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The use of cysteine proteinase inhibitors to engineer resistance against potyviruses in transgenic tobacco plants

Nature Biotechnology volume 17, pages 1223–1226 (1999)Cite this article

Abstract

As the processing mechanism of all known potyviruses involves the activity of cysteine proteinases, we asked whether constitutive expression of a rice cysteine proteinase inhibitor gene could induce resistance against two important potyviruses, tobacco etch virus (TEV) and potato virus Y (PVY), in transgenic tobacco plants. Tobacco lines expressing the foreign gene at varying levels were examined for resistance against TEV and PVY infection. There was a clear, direct correlation between the level of oryzacystatin message, inhibition of papain (a cysteine proteinase), and resistance to TEV and PVY in all lines tested. The inhibitor was ineffective against tobacco mosaic virus (TMV) infection because processing of this virus does not involve cysteine proteinases. These results show that plant cystatins can be used against different potyviruses and potentially also against other viruses, whose replication involves cysteine proteinase activity.

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Figure 1: Southern and northern blot analysis of transgenic plants.
Figure 2: Histogram showing the papain inhibition of crude extracts of transgenic plants.
Figure 3: Virus–induced symptoms in transgenic and control tobacco plants.

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Acknowledgements

The authors thank Professor S. Arai for providing us with the oryzacystatin I clone and Drs. Milton Zaitlin, Charles J. Arntzen, Robert R. Granados, and Kenneth Palmer for kindly reviewing the manuscript. We also thank Dr. Laura Silva R. and Roger. N. Beachy for providing us with TEV–infected tobacco leaves and TMV, respectively, and Norma Martinez G. for help with the papain inhibition assays.

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  1. Departamento de Química, Centro Básico Universidad Autónoma de Aguascalientes, Ave. Universidad 940, Aguascalientes, C.P. 20 100, Ags., México

    Rafael Gutierrez-Campos & Juan Antonio Torres-Acosta

  2. Departamento de Ingeniería Genética Centro de Investigación y Estudios Avanzados, Unidad Irapuato, Apartado Postal 629, Irapuato, Mexico

    Luis Jorge Saucedo-Arias & Miguel Angel Gomez-Lim

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  1. Rafael Gutierrez-Campos
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Correspondence to Miguel Angel Gomez-Lim.

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Gutierrez-Campos, R., Torres-Acosta, J., Saucedo-Arias, L. et al. The use of cysteine proteinase inhibitors to engineer resistance against potyviruses in transgenic tobacco plants. Nat Biotechnol 17, 1223–1226 (1999). https://doi.org/10.1038/70781

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