Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Research Article
  • Published:

Expression and inheritance of multiple transgenes in rice plants

Nature Biotechnology volume 16, pages 1060–1064 (1998)Cite this article

Abstract

The ability to control integration, inheritance, and expression of multiple transgenes is a prerequisite for manipulating biosynthetic pathways and complex agronomic characteristics in plants. One hundred and twenty-five independent transgenic rice plants were regenerated after cobombarding embryogenic tissues with a mixture of 14 different pUC-based plasmids. Eighty-five percent of the R0 plants contained more than two, and 17% more than nine, of the target genes. Plants containing multiple transgenes displayed normal morphologies and 63% set viable seed. Multigene cotransformation efficiency was correlated with the ratio in which the plasmids were mixed with respect to the selectable marker. All target genes had an equal chance of integration, indicating that the nature of the coding region had no effect on the efficiency of integration. Three plant lines containing 11, 10, and 9 transgenes, respectively, were analyzed for patterns of integration and inheritance until the R3 generation. Integration of multiple transgenes occurred at either one or two genetic loci, with inheritance conforming to a 3:1 Mendelian ratio. Coexpression of four marker genes was investigated until the R2 generation.

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

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1: Identification of transgenes in R0 rice plants by PCR.
Figure 2: (A) Fertile rice plants containing 12 transgenes in the greenhouse.
Figure 3: Southern blot hybridization of the hph, RTSV-CP3, and luc genes from R0, R1, and R2 plants of C27, C69, and S88.

Similar content being viewed by others

References

  1. Christou, P. 1996. Transformation technology. Trends in Plant Science 1: 423–431.

    Article  Google Scholar 

  2. Vasil, V., Castillo, A.M., Fromm, M.E., and Vasil I.K. 1992. Herbicide resistant fertile transgenic wheat plants obtained by particle bombardment of regenerable callus. Bio/Technology 10: 667–674.

    Google Scholar 

  3. Christou, P., Ford, T.L., and Kofron, M. 1991. Production of transgenic rice (Oryxa sativa L.) plants from agronomically important Indica and Japonica varieties via electric discharge particle acceleration of exogenous DNA into immature zygotic embryos. Bio/Technology 9: 957–962.

    Article  Google Scholar 

  4. Herrera-Estrella, L., Rosales, L.S., and Rivera-Bustamante, R. 1996. Transgenic plants for disease control, pp. 33–80 in Plant microbe interactions, Vol.1. Stacey, G. and Keen, N.T. (eds.). Chapman and Hall, New York.

    Google Scholar 

  5. Hayakawa, T., Zhu, Y., Itoh, K., Kimura, Y., Izawa, T., Shimamoto, K. et al. 1992. Genetically engineered rice resistant to rice stripe virus, an insect transmitted virus. Proc. Natl. Acad. Sci. USA 89: 9865 –9869.

    Article  Google Scholar 

  6. Wilson, T.M.A. 1993. Strategies to protect crop plants against viruses: pathogen-derived resistance blossoms. Proc. Natl. Acad. Sci. USA 90: 3134–3141.

    Article  Google Scholar 

  7. Ronald P.C. 1997 The molecular basis of disease resistance in rice. Plant Mol. Biol. 35: 179– 186.

    Article  Google Scholar 

  8. Zhang, S., Song, W., Chen, L., Ruan, D., Taylor, N.J., Ronald, P.C. et al. 1998. Transgenic elite Indica rice varieties resistant to Xanthomonas oryzae pv. oryzae. Molecular Breeding. In press.

  9. Fugimoto, H., Itoh, K., Yamamoto, M., Kyozuka, J., and Shimamoto, K. 1993. Insect resistant rice generated by the introduction of a modified ∂-endotoxin gene of Bacillus thuringiensis. Bio/Technology 11: 1151– 1155.

    Google Scholar 

  10. Stark, D.M., Timmerman F.P., Barry, G.F., Priess, J., and Kishore G.M. 1992. Regulation of the amount of starch in plants tissue by ADP glucose pyrophosphorylase. Science 258: 287–292.

    Article  Google Scholar 

  11. Redenbaugh, K., Berner, T., Emlay, D., Frankos, B., Hiatt, W., Houck, C. et al 1993. Regulatory issues for commercialization of tomatoes with an antisense polygalacturonase gene. In Vitro Cell Dev. Biol. 29: 17– 26.

    Article  Google Scholar 

  12. Tircoli, D.M., Carney, K.J., Russell, P.F., McMaster. J.R., Groffi, D.W., Hadden, K.C. et al. 1995. Field evaluation of transgenic squash containing single or multiple virus coat protein gene constructs for resistance to cucumber mosaic virus, watermelon virus 2 and zucchini yellow mosaic virus. Bio/Technology 13: 1458– 1473.

    Google Scholar 

  13. Li, L., Qu, R., de Kochko, A., Fauquet, C., and Beachy, R. 1993. An improved rice transformation system using the biolistic method. Plant Cell. Rep. 12: 250–255.

    Article  Google Scholar 

  14. Qu, R., de Kochko A., Zhang, L., Marmey, P., Li, L., Tian, W. et al. 1996. Analysis of a large number of independent transgenic rice plants produced by the biolistic method. In Vitro. Cell Dev. Biol. 32: 233–240.

    Article  Google Scholar 

  15. Hadi, M.Z., McMullen M.D., and Finer J.J. 1996. Transformation of 12 different plasmids into soybean via particle bombardment. Plant Cell Rep. 15: 500–505.

    Article  Google Scholar 

  16. Chen, L., Zhang, S., Beachy, R.N., and Fauquet C.M. 1998 . A protocol for consistent, large scale production of fertile transgenic rice plants. Plant Cell Reports. In press.

  17. Vickers, J.E., Graham. G.C., and Henry, R.J. 1996. A protocol for the efficient screening of putatively transformed plants for bar, the selectable marker gene, using the polymerase chain reaction. Plant Mol. Biol. Rep . 14: 363–368.

    Article  Google Scholar 

  18. Hamilton, C.M., Frary, A., Lewis, C., and Tanksley, D. 1996 Stable transfer of intact high molecular weight DNA into plant chromosomes. Proc. Natl. Acad. Sci. USA 93: 9975–9979.

    Article  Google Scholar 

  19. Zhang, S., Chen L., Qu, R., Marmey P., Beachy, R.N., and Fauquet, C. 1996 . Regeneration of fertile transgenic Indica (group 1) rice plants following microprojectile transformation of embryogenic suspension culture cells. Plant Cell Rep. 15: 465– 469.

    Article  Google Scholar 

  20. Zhang, S. 1995. Efficient plant regeneration from Indica (group 1) rice protoplasts of one advanced breeding line and three varieties. Plant Cell Rep. 15: 68–71.

    Article  Google Scholar 

  21. Ow, D.W., Wood, K.V., Deluca, M., Wet, J.R.D., Helsinki, D.R., and Howel, S.H. 1986. Transient and stable expression of the firefly luciferase gene in plants cells and transgenic plants. Science 234: 856–859.

    Article  Google Scholar 

  22. Dellaporta, S.L., Wood, J., and Hicks, J.B. 1983. A plant DNA minipreparation: version II. Plant Mol. Biol. Rep. 1: 19– 21.

    Article  Google Scholar 

  23. Sambrook, J., Fritsch, E.F., and Maniatis, T. 1989. Molecular cloning, a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.

    Google Scholar 

  24. Murashige, T. and Skoog, F. 1962. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant. 15: 473–497.

    Article  Google Scholar 

  25. Jefferson, R.A. 1987. Assaying chimeric genes in plants: The GUS gene fusion system. Plant Mol. Biol. Rep. 5: 387–405.

    Article  Google Scholar 

Download references

Acknowledgements

The authors thank S.-J. Leitner and L. Cardenas for greenhouse and culture assistance; C. Bonneau, C. Brugidou, E. Jacquot, and N. Kouassi for production of genetic constructs and S. Welsh for help with luc gene test. C.M.F., P.M., N.J.T., J.-P.B., and A.deK. were supported by the French Institute for Scientific Research for Development through Cooperation (ORSTOM) and R.N.B. by the Scripps Family Chair. Major funding for this work was provided by the Rockefeller Foundation.

Author information

Authors and Affiliations

  1. Volcani Research Institute, Tel Aviv, Israel

    Hervé Huet

Authors
  1. Lili Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Philippe Marmey
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Nigel J. Taylor
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Jean-Paul Brizard
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Celia Espinoza
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Patricia D'Cruz
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Hervé Huet
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Shiping Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Alexandre de Kochko
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Roger N. Beachy
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Claude M. Fauquet
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

International Laboratory for Tropical Agricultural Biotechnology ILTAB, (ORSTOM-TSRI), Division of Plant Biology, BCC 206, The Scripps Research Institute, La Jolla, CA 92037.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Chen, L., Marmey, P., Taylor, N. et al. Expression and inheritance of multiple transgenes in rice plants. Nat Biotechnol 16, 1060–1064 (1998). https://doi.org/10.1038/3511

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1038/3511

This article is cited by

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing