Microbiologia

URI permanente desta comunidadehttps://locus.ufv.br/handle/123456789/11840

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2008 - 200912010 - 20122

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Autor: search.filters.author.Araújo, Elza Fernandes deData inicial: 1992Tem arquivos: SimAssunto: search.filters.subject.Moniliophthora perniciosa

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Agora exibindo 1 - 3 de 3
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    Restriction enzyme improves the efficiency of genetic transformations in Moniliophthora perniciosa, the causal agent of witches’ broom disease in Theobroma cacao
    (Brazilian Archives of Biology and Technology, 2008-01) Lopes, Francis Julio Fagundes; Queiroz, Marisa Vieira de; Lima, Juliana Oliveira; Silva, Viviane Aline Oliveira; Araújo, Elza Fernandes de
    The presence of restriction enzymes in the transformation mixture improved the efficiency of transformation in Moniliophthora perniciosa. The influence of the vector shape (linear or circular), the patterns of plasmid integration in genomic sites and the influence of the promoter used to express the gene marker were also analyzed. The addition of BamHI or NotI increased the number of transformants by 3-10-fold and 3-fold, respectively, over the control without added enzyme. The use of pre-linearized plasmid did not increase the transformation efficiency in comparison with the circular plasmid. However, the frequency of multi-copy transformants increased significantly. The transformation procedure here reported resulted in better production of protoplasts and transformation efficiency. In addition, the time necessary for the detection of the first transformants and the number of insertions were reduced.
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    Development of molecular markers based on retrotransposons for the analysis of genetic variability in Moniliophthora perniciosa
    (European Journal of Plant Pathology, 2012-11) Santana, Mateus Ferreira; Araújo, Elza Fernandes de; Queiroz, Marisa Vieira de; Souza, Jorge Teodoro de; Mizubuti, Eduardo Seiti Gomide
    Moniliophthora perniciosa is a fungus that causes witches’ broom disease (WBD) in the cacao tree (Theobroma cacao). The M. perniciosa genome contains different transposable elements; this prompted an evaluation of the use of its retrotransposons as molecular markers for population studies. The inter-retrotransposon amplified polymorphism (IRAP) and retrotransposon-microsatellite amplified polymorphism (REMAP) techniques were used to study the variability of 70 M. perniciosa isolates from different geographic origins and biotypes. A total of 43 loci was amplified. Cluster analysis of different geographical regions of C biotype revealed two large groups in the state of Bahia, Brazil. Techniques using retrotransposon-based molecular markers showed advantages over previously used molecular techniques for the study of genetic variability in M. perniciosa.
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    Boto, a class II transposon in Moniliophthora perniciosa, is the first representative of the PIF/ Harbinger superfamily in a phytopathogenic fungus
    (Microbiology, 2012-10-24) Pereira, Jorge Fernando; Almeida, Ana Paula Morais Martins; Cota, Júnio; Pamphile, João Alencar; Silva, Gilvan Ferreira da; Araújo, Elza Fernandes de; Gramacho, Karina Peres; Brommonschenkel, Sérgio Hermı́nio; Pereira, Gonçalo Amarante Guimarães; Queiroz, Marisa Vieira de
    Boto, a class II transposable element, was characterized in the Moniliophthora perniciosa genome. The Boto transposase is highly similar to plant PIF-like transposases that belong to the newest class II superfamily known as PIF/Harbinger. Although Boto shares characteristics with PIF-like elements, other characteristics, such as the transposase intron position, the position and direction of the second ORF, and the footprint, indicate that Boto belongs to a novel family of the PIF/Harbinger superfamily. Southern blot analyses detected 6–12 copies of Boto in C-biotype isolates and a ubiquitous presence among the C- and S-biotypes, as well as a separation in the C-biotype isolates from Bahia State in Brazil in at least two genotypic groups, and a new insertion in the genome of a C-biotype isolate maintained in the laboratory for 6 years. In addition to PCR amplification from a specific insertion site, changes in the Boto hybridization profile after the M. perniciosa sexual cycle and detection of Boto transcripts gave further evidence of Boto activity. As an active family in the genome of M. perniciosa, Boto elements may contribute to genetic variability in this homothallic fungus. This is the first report of a PIF/Harbinger transposon in the genome of a phytopathogenic fungus.