Detection and sequence analysis of two novel co-infecting double-strand RNA mycoviruses in Ustilaginoidea virens.

Four novel double-stranded RNA molecules, named dsRNA 1 (5124 bp), dsRNA 2(1711 bp), dsRNA 3 (1423 bp) and dsRNA 4 (855 bp), were detected in strain HNHS-1 of Ustilaginoidea virens, the causal agent of rice false smut disease. Sequence analysis showed that the dsRNA1 contains two overlapping open reading frames (ORF) potentially encoding proteins with modest levels of sequence similarity to the coat protein (CP) and putative RNA-dependent RNA polymerase (RdRp), respectively, of viruses of the family Totiviridae. The deduced gene product of the ORF encoded by dsRNA2 is homologous to putative RdRp of viruses in the family Partitiviridae; the ORF encoded by dsRNA3 shares some similarity to a hypothetical protein with unknown function. It is noteworthy that the dsRNA4 lacked integrated ORFs. Isomeric viral particles of about 40 nm in diameter were observed by transmission electron microscopy in a mycelium tissue preparation of strain HNHS-1-R1, a single-spore subculture of strain HNHS-1 containing only the dsRNA1 segment. Phylogenetic analysis and examination of the organization of the two putative RdRp sequences both indicated that there are at least two novel virus species present in strain HNHS-1. We named the two novel viruses Ustilaginoidea virens RNA virus 2 and Ustilaginoidea virens partitivirus 4, respectively.

The nucleotide sequence and genome organization of two victoriviruses from the rice false smut fungus Ustilaginoidea virens.

Ustilaginoidea virens is the causal agent of false smut disease of rice. The dsRNA-Ls from a U. virens strain of Uv0901 containing three dsRNA bands were separated and sequenced. The dsRNA-Ls, with molecular weight of 5 kbp, were demonstrated to be two dsRNA segments referenced as dsRNA-L1 and dsRNA-L2. The two dsRNAs each contained two overlapped open reading frames, and showed significant similarity to those of capsid protein and RNA-directed RNA polymerase, respectively, of members of the family Totiviridae. Homology research and phylogenic analysis indicated that the dsRNA-L1 was a conspecific species of the previous reported Ustilaginoidea virens RNA virus 1, named Ustilaginoidea virens RNA virus Uv0901, and the dsRNA-L2 was a new number of the Victorivirus in the family Totiviridae, designed as Ustilaginoidea virens RNA virus 3.

Complete genome sequence and organization of a novel virus from the rice false smut fungus Ustilaginoidea virens.

In this study, three dsRNA segments from the rice false smut fungus Ustilaginoidea virens, the causal agent of a serious disease in rice, with molecular size ranging from 1.3 to 5 Kb, were isolated and named as dsRNA-L, dsRNA-M, and dsRNA-S. The complete nucleotide sequences of dsRNA-M and dsRNA-S were determined and analyzed. The dsRNA-M putatively encodes an RNA-dependent RNA polymerase, which is similar to that of the partitiviruses in the family Partitiviridae. Although the protein encoded by dsRNA-S showed less similarity to the typical coat protein of the virus in the family Partitiviridae, the structural analysis results indicated that the dsRNA-S might function as the capsid protein. We propose that the virus is Ustilaginoidea virens partitivirus 2-Uv0901, a new member, but distantly related to the newly proposed genus Gammapartitivirus with a distinct sequence pattern of capsid protein.

Detection and characterization of a novel Gammapartitivirus in the phytopathogenic fungus Colletotrichum acutatum strain HNZJ001.

Spherical virus-like particles about 40nm in diameter were observed under transmission electron microscope (TEM) and two dsRNA bands (dsRNA-1 and dsRNA-2) were detected on agarose gel after extraction from the mycelial preparation of a Colletotrichum acutatum strain HNZJ001 that isolated from an anthracnose lesion on immature pepper fruit. The complete nucleotide sequences of the dsRNAs were determined. DsRNA-1 (1762 nt) and dsRNA-2 (1381 nt) each contained a single open reading frame and potentially encoded 62 kDa and 40 kDa proteins, respectively. The 62 kDa protein showed similarity to the RNA-dependent RNA polymerase (RdRp) of partitiviruses, while the 40 kDa product had no significant similarity to any published capsid protein throughout all databases, besides of low homology with the hypothetical "capsid" protein of a few partitiviruses in fungus Ustilaginoidea virens. Genome comparison and phylogenetic analysis indicated that the virus is closely related to the mycovirus in the family Partitiviridae. The results suggested a novel two-segment dsRNA virus be detected. We name it Colletotrichum acutatum partitivirus 1 (CaPV1). RT-PCR detection, using a primer pair based on the RdRp of the dsRNA-1 showed very high efficiency of CaPV1 transmission into the progenies of the fungus. Virus curing and fungal phenotype observation for evaluation of the impact of CaPV1 in host fungus were also carried out.