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Production of siRNA targeted against TYLCV coat protein transcripts leads to silencing of its expression and resistance to the virus

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Abstract

The coat protein (CP) of Tomato yellow leaf curl virus (TYLCV), encoded by the v1 gene, is the only known component of the viral capsid. In addition, the CP plays a role in the virus transport into the host cell nucleus where viral genes are replicated and transcribed. In this study, we analyzed the effect of small interfering double-stranded RNAs (siRNAs), derived from an intron-hairpin RNA (ihpRNA) construct and targeting the v1 gene product, on CP accumulation. Transient assays involving agroinfiltration of the CP-silencing construct followed by infiltration of a fused GFP-CP (green fluorescent protein-coat protein) gene showed down-regulation of GFP expression in Nicotiana benthamiana. Some of the transgenic tomato plants (cv. Micro-Tom), expressing the siRNA targeted against the TYLCV CP gene, did not show disease symptoms 7 weeks post-inoculation with the virus, while non-transgenic control plants were infected within 2 weeks post inoculation. The present study demonstrates, for the first time, that siRNA targeted against the CP of TYLCV can confer resistance to the virus in transgenic tomato plants, thereby enabling flowering and fruit production.

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Acknowledgements

We thank Eduard Belausov for his excellent technical assistance with the confocal microscope. This work was supported by the U.S.-Israel Binational Agricultural Research and Development Fund (BARD) grant IS-3347-02 to Y. G., the Chief Scientist of the Ministry of Agriculture grant 261-0416 and the Israel Science Foundation grant 998/04 to Y.G. and A.L. We also wish to thank the Israeli Foreign Ministry for a MASHAV Fellowship awarded to P.K. This paper is a contribution from the Agricultural Research Organization, The Volcani Center, Bet Dagan, Israel, No. 17/06.

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Authors and Affiliations

  1. Department of Genetics, Agricultural Research Organization, The Volcani Center, P.O. Box 6, Bet Dagan, 50250, Israel

    Avi Zrachya, Yael Levy, Moshe Lapidot & Yedidya Gafni

  2. Department of Plant Biotechnology, School of Biotechnology, Madurai Kamaraj University, Madurai, 625021, India

    Pravin P. Kumar & Usha Ramakrishnan

  3. Department of Biological Chemistry, The Alexander Silberman Institute of Life Sciences, The Hebrew University of Jerusalem, Jerusalem, 91904, Israel

    Avi Zrachya & Abraham Loyter

  4. Department of Ornamental Horticulture, The Volcani Center, P.O. Box 6, Bet Dagan, 50250, Israel

    Tzahi Arazi

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  1. Avi Zrachya
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  2. Pravin P. Kumar
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  3. Usha Ramakrishnan
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  4. Yael Levy
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  8. Yedidya Gafni
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Correspondence to Yedidya Gafni.

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Zrachya, A., Kumar, P.P., Ramakrishnan, U. et al. Production of siRNA targeted against TYLCV coat protein transcripts leads to silencing of its expression and resistance to the virus. Transgenic Res 16, 385–398 (2007). https://doi.org/10.1007/s11248-006-9042-2

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