Development of RT-PCR for rapid detection of ssRNA ambi-like mycovirus in a root rot fungi (Armillaria spp.)

Tonka, Tomáš; Walterová, Lucie; Čurn, Vladislav

doi:10.4149/av_2022_308

Cited by 3 publications

(3 citation statements)

References 4 publications

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“…[ 50 ], and H. parviporum [ 51 ], have also been found to harbor this viral group in recent years. They have a non-segmented RNA genome of 4.5–5.0 kb with an ambisense coding nature that possesses two open reading frames (ORFs) (A and B) on each strand [ 52 ]. Moreover, in the above study, the authors also showed that fungal ambi-like viruses contain viroid-like elements that undergo rolling-circle replication and encode their own viral RdRps.…”

Section: Discussionmentioning

confidence: 99%

Comparative Analysis of Viromes Identified in Multiple Macrofungi

Zhou,

Zhang,

Deng

2024

Viruses

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Macrofungi play important roles in the soil elemental cycle of terrestrial ecosystems. Fungal viruses are common in filamentous fungi, and some of them can affect the growth and development of hosts. However, the composition and evolution of macrofungal viruses are understudied. In this study, ninety strains of Trametes versicolor, Coprinellus micaceus, Amanita strobiliformis, and Trametes hirsuta were collected in China. Four mixed pools were generated by combining equal quantities of total RNA from each strain, according to the fungal species, and then subjected to RNA sequencing. The sequences were assembled, annotated, and then used for phylogenetic analysis. Twenty novel viruses or viral fragments were characterized from the four species of macrofungi. Based on the phylogenetic analysis, most of the viral contigs were classified into ten viral families or orders: Barnaviridae, Benyviridae, Botourmiaviridae, Deltaflexiviridae, Fusariviridae, Hypoviridae, Totiviridae, Mitoviridae, Mymonaviridae, and Bunyavirales. Of these, ambi-like viruses with circular genomes were widely distributed among the studied species. Furthermore, the number and overall abundance of viruses in these four species of macrofungi (Basidiomycota) were found to be much lower than those in broad-host phytopathogenic fungi (Ascomycota: Sclerotinia sclerotiorum, and Botrytis cinerea). By employing metatranscriptomic analysis in this study, for the first time, we demonstrated the presence of multiple mycoviruses in Amanita strobiliformis, Coprinellus micaceus, Trametes hirsute, and Trametes versicolor, significantly contributing to research on mycoviruses in macrofungi.

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Section: Discussionmentioning

confidence: 99%

Comparative Analysis of Viromes Identified in Multiple Macrofungi

Zhou,

Zhang,

Deng

2024

Viruses

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“…The quality of total RNA was assessed by gel electrophoresis in 1.5% agarose gel and total RNA was stored in -80°C. LunaScript ® RT SuperMix Kit (New England Biolabs, UK) was used for synthesis of cDNA as by [45]. The success of cDNA synthesis was verified by amplification of the eukaryotic regiontranslation elongation factor 1-alpha -eEF1A (tefa) and checked by gel electrophoresis in 1.5% agarose gel [46].…”

Section: Retro Transcription (Rt) Pcr and Sanger Sequencingmentioning

confidence: 99%

Molecular Detection and Characterization of Mycoviruses in the Genus <em>Armillaria</em> in the Czech Republic

Walterová,

Botella,

Hejna

et al. 2024

Preprint

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Members of the genus Armillaria are widespread forest pathogens against which effective protection has not yet been developed. Due to their longevity and the creation of large-scale cloning of Armillaria individuals, the use of mycoviruses as biocontrol agents (BCAs) against these pathogens could be an effective alternative. This work describes the screening of double-stranded RNA banding patterns as a first approach for potential virus discovery and the use of stranded total RNA-seq for viral detection and characterization in Armillaria spp. collected in the Czech Republic. A total of five single-stranded RNA viruses were detected in Armillaria ostoyae and A. cepistipes, including the first virus of the family Tymoviridae found infecting a member the genus Armillaria and four viruses belonging to the recently described virus group "ambiviruses" with circular ambisense genome organization. Both hammerhead (HHRz) and hairpin (HpRz) ribozymes were detected in all the ambiviricot sequences. Armillaria viruses were compared by phylogenetic analysis and confirmed their specific host by direct RT-PCR, which showed the occurrence of interspecies transmission.

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“…The RNA was eluted in 30 µL of elution buffer and 1 µL of RNase inhibitor was added. The quality of total RNA was assessed by gel electrophoresis in 1.5% agarose gel and total RNA was stored at −80 • C. LunaScript ® RT SuperMix Kit (New England Biolabs, Hitchin, UK) was used for the synthesis of cDNA, as reported by [45]. The success of cDNA synthesis was verified by amplification of the eukaryotic region translation elongation factor 1-alpha-eEF1A (tefa) and checked by gel electrophoresis in 1.5% agarose gel [46].…”

Section: Retro Transcription Polymerase Chain Reaction (Rt-pcr) and S...mentioning

confidence: 99%

Characterization of Mycoviruses in Armillaria ostoyae and A. cepistipes in the Czech Republic

Walterová,

Botella,

Hejna

et al. 2024

Viruses

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Members of the genus Armillaria are widespread forest pathogens against which effective protection has not yet been developed. Due to their longevity and the creation of large-scale cloning of Armillaria individuals, the use of mycoviruses as biocontrol agents (BCAs) against these pathogens could be an effective alternative. This work describes the detection and characterization of viruses in Armillaria spp. collected in the Czech Republic through the application of stranded total RNA sequencing. A total of five single-stranded RNA viruses were detected in Armillaria ostoyae and A. cepistipes, including viruses of the family Tymoviridae and four viruses belonging to the recently described “ambivirus” group with a circular ambisense genome arrangement. Both hammerhead (HHRz) and hairpin (HpRz) ribozymes were detected in all the ambiviricot sequences. Armillaria viruses were compared through phylogenetic analysis and confirmed their specific host by direct RT-PCR. One virus appears to infect both Armillaria species, suggesting the occurrence of interspecies transmission in nature.

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Development of RT-PCR for rapid detection of ssRNA ambi-like mycovirus in a root rot fungi (Armillaria spp.)

Cited by 3 publications

References 4 publications

Comparative Analysis of Viromes Identified in Multiple Macrofungi

Comparative Analysis of Viromes Identified in Multiple Macrofungi

Molecular Detection and Characterization of Mycoviruses in the Genus <em>Armillaria</em> in the Czech Republic

Characterization of Mycoviruses in Armillaria ostoyae and A. cepistipes in the Czech Republic

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