Abstract
A full-length cDNA clone representing the large (shrunken-2) subunit of ADP-glucose pyrophosphorylase (AGP; EC 2.7.7.27) has been isolated from a cDNA library prepared from developing grain of hexaploid wheat (Triticum aestivum L., cv. Chinese Spring). The 2084-bp cDNA insert contains an open reading frame of 1566 nucleotides and primer-extension analysis indicated that the 5′ end is 10 nucleotides shorter than the mRNA. The deduced protein contains 522 amino acids (57.8 kDa) and includes a putative transit peptide of 62 amino acids (6.5 kDa). The similarity of the deduced protein to the small subunit of AGP and to other AGP genes from plants and microorganisms is discussed. Northern hybridisation shows that the Agp1 genes (encoding the small subunit in the wheat endosperm) and the Agp2 genes (encoding the large subunit in the wheat endosperm) are differentially expressed in the wheat grain. Transcripts from both gene sets accumulate to high levels in the endosperm during grain development with the majority of the expression in the endopsperm rather than the embryo and pericarp layers. Although enzyme activity is detected in developing grains prior to 10 d post anthesis, only the Agp1 genes are active at this time (the Agp2 genes are not expressed until 10 d post anthesis). The possibility that the enzyme expressed during early grain development is a homotetramer of small subunits is discussed. The Agp1 and Agp2 genes are arranged as triplicate sets of single-copy homoeoloci in wheat. The Agp2 genes are located on the long arms of chromosomes 1A, 1B and 1D, about 80 cM from the centromere. The Agp1 genes have been mapped to a position just distal to the centromere on the long arms of chromosomes 7A, 7B and 7D.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Ainsworth, C.C., Clark, J., Balsdon, J. (1993a) Expression, organisation and structure of the genes encoding the waxy protein (granule bound starch synthase) in wheat. Plant. Mol. Biol. 22, 67–82
Ainsworth, C.C., Tarvis, M.D., Clark, J.R. (1993b) Isolation and analysis of a cDNA clone encoding the small subunit of ADPglucose pyrophosphorylase from wheat. Plant. Mol. Biol. 23, 23–33
Anderson, J.M., Hnilo, J., Larson, R., Okita, T.W., Morrell, M., Preiss, J. (1989) The encoded primary sequence of a rice seed ADP-glucose pyrophosphorylase subunit and its homology to the bacterial enzyme. J. Biol. Chem. 264, 12238–12242
Baecker, P.A., Furlong, C.E., Preiss, J. (1983) Biosynthesis of bacteial glycogen. Primary structure of Escherichia coli ADP-glucose synthetase as deduced from the nucleotide sequence of the glgC gene. J. Biol. Chem. 258, 5084–5088
Bhave, M.R., Lawrence, S., Barton, C., Hannah, L.C. (1990) Identification and molecular characterisation of shrunken-2 cDNA clones of maize. Plant Cell 2, 581–588
Buchanan Wollaston (1994) Isolation of cDNA clones for genes that are expressed during leaf senescence in Brassica napus, identification of a gene encoding a senescence-specific metallothionein-like protein. Plant Physiol. 105, 839–846
Cavaner, D.R., Ray, S.C. (1991) Eukaryotic start and stop translation sites. Nucleic Acids Res. 19, 3185–3192
Charng, Y.Y., Kakefuda, G., Iglesias, A.A., Buikema, W.J., Preiss, J. (1992) Molecular-cloning and expression of the gene encoding ADP-glucose pyrophosphorylase from the cyanobacterium Anabaena-sp strain pcc7120. Plant Mol. Biol. 20, 37–47
Devos, K.M., Atkinson, M.D., Chinoy, C.N., Liu, C., Gale, M.D. (1992) RFLP-based genetic map of the homoeologous group 3 chromosomes of wheat and rye. Theor. Appl. Genet. 83, 931–939
Haugen, T.H., Ishaque, A., Preiss, J. (1979) Biosynthesis of bacterial glycogen. Characterisation of the subunit structure of Escherichia coli B glucose-1-phosphate adenylyltransferase. J. Biol. Chem. 251, 7880–7885
Iglesias, A.A., Kakefuda, G., Preiss, J. (1991) Regulatory and structural-properties of the cyanobacterial ADP-glucose pyrophosphorylases. Plant Physiol. 97, 1187–1195
Joshi, C.P. (1987) Putative polyadenylation signals in nuclear genes of higher plants, a compilation and analysis. Nucleic Acids Res. 15, 9627–9640
Keegstra, K., Olsen, L.J. (1989) Chloroplastic precursors and their transport across the envelope membranes. Annu. Rev. Plant Physiol. Plant Mol. Biol. 40, 471–501
Kleczowski, L.A., Villand, P., Lüthi, E., Olsen, O-E., Preiss, J. (1993) Insensitivity of barley endosperm ADP-glucose pyrophosphorylase to 3-phosphoglycerate and orthophosphate regulation. Plant Physiol. 101, 179–186
Klösgen, R.B., Gierl, A., Schwarz-Sommer, Z., Saedler, H. (1986) Molecular analysis of the waxy locus of maize. Mol. Gen. Genet. 203, 237–244
Lin, T-P., Caspar, T., Somerville, C., Preiss, J. (1988a) Isolation and characterisation of a starchless mutant of Arabidopsis thaliana (L.) Heynh lacking ADP-glucose pyrophosphorylase activity. Plant Physiol. 86, 1131–1135
Lin, T-P., Caspar, T., Somerville, C., Preiss, J. (1988) A starch deficient mutant of Arabidopsis thaliana with low ADP-glucose pyrophosphorylase activity lacks one of the two subunits of the enzyme. Plant Physiol. 88, 1175–1181
Maniatis, T., Fritsch, E.F., Sambrook, J. (1982) Molecular cloning: A laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York
Masojc, P., Gale, M.D. (1991) α-Amylase structural genes in rye (1991) Theor. Appl. Genet. 82, 771–776
Morell, M.K., Bloom, M., Knowles, V., Preiss, J. (1987) Subunit structure of spinach leaf ADPglucose pyrophosphorylase. Plant Physiol. 85, 182–187
Müller-Rober, B., Sonnewald, U., Willmitzer, L. (1992) Inhibition of ADP-glucose pyrophosphorylase in transgenic potatoes leads to sugar-storing tubers and influences tuber formation and expression of tuber storage protein genes. EMBO J. 11, 1229–1238
Nakamura, Y., Imamura, M. (1985) Regulation of ADP-glucose pyrophosphorylase from Chlorella vulgaris. Plant Physiol. 85, 601–605
Nakata, P.A., Greene, T.W., Anderson, J.M., Smith-White, B.J., Okita, T.W., Preiss, J. (1991) Comparison of the primary sequences of two potato tuber ADP-glucose pyrophosphorylase subunits. Plant Mol. Biol. 17, 1089–1093
Olive, M.R., Ellis, R.J., Schuch, W.W. (1989) Isolation and nucleotide sequence of cDNA clones encoding ADP-glucose pyrophosphorylase polypeptides from wheat leaf and endosperm. Plant Mol. Biol. 12, 525–538
Parsons, T.F., Preiss, J. (1978) Biosynthesis of bacterial glycogen, Isolation and characterisation of the pyridoxal-P allosteric activator site and the ADP-glucose-protected pyridoxal-P binding site of E. coli B ADP-glucose synthase. J. Biol. Chem. 253, 7638–7645
Preiss, J. (1991) Biology and molecular biology of starch synthesis and its regulation. In: Oxford surveys of plant molecular and cell biology, vol. 7, pp. 59–114, Miflin, B.J., ed. Oxford University Press
Preiss, J. (1988) Biosynthesis of starch and its regulation. In: The biochemistry of plants, vol. 14, pp. 371–423, Preiss, J., ed. Academic Press, New York
Preiss, J., Levi, C. (1980) Starch biosynthesis and degradation. In, The biochemistry of plants vol. 3, pp. 371–423, Preiss, J., ed. Academic Press, New York
Prioul, J-L., Jeanette, E., Reyss, A., Gregory, N., Giroux, M., Hannah, L.C., Causse, M. (1994) Expression of ADP-glucose pyrophosphorylase in maize (Zea mays L.) grain and source leaf during grain filling. Plant Physiol. 104, 179–187
Proudfoot, N.J., Brownlee, G.G. (1976) 3′ Non-coding region sequences in eukaryotic messenger RNA. Nature 263, 211–214
Sanger, F., Nicklen, S., Coulson, A.R. (1977) DNA sequencing with chain terminating inhibitors. Proc. Natl. Acad. Sci. USA 74, 5463–5467
Sanwall, G.G., Greenberg, E., Hardie, J., Cameron, E., Preiss, J. (1968) Regulation of starch biosynthesis in plant leaves, activation and inhibition of ADPglucose pyrophosphorylase. Plant Physiol. 43, 417–427
Sears, E. (1954) The aneuploids of common wheat. Mo. Agric. Exp. Stn. Res. Bull. 572, 1–59
Smith-White, B.J., Preiss, J. (1992) Comparison of proteins of ADP-glucose pyrophosphorylase from diverse sources. J. Mol. Evol. 34, 449–464
Smith-White, B.J., Preiss, J. (1994) Suggested mnemonics for cloned DNA corresponding to enzymes involved in starch metabolism. Plant Mol. Biol. Rep. 12, S67-S71
Stark, D.M., Timmerman, K.P., Barry, G.F., Preiss, J., Kishore, G.N. (1992) Regulation of the amount of starch in plant tissues by ADP-glucose pyrophosphorylase. Science 258, 287–292
Villand, P., Olsen, O-A., Kilian, A., Kleczkowski, L.A. (1992) ADP-glucose pyrophosphorylase large subunit cDNA from barley endosperm. Plant Physiol. 100, 1617–1618
Author information
Authors and Affiliations
Additional information
The nucleotide sequence data reported appear in the EMBL and GenBank Nucleotide Sequence Databases under the accession number Z21969.
We are very grateful to Vicky Buchanan-Wollaston for encouragement and for critical reading of the manuscript; to Prof. Curtis Hannah (University of Florida, Gainesville, USA) for kindly providing the maize shrunken-2 plasmids and to Mazhar Sheikh and Tina Mitchell for technical assistance. We are grateful to the Agriculture and Food Research Council for supporting the research.
Rights and permissions
About this article
Cite this article
Ainsworth, C., Hosein, F., Tarvis, M. et al. Adenosine diphosphate glucose pyrophosphorylase genes in wheat: differential expression and gene mapping. Planta 197, 1–10 (1995). https://doi.org/10.1007/BF00239933
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00239933