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Characterization of virus-induced gene silencing in tobacco plants infected with apple latent spherical virus

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Summary

Apple latent spherical virus (ALSV) expressing green fluorescent protein (GFP-ALSV) was used for analysis of virus-induced gene silencing (VIGS) in tobacco plants expressing GFP (GFP-tobacco). In GFP-tobacco inoculated with GFP-ALSV, small dark spots appeared on inoculated leaves at 5 days post-inoculation (dpi), then expanded, and finally covered the whole area of the leaves after 12 dpi. Most of the fluorescence of upper leaves above the 12th true leaf disappeared at 21 dpi. Thus, GFP-ALSV infection efficiently triggered VIGS of a transgene (GFP gene) in tobacco plants. Analysis of GFP-silenced leaves showed that viral RNAs and proteins accumulated in all leaves where most GFP mRNA had been degraded. The siRNAs derived from ALSV-RNAs were not detected in samples from which siRNA of GFP mRNA could be easily detected. Direct tissue blot analysis showed that the spread of GFP-ALSV always preceded the induction of VIGS in infected leaves of GFP-tobacco. GFP leaf patch tests using Nicotiana benthamiana line 16c showed that Vp20, one of the three capsid proteins, is a silencing suppressor which interferes with systemic silencing.

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Authors and Affiliations

  1. The United Graduate School of Agricultural Sciences, Iwate University, Iwate, Morioka, Japan

    H. Yaegashi, M. Isogai & N. Yoshikawa

  2. Plant Pathology Laboratory, Faculty of Agriculture, Iwate University, Iwate, Morioka, Japan

    T. Yamatsuta, M. Isogai & N. Yoshikawa

  3. The 21st Century Center of Excellence Program, Iwate University, Iwate, Morioka, Japan

    T. Takahashi & N. Yoshikawa

  4. The School of Life Sciences, Xiamen University, Xiamen, China

    C. Li

  5. Kyoto Prefectural Institute of Agricultural Biotechnology, Soraku-gun, Kyoto, Japan

    T. Kobori

  6. Graduate School of Life and Environmental Sciences, University of Osaka Prefecture, Sakai, Japan

    S. Ohki

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  1. H. Yaegashi
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  2. T. Yamatsuta
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  4. C. Li
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  6. T. Kobori
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  8. N. Yoshikawa
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Yaegashi, H., Yamatsuta, T., Takahashi, T. et al. Characterization of virus-induced gene silencing in tobacco plants infected with apple latent spherical virus. Arch Virol 152, 1839–1849 (2007). https://doi.org/10.1007/s00705-007-1011-4

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  • DOI: https://doi.org/10.1007/s00705-007-1011-4

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