The identification of a novel Pleurotus ostreatus dsRNA virus and determination of the distribution of viruses in mushroom spores.
Double-stranded RNAs and virus particles were identified in Pleurotus ostreatus strain Shin-Nong in Korea. Isometric virus particles with a diameter of 33 nm were purified, which are similar to other Pleurotus viruses reported previously. This strain contains 5 dsRNAs, 8.0, 2.5, 2.4, 2.0, and 1.8 kb in size. The virus particles contain 2 dsRNAs, designated RNA-1 (2.5 kb), and RNA-2 (2.4 kb) which is a typical pattern of Partitiviridae. A non-encapsidated dsRNA of about 8.0 kb also was identified. Partial cDNA from RNA-1 was cloned, and sequence analysis revealed that this gene codes for RdRp. The comparison of the sequence from partial cDNA clone showed 35% amino acid homology with the C-terminal end of the RdRp gene of Helicobasidum mompa virus and Rosalinia necatrix virus. Specific primers designed from the partial sequences successfully amplified RT-PCR product from the infected mycelium and a single spore culture. We used these primers to determine the pattern of distribution of viruses in spores. Of the 96 different single spore cultures generated from Shin-Nong strain, a specific RT-PCR product was identified in 25 cultures, indicating that about 26% of basidiospores contain viruses.
Kim
Yeo Jin
YJ
Department of Molecular Bioscience, Division of Bioscience and Biotechnology, Kangwon National University, Chuncheon 200-701, Republic of Korea.
Kim
Ji Yeon
JY
Kim
Ji Hye
JH
Yoon
Seon Mee
SM
Yoo
Young-Bok
YB
Yie
Se Won
SW
D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D017931:DNA Primers; D018076:DNA, Complementary / Q000235:genetics; D008969:Molecular Sequence Data; D020076:Pleurotus / Q000502:physiology / Q000821:virology*; D012313:RNA / Q000235:genetics; D012328:RNA Viruses / Q000235:genetics / Q000302:isolation & purification*; D012330:RNA, Double-Stranded / Q000737:chemistry / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017421:Sequence Analysis; D013172:Spores, Fungal / Q000235:genetics / Q000821:virology*; D014764:Viral Proteins / Q000235:genetics; D014771:Virion / Q000302:isolation & purification / Q000648:ultrastructure
D017931:DNA Primers; D018076:DNA, Complementary; D012330:RNA, Double-Stranded; D014764:Viral Proteins; D012313:RNA