Journal
VIRUS RESEARCH
Volume 197, Issue -, Pages 8-12Publisher
ELSEVIER
DOI: 10.1016/j.virusres.2014.11.016
Keywords
Lentinula edodes; dsRNA; Isogenic strain; Mycovirus
Categories
Funding
- Bio-industry Technology Development Program, Ministry for Food, Agriculture, Forestry and Fisheries, Republic of Korea
- NRF grants from MSIP [2008-0061897, NRF-2013R1A1A2012433]
- Korea Research Council of Fundamental Science & Technology (Joint Degree and Research Center for Biorefinery)
- Cooperative Research Program for Agricultural Science & Technology Development from the Rural Development Administration, Republic of Korea [PJ00999801]
- BK21 PLUS program in the Department of Bioactive Material Sciences
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This study attempted to cure the edible mushroom Lentinula edodes strain FMRI0339 of the L. edodes mycovirus (LeV) in order to obtain an isogenic virus-free fungal strain as well as a virus-infected strain for comparison. Mycelial fragmentation, followed by being spread on a plate with serial dilutions resulted in a virus-free colony. Viral absence was confirmed with gel electrophoresis after dsRNA-specific virus purification, Northern blot analysis, and PCR using reverse transcriptase (RT-PCR). Once cured, all of fungal cultures remained virus-free over the next two years. Interestingly, the viral titer of LeV varied depending on the culture condition. The titer from the plate culture showed at least a 20-fold higher concentration than that grown in the liquid culture. However, the reduced virus titer in the liquid culture was recovered by transferring the mycelia to a plate containing the same medium. In addition, oxygen-depleted culture conditions resulted in a significant decrease of viral concentration, but not to the extent seen in the submerged liquid culture. Although no discernable phenotypic changes in colony morphology were observed, virus-cured strains showed significantly higher growth rates and mycelial mass than virus-infected strains. These results indicate that LeV infection has a deleterious effect on mycelial growth. (C) 2014 Elsevier B.V. All rights reserved.
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