RESUMO
Deep sequencing of small RNAs has proved effective in the diagnosis of mycovirus infections. In this study, the presence of mycoviruses in ten isolates of the phytopathogenic fungus Fusarium circinatum was investigated by high-throughput sequencing (HTS) of small RNAs. The contigs resulting from de novo assembly of the reads were aligned to viral genome sequences. The presence of each mycovirus detected in the isolates was confirmed by RT-PCR analysis with four previously described primer pairs and seven new pairs designed on the basis of sequencing data. The findings demonstrate the potential use of HTS for reconstructing previously identified mitoviruses infecting F. circinatum.
Assuntos
Micovírus/genética , Fusarium/virologia , Genoma Viral , RNA Citoplasmático Pequeno/genética , RNA Viral/genética , Traqueófitas/microbiologia , Mapeamento de Sequências Contíguas , Micovírus/classificação , Micovírus/isolamento & purificação , Sequenciamento de Nucleotídeos em Larga Escala , Filogenia , Doenças das Plantas/microbiologia , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
Viruses of the Partitiviridae family occur at a relatively low frequency in the fungal genus Heterobasidion, but show high genetic diversity. Here, we describe four novel partitivirus species that infect three Heterobasidion species that are pathogens of conifers: H. annosum, H. parviporum and H. irregulare. We show that these viruses, designated Heterobasidion partitivirus 12 (HetPV12), HetPV13, HetPV14 and HetPV15, form a phylogenetically distinct clade together with the previously described Heterobasidion partitivirus 3 (HetPV3) found in the H. insulare species complex and Helicobasidium mompa partitivirus V70, both members of the genus Alphapartitivirus. Closely related strains of HetPV13 (over 97% polymerase identity at the nucleotide level) occur in H. annosum and H. parviporum, suggesting recent transmission of this virus species between the two fungal host species. Moreover, the occurrence of nearly identical HetPV13 strains in Finland and Poland (ca 1400km apart) indicates that the dispersal capacity of Heterobasidion partitiviruses is high. Viruses related to HetPV3 have a global distribution but only ca 2.7% overall prevalence among isolates of Heterobasidion. In three cases, these HetPV3-related viruses co-infected their hosts with distantly related partitiviruses or Heterobasidion RNA virus 6.
Assuntos
Basidiomycota/virologia , Variação Genética , Vírus de RNA/classificação , Vírus de RNA/isolamento & purificação , Análise por Conglomerados , DNA Viral/química , DNA Viral/genética , Dados de Sequência Molecular , Filogeografia , Vírus de RNA/genética , Análise de Sequência de DNA , Homologia de Sequência do Ácido NucleicoRESUMO
The cultivated and uncultivated bacterial communities of an activated sludge plant were studied. Two samples were taken and a total of 516 bacterial isolates were classified into groups using their whole-cell protein patterns. The distribution of bacteria into protein-pattern groups differed significantly between the two samples, suggesting variation in culturable bacterial flora. Partial 16S rRNA gene sequences were determined for representatives of the commonest protein-pattern groups. Most of the sequences obtained were previously unknown, but relatively closely related to known sequences of organisms belonging to the alpha, beta or gamma subclasses of the proteobacteria, the first two subclasses being predominant. This classification of bacteria isolated on a diluted nutrient-rich medium differed from recent culture-dependent studies using nutrient-rich media. The uncultivated bacterial community was studied by analyzing ten partial 16S rRNA gene sequences cloned directly from activated sludge. None of the cloned sequences was identical to those determined for culturable organisms; or to those in the GenBank database. They were, however, related to the alpha or beta subclasses of the proteobacteria, or to the gram-positive bacteria with a high G + C DNA content.
