Isolation and Characterization of a Mycovirus in Lentinula edodes

Hyo-Kyoung Won; So-Jung Park; Dong-Kyu Kim; Myeung Ju Shin; Nari Kim; Song-Hee Lee; Young-Chul Kwon; Han Kyu Ko; Hyeon-Su Ro; Hyun-Sook Lee

doi:10.1007/s12275-013-2351-2

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Research Support, Non-U.S. Gov't
Isolation and Characterization of a Mycovirus in Lentinula edodes: Hyo-Kyoung Won ¹, So-Jung Park ¹, Dong-Kyu Kim ¹, Myeung Ju Shin ¹, Nari Kim ¹, Song-Hee Lee ¹, Young-Chul Kwon ¹, Han Kyu Ko ², Hyeon-Su Ro ¹, Hyun-Sook Lee ¹; Journal of Microbiology 2013;51(1):118-122.
DOI: https://doi.org/10.1007/s12275-013-2351-2
Published online: March 2, 2013

¹Department of Microbiology and Research Institute of Life Sciences, Gyeongsang National University, Jinju 660-701, Republic of Korea, ²Forest Mushroom Research Institute, Gyeonggi 469-803, Republic of Korea

Corresponding author: Hyeon-Su Ro , Tel: +82-55-772-1321,
Hyun-Sook Lee , Tel: +82-55-772-1321,

Received: 9 July 2012 • Accepted: 12 November 2012

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Abstract

A mycovirus was isolated from an edible mushroom, Lentinula edodes, that was suffering from a severe epidemic. Fractionation of the diseased cell extract by isopycnic centrifugation with 50% CsCl revealed that the diseased mushroom was infected by Lentinula edodes spherical virus (LeSV), a new spherical virus with a diameter of 55 nm. The particle of LeSV encapsidated the 12 kb RNA genome by a 120 kDa coat protein. BLAST analysis of the partially sequenced LeSV genome showed 95% sequence identity with a putative RNAdependent RNA polymerase (RdRp) gene of the mycovirus HKB, which was previously reported as being a doublestranded RNA (dsRNA) element. In contrast to HKB, the RNA genome in LeSV is encapsidated by the 120 kDa coat protein. To confirm that the LeSV coat protein is encoded by the viral genome, the N-terminal amino acid sequence of the coat protein was determined. The resulting N-terminal amino acid sequence, N-SALDVAPVVPELYFXXLEV-C, was found to be located in the middle of the HKB ORF1, suggesting that the LeSV coat protein was indeed encoded by the virus. To detect LeSV in L. edodes, a primer set targeting the RdRp gene was designed based on the partial sequence of the LeSV genome. RT-PCR analysis showed that 56 of the 84 commercially available dikaryotic cultivars carry LeSV. The transmission pattern of the virus was determined by analysing basidiospores from LeSV-infected and LeSVfree fruiting bodies. Nine out of 10 basidiospores from the LeSV-infected cultivars contained the virus while the spores from the LeSV-free parent were free of LeSV, suggesting that vertical transmission is the primary mode of LeSV propagation.

Keywords: detection;HKB;LeSV;mushroom;mycovirus

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Isolation and Characterization of a Mycovirus in Lentinula edodes

J. Microbiol. 2013;51(1):118-122. Published online March 2, 2013

DOI: https://doi.org/10.1007/s12275-013-2351-2

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