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Reduced RNA silencing suppressor activity of onion yellow dwarf virus HC-Pro with N-terminal deletion may be complemented in mixed infection with another potyvirus in garlic

  • Viral and Viroid Diseases
  • Published:
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Abstract

We investigated virus infection rates in Japanese garlic plants and found that a potyvirus, onion yellow dwarf virus (OYDV) was always associated with another potyvirus, leek yellow stripe virus (LYSV). When the OYDV HC-Pro genes from those viral isolates were PCR-amplified and sequenced, we found that all the HC-Pros isolated from Japanese garlic lacked a long stretch of the N-terminal amino acids (~ 100 residues) including the KITC motif, suggesting possible loss of viral aphid transmission. Our phylogenetic analysis revealed that HC-Pros were divided into three major groups and that they did not branch from a single common ancestor, suggesting different mechanisms for generation of the short HC-Pros from the intact ones. Because HC-Pro is a well-known RNA silencing suppressor (RSS), we then analyzed the RSS activity of HC-Pro using the conventional agroinfiltration assay with Nicotiana benthamiana and also with onion, which is a natural host of OYDV. The results demonstrated that the short HC-Pro had reduced RSS activity compared to the long HC-Pro while both type HC-Pros were well expressed in the agroinfiltrated tissues. In addition, the tested OYDV HC-Pros exhibited more efficient RSS activity in onion than in N. benthamiana. We presume that OYDV with reduced RSS activity of HC-Pro may take advantage of mixed infection with LYSV to survive.

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Acknowledgements

We are grateful to Dr. Reika Isoda for her helpful advice on phylogenetic analyses.

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Authors and Affiliations

  1. Graduate School of Agriculture, Hokkaido University, Kita-ku, Kita 9 Nish 9, Sapporo, 060-8589, Japan

    Hangil Kim, Nana Aoki, Haruna Takahashi, Naoto Yoshida, Hanako Shimura & Chikara Masuta

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  1. Hangil Kim
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  2. Nana Aoki
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  3. Haruna Takahashi
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  4. Naoto Yoshida
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  5. Hanako Shimura
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  6. Chikara Masuta
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Correspondence to Chikara Masuta.

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10327_2020_926_MOESM1_ESM.pptx

Supplementary file1 Fig. S1. Multiple alignment of the N-terminal region of the onion yellow dwarf virus (OYDV) HC-Pro nucleotide sequences. The alignment was created using MEGA v7. Arrowheads on the left indicate the OYDV HC-Pro genes sequenced in this study. All the OYDV HC-Pro sequences in the NCBI database were included in the analysis (PPTX 652 kb)

10327_2020_926_MOESM2_ESM.pptx

Supplementary file2 Fig. S2. Agroinfiltration assay of tobacco etch virus (TEV) HC-Pro in onion epidermal cells. (a) RSS activity of TEV HC-Pro in cells. Agrobacterium carrying the 35S:GFP construct was used to coinfiltrate three onion bulb scales with the 35S:TEV-HC-Pro construct. Inocula were prepared as described for Fig. 4. Bright field (BF) and GFP (UV) images were taken using an epifluorescence microscope (Leica DMI 6000B) at 3 dpi. (b) Fold change in GFP signals in areas in onion epidermis agroinfiltrated with different HC-Pro isolates. GFP signal intensities were quantified as described for Fig. 4. Means (±SE) among isolates were analyzed for significant differences using Student’s t-test (*P < 0.05) (PPTX 787 kb)

10327_2020_926_MOESM3_ESM.pdf

Supplementary file3 Fig. S3 Prediction of 3D structures of onion yellow dwarf virus (OYDV) HC-Pro proteins. Ab initio structural prediction of OYDV HC-Pro used in this study was done using the I-TASSER v5.1 server (https://zhanglab.ccmb.med.umich.edu/I-TASSER/), and the prediction models were visualized using UCSF chimera program v1.9. The two blue arrowheads in China HC-Pro indicate the start and end sites of the unique long α-helix motif. The schematic structures above each model show a simplified image of the HC-Pro proteins. Dotted boxes represent the deletion in the HC-Pro N-terminal region (PDF 37 kb)

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Kim, H., Aoki, N., Takahashi, H. et al. Reduced RNA silencing suppressor activity of onion yellow dwarf virus HC-Pro with N-terminal deletion may be complemented in mixed infection with another potyvirus in garlic. J Gen Plant Pathol 86, 300–309 (2020). https://doi.org/10.1007/s10327-020-00926-2

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