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

Avi Zrachya; Pravin P. Kumar; Usha Ramakrishnan; Yael Levy; Abraham Loyter; Tzahi Arazi; Moshe Lapidot; Yedidya Gafni

doi:10.1007/s11248-006-9042-2

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

Avi Zrachya, Pravin P. Kumar, Usha Ramakrishnan, Yael Levy, Abraham Loyter, Tzahi Arazi, Moshe Lapidot, Yedidya Gafni

Research output: Contribution to journal › Article › peer-review

72 Scopus citations

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.

Original language	English
Pages (from-to)	385-398
Number of pages	14
Journal	Transgenic Research
Volume	16
Issue number	3
DOIs	https://doi.org/10.1007/s11248-006-9042-2
State	Published - Jun 2007
Externally published	Yes

Bibliographical note

Funding Information:
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.

Funding

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.

Funders	Funder number
Agricultural Research Organization
Israeli Foreign Ministry
The Volcani Center	17/06
United States - Israel Binational Agricultural Research and Development Fund	IS-3347-02
Israel Science Foundation	998/04
Ministry of Agriculture and Rural Development	261-0416
Office of the Chief Scientist, Ministry of Economy

Keywords

Geminiviruses
PTGS
Virus resistance

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10.1007/s11248-006-9042-2

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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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note = "Funding Information: 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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AU - Loyter, Abraham

AU - Arazi, Tzahi

AU - Lapidot, Moshe

AU - Gafni, Yedidya

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

Abstract

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Keywords

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