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1.
J Virol ; 89(23): 11926-34, 2015 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-26378173

RESUMEN

UNLABELLED: The selective accumulation of both DNA components of a bipartite geminivirus, Abutilon mosaic virus, was recorded during early systemic infection of Nicotiana benthamiana plants. Purified nuclei were diagnosed for viral DNA using hybridization specific for DNA A or DNA B to detect these individual genome components either alone or both simultaneously by dual-color staining. Although this virus needs both components for symptomatic infection, DNA A alone was transported to upper leaves, where it was imported into phloem nuclei and replicated autonomously. The coinfection with DNA A and DNA B revealed an independent spread of both molecules, which resulted in a stochastic distribution of DNA A- and DNA A/B-infected nuclei. A population genetics evaluation of the respective frequencies was compared to a model computation. This elucidated a surprisingly simple relationship between the initial frequencies of the viral DNA components and the number of susceptible cells during the course of early systemic infection. IMPORTANCE: For bipartite begomoviruses, DNA B-independent long-distance spread of DNA A has been described before, but it has never been shown whether viral DNA A alone invades nuclei of systemic tissues and replicates therein. This is demonstrated now for the first time. During infection with DNA A and DNA B, a similar solitary spread of DNA A can be recognized at early stages. We describe a population genetics model of how the hit probabilities of DNA A and DNA B for susceptible cells determine the relative frequencies of either genome component during the course of infection.


Asunto(s)
Núcleo Celular/virología , ADN Viral/aislamiento & purificación , Geminiviridae/genética , Nicotiana/virología , Cartilla de ADN/genética , ADN Viral/clasificación , Genética de Población , Hibridación in Situ , Modelos Genéticos , Nicotiana/citología
2.
Microbiology (Reading) ; 155(Pt 12): 4069-4083, 2009 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-19744988

RESUMEN

Pseudomonas putida GJ31 has been reported to grow on chlorobenzene using a meta-cleavage pathway with chlorocatechol 2,3-dioxygenase (CbzE) as a key enzyme. The CbzE-encoding gene was found to be localized on the 180 kb plasmid pKW1 in a cbzTEXGS cluster, which is flanked by transposases and encodes only a partial (chloro)catechol meta-cleavage pathway comprising ferredoxin reductase, chlorocatechol 2,3-dioxygenase, an unknown protein, 2-hydroxymuconic semialdehyde dehydrogenase and glutathione S-transferase. Downstream of cbzTEXGS are located cbzJ, encoding a novel type of 2-hydroxypent-2,4-dienoate hydratase, and a transposon region highly similar to Tn5501. Upstream of cbzTEXGS, traNEOFG transfer genes were found. The search for gene clusters possibly completing the (chloro)catechol metabolic pathway of GJ31 revealed the presence of two additional catabolic gene clusters on pKW1. The mhpRBCDFETP cluster encodes enzymes for the dissimilation of 2,3-dihydroxyphenylpropionate in a novel arrangement characterized by the absence of a gene encoding 3-(3-hydroxyphenyl)propionate monooxygenase and the presence of a GntR-type regulator, whereas the nahINLOMKJ cluster encodes part of the naphthalene metabolic pathway. Transcription studies supported their possible involvement in chlorobenzene degradation. The upper pathway cluster, comprising genes encoding a chlorobenzene dioxygenase and a chlorobenzene dihydrodiol dehydrogenase, was localized on the chromosome. A high level of transcription in response to chlorobenzene revealed it to be crucial for chlorobenzene degradation. The chlorobenzene degradation pathway in strain GJ31 is thus a mosaic encoded by four gene clusters.


Asunto(s)
Clorobencenos/metabolismo , Genes Bacterianos , Familia de Multigenes , Pseudomonas putida/genética , Pseudomonas putida/metabolismo , Secuencia de Bases , Biodegradación Ambiental , Cromosomas Bacterianos/genética , Clonación Molecular , Cartilla de ADN/genética , ADN Bacteriano/genética , Dioxigenasas/genética , Dioxigenasas/metabolismo , Datos de Secuencia Molecular , Filogenia , Plásmidos/genética , Pseudomonas putida/crecimiento & desarrollo
3.
Virology ; 336(2): 240-50, 2005 Jun 05.
Artículo en Inglés | MEDLINE | ID: mdl-15892965

RESUMEN

The biochemical and genetic features of Poinsettia latent virus (PnLV, formerly named Poinsettia cryptic virus), which is spread worldwide in commercial cultivars of Euphorbia pulcherrima without inducing symptoms, have been determined using virus-purification, immunological techniques, electron microscopy, cloning, and sequencing. PnLV was found to be a chimeric virus with one 4652 bases, plus strand RNA showing a close relationship to poleroviruses within the first three quarters of its genome but to sobemoviruses in the last quarter. Thus, we propose to classify this virus as "polemovirus". Similarities of protein and nucleic acid sequences at the 5' and extreme 3' end of its RNA suggest a replication mode like that of poleroviruses, whereas the coat protein sequence is closely related to that of sobemoviruses. Consistent with these results, PnLV forms stable icosahedra of 34 nm in diameter. The consequences for the taxonomy of PnLV and for gardeners' practice are discussed.


Asunto(s)
Euphorbiaceae/virología , Virus ARN/clasificación , Recombinación Genética , Secuencia de Bases , Proteínas de la Cápside/genética , Clonación Molecular , Luteovirus/fisiología , Datos de Secuencia Molecular , Conformación de Ácido Nucleico , Virus ARN/química , Virus ARN/fisiología , ARN Viral/química , Alineación de Secuencia , Tombusviridae/fisiología , Replicación Viral
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