Chinese Science Bulletin

, Volume 49, Issue 19, pp 2053–2057 | Cite as

Expression of lysine-rich protein gene and analysis of lysine content in transgenic wheat

  • Chaomin Meng
  • Xüqing Chen
  • Rongqi Liang
  • Fengping Yang
  • Liquan Zhang
  • Xiaodong Zhang
  • Tianyou Chen
  • S. S. M. Sun


Expression vector pBPC102, which carries winged bean lysine-rich protein (wblrp) gene and dihydropicolinate synthase (DHDPS) gene, was transferred into hexaploid winter wheat cv. Jinghua No.1, Jing411, You899 and Yangnong15 expiants of immature inflorescence and immature embryos by particle bombardment. More than 100 transgenic plants were obtained under the selection of s-(2-aminoethyl)-L-cysteine (AEC). Confirmed transgenic plants of T0 and T1 generation by PCR and PCR-Southern blotting analyses showed successful integration of wblrp gene into wheat genome. Analysis of transgenic plant lines of T2 by Northern dot-blotting showed good expression of wblrp gene in offspring seed. The content of free lysine in leaves, contents of bound lysine and total proteins in seeds of T2 transgenic wheat lines were determined and analyzed. Among 34 tested transgenic lines, levels of free lysine content in leaves of 9 transgenic lines are 2–3times higher than un-transformed wild-type cultivars. Among 17 analyzed transgenic lines, bound lysine content of 4 transgenic lines is more than 10% higher than that of wild-type cultivars. Our research suggests that introducing wblrp gene into wheat is an effective way to improve its nutrition quality.


wheat (Triticum aestivum L.) lysine-rich protein transgenic expression nutrition quality 


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  1. 1.
    Zhang, X. J., Liu, J. Q., Zhao Q. et al., Transfer of high lysine-rich gene into maize by microprojectile bombardment and detection of transgenic plants, J. Agri. Biotech. (in Chinese with English abstract), 1999, 7(4): 363–367.Google Scholar
  2. 2.
    Sun, X. H., Ao, G. M., Yu, J. J. et al., Transfer of high lysine-rich gene into maize inbred lines and the detection of transgenic plants, J. Agri. Biotech. (in Chinese with English abstract), 2001, 9(2): 156–158.Google Scholar
  3. 3.
    Gao, Y. F., Jing, Y. X., Shen, S. H. et al., Transfer of Lysine-rich protein gene into rice and production of fertile transgenic plants, Acta Botanica Sinica, 2001, 43 (5): 506–511.Google Scholar
  4. 4.
    Shaul, O., Galili, G., Increased lysine biosynthesis in tobacco plants that express high levels of bacterial dibydrodipicolinate syntbase in their chloroplssts, Plant J., 1992, 11 (2): 203–209.CrossRefGoogle Scholar
  5. 5.
    Brinch-pedersen, H., Galili, G., Knuden, S. et al., Engineering of the aspartate family biosynthetic pathway in barley (Hordeum vulgare L.) by transformation with heterologous genes encoding feed-back-insensitive aspartate kinase and dihydrodipicolinate synthase, Plant Mol. Biol., 1996, 32 (4): 611–620.PubMedCrossRefGoogle Scholar
  6. 6.
    Vasil, V., Castillo, A. M., Fromm, M. et al., Herbicide resistant fertile transgenic wheat plant obtained by microprojectile bombardment of regenerable embryogenic calls, Bio /Technology, 1992, 10: 667–674.Google Scholar
  7. 7.
    Zhang, X. D., Liang, R. Q., Cheng, X. Q. et al., Transgene inheritance and quality improvement by expressing novel HMW glutenin subunit (HMW-GS) genes in winter wheat, Chinese Science Bulletin, 2003, 48(8): 771–776.CrossRefGoogle Scholar
  8. 8.
    Sambrook, J., Fritsch, E. F., Maniatis, T., Molecular Cloning—A Laboratory Manual, 2nd ed., New York: Cold Spring Harbor Laboratory Press, 1989.Google Scholar
  9. 9.
    Lassenr, M. W., Peterson, P., Yoder, J. I., Simultaneous amplification of multiple DNA fragments by polymerase chain reaction in the analysis of transgenic plants and their progeny, Plant Molec. Biol. Rep., 1989, 7: 116–128.CrossRefGoogle Scholar
  10. 10.
    Ning, Z. X., Manual for Food Composition Analysis (in Chinese), Chinese Light Industry Press, 1998, 124–126.Google Scholar
  11. 11.
    He, Z. X., Zhang, D. Q., Chemistry of Food Protection and Technique Check Measure (in Chinese), Chinese Light Industry Press, 1998, 142–143.Google Scholar
  12. 12.
    Mertz, E. T., Bates, L. S., Nelson, O. E., Mutant gene that changes protein composition and increase lysine content of maize endosperm, Science, 1964, 145: 279–280.PubMedCrossRefGoogle Scholar
  13. 13.
    Munch, L., The case of high-lysine barley breeding, C.A.B. international, 1992: 573–596.Google Scholar
  14. 14.
    Xu, Q. F., Li, L. C., Chen, X. et al., Study on the obtaining of transgenic wheats with GNA alien gene by biolistic particle, Scientia Agricultura Sinica (in Chinese with English abstract), 2001, 34 (1): 1–4.Google Scholar
  15. 15.
    Takumi, S., Shimada, T., Variation in transformation frequencies among six common wheat cultivars through particle bombardment of scutellar tissues, Genes Genet. Syst., 1997, 72 (2): 63–69.PubMedCrossRefGoogle Scholar
  16. 16.
    Shimoni, Y., Blechl, A. E., Anderson, O. D. et al., A recombinant protein of two high molecular weight glutenins alters gluten polymer formation in transgenic wheat, J. Biol. Chem., 1997, 272 (24): 15488–15495.PubMedCrossRefGoogle Scholar
  17. 17.
    Barro, F., Rooke, L., Bekes, F. et al., Transformation of wheat with high molecular weight subunit genes results in improved functional properties, Nat. Biotechnol., 1997, 15(12): 1295–1299.PubMedCrossRefGoogle Scholar
  18. 18.
    Cheng, M., Joyce, E. F., Pang, S. Z. et al., Genetic transformation of wheat mediated by Agrobacterium tumefacien, Plant Physiol., 1997, 115: 971–980.PubMedGoogle Scholar
  19. 19.
    Weir, B., Gu, X., Wang, M. B. et al., Agrobacterium tumefaciens-mediated transformation of wheat using suspension cell as a model system and green fluorescent protein as a visual market, Aust. J. Plant Physiol., 2001, 28: 807–818.Google Scholar
  20. 20.
    Huang, Y. H., Zhou, M. P., Ye, X. G. et al., Study on the development of transgenic wheat mediated by Agrobacterium tumefaciens, Acta Agromomica Sinica (in Chinese with English abstract), 2002, 28(4): 510–515.Google Scholar
  21. 21.
    Soger, E., Williams, S., Christou, P. et al., Expression of the insecticidal lectin from snowdarop (Galathus nivalis agglutinin; GNA) intransgenic wheat plants: Effect on predation by grain aphid sitobion avenae, Molecular Breeding, 1999, 5(1): 65–73.CrossRefGoogle Scholar
  22. 22.
    Alpeter, F., Diaz, I., McAuslane, H. et al., Increased insect resistance in transgenic wheat stably expressing trypsin inhibitor CME, Molec. Breed., 1999, 5: 53–63.CrossRefGoogle Scholar
  23. 23.
    Sivamani, E., Brey, C. W., Talbert, L. E. et al., Resistance to wheat streak mosaic virus in transgenic wheat engineered with the viral coat protein gene, Transg. Res., 2002, 11: 31–41.CrossRefGoogle Scholar
  24. 24.
    Hou, W. S., Guo, S. D., Lu, M., Production of transgenic wheat plants with cryla gene via microprojection bombardment, Scientia Agricultura Sinica (in Chinese with English abstract), 2003, 36 (5): 469–472.Google Scholar
  25. 25.
    Xu, H. J., Pang, J. L., Ye, X. G. et al., Study on the gene transferring of Nib8 into wheat for its resistance to the yellow mosaic virus by bombardment, Acta Aggronomica Sinica (in Chinese with English abstract), 2001, 27(6): 688–693.Google Scholar
  26. 26.
    Sun, S. S. M., Xing, L. W., Jing, X. Y. et al., Lysine rich protein from winged bean, U.S.A. patent application no.08/964,722. 1998.Google Scholar
  27. 27.
    Yasuyuki, Y., Rosarin, K., Takashi, H. et al., Growth and aspartate kinase activity in wheat cell suspension culture: Effects of lysine analogs and asparatate-derived amino acids, Plant Cell Physiol., 1986, 27(4): 607–617.Google Scholar

Copyright information

© Science in China Press 2004

Authors and Affiliations

  • Chaomin Meng
    • 1
    • 2
  • Xüqing Chen
    • 1
  • Rongqi Liang
    • 1
  • Fengping Yang
    • 1
  • Liquan Zhang
    • 1
  • Xiaodong Zhang
    • 1
  • Tianyou Chen
    • 2
  • S. S. M. Sun
    • 3
  1. 1.Beijing Agro-Biotechnology Research CenterBeijingChina
  2. 2.College of AgronomyNorthwest Sci-Tech University of Agriculture and ForestryYanglingChina
  3. 3.Department of BiologyThe Chinese University of Hong KongHong KongChina

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