Skip to main content
Log in

Characterisation of the wheat Mr 15000 “grain-softness protein” and analysis of the relationship between its accumulation in the whole seed and grain softness

  • Published:
Theoretical and Applied Genetics Aims and scope Submit manuscript

Abstract

The Mr 15000 protein associated with water-washed wheat starch granules from soft wheats was shown to be heterogeneous: it could be divided into a fraction containing one or moreα-amylase inhibitor subunits and a fraction largely composed of a previously uncharacterised polypeptide(s) referred to as the “grainsoftness protein” (GSP). The major N-terminal sequence and sequences of peptides derived from protease digests of GSP are reported. An antiserum specific for GSP was used to show that GSP accumulated in both hard and soft wheat grains, but the GSP in soft grains associated more strongly with starch granules than the GSP in hard grains. A positive correlation between grain softness and accumulation of GSP in the seed was demonstrated for a range of cultivars. This differs from the qualitative relationship, based on the isolated starch fraction, between GSP and grain softness that has already been reported. Analysis of wholemeal extracts with the antiserum demonstrated that the accumulation of GSP in the seed was dependent on the short arm of chromosome 5D, which also encodes theHa locus. In addition, examination of near-isogenic lines differing in hardness indicated that the gene(s) controlling GSP was (were) linked with theHa locus. The findings indicate that GSP may be the product of theHa locus and thus be the major factor that determines the milling characteristics of bread wheats.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Barlow KK, Buttrose MS, Simmonds DH, Vesk M (1973) The nature of the starch-protein interface in wheat endosperm. Cereal Chem 50:443–454

    Google Scholar 

  • Biffen RH (1908) On the inheritance of strength in wheat. J Agric Sci 3:86–101

    Google Scholar 

  • Carpenter HC, Skerritt JH, Wrigley CW, Margolis J (1986) A device for preparative elution electrophoresis using a polyacrylamide-gel slab. Electrophoresis 7:221–226

    Google Scholar 

  • Christie GE, Platt T (1980) Gene structure in the tryptophan operon ofEscherichia coli: nucleotide sequence oftrpC and the flanking intercistronic regions. J Mol Biol 142:519–530

    Google Scholar 

  • Coombs RRA, Kieffer M, Fraser DR, Frazier PJ (1983) Naturally developing antibodies to wheat gliadin fractions and to other cereal antigens in rabbits, rats and guinea pigs on normal laboratory diets. Int Archs Allergy Appl Immun 70:200–204

    Google Scholar 

  • Devereaux J, Haeberli P, Marques P (1984) Sequence analysis software package of the genetics computer group. University of Wisconsin Biotechnology Centre

  • Gallagher S, Winston SE, Fuller SA, Hurrell JGR (1992) Immunoblotting and immunodetection. In: Ausabel, FM, Brent R, Kingston RE, Moore DD, Seidman JG, Smith JA, Stuhl K (eds) Current protocols in molecular biology. Greene Publishing and Wiley Interscience, New York, pp 10.8.1–10.8.16

    Google Scholar 

  • García-Maroto F, Marana C, Mena M, Garcia-Olmedo F, Carbonero P (1990) Cloning of the cDNA and chromosomal location of genes encoding the three types of subunits of the wheat tetrameric inhibitor of insectα-amylase. Plant Mol Biol 14:845–853

    Google Scholar 

  • García-Olmedo F, Salcedo G, Sanchez-Monge R, Gomez L, Royo J, Carbonero P (1987) Plant proteinaceous inhibitors of proteinases andα-amylases. Oxford Surveys Plant Mol Cell Biol 4:275–334

    Google Scholar 

  • Glenn GM, Saunders RM (1990) Physical and structural properties of wheat endosperm associated with grain texture. Cereal Chem 67:176–181

    Google Scholar 

  • Görg A, Postel W, Gunther S (1988) The current state of 2-dimensional electrophoresis with immobilised pH gradients. Electrophoresis 9:531–546

    Google Scholar 

  • Greenwell P, Schofield JD (1986) A starch-granule protein associated with endosperm softness in wheat. Cereal Chem 63:379–380

    Google Scholar 

  • Greenwell P, Schofield JD (1989) The chemical basis of grain hardness and softness. In wheat end-use properties. Proc 1989 Int Assoc Cereal Sci Technol Symp Helsinki. Helsinki Press, pp 59–71

  • Greer EN, Hinton JJC (1950) The two types of wheat endosperm. Nature 13:746–748

    Google Scholar 

  • Greer EN, Hinton JJC, Jones CR, Kent NL (1951) The occurrence of endosperm cells in wheat flour. Cereal Chem 28:58–67

    Google Scholar 

  • Hashimoto F, Horigome T, Kanbayashi M, Yoshida K, Sugano H (1983) An improved method for separation of low-molecular-weight polypeptides by electrophoresis in sodium docecyl sulphate-polyacrylamide gel. Anal Biochem 129:192–199

    Google Scholar 

  • Jolly C (1991) The biochemistry and molecular genetics of grain softness and hardness in wheat,Triticum aestivum. PhD thesis, Macquarie University

  • Jolly C, Rahman S, Kortt AA, Higgins TJV (1990) Characterisation of grain-softness protein, a marker of endosperm texture in wheat. Proc 40th Aust Cereal Chem Conf. Albury, Australia, pp 92–95

  • Kashlan N, Richardson M (1981) The complete amino-acid sequence of a major wheat protein inhibitor ofα-amylase. Phytochemistry 20:1781–1781

    Google Scholar 

  • Konik CN, Miskle DM, Gras PW (1991) Optimisation of small scale tests for predicting noodle quality. Proc Cereals Int Conf. Brisbane, Australia, pp 271–275

  • Kortt AA, Caldwell JB, Lilley GG, Higgins TJ, Spencer D (1991) Amino-acid cDNA sequences of a methionine-rich 2S protein from sunflower seed (Helianthus annus L.). Eur J Biochem 195:329–334

    Google Scholar 

  • Kyshe-Andersen J (1984) Electoblotting of multiple gels: a simple apparatus without buffer tank for rapid transfer of proteins from polyacrylamide to nitrocellulose. J Biochem Biophys Methods 10:203–209

    Google Scholar 

  • Law CN, Young CF, Brown JWS, Snape JW, Worland AJ (1978) The study of grain-protein control in wheat using wholechromosome substitution lines. In: Seed protein improvement by nuclear techniques Int Atomic Energy Agency, Vienna, pp 483–502

    Google Scholar 

  • Mattern PJ, Morris R, Schmidt JW, Johnson VA (1973) Locations of genes for kernel properties in the wheat variety “Cheyenne” using chromosome substitution lines. Proc 4th Int Wheat Genet Symp., pp 703–707, Columbus, MO

  • Morrison WR, Wylie LJ, Law CN (1984) The effect of the group-5 chromosomes on the free and total galactosylglycerides in wheat endosperm J Cereal Sci 2:145–152

    Google Scholar 

  • Morrison WR, Law CN, Wylie LJ, Coventry AM, Seekings J (1989) The effect of group-5 chromosomes on the free polar lipids and breadmaking quality of wheat. J Cereal Sci 9:41–51

    Google Scholar 

  • Morrison WR, Greenwell P, Law CN, Sulaiman BD (1992) Occurrence of friabilin, a low molecular weight protein associated with grain softness, on starch granules isolated from some wheats and related species. J Cereal Sci 15:143–149

    Google Scholar 

  • Oda S, Komae K, Yasui Y (1992) Relation between starch granule protein and endosperm softness in Japanese wheat (Triticum aestivum L.) cultivars. Japan J Breed 42:161–165

    Google Scholar 

  • Parish JA, Halse NJ (1968) Effects of light, temperature, and rate of desiccation on translucency in wheat grain. Aust J Agric Res 19:365–372

    Google Scholar 

  • Schofield JD, Greenwell P (1987) Wheat starch-granule proteins and their technological significance. In: Morton ID (ed) Cereals in a European context. Ellis Horwood, Chichester, pp 407–420

    Google Scholar 

  • Simmonds DH (1972) The ultrastructure of the mature wheat endosperm. Cereal Chem 49:212–222

    Google Scholar 

  • Simmonds DH, Barlow KK, Wrigley CW (1973) The biochemical basis of grain hardness in wheat. Cereal Chem 50:553–563

    Google Scholar 

  • Singh NK, Donovan GR, Batey IL, MacRitchie F (1990) Use of sonication and size-exclusion high-performance liquid chromatography in the study of wheat flour proteins. I. Dissolution of total proteins in the absence of reducing agents. Cereal Chem 67:150–160

    Google Scholar 

  • Skerritt JH, Frend AJ, Robson LG, Greenwell P (1990) Immunological homologies between wheat, gluten and starchgranule proteins. J Cereal Sci 123–136

  • Spencer D, Higgins TJV, Button S, Davey R (1980) Pulse-labelling studies on protein synthesis in developing pea seeds and evidence of a precursor form of the legumin small subunit. Plant Physiol 66:510–515

    Google Scholar 

  • Stenvert NL, Kingswood K (1977) The influence of the physical structure of the protein matrix on wheat hardness. J Sci Food Agric 28:11–19

    Google Scholar 

  • Sulaiman D, Morrison WR (1990) Proteins associated with the surface of wheat starch-granules purified by centrifuging through caesium chloride. J Cereal Sci 12:53–61

    Google Scholar 

  • Symes KJ (1965) The inheritance of grain hardness in wheat as measured by the particle-size index. Aust J Agric Res 16:113–123

    Google Scholar 

  • Symes KJ (1969) Influence of a gene causing hardness on the milling and baking quality of two wheats. Aust J Agric Res 20:971–979

    Google Scholar 

  • Williams PC (1979) Screening wheat for protein and hardness by near infrared reflectance spectroscopy. Cereal Chem 56:169–172

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Additional information

Communicated by G. Wenzel

Rights and permissions

Reprints and permissions

About this article

Cite this article

Jolly, C.J., Rahman, S., Kortt, A.A. et al. Characterisation of the wheat Mr 15000 “grain-softness protein” and analysis of the relationship between its accumulation in the whole seed and grain softness. Theoret. Appl. Genetics 86, 589–597 (1993). https://doi.org/10.1007/BF00838714

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00838714

Key words

Navigation