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:

Increased baculovirus susceptibility of armyworm larvae feeding on transgenic rice plants expressing an entomopoxvirus gene

Abstract

We have introduced an entomopoxvirus gene encoding a virus enhancing factor (EF) into rice, which resulted in high-level accumulation of the EF in the transgenic plants. The introduced gene was stably inherited in the progeny of the primary transformants, as shown by analysis of their genomic DNA. Bioassays for insect susceptibility to baculovirus infection showed that armyworm larvae feeding on the transgenic rice had increased susceptibility to a Nucleopolyhedrovirus. Thus, introduction of the EF gene into plants can be used as a strategy to increase the effectiveness of baculoviruses in insect pest management.

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

Access options

Buy this article

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

Figure 1: Diagram of expression vector pEF120 used for rice transformation.
Figure 2: Analysis of transgene expression.

Similar content being viewed by others

References

  1. Miller, L.K. & Lu, A. in The baculoviruses. (ed. Miller, L.K.) 217–235 (Plenum, New York; 1997).

    Google Scholar 

  2. Murphy F.A. (ed.) Virus taxonomy (Springer-Verlag, Berlin; 1995), pp. 104–113.

    Google Scholar 

  3. Hunter-Fujita, F.R., Entwistle, P.F., Evance, H.F. & Crook, N.E. Insect viruses and pest management 620, (John Wiley & Sons,C hichester, UK; 1998).

    Google Scholar 

  4. Blak, B.C., Brennan, A., Dierks, P.M., & Gard, I.E. in The baculoviruses. (ed. Miller, L.K.) 341–387 (Plenum, New York; 1997).

    Book  Google Scholar 

  5. Dougherty, E.M., Guthrie, K.P. & Shapiro, M. Optical brighteners provide baculovirus activity enhancement and UV radiation protection. Biol. Control 7, 71–74 (1996).

    Article  Google Scholar 

  6. Shapiro, M., Robertson, J.L., & Weep, R.E. 1994.Effect of neem extract upon the gypsy moth (Lepidoptera:Lymantriidae) and its nuclear polyhedrosis virus. J. Econ. Entomol. 87, 356–360.

    Article  Google Scholar 

  7. Tanada, Y. A synopsis of studies on the synergistic property of an insect baculovirus: a tribute to Edward A. Steinhaus. J. Invertebr. Pathol. 45, 125–138 (1985).

    Article  Google Scholar 

  8. Xu, J. & Hukuhara, T. Enhanced infection of a nuclear polyhedrosis virus in larvae of the armyworm, Pseudaletia separata, by a factor in the spheroids of an entomopoxvirus. J. Invertebr. Pathol. 60, 259–264 (1992).

    Article  Google Scholar 

  9. Xu, J. & Hukuhara, T. Biochemical properties of an enhancing factor of an entomopoxvirus. J. Invertebr. Pathol. 63, 14–18 (1994).

    Article  CAS  Google Scholar 

  10. Hayakawa, T., Xu, J. & Hukuhara, T. Cloning and sequencing of the gene for an enhancing factor from Pseudaletia separata entomopoxvirus. Gene 177, 269–270 (1996).

    Article  CAS  Google Scholar 

  11. Pfadt, R.E. in Fundamentals of applied entomology (ed. Pfadt, R.E.) 247–281 (Macmillan, New York; 1985).

    Google Scholar 

  12. Hayakawa, T. Genetically modified rice plants and their risk assessment prior to environmental release. Recent Research Developments in Agriculture and Biological Chemistry 1, 51–58 (1997).

    Google Scholar 

  13. Tomita, M., Takahashi, Y., Hukuhara, T. & Miyajima, S. Serological relationship between inclusion body proteins and a virus enhancing factor of an entomopoxvirus, Appl. Entomol. Zool. 33, 277–280 (1998).

    Article  CAS  Google Scholar 

  14. Fujimoto, H., Itoh, K., Yamamoto, M., Kyozuka, J. & Shimamoto, K. Insect resistant rice generated by introduction of a modified δ-endotoxin gene of Bacillus thuringiensis. Bio/Technology 11, 1151–1155 (1993).

    CAS  PubMed  Google Scholar 

  15. Hayakawa, T. et al. Genetically engineered rice resistant to rice stripe virus, an insect-transmitted virus. Proc. Natl. Acad. Sci. USA 89, 9865–9869 (1992).

    Article  CAS  Google Scholar 

  16. Wijonarko, A. & Hukuhara, T. Detection of a virus enhancing factor in the spheroid, spindle, and virion of an entomopoxvirus. J. Invertebr. Pathol. 72, 82–86 (1998).

    Article  CAS  Google Scholar 

  17. Mitsuhashi, W., Furuta, Y. & Sato, M. The spindles of an entomopoxvirus of Coleoptera (Anomala cuprea) strongly enhance the infectivity of a nucleopolyhedrovirus in Lepidoptera (Bombyx mori). J. Invertebr. Pathol. 71, 186–188 (1998).

    Article  CAS  Google Scholar 

  18. Jaques, R.P. in Viral insecticides for biological control. (eds Maramorosch, K. & Sherman, K.E.) 285–360 (Academic Press, New York; 1985).

    Book  Google Scholar 

  19. Kalmakoff, J. & Crawford, A.M. in Microbial and viral pesticides (ed. Kurstak, E.). 435–448 (Marcel Dekker, New York; 1982).

    Google Scholar 

  20. Ohta, S., Mita, S., Hattori, T. & Nakamura, K. Construction and expression in tobacco of β-glucuronidase (GUS) reporter gene containing the intron within the coding sequence. Plant Cell Physiol. 31, 805–813 (1990).

    CAS  Google Scholar 

  21. Zhu, Y., Hayakawa, T., Toriyama, S. & Takahashi, M. Complete nucleotide sequence of RNA 3 of rice stripe virus: an ambisense coding strategy. J. Gen. Virol. 72, 763–767 (1991).

    Article  CAS  Google Scholar 

  22. Kyozuka, J., Hayashi, Y. & Shimamoto, K. High frequency plant regeneration from rice protoplasts by novel nurse culture methods. Mol. Gen. Genet. 206, 408–413 (1987).

    Article  CAS  Google Scholar 

  23. Shimamoto, K., Terada, R., Izawa, T. & Fujimoto, H. Fertile transgenic rice plants regenerated from transformed protoplasts. Nature 338, 274–276 (1989).

    Article  CAS  Google Scholar 

  24. Baba, A., Hasezawa, S. & Shono, K. Cultivation of rice protoplasts and their transformation mediated by Agrobacterium spheroplasts. Plant Cell Physiol. 27, 463–472 (1986).

    CAS  Google Scholar 

  25. Ohira, K., Ojima, K. & Fujiwara, A. Studies on the nutrition of rice cell culture: A simple, defined medium for rapid growth in suspension culture. Plant Cell Physiol. 14, 1113–1121 (1973).

    CAS  Google Scholar 

  26. Richards, E. in Current protocols in molecular biology (eds. Ausubel, F.M. et al. ). 2.3.1–2.3.3. (Wiley, New York; 1987).

    Google Scholar 

  27. Chomczynski, P. & Sacchi, N. Single step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal. Biochem. 162,156–159 (1987).

    Article  CAS  Google Scholar 

  28. Bradford, M.M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72, 248–254 (1976).

    Article  CAS  Google Scholar 

  29. Tanada, Y., Hukuhara, T. & Chang, G.Y. A strain of nuclear-polyhedrosis virus causing extensive cellular hypertrophy. J. Invertebr. Pathol. 13, 394–409 (1969).

    Article  Google Scholar 

  30. Yamamoto, T. & Tanada, Y. Biochemical properties of viral envelopes of insect baculoviruses and their role in infectivity. J. Invertebr. Pathol. 31, 48–56 (1978).

    Article  CAS  Google Scholar 

  31. Hukuhara, T., Yano, K., Xu, J., Tomita, M. & Miyajima, S. Detection of a virus enhancing factor in inclusion bodies of an entomopoxvirus by immunoelectron microscopy. J. Invertebr. Pathol. 65, 315–317 (1995).

    Article  Google Scholar 

  32. Berkeson, J. Estimate of the integrated normal curve by minimum normit chi-square with particular reference to bioassay. J. Am. Statist. Assoc. 50, 529–549 (1955).

    Article  Google Scholar 

Download references

Acknowledgements

We thank Mr. Y. Hayashi at Plantech Research Institute for his cooperation in transformation experiments. This research was supported in part by a grant from the Agriculture, Forestry and Fisheries Technical Information Society, Japan, and the Grant-in-Aid for Scientific Research from the Ministry of Education, Japan.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Tosihiko Hukuhara.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Hukuhara, T., Hayakawa, T. & Wijonarko, A. Increased baculovirus susceptibility of armyworm larvae feeding on transgenic rice plants expressing an entomopoxvirus gene. Nat Biotechnol 17, 1122–1124 (1999). https://doi.org/10.1038/15110

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1038/15110

This article is cited by

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