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Estimation of the functions of viral RNA silencing suppressors by apple latent spherical virus vector

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Abstract

Apple latent spherical virus (ALSV) is a latent virus with wide host range of plant species. In the present study, we prepared ALSV vectors expressing RNA silencing suppressors (RSSs) from eight plant viruses: P19 of carnation Italian ring spot virus (tombusvirus), 2b of peanut stunt virus (cucumovirus), NSs of tomato spotted wilt virus (tospovirus), HC-Pro of bean yellow mosaic virus (potyvirus), γb of barley stripe mosaic virus (hordeivirus), P15 of peanut clump virus (pecluvirus), P1 of rice yellow mottle virus (sobemovirus), or P21 of beet yellows virus (closterovirus). These vectors were inoculated to Nicotiana benthamiana to investigate the effects of RSSs on the virulence and accumulation of ALSV. Among the vectors, ALSV expressing NSs (ALSV-NSs) developed severe mosaic symptoms in newly developed leaves followed by plant death. Infection of ALSV-γb induced characteristic concentric ringspot symptoms on leaves, and plants infected with ALSV-HC-Pro showed mosaic and dwarf symptoms. Infection of the other five ALSV vectors did not show symptoms. ELISA and immunoblot assay indicated that virus titer increased in leaves infected with ALSV-NSs, γb, HC-Pro, or P19. RT-qPCR indicated that the amount of ALSV in plants infected with ALSV-NSs was increased by approximately 45 times compared with that of wtALSV without expression of any RSS. When ALSV-P19, NSs, or HC-Pro was inoculated to Cucumis sativus plants, none of these ALSV vectors induced symptoms, but accumulation of ALSV in plants infected with ALSV-NSs was increased, suggesting that functions of RSSs on virulence and accumulation of ALSV depend on host species.

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Acknowledgements

This work was supported by the Programme for Promotion of Basic and Applied Researches for Innovations in Bio-oriented Industry from the Ministry of Agriculture, Forestry, and Fisheries, Japan

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Authors and Affiliations

  1. Faculty of Agriculture, Iwate University, Ueda 3-18-8, Morioka, Iwate, 020-8550, Japan

    Chunjiang Li, Makoto Ito & Nobuyuki Yoshikawa

  2. Agri-Innovation Center, Iwate University, Ueda 3-18-8, Morioka, Iwate, 020-8550, Japan

    Ichiro Kasajima & Nobuyuki Yoshikawa

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  1. Chunjiang Li
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  2. Makoto Ito
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  3. Ichiro Kasajima
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  4. Nobuyuki Yoshikawa
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CL and NY designed the study; CL and MI performed the experiments; IK analyzed the data; CL, IK, and NY drafted the manuscript; and all authors approved the final version.

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Correspondence to Nobuyuki Yoshikawa.

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Li, C., Ito, M., Kasajima, I. et al. Estimation of the functions of viral RNA silencing suppressors by apple latent spherical virus vector. Virus Genes 56, 67–77 (2020). https://doi.org/10.1007/s11262-019-01708-5

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