First published online October 15, 2002; 10.1104/pp.010363
Plant Physiol, November 2002, Vol. 130, pp. 1464-1475
Characterization of the Genes Encoding the Cytosolic and
Plastidial Forms of ADP-Glucose Pyrophosphorylase in Wheat
Endosperm1
Rachel A.
Burton,2
Philip E.
Johnson,2
Diane M.
Beckles,
Geoffrey B.
Fincher,
Helen L.
Jenner,
Mike J.
Naldrett, and
Kay
Denyer*
John Innes Centre, Norwich Research Park, Colney, Norfolk NR4 7UH,
United Kingdom (P.E.J., H.L.J., M.J.N., K.D.); Department of Plant
Science, Waite Campus, University of Adelaide, Glen Osmond South
Australia 5064, Australia (R.B., G.B.F.); and DuPont Agricultural
Products, Newark, Delaware 19711-6104 (D.M.B.)
In most species, the synthesis of ADP-glucose (Glc) by the
enzyme ADP-Glc pyrophosphorylase (AGPase) occurs entirely within the
plastids in all tissues so far examined. However, in the endosperm of
many, if not all grasses, a second form of AGPase synthesizes ADP-Glc
outside the plastid, presumably in the cytosol. In this paper, we show
that in the endosperm of wheat (Triticum aestivum), the
cytosolic form accounts for most of the AGPase activity. Using a
combination of molecular and biochemical approaches to identify the
cytosolic and plastidial protein components of wheat endosperm AGPase
we show that the large and small subunits of the cytosolic enzyme are
encoded by genes previously thought to encode plastidial subunits, and
that a gene, Ta.AGP.S.1, which encodes the small subunit of the
cytosolic form of AGPase, also gives rise to a second transcript by the
use of an alternate first exon. This second transcript encodes an
AGPase small subunit with a transit peptide. However, we could not find
a plastidial small subunit protein corresponding to this transcript.
The protein sequence of the purified plastidial small subunit does not
match precisely to that encoded by Ta.AGP.S.1 or to the predicted
sequences of any other known gene from wheat or barley (Hordeum
vulgare). Instead, the protein sequence is most similar
to those of the plastidial small subunits from chickpea (Cicer
arietinum) and maize (Zea mays) and rice
(Oryza sativa) seeds. These data suggest that the gene encoding the major plastidial small subunit of AGPase in wheat
endosperm has yet to be identified.
1
This work was supported by the Biotechnology and
Biological Sciences Research Council (UK; competitive strategic grant
to the John Innes Centre), by DuPont Agricultural Products, and by the
Australian Grains Research and Development Corporation.
2
These authors contributed equally to the paper.
*
Corresponding author; e-mail kay.denyer{at}bbsrc.ac.uk; fax
44-1603-450045.
© 2002 American Society of Plant Biologists
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