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Japanese star anise ringspot-associated virus is a distinct emaravirus transmitted by the eriophyid mite (the family Diptilomiopidae)

  • Viral and Viroid Diseases
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Abstract

Japanese star anise (Illicium anisatum L., JSA) is seriously damaged by a ringspot disease in Japan. Herein, to determine the causal agent using high-throughput sequencing, we discovered viral RNAs associated with JSA ringspot disease. We then determined the complete or near-complete nucleotide sequences of these RNAs using Sanger sequencing and RACE. The complementary strand of viral RNAs 1, 2, 3, 4, and 5 encoded a single protein, which shared sequence identity with P1 (RNA-dependent RNA polymerase), P2 (glycoprotein precursor), P3 (nucleocapsid protein), P4 (movement protein), and a protein with unknown function of emaraviruses (genus Emaravirus), respectively; however, the highest amino acid sequence identity for the P1–P5 proteins between JSARaV and known emaraviruses was 41.9%, 30.0%, 30.1%, 52.2%, and 38.0%, respectively, all of which were lower than the species demarcation criterion. Furthermore, RNA segments harbored conserved 12-nt terminal sequences at the 5′- and 3′-termini, and a high complementarity of approximately 20 nt in 5′- and 3′-terminal sequences. Transmission electron microscopy confirmed the presence of virus-like particles. JSA ringspot disease was found to be transmitted by an eriophyid mite (subclass Acari, superfamily Eriophyoidea) that belongs to the family Diptilomiopidae. Taken together, these results identified the virus responsible for the ringspot disease of JSA as a new member of the genus, Emaravirus, which we named as the Japanese star anise ringspot-associated virus (JSARaV). Moreover, this is the first report noting that eriophyid mites of the family Diptilomiopidae are capable of transmitting emaravirus.

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Acknowledgements

This work was supported by the Fund for Objective Basic Research provided by CARC/NARO to K.K. and H.Y.

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Correspondence to Yoshifumi Shimomoto.

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10327_2021_1038_MOESM1_ESM.docx

Supplementary file1. Supplementary Fig. S1 Detection of five RNA segments of Japanese star anise ringspot-associated virus (JSARaV) from symptomatic Japanese star anise leaves by RT-PCR. RT-PCR primers JSA1 and JSA2 were used to detect JSARaV RNA1, JSARaV-R2-1800fw, and JSARaV-R2-2247rv to detect RNA2, JSARaV-R3-1200fw, and JSARaV-R3-1689rv to detect RNA3, JSARaV-R4-1200fw, and JSARaV-R4-1500rv to detect RNA4, and JSARaV-R5-600fw and JSARaV-R5-900rv to detect RNA5 (Supplementary Table S1). M, 1 kb Plus DNA ladder (New England Biolabs, Ipswich, MA, USA). 1–6, Symptomatic leaves collected from different fields in Kochi and Shizuoka Prefectures, Japan; 7–9, asymptomatic leaves from different fields in Kochi Prefecture. (DOCX 335 kb)

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Shimomoto, Y., Okada, T., Ikeda, K. et al. Japanese star anise ringspot-associated virus is a distinct emaravirus transmitted by the eriophyid mite (the family Diptilomiopidae). J Gen Plant Pathol 88, 69–80 (2022). https://doi.org/10.1007/s10327-021-01038-1

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