Curing viruses in Pleurotus ostreatus by growth on a limited nutrient medium containing cAMP and rifamycin

Kwon, Young Ho; Jeong, Dong-Kyu; Gim, Sang-Ah; Ro, Hyeon Su; Lee, H.S.

doi:10.1016/j.jviromet.2012.06.002

Cited by 14 publications

(13 citation statements)

References 16 publications

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“…Fruiting bodies of the healthy-looking P. ostreatus strains show typical phenotypes of oyster mushroom, such as a long and thick stipe, and a middle slate-gray pileus with a low funnel-shape [ 6 16 ]. Abnormal mycelial cells were obtained from the malformed fruiting body of P. ostreatus ASI2792 strain, comparing with a normal fruiting body, and were maintained on PDA with periodic transfer as described in Materials and Methods.…”

Section: Resultsmentioning

confidence: 99%

“…Abnormal mycelial cells were obtained from the malformed fruiting body of P. ostreatus ASI2792 strain, comparing with a normal fruiting body, and were maintained on PDA with periodic transfer as described in Materials and Methods. The symptoms of mycoviruses infection in P. ostreatus are reduced mycelial growth, fruiting body abnormalities, and decreased fruiting body yield [ 9 10 11 12 16 ]. For that reason, a dsRNA purified with CC41 was visualized by 1.0% agarose gel electrophoresis and was confirmed to be resistant to DNase and RNase A at a high-salt concentration, but disappeared upon exposure to dsRNA-specific RNase A at low-salt concentrations to verify the presence of about 8.0-kb dsRNA fragment (data not shown).…”

Section: Resultsmentioning

confidence: 99%

“…Although curing methods of virus-infected fungi have been attempted using cycloheximide or ribavirin treatment, hyphal tip cut, single spore or other subculture, protoplast regeneration, or incubation with temperature stress, these methods have not always resulted in mycovirus curing. Further, these stresses do damage to fungal growth as circumstances require, thus, these virus-cured strains are unsuitable for the comparison group to research on fungus-mycovirus interactions [ 13 14 15 16 17 ]. The appropriate virus-curing method was previously developed to research fungal symptoms in response to viral infection using the mycelial fragmentation method followed by single colony isolation [ 18 19 ].…”

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Viral Effects of a dsRNA Mycovirus (PoV-ASI2792) on the Vegetative Growth of the Edible Mushroom Pleurotus ostreatus

et al. 2016

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A double-stranded RNA (dsRNA) mycovirus was detected in malformed fruiting bodies of Pleurotus ostreatus strain ASI2792, one of bottle cultivated commercial strains of the edible oyster mushroom. The partial RNA-dependent RNA polymerase (RdRp) gene of the P. ostreatus ASI2792 mycovirus (PoV-ASI2792) was cloned, and a cDNA sequences alignment revealed that the sequence was identical to the RdRp gene of a known PoSV found in the P. ostreatus strain. To investigate the symptoms of PoV-ASI2792 infection by comparing the isogenic virus-free P. ostreatus strains with a virus-infected strain, isogenic virus-cured P. ostreatus strains were obtained by the mycelial fragmentation method for virus curing. The absence of virus was verified with gel electrophoresis after dsRNA-specific virus purification and Northern blot analysis using a partial RdRp cDNA of PoV-ASI2792. The growth rate and mycelial dry weight of virus-infected P. ostreatus strain with PoV-ASI2792 mycovirus were compared to those of three virus-free isogenic strains on 10 different media. The virus-cured strains showed distinctly higher mycelial growth rates and dry weights on all kinds of experimental culture media, with at least a 2.2-fold higher mycelial growth rate on mushroom complete media (MCM) and Hamada media, and a 2.7-fold higher mycelial dry weight on MCM and yeastmalt-glucose agar media than those of the virus-infected strain. These results suggest that the infection of PoV mycovirus has a deleterious effect on the vegetative growth of P. ostreatus.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 99%

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Viral Effects of a dsRNA Mycovirus (PoV-ASI2792) on the Vegetative Growth of the Edible Mushroom Pleurotus ostreatus

et al. 2016

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“…Firstly, viruses could be introduced into virus-free strains by using the techniques like pairing-culture [ 3 ], virion transfection [ 21 ], and construction of infectious cDNA clones [ 24 ]. On the other hand, some investigations, including sequential hyphal tip isolation [ 25 ], protoplasts/small mycelial fragments regeneration [ 26 ], treatment of cycloheximide [ 27 , 28 ], or cAMP-rifamycin [ 29 ], and single spore isolation [ 20 ] were also explored to cure the viruses in their original strains. Therefore, similar experiments will also be carried out to elucidate the role of BcMyV1 on B. cinerea biology in the future.…”

Section: Discussionmentioning

confidence: 99%

Molecular Characterization and Geographic Distribution of a Mymonavirus in the Population of Botrytis cinerea

Hao

2018

Viruses

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Here, we characterized a negative single-stranded (−ss)RNA mycovirus, Botrytis cinerea mymonavirus 1 (BcMyV1), isolated from the phytopathogenic fungus Botrytis cinerea. The genome of BcMyV1 is 7863 nt in length, possessing three open reading frames (ORF1–3). The ORF1 encodes a large polypeptide containing a conserved mononegaviral RNA-dependent RNA polymerase (RdRp) domain showing homology to the protein L of mymonaviruses, whereas the possible functions of the remaining two ORFs are still unknown. The internal cDNA sequence (10-7829) of BcMyV1 was 97.9% identical to the full-length cDNA sequence of Sclerotinia sclerotiorum negative stranded RNA virus 7 (SsNSRV7), a virus-like contig obtained from Sclerotinia sclerotiorum metatranscriptomes, indicating BcMyV1 should be a strain of SsNSRV7. Phylogenetic analysis based on RdRp domains showed that BcMyV1 was clustered with the viruses in the family Mymonaviridae, suggesting it is a member of Mymonaviridae. BcMyV1 may be widely distributed in regions where B. cinerea occurs in China and even over the world, although it infected only 0.8% of tested B. cinerea strains.

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“…However, studies of mycovirus-host interactions have been insufficient to reveal the mechanisms involved in infection of P. ostreatus because it is difficult to secure definite isogenic virus-free or -cured strains to compare to virus-infected strains 3 . Because different genotypes of the same fungal species can produce inconsistent data with regard to identical mycovirus strains, various approaches have been used to obtain isogenic virus-cured strains, such as cycloheximide or ribavirin treatment, hyphal tip cutting, single-sporing, protoplast regeneration, or thermal stress treatment [11][12][13][14] . Even when the virus is successfully cured from a virus-infected fungal cell, impairments caused by the treatment make the cured fungal cell inappropriate for investigations of fungal-mycoviral interactions.…”

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confidence: 99%

The PoV mycovirus affects extracellular enzyme expression and fruiting body yield in the oyster mushroom, Pleurotus ostreatus

Song

Kim

et al. 2020

Sci Rep

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transcript with at least three independent preparations of the same RNA sample. The cytochrome C gene (JGI gene ID: no. 1113744) or actin gene (JGI gene ID: no. 1087906) of P. ostreatus was used as the reference gene 30. Statistical analyses. Mycelial colony diameter, fruiting body yield, and area and color intensity of the brown oxidation region were analyzed via one-way analysis of variance using SPSS software (version 18; IBM Corporation, Armonk, NY, USA), with p < 0.05 considered to indicate significant differences between constructs. When a significant F-value was obtained, Duncan's multiple range test was used to test the significance of the effect (p < 0.05) between virus-cured and virus-infected strains. Relative gene expression levels obtained using qRT-PCR were analyzed using the same statistical test.

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Curing viruses in Pleurotus ostreatus by growth on a limited nutrient medium containing cAMP and rifamycin

Cited by 14 publications

References 16 publications

Viral Effects of a dsRNA Mycovirus (PoV-ASI2792) on the Vegetative Growth of the Edible Mushroom Pleurotus ostreatus

Viral Effects of a dsRNA Mycovirus (PoV-ASI2792) on the Vegetative Growth of the Edible Mushroom Pleurotus ostreatus

Molecular Characterization and Geographic Distribution of a Mymonavirus in the Population of Botrytis cinerea

The PoV mycovirus affects extracellular enzyme expression and fruiting body yield in the oyster mushroom, Pleurotus ostreatus

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