Protein and Activity during Vegetative and Reproductive Development

Susan Malone1,2, Zhi-Hui Chen1,3, Ahmad R. Bahrami1,4, Robert P. Walker1, Julie E. Gray1 and Richard C. Leegood1,*

1Robert Hill Institute, University of Sheffield, Sheffield S10 2TN, UK

*Corresponding author: E-mail, r.leegood{at}sheffield.ac.uk; Fax, +44-114-2220050.


  Abstract

The aim of this work was to investigate the occurrence of phosphoenolpyruvate carboxykinase (PEPCK) in different tissues of Arabidopsis thaliana throughout its vegetative and reproductive growth. The A. thaliana genome contains two PEPCK genes (PCK1 and PCK2), and these are predicted to generate 73,404 and 72,891 Da protein products, respectively. Both genes were transcribed in a range of tissues; however, PCK1 mRNA appeared to be more abundant and was present in a wider range of tissues. PEPCK protein was present in flowers, fruit, developing seed, germinating seed, leaves, stems and roots. Two PEPCK polypeptides, of ~74 and ~73 kDa were detected by immunoblotting, and these may arise from PCK1 and PCK2, respectively. PEPCK was abundant in cotyledons during post-germinative growth, and this is consistent with its well established role in gluconeogenesis. PEPCK was also abundant in sink tissues, such as young leaves, in developing flowers, fruit and seed. Immunohistochemistry and in situ hybridization showed that PEPCK was present in the nectaries, stigma, endocarp of the fruit wall and in tissues involved in the transfer of assimilates to the developing ovules and seeds, such as the vasculature and seed coat. The potential functions of PEPCK in A. thaliana are discussed.

Keywords: Arabidopsis thaliana - Development - Flower - Phosphoenolpyruvate carboxykinase — Seed

Abbreviations: OAA, oxaloacetate; PEPCK, phosphoenolpyruvate carboxykinase; RT–PCR, reverse transcription–PCR.

2Present address: Faculty of Medicine, Imperial College, London SW7 2AZ, UK.

3Present address: Division of Cell and Developmental Biology, School of Life Sciences, University of Dundee, Dundee DD1 5EH, UK.

4Present address: Department of Biology, Ferdowsi University of Mashhad, Mashhad, Iran.

(Received November 21, 2006; Accepted January 19, 2007)
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