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.

  • Article
  • Published:

Engineering hypervirulence in a mycoherbicidal fungus for efficient weed control

Nature Biotechnology volume 20, pages 1035–1039 (2002)Cite this article

Abstract

Agents proposed for biocontrol of major weeds in arable row-crop agriculture have not met expectations because an evolutionary balance has developed between microorganism and weed, even when the mycoherbicide is used inundatively at very high levels (>104 spores/cm2). Sufficient virulence can be achieved by transferring genes to the microorganism, tipping the evolutionary balance. Virulence was increased ninefold and was more rapidly effected; furthermore, the requirement for a long duration at high humidity was decreased by introducing NEP1 encoding a phytotoxic protein, to an Abutilon theophrasti–specific, weakly mycoherbicidal strain of Colletotrichum coccodes. The parent strain was at best infective on juvenile cotyledons of this intransigent weed. The transgenic strain was lethal through the three-leaf stage, a sufficient time window to control this asynchronously germinating weed. Strategies of coupling virulence genes with fail-safe mechanisms to prevent spread (due to broadened host range) and to mitigate transgene introgression into crop pathogens could be very useful in the biocontrol of major weeds in row crops.

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: The presence of NEP1 and its product in transformants.
Figure 2: Greater efficiency of NEP1 Colletotrichum inoculum in controlling Abutilon theophrasti seedlings treated at the cotyledonary stage of development.
Figure 3: The ability of NEP1 Colletotrichum coccodes to control Abutilon plants successfully through the stage of three true leaves.
Figure 4: Shortening of dew period requirements for successful disease establishment using NEP1 Colletotrichum.

Similar content being viewed by others

Accession codes

Accessions

GenBank/EMBL/DDBJ

References

  1. Boyette, C.D., Quimby, P.C., Connick, W.J., Daigle, D.J. & Fulgham, F.E. Progress in the production, formulation and application of mycoherbicides. in Microbial Control of Weeds (ed. TeBeest, D.O.) 209–222 (Chapman and Hall, New York, 1991).

    Chapter  Google Scholar 

  2. Quimby, P.C., Zidack, N.K., Boyette, C.D. & Grey, W.E. A simple method for stabilizing and granulating fungi. Biocont. Sci. Tech. 9, 5–8 (1999).

    Article  Google Scholar 

  3. Kistler, H.C. Genetic manipulation of plant pathogenic fungi. in Microbial Control of Weeds (ed. TeBeest, D.O.) 151–170 (Chapman and Hall, New York, 1991).

    Google Scholar 

  4. Gressel, J. Molecular Biology of Weed Control (Taylor and Francis, London, 2002).

    Google Scholar 

  5. Gressel, J. Potential failsafe mechanisms against the spread and introgression of transgenic hypervirulent biocontrol fungi. Trends Biotechnol. 19, 149–154 (2001).

    Article  CAS  Google Scholar 

  6. Cohen, B., Amsellem, Z., Maor, R., Sharon, A. & Gressel, J. Transgenically-enhanced expression of indole-3-acetic acid (IAA) confers hypervirulence to plant pathogens. Phytopathology 92, 590–596 (2002).

    Article  CAS  Google Scholar 

  7. Brandl, M.T. & Lindow, S.E. Contribution of indole-3-acetic acid production to the epiphytic fitness of Erwinia herbicola. Appl. Environ. Microbiol. 64, 3256–3263 (1998).

    CAS  PubMed  PubMed Central  Google Scholar 

  8. Manulis, S., Haviv-Chesner, A., Brandl, M.T., Lindow, S.E. & Barash, I. Differential involvement of indole-3-acetic acid biosynthesis pathways in pathogenicity and epiphytic fitness of Erwinia herbicola pv gypsophilae. Mol. Plant–Microbe Interact. 11, 634–642 (1998).

    Article  CAS  Google Scholar 

  9. Gaudin, V., Vrain, T. & Jouanin, L. Bacterial genes modifying hormonal balances in plants. Plant Physiol. Biochem. 32, 11–29 (1994).

    CAS  Google Scholar 

  10. Bailey, B., Jennings, J. & Anderson, J. The 24-kDa protein from Fusarium oxysporum f.sp. erythroxyli: occurrence in related fungi and the effect of growth medium on its production. Can. J. Microbiol. 43, 45–55 (1997).

    Article  CAS  Google Scholar 

  11. Nelson, A.J., Apel-Birkhold, P.C. & Bailey, B.A. Sequence announcements. Plant Mol. Biol. 38, 911–912 (1998).

    Article  Google Scholar 

  12. Jennings, J.C. et al. Induction of defense responses in tobacco by the protein Nep1 from Fusarium oxysporum. Plant Sci. 161, 891–899 (2001).

    Article  CAS  Google Scholar 

  13. Jennings, J.C., Apel-Birkhold, P.C., Bailey, B.A. & Anderson, J.D. Induction of ethylene biosynthesis and necrosis in weed leaves by a Fusarium oxysporum protein. Weed Sci. 48, 7–14 (2000).

    Article  CAS  Google Scholar 

  14. Bailey, B., Collins, R. & Anderson, J. Factors influencing the herbicidal activity of Nep1, a fungal protein that induces the hypersensitive response in Centaurea maculosa. Weed Sci. 48, 776–785 (2000).

    Article  CAS  Google Scholar 

  15. Bailey, B.A. et al. Nep1 protein from Fusarium oxysporum enhances biological control of opium poppy by Pleospora papaveracea. Phytopathology 90, 812–818 (2000).

    Article  CAS  Google Scholar 

  16. Bailey, B.A., Apel-Birkhold, P.C. & Luster, D.G. Expression of NEP1 by Fusarium oxysporum f. sp. erythroxyli after gene replacement and overexpression using PEG-mediated transformation. Phytopathology 92, 833–841 (2002).

    Article  CAS  Google Scholar 

  17. Wymore, L.A., Poirier, C., Watson, A.K. & Gotleib, A.R. Colletotrichum coccodes, a potential bioherbicide for control of velvetleaf (Abutilon theophrasti). Plant Dis. 72 534–538 (1988).

    Article  Google Scholar 

  18. Vurro, M. Microbial toxins in biocontrol enhancement strategies. in Enhancing Biocontrol Agents and Handling Risks (eds Vurro, M. et al.) 28–37 (IOS Press, Amsterdam, 2001).

    Google Scholar 

  19. Quimby, P.C. et al. Enhancing biological control through superior formulations: a worthy goal, but still a work in progress. in Enhancing Biocontrol Agents and Handling Risks (eds Vurro, M. et al.) 86–95 (IOS Press, Amsterdam, 2001).

    Google Scholar 

  20. Perfect, S.E., Hughes, H.B., O'Connell, R.J. & Green, J.R. Colletotrichum—a model genus for studies on pathology and fungal–plant interactions. Fungal Genet. Biol. 27, 186–198 (1999).

    Article  CAS  Google Scholar 

  21. Amsellem, Z., Zidack, N.K., Quimby, P.C. & Gressel, J. Long-term dry preservation of active mycelia of two mycoherbicidal organisms. Crop Protect. 18, 643–649 (1999).

    Article  Google Scholar 

  22. Sands, D.C. & Miller, R.V. Altering the host range of mycoherbicides by genetic manipulation. in Pest Control with Enhanced Environmental Safety (eds Duke, S.O., Menn, J.J. & Plimmer, J.R.) 101–109 (American Chemical Society, Washington, DC, 1993).

    Chapter  Google Scholar 

  23. Sands, D.C. & Pilgeram, A. Enhancing the efficacy of biocontrol agents against weeds. in Enhancing Biocontrol Agents and Handling Risks (eds Vurro, M. et al.) 3–13 (IOS Press, Amsterdam, 2001).

    Google Scholar 

  24. DiTommaso, A. & Watson, A.K. Effect of the fungal pathogen, Colletotrichum coccodes, on Abutilon theophrasti height hierarchy development. J. Appl. Ecol. 34, 518–529 (1997).

    Article  Google Scholar 

  25. Watson, A.K., Gotlieb, A.R. & Wymore, L.A. Interactions between a mycoherbicide Colletotrichum coccodes, and herbicides for the control of velvetleaf (Abutilon theophrasti Medic.). Weed Sci. Soc. Amer. Abstracts 26, 143 (1986).

  26. Wymore, L.A. & Watson, A.K. Interaction between a velvetleaf isolate of Colletotricum coccodes and thidiazuron for velvetleaf (Abutilon theophrasti) control in the field. Weed Sci. 37, 478–483 (1989).

    CAS  Google Scholar 

  27. Khan, R. & Straney, D.C. Regulatory signals influencing expression of the PDA1 gene of Nectria haematococca MPVI in culture and during pathogenesis of pea. Mol. Plant–Microbe Interact. 12, 733–742 (1999).

    Article  CAS  Google Scholar 

  28. Jennings, J.C., Apel-Birkhold, P.C., Bailey, B.A. & Anderson, J.D. Induction of ethylene biosynthesis and necrosis in weed leaves by a Fusarium oxysporum protein. Weed Sci. 48, 7–14 (2000).

    Article  CAS  Google Scholar 

  29. Turgeon, B.G., Garber, R.C. & Yoder, O.C. Development of a fungal transformation system based on selection of sequences with promoter activity. Mol. Cell Biol. 7, 3297–3305 (1987).

    Article  CAS  Google Scholar 

  30. Daboussi, M.J. et al. Transformation of seven species of filamentous fungi using the nitrate reductase gene of Aspergillus nidulans. Curr. Genet. 15, 453–456 (1989).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

The assistance of Hila Elifantz is gratefully acknowledged. Bryan Bailey (USDA, Beltsville MD) provided the plasmid containing the NEP1 gene and the Nep1 antiserum as well as a considerable amount of useful unpublished background information. Alan K. Watson, McGill University, kindly provided the untransformed organism and unpublished information on host specificity. This research was supported by a DFG trilateral Israel-German-Palestinian project. Jonathan Gressel holds the Gilbert de Botton Chair of Plant Sciences.

Author information

Author notes

  1. Barry A. Cohen

    Present address: CBD Technologies, Ltd., Park Tamar, Rehovot, Israel

Authors and Affiliations

  1. Plant Sciences, Weizmann Institute of Science, Rehovot, 76100, Israel

    Ziva Amsellem, Barry A. Cohen & Jonathan Gressel

Authors
  1. Ziva Amsellem
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Barry A. Cohen
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Jonathan Gressel
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Jonathan Gressel.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Amsellem, Z., Cohen, B. & Gressel, J. Engineering hypervirulence in a mycoherbicidal fungus for efficient weed control. Nat Biotechnol 20, 1035–1039 (2002). https://doi.org/10.1038/nbt743

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nbt743

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