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JXB Advance Access published online on March 31, 2003
Journal of Experimental Botany, doi:10.1093/jxb/erg151
© 2003 by Oxford University Press
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Received August 26, 2002; accepted February 12, 2003
© 2003 Society for Experimental Biology

RESEARCH PAPER

Molecular and biochemical characterization of cytosolic phosphoglucomutase in wheat endosperm (Triticum aestivum L. cv. Axona)

Emma J. Davies 1, Ian J. Tetlow 2, Caroline G. Bowsher 1, Michael J. Emes 2*

1 School of Biological Sciences, 3.614 Stopford Building, Oxford Road, University of Manchester, Manchester M13 9PT, UK
2 Department of Botany, College of Biological Sciences, University of Guelph, Guelph, Ontario, N1G 4C4, Canada

* To whom correspondence should be addressed. E-mail: memes{at}uoguelph.ca.


  Abstract

Evidence from a number of plant tissues suggests that phosphoglucomutase (PGM) is present in both the cytosol and the plastid. The cytosolic and plastidic isoforms of PGM have been partially purified from wheat endosperm (Triticum aestivum L. cv. Axona). Both isoforms required glucose 1,6-bisphosphate for their activity with Ka values of 4.5 µM and 3.8 µM for cytosolic and plastidic isoforms, respectively, and followed normal Michaelis-Menten kinetics with glucose 1-phosphate as the substrate with Km values of 0.1 mM and 0.12 mM for the cytosolic and plastidic isoforms, respectively. A cDNA clone was isolated from wheat endosperm that encodes the cytosolic isoform of PGM. The deduced amino acid sequence shows significant homology to PGMs from eukaryotic and prokaryotic sources. PGM activity was measured in whole cell extracts and in amyloplasts isolated during the development of wheat endosperm. Results indicate an approximate 80% reduction in measurable activity of plastidial and cytosolic PGM between 8 d and 30 d post-anthesis. Northern analysis showed a reduction in cytosolic PGM mRNA accumulation during the same period of development. The implications of the changes in PGM activity during the synthesis of starch in developing endosperm are discussed.

Key words: Phosphoglucomutase, starch synthesis, Triticum aestivum.


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