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      Occurrence of dsRNA Mycovirus (LeV-FMRI0339) in the Edible Mushroom Lentinula edodes and Meiotic Stability of LeV-FMRI0339 among Monokaryotic Progeny

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          Abstract

          dsRNA was found in malformed cultures of Lentinula edodes strain FMRI0339, one of the three most popular sawdust cultivated commercial strains of shiitake, and was also found in healthy-looking fruiting bodies and actively growing mycelia. Cloning of the partial genome of the dsRNA revealed the presence of the RdRp sequence of a novel L. edodes mycovirus (LeV), and sequence comparison of the cloned amplicon showed identical sequences sequence to known RNA-dependent RNA polymerase genes of LeV found in strain HKA. The meiotic stability of dsRNA was examined by measuring the ratio of the presence of dsRNA among sexual monokaryotic progeny. More than 40% of the monokaryotic progeny still contained the dsRNA, indicating the persistence of dsRNA during sexual reproduction. Comparing the mycelia growth of monokaryotic progeny suggested that there appeared to be a tendency toward a lower frequency of virus incidence in actively growing progeny.

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          Mycoviruses of filamentous fungi and their relevance to plant pathology.

          Mycoviruses (fungal viruses) are reviewed with emphasis on plant pathogenic fungi. Based on the presence of virus-like particles and unencapsidated dsRNAs, mycoviruses are common in all major fungal groups. Over 80 mycovirus species have been officially recognized from ten virus families, but a paucity of nucleic acid sequence data makes assignment of many reported mycoviruses difficult. Although most of the particle types recognized to date are isometric, a variety of morphologies have been found and, additionally, many apparently unencapsidated dsRNAs have been reported. Until recently, most characterized mycoviruses have dsRNA genomes, but ssRNA mycoviruses now constitute about one-third of the total. Two hypotheses for the origin of mycoviruses of plant pathogens are discussed: the first that they are of unknown but ancient origin and have coevolved along with their hosts, the second that they have relatively recently moved from a fungal plant host into the fungus. Although mycoviruses are typically readily transmitted through asexual spores, transmission through sexual spores varies with the host fungus. Evidence for natural horizontal transmission has been found. Typically, mycoviruses are apparently symptomless (cryptic) but beneficial effects on the host fungus have been reported. Of more practical interest to plant pathologists are those viruses that confer a hypovirulent phenotype, and the scope for using such viruses as biocontrol agents is reviewed. New tools are being developed based on host genome studies that will help to address the intellectual challenge of understanding the fungal-virus interactions and the practical challenge of manipulating this relationship to develop novel biocontrol agents for important plant pathogens.
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            Origin, Adaptation and Evolutionary Pathways of Fungal Viruses

            Fungal viruses or mycoviruses are widespread in fungi and are believed to be of ancient origin. They have evolved in concert with their hosts and are usually associated with symptomless infections. Mycoviruses are transmitted intracellularly during cell division, sporogenesis and cell fusion, and they lack an extracellular phase to their life cycles. Their natural host ranges are limited to individuals within the same or closely related vegetative compatibility groups. Typically, fungal viruses are isometric particles 25–50 nm in diameter, and possess dsRNA genomes. The best characterized of these belong to the family Totiviridae whose members have simple undivided dsRNA genomes comprised of a coat protein (CP) gene and an RNA dependent RNA polymerase (RDRP) gene. A recently characterized totivirus infecting a filamentous fungus was found to be more closely related to protozoan totiviruses than to yeast totiviruses suggesting these viruses existed prior to the divergence of fungi and protozoa. Although the dsRNA viruses at large are polyphyletic, based on RDRP sequence comparisons, the totiviruses are monophyletic. The theory of a cellular self-replicating mRNA as the origin of totiviruses is attractive because of their apparent ancient origin, the close relationships among their RDRPs, genome simplicity and the ability to use host proteins efficiently. Mycoviruses with bipartite genomes (partitiviruses), like the totiviruses, have simple genomes, but the CP and RDRP genes are on separate dsRNA segments. Because of RDRP sequence similarity, the partitiviruses are probably derived from a totivirus ancestor. The mycoviruses with unencapsidated dsRNA-like genomes (hypoviruses) and those with bacilliform (+) strand RNA genomes (barnaviruses) have more complex genomes and appear to have common ancestry with plant (+) strand RNA viruses in supergroup 1 with potyvirus and sobemovirus lineages, respectively. The La France isometric virus (LIV), an unclassified virus with multipartite dsRNA genome, is associated with a severe die-back disease of the cultivated mushroom. LIV appears to be of recent origin since it differs from its host in codon usage.
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              Hypoviruses and chestnut blight: exploiting viruses to understand and modulate fungal pathogenesis.

              Fungal viruses are considered unconventional because they lack an extracellular route of infection and persistently infect their hosts, often in the absence of apparent symptoms. Because mycoviruses are limited to intracellular modes of transmission, they can be considered as intrinsic fungal genetic elements. Such long-term genetic interactions, even involving apparently asymptomatic mycoviruses, are likely to have an impact on fungal ecology and evolution. One of the clearest examples supporting this view is the phenomenon of hypovirulence (virulence attenuation) observed for strains of the chestnut blight fungus, Cryphonectria parasitica, harboring members of the virus family Hypoviridae. The goal of this chapter is to document recent advances in hypovirus molecular genetics and to provide examples of how that progress is leading to the identification of virus-encoded determinants responsible for altering fungal host phenotype, insights into essential and dispensable elements of hypovirus replication, revelations concerning the role of G-protein signaling in fungal pathogenesis, and new avenues for enhancing biological control potential.
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                Author and article information

                Journal
                Plant Pathol J
                Plant Pathol. J
                PPJ
                The Plant Pathology Journal
                Korean Society of Plant Pathology
                1598-2254
                2093-9280
                December 2013
                : 29
                : 4
                : 460-464
                Affiliations
                [1 ]Department of Bio-Environmental Chemistry, Wonkwang University, Iksan, Chonbuk 570-749, Korea
                [2 ]Department of Molecular Biology, Department of Bioactive Material Sciences, Center for Fungal Pathogenesis, Chonbuk National University, Jeonju, Chonbuk 561-756, Korea
                [3 ]Forest Mushroom Research Institute, Yeoju, Gyeonggi 469-803, Korea
                Author notes
                [* ]Corresponding author. Phone) +82-63-270-3440, FAX) +82-63-270-4312, E-mail) dhkim@ 123456jbnu.ac.kr
                Article
                ppj-29-460
                10.5423/PPJ.NT.03.2013.0037
                4174826
                25288977
                2b4d935f-973a-48c7-b90d-384c0c0e314e
                ©The Korean Society of Plant Pathology

                This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License ( http://creativecommons.org/licenses/by-nc/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

                History
                : 30 March 2013
                : 22 May 2013
                : 30 June 2013
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                dsrna,lentinula edodes,meiotic stability,mycovirus
                dsrna, lentinula edodes, meiotic stability, mycovirus

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