Skip to main content

Sequence heterogeneity and differential expression of the α-Amy2 gene family in wheat

Summary

The α-Amy2 genes of wheat are a multigene family which is expressed in the aleurone cells of germinating grain under control of the plant hormone gibberellin. A subset of the genes are also expressed in developing grain. Comparison of five genomic clones containing α-Amy2 genes, using DNA sequence analysis and Southern hybridisation, showed that the extent of similarity between genes differed. Two of the most heterogeneous genes compared were located to the same group 7 chromosome while the most similar genes α-Amy2/54 and α-Amy2/8 were located to different ones; hence sequence variation could not be correlated to the ancestry of the α-Amy2 genes during the separate existence of the constituent genomes of hexaploid wheat. Expression of the cloned genes was measured using an S1 nuclease protection assay and this identified α-Amy2/54 and α-Amy2/8 as part of the subset of α-Amy2 genes expressed in both the developing grain and in aleurone cells. Comparison of the 5′ upstream regions of all five genes showed high similarity, with the exception of one gene, up to-280 nucleotides from the transcriptional start, while similarity between α-Amy2/54 and α-Amy2/8 extended a further 90 bp upstream of this point. It is suggested that regulatory elements responsible for tissue specificity and gibberellin regulation may be located within these regions of similarity.

This is a preview of subscription content, access via your institution.

References

  • Baulcombe DC, Buffard D (1983) Gibberellic acid-regulated expression of α-amylase and six other genes in wheat aleurone layers. Planta 157:493–501

    Google Scholar 

  • Baulcombe DC, Huttly AK, Martienssen RA, Barker RF, Jarvis MG (1987) A novel wheat α-amylase gene (α-Amy3). Mol Gen Genet 209:33–40

    Google Scholar 

  • Biggin MD, Gibson T, Hong GF (1983) Buffer gradient gels and 35S label as an aid to rapid DNA sequence determination. Proc Natl Acad Sci USA 80:3963–3965

    Google Scholar 

  • Brown JWS (1986) A catalogue of splice junction and putative branch point sequences from plant introns. Nucleic Acids Res 14:9549–9559

    Google Scholar 

  • Daussant J, Renard HA (1986) Development of different α-amylase isozymes having high and low isoelectric points during early stages of kernel development in wheat. J Cereal Sci 5:13–19

    Google Scholar 

  • Devereux J, Haeberli P, Smithies O (1984) A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res 12:387–395

    Google Scholar 

  • Gale MD, Ainsworth CC (1984) The relationship between α-amylase species found in developing and germinating wheat grain. Biochem Genet 22:1031–1036

    Google Scholar 

  • Gale MD, Law CN, Chojecki AJ, Kempton RA (1983) Genetic control of α-amylase production in wheat. Theor Appl Genet 64:209–316

    Google Scholar 

  • Ho T-HD, Nolan RC, Shute DE (1981) Characterization of a gibberellin-insensitive dwarf wheat, D6899. Plant Physiol 67:1026–1031

    Google Scholar 

  • Huang J-K, Swegle M, Dandekar AM, Muthukrishnan S (1984) Expression and regulation of α-amylase gene family in barley aleurones. J Mol Appl Genet 2:579–588

    Google Scholar 

  • Knox CAP, Sonthayanon B, Chandra GR, Muthukrishnan S (1987) Structure and organisation of two divergent α-amylase genes from barley. Plant Mol Biol 9:3–17

    Google Scholar 

  • Lazarus CM, Baulcombe DC, Martienssen RA (1985) α-Amylase genes of wheat are two multigene families which are differentially expressed. Plant Mol Biol 5:13–24

    Google Scholar 

  • Maniatis T, Fritsch EF, Sambrook J (1982) In: Molecular cloning: A laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York

    Google Scholar 

  • Martienssen RA (1986) The molecular genetics of α-amylase gene families in wheat. PhD thesis, University of Cambridge, UK

    Google Scholar 

  • Messing J, Geraghty D, Heidecker G, Hu N-T, Kridl J, Rubenstein I (1983) Plant gene structure. In: Kosinge T, Meredith CP, Hollaender A (eds) Genetic engineering of plants. Plenum Publishing Corp, New York, pp 211–227

    Google Scholar 

  • Rigby PWJ, Dieckmann M, Rhodes C, Berg P (1977) Labelling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol 113:237–251

    Google Scholar 

  • Sanger F, Nicklen S, Coulsen AR (1977) DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467

    Google Scholar 

  • Sargent JG (1980) α-Amylase isoenzymes and starch degradation. Cereal Res Commun 8:77–86

    Google Scholar 

  • Staden R (1982) Automation of the computer handling, of gel reading data produced by the shotgun method of DNA sequencing. Nucleic Acids Res 10:4731–4751

    Google Scholar 

  • Thompson RD, Bartels D, Harberd NP, Flavell RB (1983) Characterisation of the multigene family coding for HMW glutenin subunits in wheat using cDNA clones. Theor Appl Genet 67:87–96

    Google Scholar 

  • von Heijne G (1984) Analysis of the distribution of charged residues in the N-terminal region of signal sequences: implications for protein export in prokaryotic and eukaryotic cells. EMBO J 3:2315–2318

    Google Scholar 

  • Whittier RF, Dean DA, Rogers JC (1987) Nucleotide sequence analysis of α-amylase and thiol protease genes that are hormonally regulated in barley aleurone cells. Nucleic Acids Res 15:2515–2535

    Google Scholar 

  • Young RA, Hagenbuchle O, Schibler U (1981) A single mouse α-amylase gene specifies two different tissue-specific mRNAs. Cell 23:451–458

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Additional information

Communicated by H. Saedler

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Huttly, A.K., Martienssen, R.A. & Baulcombe, D.C. Sequence heterogeneity and differential expression of the α-Amy2 gene family in wheat. Mol Gen Genet 214, 232–240 (1988). https://doi.org/10.1007/BF00337716

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00337716

Key words

  • α-Amylase
  • Chromosome location
  • Promoter sequence comparisons
  • Sequence heterogeneity
  • Tissuespecific expression