Plant Molecular Biology

, Volume 61, Issue 3, pp 525–535

High Lysine and High Tryptophan Transgenic Maize Resulting from the Reduction of Both 19- and 22-kD α-zeins

  • Shihshieh Huang
  • Alessandra Frizzi
  • Cheryl A. Florida
  • Diane E. Kruger
  • Michael H. Luethy
Article

Abstract

The major maize seed storage proteins, zeins, are deficient in lysine and tryptophan content, which contribute to the poor nutritional quality of corn. Whether through the identification of mutations or genetic engineering, kernels with reduced levels of zein proteins have been shown to have increased levels of lysine and tryptophan. It has been hypothesized that these increases are due to the reduction of lysine-poor zeins and a pleiotropic increase in the lysine-rich non-zein proteins. By transforming maize with constructs expressing chimeric double-stranded RNA, kernels derived from stable transgenic plants displayed significant declines in the accumulation of both 19- and 22-kD α-zeins, which resulted in higher lysine and tryptophan content than previously reported for kernels with reduced zein levels. The observation that lysine and tryptophan content is correlated with the protein levels measured in transgenic maize kernels is consistent with the hypothesis that a pleiotropic increase in non-zein proteins is contributing to an improved amino acid balance. In addition, a large increase in accumulation of free amino acids, consisting predominantly of asparagine, asparate and glutamate, was observed in the zein reduction kernels.

Keywords

gene suppression high lysine high tryptophan maize nutritional quality maize quality trait zein reduction 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Adams, W.R., Huang, S., Kriz, A.L., Luethy, M.H. 2004Matrix-assisted laser desorption ionization time-of-flight mass spectrometry analysis of zeins in mature maize kernelsJ. Agric. Food Chem.5218421849PubMedCrossRefGoogle Scholar
  2. Allen, R.S., Millgate, A.G., Chitty, J.A., Thisleton, J., Miller, J.A.C., Fist, A.J., Gerlach, W.L., Larkin, P.J. 2004RNAi-mediated replacement of morphine with the non-narcotic alkaloid reticuline in opium poppyNature Biotechnol.2215591566CrossRefGoogle Scholar
  3. Armstrong, C.L. and Rout, J.R. 2001. A novel Agrobacterium-mediated plant transformation method. WO Patent 0109302Google Scholar
  4. Coleman, C.E., Larkins, B.A. 1999

    The prolamins of maize

    Shewry, P.R.Casey, R. eds. Seed ProteinsKluwer Academic PublishersThe Netherlands109139
    Google Scholar
  5. Dellaporta, S. 1994

    Plant DNA miniprep and microprep

    Freeling, M.Walbot, V. eds. The Maize HandbookSpringer-VerlagNew York522525
    Google Scholar
  6. Dembinski, E., Bany, S., Raczynska-Bojanowska, K. 1995Asparagine and glutamine in the leaves of high and low protein maizeActa Physiol. Plant17361365Google Scholar
  7. Dyer, D.J., Feng, P. 1997NIR destined to be major analytical influenceFeedstuffs691625Google Scholar
  8. Glassman, K.F., Gordon-Kamm, W.J., Kinney, K.J., Lowe, K.S., Nichols, S.E. and Stecca, K.L. 2003. Recombinant constructs and their use in reducing gene expression. US Pat. Appl. Pub. No. US 2003/0036197 A1Google Scholar
  9. Habben, J.E., Moro, G.L., Hunter, B.G., Hamaker, B.R., Larkins, B.A. 1995Elongation factor 1 alpha concentration is highly correlated with the lysine content of maize endospermProc. Natl. Acad. Sci. U.S.A.9286408644PubMedCrossRefGoogle Scholar
  10. Harper, A.E., Miller, R.H., Block, K.P. 1984Branched-chain amino acid metabolismAnn. Rev. Nutr.4409454CrossRefGoogle Scholar
  11. Helliwell, C.A., Wesley, S.V., Wielopolska, A.J., Waterhouse, P.M. 2002High-throughout vectors for efficient gene silencing in plantsFunct. Plant Biol.2912171225CrossRefGoogle Scholar
  12. Huang, S., Adams, W.R., Zhou, Q., Malloy, K.P., Voyles, D.A., Anthony, J., Kriz, A.L., Luethy, M.H. 2004aImproving nutritional quality of maize proteins by expressing sense and antisense zein genesJ. Agric. Food Chem.5219581964CrossRefGoogle Scholar
  13. Huang, S., Gilbertson, L.A., Adams, T.H., Malloy, K.P., Reisenbigler, E.K., Birr, D.H., Snyder, M.W., Zhang, Q., Luethy, M.H. 2004bGeneration of marker-free transgenic maize by regular two-border Agrobacterium transformation vectorsTrans. Res.13451461CrossRefGoogle Scholar
  14. Huang, S., Kruger, D.E., Frizzi., A., D’Ordine, R.L., Florida, C.A., Adams, W.R., Brown, W.E., Luethy, M.H. 2005High lysine corn produced by the combination of enhanced lysine biosynthesis and reduced zein accumulationPlant Biotechnol.3555569CrossRefGoogle Scholar
  15. Lohaus, G., Büker, M., Hussmann, M., Soave, C., Heldt, H.-W. 1998Transport of amino acids with special emphasis on the synthesis and transport of asparagine in the Illinois Low Protein and Illinois High Protein strains of maizePlanta205181188CrossRefGoogle Scholar
  16. Mertz, E.T., Bates, L.S., Nelson, O.E. 1964Mutant gene that changes protein composition and increases lysine content of maize endospermScience145279280PubMedGoogle Scholar
  17. Nelson, O.E., Mertz, E.T., Bates, L.S. 1965Second mutant gene affecting the amino acid pattern of maize endosperm proteinsScience15014691470Google Scholar
  18. Prasanna, B.M., Vasal, S.K., Kassahun, B., Singh, N.N. 2001Quality protein maizeCurr. Sci.8113081319Google Scholar
  19. Puckett, J.L., Kriz, A.L. 1991Globulin gene expression in opaque-2 and floury-2 mutant maize embryosMaydica36161167Google Scholar
  20. Segal, G., Song, R., Messing, J. 2003A new opaque variant of maize by a single dominant RNA-interference-inducing transgeneGenetics165387397PubMedGoogle Scholar
  21. Wang, X., Larkins, B.A. 2001Genetic analysis of amino acid accumulation in opaque-2 maize endospermPlant Physiol.12517661777PubMedCrossRefGoogle Scholar
  22. Wesley, S.V., Helliwell, C.A., Smith, N.A., Wang, M.-B., Rouse, D.T., Liu, Q., Gooding, P.S., Singh, S.P., Abbott, D., Stoutjesdijk, P.A., Robinson, S.P., Gleave, A.P., Green, A.G., Waterhouse, P.M. 2001Construct design for efficient, effective and high-throughput gene silencing in plantsPlant J.27581590PubMedCrossRefGoogle Scholar
  23. Woo, Y.-M., Hu, D.W.-N., Larkins, B.A., Jung, R. 2001Genomics analysis of genes expressed in maize endosperm identifies novel seed proteins and clarifies patterns of zein gene expressionPlant Cell1322972317PubMedCrossRefGoogle Scholar

Copyright information

© Springer 2006

Authors and Affiliations

  • Shihshieh Huang
    • 1
  • Alessandra Frizzi
    • 1
  • Cheryl A. Florida
    • 2
  • Diane E. Kruger
    • 1
  • Michael H. Luethy
    • 2
  1. 1.Mystic ResearchMonsanto CompanyMysticUSA
  2. 2.Chesterfield Village CampusMonsanto CompanyChesterfieldUSA

Personalised recommendations